Biodegradable frame for soft tissue regeneration and use thereof

FIELD: medicine.

SUBSTANCE: there are described new reinforced biodegradable frames for soft tissue regeneration; there are also described methods for living tissue support, extension and regeneration, wherein the reinforced biodegradable frame is applied for relieving the symptoms requiring high durability and stability apart from patient's soft tissue regeneration. What is described is using the frames together with cells or tissue explants for soft tissue regeneration in treating medical prolapsed, e.g. rectal or pelvic prolapse, or hernia.

EFFECT: frames are adequately durable to be applicable for implantation accompanying the medical conditions requiring the structural support of the injured tissues.

14 cl, 19 dwg, 2 tbl, 8 ex

 

The scope of the invention

This invention relates to new reinforced biodegradable frame for the regeneration of soft tissues, as well as ways to support the expansion and regeneration of living tissue, where the reinforced biodegradable frame is used to treat symptoms that require increased strength and stability in addition to the necessary regeneration of living tissue of the patient. This invention also relates to the application of frames with cells or tissue explants for regeneration of soft tissues, such as in the treatment of medical prolapse, for example, or rectal prolapse, pelvic, or hernia, or incontinence of urine.

Background of the invention

Frames are structures, such as synthetic polymeric structure used for the organization, growth and differentiation of cells in the formation of new functional tissue at the site of tissue defect, wound, used in combination with surgical intervention.

To achieve the target tissue reconstruction frames must meet certain specific requirements. High porosity and an adequate pore size necessary to facilitate cell growth and diffusion throughout the structure of both cells and nutrients. The ability to biodegradation is significant, so it is to need so the frames were absorbed by surrounding tissues without the need for surgical removal.

Many different materials (natural and synthetic, biodegradable and permanent) have been investigated for use as frames. Most of these materials have been known in the field of medicine before the advent of tissue engineering as a research topic, being used as absorbable stitches. Examples of such materials are collagen or some linear aliphatic polyesters.

Conditions such as stress incontinence and prolapse of the pelvic organs (POP), are symptoms women experience as the result of a multiple birth, muscle weakness due to aging and hormonal deficiency. However, the same symptoms occur in young inactive patients who have never given birth. Since the 1980s, the use of synthetic mesh made of polypropylene, was the preferred treatment. Examples of such grids are Prolene (Ethicon), Polyform (Boston Scientific) and Pelvitex (Bard). In recent years, the increased number of side effects noted in up to 10% of cases. Vaginal erosion and vaginal reduction are some of the most serious ["Rising use of synthetic mesh in transvaginal pelvic reconstructive surgery: A review of the risk of vaginal erosion". E. Mistrangelo et al., J. Minimally Inasive Gynecology, 007, 4, p.564-69].

To overcome these side effects was developed lightweight version (less material) from the traditional grid and several made partially degradable by combining polypropylene with degradable synthetic-like polymer by polylactides (Ultrapro, Ethicon). Cook Inc. has xenotransplantation approach, which is fully biodegradable and is based on cell-free extracellular matrix of the small intestine of the pig [Mantovani F., Trinchieri, A., Castelnuovo C., Romano A. L., Pisani E. "Reconstructive urethroplasty using porcine acellular matrix."Eur Urol2003;44:600-602].

U.S. patent No. 2009024162 relates to absorbable composite medical components, such as a surgical mesh and braided suture, which demonstrate two or more of the resorption profile/biodegradation and preservation of tensile strength.

International publication WO 08083394 relates to power grids for retropubically implant for treating urinary incontinence and/or disorders of the pelvic floor and related applications.

International publication WO 08042057 refers to the components for tissue strengthening, in particular, to components having both macroporous and microporous structure to allow cell growth and tissue integration.

International publication WO 2006044881 and WO 07117238 relate to multi-layer material containing the first is the first absorbable nonwoven fabric and the second absorbable woven or knitted material, and method of its manufacture.

European patent 1674048 refers to rasskazyvaemoe polymeric mesh implant, which is designed for use in reconstruction of soft tissue defects. Mesh implant includes at least first and second material, where the second material mainly decomposes at a later point in time than the first material after the time of implantation.

Patent document U.S. No. 20080241213 relates to a biocompatible tissue implant, which may be bioabsorbable, and made from a biocompatible polymeric foam. Tissue implant also includes a biocompatible reinforcing element. The polymer foam and the reinforcing element are soluble in a common solvent.

International publication WO 0222184 refers to tissue engineered prostheses made from recycled tissue matrices obtained from native tissues, which are biocompatible with the patient or host to which they are implanted.

Patent document U.S. No. 2002062152 relates to medical camarasaurus the implant, in particular, for the increment of the cruciate ligament, is constructed as a composite structure in textile design from the at least two biocompatible polymeric materials that are different in their chemical composition and/or poly the agreement structure and which are biodegradable implant, with a given initial tensile strength and different decomposition behavior of polymers and/or textile design, chosen so that the tensile strength decreases during decomposition.

International publication WO 06020922 relates to absorbable polylactide contractile polymer barrier membranes scar tissue and reveals the ways of their application.

European patent 1216717 relates to a bioabsorbable, porous, reinforced tissue implants designed components for use in the recovery of soft tissue injuries, such as damage to the pelvic floor, and methods of making such components.

The purpose of the invention

The purpose of embodiments of the present invention is to provide a support for the regeneration of soft tissue by providing a fully degradable scaffold for germination or re-growing cells, grown in vitro, cells/tissues collected in the operating room, either germination or re-growing cells from the surrounding tissue.

Accordingly, presented frames with good properties with respect to tissue reconstruction, which at the same time strong enough to be suitable for implantation in medical conditions that require structural support, for example, with the centres of tissues, which require surgical intervention.

Brief description of the invention

Data author(s), it was found that reinforced porous framework facilitates manipulation at the time of operation, i.e. during surgery. Frames made according to this invention, provide sufficient strength in the process of manipulation in combination with the properties of stimulating regeneration of tissue of a patient, which requires implant, and they are strong enough to provide sufficient structural support to the recovery area.

It should be understood that surgical implants, being optimized with respect to the properties, ensuring the regeneration of soft tissues in a patient, it is not always optimal to manipulate or to provide sufficient support at the site of the implant. This can be especially true when medical conditions, including damage to the structural support of the soft tissues, such as in medical prolapse, for example, prolapse of pelvic organs, stress incontinence or hernias.

These inventors found that the structural support and reinforcement can be attached to the implant without compromising the ability to stimulate the regeneration of tissue of the patient at the site of damage.

Thus, in pervomayka this invention relates to a biodegradable surgical implant for support, capacity and regeneration of living tissue in a subject, including:

a) synthetic biodegradable homogenous layer frame;

b) one or more biodegradable reinforcing details.

In the second aspect, the invention relates to a method for support, enhancement and regeneration of living tissue in the subject, and the method includes the implantation of the biodegradable surgical implant comprising a synthetic biodegradable frame together with a sample of autologous cells or tissue explants, a specified subject in the section where you want to support, increase and regeneration of living tissue.

In the third aspect of this invention relates to a method for preparing a biodegradable surgical implant comprising a synthetic biodegradable frame and autologous cells or tissue explants of the subject, suitable for the support, enhancement and regeneration of living tissue in the specified entity with the specified method comprises ex vivo application of the sample indicated autologous cells or tissue explants on or in the specified biodegradable surgical implant comprising a synthetic biodegradable frame, prior to implantation specified subject to the area where you want to support, increase and regeneration of living tissue.

The following aspect of this invention relative to the tsya to the biodegradable surgical implant, including synthetic biodegradable scaffold for use in the method of support, enhancement and regeneration of living tissue in the subject, and the method includes the implantation of the specified biodegradable surgical implant comprising a synthetic biodegradable frame together with a sample of autologous cells or tissue explants, a specified subject in the section where you want to support, increase and regeneration of living tissue.

The following aspect of this invention relates to a biodegradable surgical implant comprising a synthetic biodegradable frame, for use in the method of support, enhancement and regeneration of living tissue in the subject, and the method includes the steps of (i) screening a tissue sample from the subject; (ii) crushing or destruction of the tissue sample; (iii) the implantation of the frame and shredded tissue sample of the subject.

The following aspect of this invention relates to a kit including:

a) a biodegradable surgical implant comprising a synthetic biodegradable frame;

b) a sample of autologous cells or tissue explants; and

c) optionally instructions for use of the method of support, enhancement and regeneration of living tissue in a subject, for example, a subject with a medical prolapse, for example, rectal and is and prolapse of pelvic organs, or hernia, and this method includes the implantation of the specified biodegradable surgical implant with autologous sample of cells or tissue explants specified subject to the area where you want to support, increase and regeneration of living tissue.

The following aspect of this invention relates to a kit including:

a) synthetic biodegradable frame; and

b) component, suitable for crushing or destruction of the tissue sample.

In the following aspect, the invention relates to a method for support, enhancement and regeneration of living tissue in the subject with the medical prolapse, such as prolapse of the pelvic organs, or hernia, and the method includes the implantation of the biodegradable surgical implant comprising a synthetic biodegradable homogenous layer frame together with the sample of cells or tissue explants, subject to section prolapse or hernia.

In the following aspect, the invention relates to a method for support, enhancement and regeneration of living tissue in a subject, the method including the implantation of the biodegradable surgical implant for support, enhancement and regeneration of living tissue in a subject, including:

a) synthetic biodegradable homogenous layer frame,

b) one or more biodegradable reinforcing ale is now;

characterized in that the synthetic biodegradable homogenous layer frame is hydrophilic in the subject.

The following aspect of this invention relates to a method for preparing a biodegradable surgical implant according to this invention in a way that simultaneously includes sequential steps:

a) preparation of synthetic biodegradable homogenous layer frame;

b) preparation and inclusion of one or more biodegradable reinforcing elements in the synthetic biodegradable homogenous layer frame;

c) optional inclusion of one or more components, as defined in this document.

The following aspect of this invention relates to a kit including:

a) a biodegradable surgical implant according to this invention;

b) a sample of cells or tissue explants and

c) optionally instructions for use of the method of support, enhancement and regeneration of living tissue in the subject with the medical prolapse, such as prolapse of the pelvic organs, or hernia, and the method includes the implantation of the biodegradable surgical implant together with a sample of cells or tissue explants on a specified subject area prolapse or hernia.

The following aspect of this invention relates to an implant according to this the invention for use in medicine.

The following aspect of this invention relates to an implant according to this invention for use in the treatment of diseases associated with prolapse of the pelvic organs and hernia.

Captions to figures

Fig.1: SEM image of the cross section of the frame, made by lyophilization. The orientation of the material in the direction of freezing.

Fig.2a: 40×40 mm frames. Left: not modified. Right: welded along the edges for extra strength.

Fig 2b: the Frame is welded in the form of a mesh structure for added strength.

Fig.3: Frame welded to the substrate from electrospraying PLGA.

Fig.4: Dried structure, which is enhanced by the inclusion of a grid of suture material.

Fig.5: Illustration depicts the various patterns that can be used to enhance the framework by incorporating biodegradable threads.

Fig.6: the elasticity of the frame. This figure shows that when the frame is made of mPEG-PLGA is dry, it is hard. On the other hand, as soon as it becomes wet, it becomes very flexible. This compares with polypropylene mesh, which is no less hard after exposure to water.

Figure 7: Mesh with 200, 300 and 400 μm, the specific surface soldered to the washers 8 mm specific surface.

Figure 8: The high pressure threads not shown).

Figure 9: Valve; biodegradable surgical implant comprising a frame.

Figure 10: Left: the cells are marked. Right: the valve is closed and need not be sewn.

Figure 11: Full or segmented valves.

Figure 12: a Tube; a biodegradable surgical implant comprising a frame, designed in the form of a tube.

Figure 13: the Pocket; biodegradable surgical implant comprising a frame, designed in the form of a pocket.

Figure 14: Absorbent 3D frame, welded to the substrate material.

Figure 15: Wetting the E-spun layers of blood (15 minutes). Left: PCL coaxially covered with MPEG-PLGA 2-30 50DL. Right: simple PCL.

Figure 16: A: a Porous spongy structure of MPEG-PLGA. The dashed line marks the boundary between the surface and cross-section of the implant. B: knitted Vicryl mesh structure. Digital dark-field image of stereomicroscope at 10x magnification. Scale range: 1,0 mm

Figure 17: MPEG-PLGA combined with fragmented muscle fibers after 3 weeks. Muscle tissue is located beneath the implant.

Figure 18: MPEG-PLGA combined with fragmented muscle fibers 8 weeks. Muscle tissue is where the implant and fragmented muscle fibers have been previously implanted.

Figure 19: biodegradable surgical implant shoulder and/or extensions to attach to the structures in the pelvis.

Detailed disclosure of the invention

In this context, the expression "biodegradable", "bioabsorbable" or simply "degradable" as used herein refers to the polymer, which disappears within a certain period of time after introduced into a biological system, which can be in vivo (e.g., human body), as in this invention, or in vitro (in culture with cells); the mechanism by which it disappears, can vary, it can be subjected to hydrolysis, decomposed, bioresour, bioassay, biopower, borishade, dissolved or otherwise disappears from the biological system. When used in a clinical context, this is a significant clinical advantage, because nothing must be removed from the restoration area. Thus, the newly formed tissue is not disturbed or subjected to a load due to the presence or even the removal of the temporary frame. In some embodiments, the implement frame is decomposed within 1 day to 4 years, for example, from 1 day to one year, for example, within 2 to 6 months.

The terms “biocompatible” refers to a composition or compound which, when inserted into the body of a mammal, for example, in the patient's body, in particular, when inserted into the site of a defect, does not lead to substantial toxicness is or deleterious immune response in the individual.

When the expression "approximately" is used herein in combination with a specific value or range of values, this expression is used to denote a range of values, and the actual relative values.

The expression "the cultivation ofin vitro", as used herein, refers to the stage of the method according to this invention, where a sample of cells or tissue explants maintained in terms ofin vitro, i.e., under conditions of controlled environment outside of a living mammal. Alternatively, a qualified professional may use phrases such as "cells grown" or "cells proliferatory"in vitroalso in the meaning of "cultivation".

The term "elongation" as used herein, refers to the % elongation, where the polymer frame or reinforced surgical implant according to this invention will be interrupted, as measured in the case described in example 3.

The term "tensile strength" as used herein refers to the strength polymer frame or reinforced surgical implant according to this invention, as measured in N/m2or in pounds per square inch in the case described in example 3.

The phrase "vertical porous structure", as used herein, refers koristi the frame structure of the polymer, used according to this invention, in which the pores are mainly oriented in the vertical direction to the layer frame. This will help to absorb fluid and cells at the site of implantation.

The expression "interconnected pores" as used herein, refers to the type of polymer used in this invention, which has a porous structure with holes between the individual pores, such as holes in a horizontal direction between the individual pores mainly with vertical orientation. This will allow the cells to migrate in either direction through the frame polymeric material.

The term "fabric" as used herein refers to a solid living tissue that is part of a living individual mammal, such as man. The fabric can be solid tissue (e.g.,bone, joint and cartilage or soft tissue, including tendons, ligaments, fascia, fibrous tissues, fat, synovial membrane and muscles, nerves and blood vessels.

In specific aspects, the sample of cells or tissue explants, for example, a sample of body fluid, optionally mixed with culture medium, placed on the surface or at least together with the frame, usually on the cultural Cup or flask. A sample of cells or tissue is of explantation can be placed together with the component, which promotes cell adhesion, re-growth and/or germination through the frame.

In another aspect of the muscle biopsy is placed in the container with an appropriate buffer, for example, the cellular environment, PBS, etc. Cells and muscle fibers extracted from the biopsies using shredder tissue (e.g., Sigma-Aldrich). Then the suspension of the muscles is applied to the surface of the frame before or simultaneously with implantation.

Suspension of the muscles used according to aspects of the present invention, usually planted with a density in the range of 1-100 mg suspension muscles on cm2frame of formation.

In another aspect of the muscle fibers differ from a biopsy or dissection muscles using, for example, scalpels, or by dissolving the muscles with the use of enzymatic treatment, for example, collagenase, to obtain the individual fibers with the satellite cells. These fibers are bonded to the surface of the frame before implantation.

Accordingly, tissue explants from the muscle tissue can be from muscle, cut in muscle puree, for example, using scalpels, or where muscle fibers isolated from the rest of the tissue by mechanical or enzymatic means, or where the muscle shredded muscle suspension, all of which include a population of cultured myoblasts and fibroblasts, and/or muscle precursor cells in Kacha is TBE satellite cells.

It should be understood that as soon as a sample of body fluid has been applied to synthetic biodegradable carcass, cellsin situon site medical use, or alternatively the cells contained in the sample of body fluid, the opportunity to migrate and/or to grow through the frame for the formation of new tissue, such as a new connection and/or muscle tissue. In one embodiment, a component that promotes cell adhesion and/or germination, at the same time is applied to the frame.

Frame

Synthetic biodegradable frame used in this invention is a porous structure that encourages and facilitates the growth of tissue and cells. The frame is made of a biocompatible, biodegradable materials and is used in the implant for holding the organization, growth and differentiation of cells in the process of forming functional tissue at the site of damage in a patient.

In most aspects of this invention, the synthetic biodegradable frame completely or partially decomposedin situon site medical use in the period up to about 48 months, for example, in the period up to 36 months, for example, in the period up to about 24 months, for example, in the period up to priblisitelno months for example, during the period until approximately 10 months, for example, in the period up to approximately 9 months, for example, in the period up to approximately 6 months, for example, in the period up to approximately 5 months, for example, in the period up to about 4 months, for example, in the period up to about 3 months, for example, in the period up to about 2 months, for example, in the period up to about 1 month, as measured after medical applications.

In some important aspects of this invention, the synthetic biodegradable frame is not fully or partially decomposedin situon the site of surgical applications before the end of the period of about 1 month, for example, after a period of approximately 2 months, for example, after a period of approximately 3 months, for example, after a period of approximately 4 months, for example, after a period of approximately 5 months, for example, during a period of approximately 6 months, for example, after a period of approximately 9 months, for example, during a period of approximately 12 months, for example, during a period of approximately 24 months, for example, during the period of approximately 36 months, as measured after medical applications.

The phrase "wholly or partly decomposedin situ" refers to synthetic biodegradable is in the frame, which decomposed on site medical use under the action of the internal components of the body or the external components of the frame or sample of body fluid applied to the frame. This action may be endogenous enzymatic activity of body fluids or alternative activity of the compounds added to the frame.

In some embodiments, the implementation of synthetic biodegradable frame is decomposed to the level of at least about 50%, for example at least about 60%, for example at least about 70%, for example at least about 70%, for example at least about 80%, for example at least about 90%, for example at least about 100% during this time period.

It should be understood that the frame material is typical of the level of decomposition can be chosen to match the time needed to provide sufficient support and strengthen on-site medical use up until native tissue of the patient will not provide the necessary support and strength.

In some embodiments, the implementation of synthetic biodegradable frame is selected to be partially or fully degradable cellular decomposition, i.e., decomposed under the action of cellular enzymes, for example, the enzymatic action of the body fluids of the patient.

The following is the duty to regulate to understand what frame material, sensitive to cellular decomposition, can be selected to match the specific and appropriate period of decay.

In some embodiments, the implementation of synthetic biodegradable frame is sterilized by application of radiation, such as beta-irradiation, or plasma sterilization.

Synthetic biodegradable frame before implantation can be trimmed or adjusted in size to suit the needs of the defect, respectively, the frame can be cut to a particular shape or form to suit site specific defect and/or the desired shape/form new tissue.

Synthetic biodegradable scaffold could be applied in one or more layers, in the form of fibers, woven and/or nonwoven materials, for example, with a porous structure.

In some embodiments, the implement frame is biocompatible.

In one embodiment, the frame includes a polymer, which can be selected from the group consisting of: collagen, alginate, polylactic acid (PLA), polyglycolic acid (PGA), MPEG-PLGA or PLGA.

In one embodiment, the frame includes a polymer, which can be selected from the group consisting of: 1) Homo - or copolymers: glycolide, L-lactide, DL-lactide, mizolastine, e-caprolactone, 1,4-dioxane-2-it, d-Valerya is she, β-butyrolactone, g-butyrolactone, e-decalactone, 1,4-dioxan-2-it, 1, 5-dioxan-2-it, 1,5,8,12-tetrachloroethylene-7-14-Dion, 6,6-dimethyl-1,4-dioxane-2-she and trimethylhexanoate, 2) block copolymers of mono - or bifunctional polyethylene glycol and polymers 1) above; 3) block copolymers of mono - or bifunctional polyalkyleneglycol and polymers 1) above; 4) mixtures of the above polymers and 5) polyanhydrides and polyarteritis.

In some embodiments, the implementation of the framework has the ability to be hydrophilic. Accordingly, the frame is wetted in water, isotonic buffers and/or blood and other body fluids.

In one embodiment, the frame mainly consists of or includes a polymer or polymers with a molecular weight of, for example, an average molecular weight of more than about 1 kDa, for example, from about 1 kDa to about 1 million kDa, for example from 25 kDa to 100 kDa.

The frame is preferably made in the form of a layer, which is suitable for implantation in the diaphragm, the abdominal cavity or region of the pelvic floor.

Frame layer may be selected from the group consisting of a membrane, for example, the porous membrane layer, for example, porous layer, the layer of fibers, the layer may have a different two-dimensional shapes, for example, an implant made on C is KAZ, to insert to the site of the defect, for example, to match the surgical reconstruction of the fascia of the body of a mammal, the foam layer may be woven or non-woven, liofilizirovannam polymer, for example, liofilizirovannami polymer layers, or any combination thereof.

Alternatively, the frame can be custom three-dimensional structure of the desired shape suitable for implantation at the site, requiring implantation.

Accordingly, the frames can be of any type and size and any thickness of the frame, for example, varying from thin membranes, to frames with a thickness of several millimeters, for example, in the range from approximately 0.1 mm to 6 mm, for example, in the range from about 0.2 mm to 6 mm, for example, in the range from about 0.5 mm to 6 mm

In one embodiment, the frame is in the form of a layer, which can be pre-cut or trimmed to size to fit the defect. This frame may be of a thickness of, for example, from about 0.2 mm to 6 mm

The pores of the skeleton can be partially occupied by a component which facilitates the cell adhesion and/or sprouting for tissue regeneration, for example, a component selected from the group consisting of estrogen, derived estrogen, ECM powder, thrombin, chondroitin sulfate, hyaluronan, heparinase the ATA, heparan sulfate, dermatosurgery, growth factors, fibrin, fibronectin, elastin, collagen, gelatin and aggrecan. Alternatively, the components may be fully or partially included or built into the frame.

As discussed above, the frames may consist of or include any suitable biologically acceptable material, however, in the preferred embodiment, the frame includes a compound selected from the group consisting of: polylactide (PLA), polycaprolactone (PCL), polyglycolide (PGA), a copolymer(D,L-lactide and glycolide) (PLGA), MPEG-PLGA (methoxypolyethyleneglycol)-copolymer(D,L-lactide and glycolide), polyhydroxybutyrate in General. In this regard, the frame, excluding the pore space and any additional components, for example, those that promote cell adhesion and/or sprouting for tissue regeneration, can include at least 50%, such as at least 60%, at least 70%, at least 80% or at least 90% of one or more polymers proposed in this document, including mixtures of polymers.

In some embodiments, the implementation of the framework and a reinforcing member made of polycaprolactone (PCL), for example, electroplating PCL copolymers caprolactone and lactide or biodegradable polyurethanes.

PLGA and MPEG-PLGA are suitable frame materials.

In some aspects of this invention, the synthetic biodegradable frame is a frame made according to the method, opened in international publication WO 07/101443, the method which includes the steps:

(a) dissolving a polymer, as defined herein, in a non-aqueous solvent to obtain a polymer solution;

(b) freezing the solution obtained in step (a) to get the frozen polymer solution; and

(c) freeze-drying the frozen polymer solution obtained in step (b), so as to obtain a biodegradable porous material.

Non-aqueous solvent used in the method as disclosed in international publication WO 07/101443, should be selected relative to the melting temperature so that it could be conveniently frozen. Typical examples of this are dioxane (tPL12°C) and dimethylcarbonate (tPL4°C).

In one embodiment, the method as disclosed in international publication WO 07/101443, the polymer solution after step (a) above is poured or cast in a suitable mold. Thus, it is possible to obtain a three-dimensional shape of the material, specially designed for a particular application.

In a variant of the implementation where the particles are components of the extracellular matrix used in the methods according to this invention, these extracellular matrix components can be dispersed in the solution obtained in step (a), before the solution (dispersion) is frozen, as defined in step (b).

Components of the extracellular matrix may, for example, be dissolved in a suitable solvent and then added to the solution obtained in step (a). By mixing with the solvent of step (a), i.e., a solvent for the polymer, as defined herein, components of the extracellular matrix will likely be deposited to form the dispersion.

In one aspect, the biodegradable porous material obtained in step (c), in a subsequent step, is immersed in a solution of fractions (e.g., hyaluronan), and then again liabilitiesa.

In some alternative embodiments, the implementation of the materials are present in the form of fibers or fibrous structures made from polymer defined in this document, possibly in combination with components of the extracellular matrix. Fibers or fibrous materials can be prepared using methods known to the person skilled in the art, for example, by molding from a melt, electroprecision, extrusion and so on

In preferably the x options exercise of synthetic biodegradable frame is biocompatible. Even if the frame structure according to this invention is biodegradable decomposition framework can still be present at the site of the original implant. Accordingly, again, this can be the advantage of using biocompatible frame material.

Porous frame material can be prepared by known methods, for example, as disclosed inAntonios G. Mikos, Amy J. Thorsen, Lisa A Cherwonka, Yuan Bao & Robert Langer. Preparation and characterization of poly(L-lactide) foams.Polymer35, 1068-1077 (1994). However, one frequently used method for the preparation of porous materials is lyophilization.

In some embodiments, the implementation of the framework has a porosity in the range from 20% to 99%, for example at least 50%, for example, from 50 to 95% or from 75% to 95% or 99%.

A high degree of porosity can be obtained by lyophilization.

In some embodiments, the implementation of the surgical implant according to this invention does not include a biological polymer, i.e., a biopolymer, e.g., protein, polysaccharide, polyisoprene, lignin, polyphosphate or polyhydroxyalkanoate.

In other embodiments, the implement frame further includes a biological polymer, i.e., a biopolymer, for example, a polypeptide, a protein, a polysaccharide, lignin, polyphosphate or polyhydroxyalkanoate (for example, as described in U.S. patent No. 6495152). Suitable biopolymer may be selected from the group consisting of gelatin, hyaluronan, hyaluronic acid, chondroitin sulphate, dermatosurgery, collagen, such as collagen type I and/or type II, alginate, chitin, chitosan, keratin, silk, elastin, cellulose and their derivatives.

The frame can be prepared by lyophilization of a solution comprising the compound, such as those listed above in solution.

Components of the extracellular matrix can be added either as particles, which are heterogeneously dispersed or as a surface coating. The concentration of components of the extracellular matrix relative to the synthetic polymer is usually in the range of 0.5-15% (wt./weight), for example, below 10% (wt./weight.). Moreover, the concentration of extracellular matrix components is preferably at most 0.3% (wt./volume), for example,most of 0.2 (wt./volume), relative to the volume of the material.

The type of frames used in the context of this invention should be a frame that is not acting as a foreign body in mammals (including humans), so that there may be a lack of immunity or low immunity, and frameworks used in this context should not be toxic or significantly harmful to the body in which they are placed. Preferably, the frames did not contain any of libmissioncontrol or any other harmful impurities.

Cells or tissue explants used in the frame, can be embedded in the hydrogel and can be capable of placement on the frame before placing the frame in the target area. The frame may be hydrophilic so that the cellular material was quickly absorbed by the frame. In other suitable embodiments, the implementation of the cellular material is placed in a pocket under the flap or in the tube material of the frame.

The frame may be hydrophilic, i.e. capable of within 5 minutes, for example, for 2 minutes at 30°C to absorb at less than a small amount of water or aqueous solution (for example, composition of cell suspension), for example, to absorb at least 1%, such as at least 2%, for example at least 5%, such as at least 10%, for example at least 20%, e.g. at least 30%, such as at least 50% water from the volume of the frame (or equivalent aqueous solution) when placed in an aqueous solution, such as physiological environment, buffer, water, blood or other body fluids, it is particularly useful that the frame can absorb a specified amount of cell suspension in its porous structure, thereby providing a relatively uniform distribution of cells, for example, endogenous cells or in vitro caused to cells or tissue explants around Karka is, as soon as it is inserted and fixed at the site of the defect.

In other embodiments, the implement frame material is defined as "hydrophilic" method, where a drop of plasma or blood is placed on the upper part of the reservoir frame material; observe the lower part of the reservoir frame material, and the layer frame material is hydrophilic, if the flow of fluid found within 15 minutes.

In some embodiments, the implementation of the biodegradable polymer is at least partially hydrophilic, i.e., has a component of the polymer, which may reasonably be considered to be hydrophilic, for example, the MPEG portion of the copolymer MPEG-PLGA.

The expression "hydrophilic" is used interchangeably with the expression "polar".

One way to improve hydrophilicity skeleton of the polymer is pre-processing tool that facilitates the absorption of endogenous cells at the site of the implant or cells applied to the frame prior to implantation, for example, anionic, cationic, non-ionic surfactants or amphiphilic surfactants, buffers or salts. Hydrating agents may also be used in combination with hydrophilic frames to further improve cell penetration into the porous structure.

Biocompatible frame of the present invention outstanging polyesters. The inclusion of a balanced hydrophilic block polymer biocompatibility of the polymer can be improved, because it improves the wettability characteristics of the material, and the initial adhesion of cells disrupted for non-polar materials.

In one important aspect of this invention, the frame is biodegradable.

In some embodiments, the implement frame is porous, for example,has a porosity of at least 25%, 50%, for example,in the range of 50-99,5%. Porosity can be measured by any method known in this field, for example, by comparison of pore volume compared to the volume of the solid skeleton. This can be done by determination of density frame compared to non-porous sample of the same composition as that of the frame. Alternatively, it may be used mercury intrusion parametria or BET.

In a very interesting embodiment of the present invention the biocompatible cage according to this invention consists of or includes one or more polymers selected from the group comprising poly(L-lactic acid) (PLLA), poly(D/L-lactic acid) (PDLLA), poly(caprolactone) (PCL) and the copolymer (lactic and glycolic acid) (PLGA), and their derivatives, in particular derivatives which contain the appropriate chain of the polymer with the addition of substitutional groups or songs that gain is live hydrophilic nature of the polymer, for example, MPEG or PEG. The examples presented in this document and include the group of polymers MPEG-PLGA.

In one embodiment, the frame consists of or contains a synthetic polymer.

The polymers used in the preparation of skeleton

International publication WO 07/101443 discloses suitable polymers for use as frame materials in this invention and methods for their preparation.

Suitable biodegradable polymers for use in the method of this invention consist of polyalkyleneglycols residue and one or two residues of the copolymer(lactic and glycolic acid).

Therefore, in one aspect of the present invention the frame is made from or contains or includes a polymer of the General formula:

A-O-(CHR1CHR2O)n-B

where

A is a residue of a copolymer(lactide and glycolide) with a molecular weight of at least 4000 g/mol, the molar ratio of (i) lactide links and (ii) glycosidic links in the remainder of the copolymer(lactide and glycolide) in the range from 80:20 to 10:90, in particular from 70:30 to 10:90, 60:40 to 40:60, for example, approximately 50:50, for example, 50:50.

B is either a remainder of the copolymer (lactide and glycolide), as defined for A, or selected from the group consisting of hydrogen, C1-6-alkyl and protective groups of hydroxyl,

one and the R 1and R2in each -(CHR1CHR2O)- link selected from hydrogen and methyl, and the other of R1and R2in the same -(CHR1CHR2O)- the link is hydrogen,

n represents the average number of -(CHR1CHR2O)- links in the polymer chain and is an integer in the range of 10-1000, in particular, 16-250,

the molar ratio (iii) polyalkyleneglycol links -(CHR1CHR2O)- total number of (i) lactide units and (ii) glycolide units in the residue(remainder) of the copolymer(lactide and glycolide) is at most 20:80, and where the molecular weight of the copolymer is at least 10000 g/mol, preferably at least 15000 g/mol, or even at least 20,000 g/mol.

Therefore, the polymers for use in the method of this invention can be deblocage type or reblockage type.

In some important aspects of this invention, the synthetic biodegradable frame is designed to have a specific rate of decompositionin vitro.This can be achieved by changing the individual components (or the ratio of individual components in the polymer.

In some embodiments, the implementation time of decomposition varies G-L-ratio and molecular weight of MPEG-PLGA polymers. Time decomposition of copolymers of DL-lactide and glycolide can be changed by changing the mole is REGO ratio of lactide and glycolide. Clean polyglycolide has a degradation time of 6-12 months, poly(D,L-lactide) - 12-16 months, the copolymer(D,L-lactide and glycolide) at a molar ratio of 85:15 is 2-4 months. The shortest decomposition obtained with a molar ratio of 50:50 for a 1-2 month. You can also change the time of decomposition by changing the molecular weight, but this effect is small compared to the variations that are possible with ratio L:G. Theoretically, you can get significantly more rapid decomposition of materials with very low molecular weights, but they have mechanical properties that preclude their use for most medical components.

In one specific embodiment, As in the above formula is the residue of a copolymer(lactide and glycolide) with a molecular weight of at least 4000 g/mol, the molar ratio (i) lactide units and (ii) glycolide units in the remainder of the copolymer(lactide and glycolide) in the range of about 50:50 molar ratio.

The porosity of the polymer can be at least 50%, for example, be in the range of 50-99,5%.

It is clear that the polymer for use in the method of the present invention includes one or two of rest, that is the balance(remainder) of the copolymer(lactide and glycolide). It is established that these residues should have a molecular weight of at least 4000 g/mol, more specifically, p is at least 5000 g/mol, or even at least 8000 g/mol.

The copolymer(lactide and glycolide) can be decomposed under physiological conditions, for example, in body fluids and tissues. However, because the molecular weight of these residues (and other requirements outlined in this document) it is believed that the decomposition will be slow enough to materials and articles made of polymer, were able to perform their role to the full decomposition of the polymer.

The expression "copolymer(lactide and glycolide) covers a number of variants of the polymer, for example, the copolymer(stat.-lactide-glycolide), the copolymer(DL-lactide and glycolide), the copolymer(mesolectal and glycolide), the copolymer(L-lactide and glycolide), the copolymer(D-lactide and glycolide), the sequence of lactide/glycolide in PLGA can be either static, gradient or block, and lactide can be L-lactide, DL-lactide or D-lactide.

Preferably the copolymer(lactide and glycolide) is a copolymer(random lactide-glycolide) or copolymer(deg.-lactide-glycolide).

Another important feature is that the molar ratio (i) lactide units and (ii) glycolide units in the residue(remainder) of the copolymer(lactide and glycolide) must be in the range from 80:20 to 10:90, in particular from 70:30 to 10:90.

In General, it was noted that the best results are obtained for polymers, where the molar ratio (i) lactide units and (ii) CH is kolednik units in the residue(remainder) of the copolymer(lactide and glycolide) is 70:20 or less. However, good results were also observed when the polymer had an appropriate molar ratio up to 80:20, with the proviso that the molar ratio (iii) polyalkyleneglycol units -(CHR1CHR2O)- total number of (i) lactide units and (ii) glycolide units in the residue(remainder) of the copolymer(lactide and glycolide) was at most 8:92.

As mentioned above, B is either a remainder of the copolymer(lactide and glycolide), as defined for A, or selected from the group consisting of hydrogen, C1-6-alkyl and protective hydroxyl groups.

In one embodiment, B is a residue of a copolymer(lactide and glycolide), as defined for A, i.e., polymer triblocal type.

In another embodiment, B is selected from the group consisting of hydrogen, C1-6-alkyl and protective hydroxyl groups, i.e. the polymer deblocage type.

Most typically (in this embodiment) is1-6-alkyl, for example methyl, ethyl, 1-propyl, 2-propyl, 1-butyl, tert-butyl, 1-pentyl and so on, most preferably, methyl. In the case when b is hydrogen, i.e., corresponds to the end HE group, the polymer typically get with the application of the protective group of the hydroxyl as B. "Protective hydroxyl groups are groups that can be removed after synthesis of the polymer, for example, the ay hydrogenolysis, hydrolysis, or other suitable means without destroying the polymer, thus, PEG-part remains free hydroxyl group, see, for example, textbooks that describe known from the prior art procedures, such as described by Greene, T. W. and Wuts, P. G. M. (Protecting Groups in Organic Synthesis, third or subsequent editions). Especially useful are the examples benzyl, tetrahydropyranyl, methoxymethyl and benzyloxycarbonyl. Such protective hydroxyl groups can be removed to obtain a polymer, where a is hydrogen.

One of R1and R2within each -(CHR1CHR2O)- link selected from hydrogen and methyl, and the other of R1and R2in the same -(CHR1CHR2O)- the link is hydrogen. Therefore, -(CHR1CHR2O)n- the balance can be polyethylene glycol, polypropyleneglycol, or a copolymer(ethylene glycol-propylene glycol). Preferably, the residue is -(CHR1CHR2O)n- is a polyethylene glycol, i.e., both R1and R2in each unit are hydrogen.

n represents the average number of units -(CHR1CHR2O) in the polymer chain and is an integer in the range of 10-1000, in particular, 16-250. It should be understood that n represents the average units -(CHR1CHR2O)- in the total number of polymer molecules. It will be obvious to the specification of the sheet in this area. Molecular weight polyalkyleneglycols balance (-(CHR1CHR2O)n-) typically is in the range 750-10000 g/mol, for example, 750-5000 g/mol.

Balance -(CHR1CHR2O)n- typically is non-biodegradable under physiological conditions, but, on the other hand, can be secreted in vivo, for example, from the human body.

The molar ratio (iii) polyalkyleneglycol units -(CHR1CHR2O)- total number of (i) lactide units and (ii) glycolide units in the residue(remainder) of the copolymer(lactide and glycolide) also plays a role and should be at most 20:80. More typically, the ratio is at most 18:82, for example, at most 16:84, preferably at most 14:86, or at most 12:88, in particular at most 10:90, or even at most 8:92. Often the ratio is in the range from 0.5:99,5 to 18:82, for example, in the range from 1:99 to 16:84, preferably in the range of from 1:99 to 14:86, or in the range from 1:99 to 12:88, in particular in the range from 2:98 to 10:90, or even in the range of from 2:98 to 8:92.

It is believed that the molecular weight of the copolymer is not particularly important, provided that it is at least 10000 g/mol. However, preferably the molecular weight is at least 15000 g/mol. "Molecular weight" should be understood as the average molecular weight of the polymer, as the specialist will be clear, the molecular weight of the polymer molecules in the total number of polymer molecules will be presented values distributed around the mean, for example, is represented by a Gaussian distribution. More typically, the molecular weight is in the range 10000-1000000 g/mol, for example, 15000-250000 g/mol or 20000-200000 g/mol. A particularly interesting polymers, as identified, are those which have a molecular weight of at least 20,000 g/mol, for example, at least 30,000 g/mol.

The polymer structure can be illustrated as follows (where R is selected from hydrogen, C1-6-alkyl and protective groups of hydroxyl, n is as defined above, and m, p and ran chosen so that you meet the above-mentioned provisions for the remainder(residue) of the copolymer(lactide and glycolide)):

(I)

the polymer deblocage type

(II)

the polymer triblocal type.

Note that for each of the above polymeric structures (I) and (II) units of lactide and glycolide represented by R and m, can be randomly distributed, depending on the starting materials and reaction conditions.

Also, take into account that lactide units can be either D/L or L, or D usually D/L or L.

As mentioned above, the balance(remainder) of the copolymer(lactide and glycolide), i.e., the remainder (the action) complex of the polyester, is/are hydrolytically degradable under physiological conditions, and polyalkyleneglycols the remainder is secreted, for example, from a mammalian organism. The Biodegradability can be estimated as described in the experimental section.

The polymers can, in principle, be obtained in accordance with the basic rules, well-known specialist in this field.

In principle, the polymer, where B is the residue of A (polymers deblocage type), can be obtained in the following way:

.

In principle, the polymer, where B is the residue of A (polymers triblocal type), can be obtained in the following way:

.

Unless special conditions apply, the distribution lactide units and glycolide units will be randomly distributed or gradient in each residue of the copolymer(lactide and glycolide).

Preferably, the ratio glycolide units and lactide units present in the polymer used in the frame is between the upper limit of approximately 80:20 and the lower limit of approximately 10:90, and a more preferred range is from about 60:40 to 40:60.

Preferably, the upper limit of the content of PEG is at most approximately 20 mol %, for example at most about 15 mole percent, for example, 1-15 mol %, preferably from 4 to 9 mol %, for example, about 6 mol %.

Synthesis of polymers according to this invention is additionally illustrated in international patent application publication WO 07/101443, the contents of which are incorporated herein by reference in its entirety.

In some embodiments, the implement frame, the polymer used in this invention has a vertical porous structure. In some embodiments, the implementation of vertical open porous structure has a significant number of holes in a horizontal direction between the individual pores, i.e. interconnected pores.

A reinforcing member implant

As mentioned in other places, biodegradable frame used in the implant of the present invention, is enhanced with the aim of providing the implant with the desired gain for the convenience of the implant. Accordingly, the reinforcing element may have a higher tensile strength than the frame used in the implant of the present invention.

A reinforcing member may be in the form of a second polymer different from the polymer frame. In this aspect, the reinforcing element may have the time of decomposition, which differs from the time of decomposition of the carcass.

Alternative and private aspects reinforce the second element is made from the same polymer, as the frame. In these aspects of durability is ensured by the presence of the polymer in the form of fibers or fibrous material obtained by techniques known to experts in the art, for example, by molding from a melt, electroprecizia, extrusion and other Alternative, the strength is provided by means of a connecting seams frame a polymeric material. The reinforcing element may have the time of decomposition, which differs from the time of decomposition of the carcass, but it may also be similar or close to it.

The density and volume of the reinforcing element must be sufficient to provide the necessary gain for the convenience of handling the implant and functionality. In some embodiments, implementation volume % should be sufficient to ensure a staple in the implant without fracture of the implant. However,% by volume should not be so great as to discourage flexibility or compromise the ability of the implant to support tissue regeneration. Accordingly, in some embodiments, implementation volume % of a reinforcing element is in the range of from less than about 12%, for example less than about 10%, such as less than about 8%.

"Volume %" reinforcing element can be evaluated using the analysis shows the I. The amount of gain as a percentage of the total volume occupied by the reinforcing element(elements).

In some embodiments, the implementation of enhanced implant according to this invention is elastic when wetted to saturation with liquid, as measured by the elasticity analysis, which is described in example 2.

Accordingly, the term "elastic" as used herein refers to the ability of the implant or of the frame in the size of 1-2 cm2to bend when it is taken with tweezers.

Polymers suitable for use as a reinforcing element is a polymer made of a polymer-based copolymer(lactide and glycolide) PLGA, for example, a polymer, where the molar ratio (i) of the elementary units of lactide and (ii) of the elementary parts of glycolide in the remainder of the copolymer(lactide and glycolide) in the range from 30:70 to 10:90, for example, in the range from 20:80 to 10:90, for example, about 10:90. Alternatively, you can apply polycaprolacton, polylactide, copolymers of caprolactone and lactide or biodegradable polyurethanes.

In some embodiments, the implementation of the polymer for use as a reinforcing element or combined frame and a reinforcing element surgical implant is mainly hydrophobic.

In particular, the polymers suitable for use as usilivaya the element, will decompose more slowly than synthetic biodegradable homogenous layer frame. Typically, the polymers suitable for use as a reinforcing element, will completely decompose within 2-48 months, for example, within 2-36 months, for example, for 2-24 months, for example, within 2-12 months of usein situ.

The frame may be strengthened to facilitate the doctors to deal with him in the operating room. You can apply different ways. An example of a reinforced implant according to this invention can be porous frame with welded edges and/or with welded structure on the frame. These welds provide the carcass reinforcement and can also be used as a line of incision by the surgeon, if he wants to form a framework for the defect.

The frame may be reinforced by attaching a frame to a non-woven membrane, which can be obtained by electroprecizia. The membrane preferably should be much thinner frame. The membrane can be placed in the upper part of the frame or in the center of the frame.

The alternative frame may be strengthened by the inclusion of biodegradable thread seams. Threads can be either welded, or connected at the intersection. The squares in the grid in some embodiments, the implementation have a size of at least 1 cm2. In another embodiment, the iti have no intersections, and amplification occurs due to, for example, "lithobraking" spiral seam inside the frame. Basically, the time decay for longer threads than for the frame.

Illustrative examples of figure 5 depict various patterns that can be applied to enhance the framework by incorporating biodegradable threads.

In some specific embodiments, the implementation of enhanced implant according to this invention is enhanced by the presence of a combination of biodegradable filiform welded seams and edges and/or welded structure.

Cells and other components that can be applied to the frame

In some embodiments, the implementation of this invention, the synthetic biodegradable frame is injected with a component which facilitates the cell adhesion and/or germination, for the formation of tissue inside synthetic biodegradable frame, for example, a component of the extracellular matrix of any suitable fabric, for example, components of the extracellular matrix of the bladder, bowel, skin or muscle.

In some embodiments, the implementation of this invention, the synthetic biodegradable frame injected with components and/or cells obtained from the blood, which facilitates the cell adhesion and/or germination for the formation of tissue inside synthetic biodegradable frame.

Used the second in this document "components and/or cells, derived from blood" refers to any component or cell, for example, platelets, leukocytes, serum proteins, etc. that can be obtained from a blood sample.

Accordingly, in some embodiments, the implementation of this invention, the synthetic biodegradable frame is injected with a component which facilitates the cell adhesion and/or germination, for the education of the patient tissue in situ inside synthetic biodegradable frame, for example, a component selected from the group consisting of: estrogen, derived estrogen, thrombin, powder ECM (extracellular matrix), chondroitin sulfate, hyaluronan, hyaluronic acid (ON), heparansulfate, heparan sulfate, dermatosurgery, growth factors, for example, insulin-like growth factors (IGF), such as IGF-1 or IGF-2, or transforming growth factors (TGF) such as TGF-alpha or TGF-beta, or fibroblast growth factors (FGF) such as FGF-1 and FGF-2, or platelet-derived growth factors (PDGF), such as PDGF-AA, PDGF-BB or PDGF-AB, or mechanical growth factor (MGF), or nerve growth factor (NGF), or human growth hormone (HGH); fibrin, fibronectin, elastin, collagen, for example, collagen type I and/or type II, type III, type IV, type V or type VII, gelatin and aggrecan, or any other suitable components of the extracellular matrix.

In one individual in whom the version of the implementation in synthetic biodegradable frame included hyaluronic acid. In one embodiment, the hyaluronic acid is present in the synthetic biodegradable frame in a ratio of from about 0.1 to about 15 wt.%.

The following specific embodiment, the synthetic biodegradable frame included frame dermatologit. In one embodiment, dermatologit present in synthetic biodegradable frame in a ratio of from about 0.1 to about 15 wt.%.

Discussed above, compounds that enhance cell migration and/or tissue regeneration, can be added to the treatment to the porous frame structure in the form of pure compounds or in the form of solutions. Alternatively, you can add them coated or encapsulated in the form of nano - or microparticles.

In some embodiments, the implementation of this invention, the synthetic biodegradable frame is injected with a suspension of cells or tissue from mammals, for example, human stem cells or other cells or tissues, e.g. muscle cells, fibroblasts and endothelial cells or muscle tissue, for example, cells or tissues obtained from smooth, skeletal or cardiac muscle. Usually, this can be a suspension of muscle tissue, for example, biopsies or dedicated muscle fibers obtained from the patient. Alternatively it can bitmachine cells or components, derived from muscle cells, proliferatingin vitro.Muscle suspension can be applied to the surface of the frame prior to implantation or at the same time. Muscle suspension used according to aspects of the present invention, typically seeded with a density in the range of 1-100 mg muscle suspension cm2frame layer. Muscle fibers are separated from the biopsy or dissection of the muscles by using, for example, scalpels, or dissolution muscles by enzymatic treatment, for example, collagenase, obtaining the individual fibers with the satellite cells.

In other embodiments, the implementation of this invention, the synthetic biodegradable frame is injected with a suspension of components produced by muscle cells, together with these muscle cells.

In one embodiment, a cell suspension or tissue of a mammal derived or descended from the living species of mammal, which requires the exercise of medical applications, i.e.it is autogenic.

Cells or tissue can also be homologous, i.e., compatible with the tissue to which they are applied, or can be obtained from multipotent or even pluripotent stem cells, for example, in the form of allogeneic cells. In one embodiment, the cells or tissue are not autologous. In one VA is iante implementation of non-homologous cells. In one embodiment, cells may be allogeneic, from another specie, or xenogenic, that is obtained from a species different from the treated organism. Allogeneic cells can be differentiated cells, cells predecessors or cells derived from multipotent (e.g., embryonic, or a combination of embryonic and adult specialized cells or cells), pluripotent stem cells derived from umbilical cord blood, adult stem cells and others), constructed cells either by replacement, insertion, or adding genes from other cells or gene constructs, the application of the transfer of the nucleus of differentiated cells into embryonic stem cells or multipotent stem cells, for example, stem cells derived from umbilical cells.

In one embodiment, the method of the present invention also encompasses the use of stem cells and cells derived from stem cells, and the cells can be preferably obtained from the same species as exposed individual treatment of a mammal, for example, human stem cells or stem from them cells.

The mammalian cells used in this invention can be supplied in the form of a cell suspension or tissue perfo is tatov. Tissue explants can be directly taken from other parts of the species of mammal and, therefore, can be in the form of tissue transplants, for example, the graft of muscle tissue taken from the large muscles of the mammal.

Cells of smooth or skeletal muscles of the person or, alternatively, fibroblasts and other connective tissue cell types, introduced in synthetic biodegradable frame will be particularly preferred. However, it is assumed that it is also possible to use stem cells or any other suitable precursor cells that can become or to make muscle and/or connective tissue cells. Usually cells are used in this application, there are a sufficient number of cells in order to cause regeneration or recovery of the target tissue or defect, for example, from approximately 0.1×104up to approximately 10×106cells/cm2or 0.1×106cells/cm2up to approximately 10×106cells/cm2.

In some embodiments, the implementation of the muscle cells used in this invention are in the form of cell suspensions or tissue explants.

In some embodiments, the implementation of mammalian cells applied on synthetic biodegradable frame according to this invention, the nano is drawn in the amount of from about 0.1×10 4cells to approximately 10×106cells / cm2synthetic biodegradable frame.

In some embodiments, the implementation of mammalian cells or tissue explants put on synthetic biodegradable frame according to this invention during a medical application, for example, during surgical operations. It should be understood that the surgeon can take tissue explants for use in the methods of the present invention before or during the operation.

In some embodiments, the implementation of mammalian cells or tissue explants are cultivated in synthetic biodegradable frame to medical applications, for example, operations for at least 1 day, at least 3 days, at least 1 week, for example, at least 2 weeks, for example, at least 3 weeks, for example, at least 6 weeks.

Surgical technique and patient

“A living individual mammal” means any living individual mammal, suitable for blending synthetic reinforced biodegradable frame according to this invention, and is, preferably, a human being, typically, the patient. However, the methods of the present invention may also be applicable to other mammals such as domestic animals, including dogs, cats and horses.

Methods of blending synth is political biodegradable frame with high strength according to this invention may be made as, or during a surgical method of operation, for example, a method of endoscopic, laparoscopic or other minimally invasive surgery, or conventional or open surgery.

In specific aspects of this invention, the overlay reinforced synthetic biodegradable frame according to this invention can be used with any medical condition requiring reconstructive surgery that require strengthening in the area of surgical operation.

In specific aspects of this invention, the overlay reinforced synthetic biodegradable frame according to this invention is used during the surgery prolapse, for example, prolapse of pelvic organs, also called pelvic reconstructive surgery or surgery of stress incontinence.

It should be understood that reinforced synthetic biodegradable frame according to this invention can be used in reconstructive surgery involving the diaphragm, the area of the pelvic floor and the abdominal cavity. However, it is assumed that reinforced synthetic biodegradable frame according to this invention can be applied in surgical reconstructions of other fascial components of the body of a mammal, including a dense fibrous connective tissue that permeates and surrounds muscles, bones, organs, nerves and blood SOS the water body.

Accordingly, reinforced synthetic biodegradable frame according to this invention can be applied in the treatment of increased pressure in any anatomical cavity, constrictive pericarditis, hemopneumothorax, hemothorax, injury of the Dura and various hernias, including a framework for prevention of hernias in all surgical operations on the abdominal cavity.

Hernias are medical conditions for which you can be shown the implant according to this invention. Used in this document, the term “hernia” includes abdominal hernia, diaphragmatic hernia and hiatal hernia repair (for example, paraesophageal hernia of the stomach), pelvic hernia, for example, locking hernia, ventral hernia, herniated nucleus pulposus herniation, intracranial hernia and hernia Spigelia line.

Types of surgical operations usually associated with pelvic reconstructive surgery includes laparoscopic assisted vaginal hysterectomy, complete laparoscopic hysterectomy, vaginal hysterectomy, laparoscopic suspension of the vaginal vault, laparoscopic sacrocolpopexy, laser vaginal reconstruction, construction laser vaginoplasty, vaginal approach to the treatment of prolapse introduction grid, procedures maintain and lapar scopacasa paravaginal repair.

Used in this document the expression "prolapse of pelvic organs" refers to any medical condition, including prolapse through the wall of the pelvis. Other expressions used and included in this definition, is uterine prolapse, genital prolapse, uterine-vaginal prolapse, pelvic relaxation, dysfunction of the pelvic floor, urinary prolapse, prolapse of vaginal walls, cystocele, and prolapse of the bladder, urethrocele, enterocele, rectocele, vaginal vault prolapse, prolapse of the small intestine, a prolapsed uterus or prolapse of the urethra.

One important aspect of this invention relates to a method of treatment or partial withdrawal symptoms of prolapse of the connective tissue, for example, prolapse of pelvic organs, living species of mammal, e.g. human, the method includes the step of blending synthetic biodegradable frame with high strength according to this invention to the site of the defect or place required surgery.

As described above, another important aspect of this invention relates to synthetic biodegradable frame with the reinforcing element(elements) according to this invention; for use as an implant.

In one embodiment, this reinforced synthetic biodegradable frame according to this invention for use in the icenii or partial withdrawal symptoms defect of connective tissue in live animals of the mammal, for example, human.

In some specific embodiments, the implementation of the cells receive from the living species of mammal in operation and applied to reinforced synthetic biodegradable frame before and/or simultaneously and/or after the application of a reinforced synthetic biodegradable scaffold at the site of the defect. The inventors of the present invention, it is expected that this will contribute to the absorption and tolerability of a reinforced synthetic biodegradable frame, and increase the growth and reconstruction of tissues of living animals of the mammal at the site of the operation, and, thus, will increase the speed of recovery under treatment of a mammal, for example, the patient is human. In some embodiments, the implementation of the muscle cells are in suspension, which is applied on the surface or in the frame in connection with the operation and implantation of the implant according to this invention. Used muscle, the suspension is usually sown with a density in the range of 1-100 mg muscle suspension cm2frame layer. Muscle fibers can be distinguished from biopsy or dissection of the muscles by using, for example, scalpels, or dissolution muscles using an enzymatic treatment, for example, collagenase, obtaining the individual fibers with the satellite cells. It should be understood that muscle, the drug can be taken from the same with the constituent of the patient, which receives the implant, i.e., autologous preparation.

In some embodiments, the implementation process of this invention perform as part of a surgical operation, for example, endoscopic, laparoscopic or other minimally invasive surgical procedures, as well as normal or extensive open surgery.

In some embodiments, the implementation process of this invention perform as part of a reconstructive surgery.

Synthetic biodegradable reinforced frame according to this invention can be attached to the fascia sutures, pins and/or fabric glue different types. Preferably such means of attachment are also biodegradable.

Kit

As described elsewhere, this invention also provides a kit for the treatment or partial withdrawal symptoms of prolapse in the living species of mammal, the kit includes a reinforced synthetic biodegradable frame and instructions for use of this reinforced synthetic biodegradable frame.

Also provided kits of parts to support the expansion and regeneration of living tissue in a subject, for example, a subject with a medical prolapse, for example, or rectal prolapse of pelvic organs, or hernia, this set includes the t biodegradable surgical implant, including synthetic biodegradable frame, and the component, suitable for crushing or destruction of the tissue sample, or, alternatively, a sample of autologous cells or tissue explants for use in the methods of the present invention.

Specific embodiments of this invention

In some embodiments of this invention, the synthetic biodegradable homogenous layer frame is hydrophilic.

In some embodiments of this invention, the synthetic biodegradable homogenous layer frame has the ability within 5 minutes, for example, for 2 minutes at 30°C, to absorb water in an amount of at least 10%, for example at least 20%, e.g. at least 30%, such as at least 50% of the frame.

In some embodiments, the implementation of the biodegradable surgical implant according to this invention has a volume % of the specified reinforcing element is less than 40%, e.g. less than 30%, e.g. less than 20%, such as less than 15%, such as less than 12% of the implant.

It should be clear that the balance of strength, elasticity and Biodegradability combination of a reinforcing element and the frame material will be required depending on the specific indications treated implant. Accordingly, the higher is rocheste will be required to restore pelvic, than, for example, for the treatment of urinary incontinence.

In some embodiments of this invention, the synthetic biodegradable homogenous layer frame shows the elongation at break in the range of about 10-200%, for example, in the range of about 30-100%, for example, in the range of about 30-70%, for example, in the range of about 30-60%.

In some embodiments of this invention a surgical implant shows the elongation at break in the range of about 20-1000%, for example, in the range of about 20-800%, for example, in the range of about 20-500%, for example, in the range of about 20-400%, for example, in the range of about 20-300%.

In some embodiments of this invention, the synthetic biodegradable homogenous layer frame shows the tensile strength in the range of about 5-40 pounds per square inch, for example, in the range of about 8-30 pounds per square inch, for example, in the range of about 8 to 20 pounds per square inch, for example, in the range of about 8-16 pounds per square inch, for example, in the range of about 8-14 pounds per square inch.

In some embodiments of this invention a surgical implant demonstrates within the tensile strength in the range of about 300-50000 pounds per square inch, for example, in the range of about 500-30000 pounds per square inch, for example, in the range of about 1000-20000 pounds per square inch, for example, in the range of about 1000-10000 pounds per square inch, for example, in the range of about 5000-10000 psi, or in the range of about 1000-8000 pounds per square inch.

In some embodiments of this invention the frame, the material exhibits a tensile strength in the range of about 300-50000 pounds per square inch, for example, in the range of about 500-30000 pounds per square inch, for example, in the range of about 1000-20000 pounds per square inch, for example, in the range of about 1000-10000 pounds per square inch, for example, in the range of about 1000-8000 pounds per square inch.

In some embodiments of this invention, the synthetic biodegradable homogenous layer frame demonstrates flexibility when wetted to saturation with liquid.

In some embodiments of this invention, the synthetic biodegradable homogenous layer frame has otkrytostju structure with a size in the range of 30-200 μm.

In some embodiments of this invention, the synthetic biodegradable homogenous layer frame is basically the om verticale.pamintul structure.

In some embodiments of this invention, the synthetic biodegradable homogenous layer frame has otkrytostju structure with interconnected pores.

In some embodiments of this invention, the synthetic biodegradable homogenous layer frame obtained by lyophilization.

In some embodiments, the implementation of the present invention, the biodegradable reinforcing element is based fibers and/or filaments with a thickness of approximately 10 nm-1000 μm, for example, in the range of about 10 nm to 800 μm, for example, in the range of about 10 nm-500 μm.

In some embodiments, the implementation of the present invention, the biodegradable reinforcing element is a layer made of a woven material, knitted material, mesh, non-woven felt of filaments or staple fibers.

In some embodiments, the implementation of the present invention, the biodegradable reinforcing element is a layer made of a woven material, knitted material, mesh, non-woven felt of filaments or staple fibers, where the layer has a thickness of 30 μm-5 mm, for example, 3-5 mm, for example, 1-4 mm.

In some embodiments of this invention, the synthetic biodegradable homogenous layer frame is completely biodegradable within 1-48 months, for example, 4-36, for example, 6-24 and the and 1-12 months in situ application.

In some embodiments, the implementation of the present invention, the biodegradable reinforcing element promotes cell attachment and germination of cells derived from living tissue of a specified entity or from the application of cellular or tissue explants.

In some embodiments, the implementation of the present invention, the biodegradable reinforcing element is completely biodegradable within 1-12 months, for example, in the range of 2-12 monthsin situapplication.

In some embodiments, the implementation of the present invention reinforced biodegradable element is made of polymer-based copolymer(lactide and glycolide) PLGA, e.g., polymer, where the molar ratio (i) of the elementary units of lactide and (ii) of the elementary parts of glycolide in the remainder of the copolymer(lactide and glycolide) is in the range from 90:10 to 10:90, for example, in the range from 80:20 to 10:90, for example, about 10:90.

In some embodiments of this invention, the synthetic biodegradable homogenous layer frame is a polymer with the General formula:

A-O-(CHR1CHR2O)n-B

where

A is a residue of a copolymer(lactide and glycolide) with a molecular weight of at least 4000 g/mol, the molar ratio (i) of the elementary units of lactide and (ii) of the elementary parts of glycolide in the rest of the sprinklers is the measure(lactide and glycolide) is in the range from 80:20 to 10:90;

B is either a remainder of the copolymer(lactide and glycolide), as defined for A, or it is selected from the group consisting of hydrogen, C1-6-alkyl and protective groups of hydroxyl,

one of R1and R2within each elementary group -(CHR1CHR2O)- is selected from hydrogen and methyl, and the other of R1and R2within the same elementary link -(CHR1CHR2O) is hydrogen,

n represents the average number of elementary units -(CHR1CHR2O) in the polymer chain and is an integer in the range 10-1000; and where the

the molar ratio of (iii) polyalkyleneglycol elementary links(CHR1CHR2O) to the combined amount of (i) the elementary units of lactide and (ii) of the elementary parts of glycolide in the residue(remainder) of the copolymer(lactide or glycolide) is at most 20:80;

and where the molecular weight of the copolymer is at least 10000 g/mol, preferably at least 15000 g/mol.

In some embodiments, the implementation of the present invention as R1 and R2 in each elementary element is hydrogen.

In some embodiments of this invention, B is a residue of a copolymer(lactide or glycolide), as defined for A.

In some embodiments of this invention, B represents a C1-6-alkyl.

In some of the options which the implementation of the present invention B is a protective group of hydroxyl.

In some embodiments of this invention, B is a hydroxy-group.

In some embodiments of this invention, the weight percentage (iii) polyalkyleneglycol elementary links(CHR1CHR2O) to the combined amount of (i) the elementary units of lactide and (ii) of the elementary parts of glycolide in the residue(remainder) of the copolymer(lactide or glycolide) is in the range of 4%-10% wt./weight.

In some embodiments of this invention, the synthetic biodegradable homogenous layer frame obtained by lyophilization of a solution comprising a biodegradable polymer in solution.

In some embodiments of this invention, a reinforcing member made of biodegradable fibers and/or threads.

In some embodiments of this invention the reinforcing element looks like a structure selected from the group consisting of triangles, circles, connecting waves, mesoeconomics waves and overlapping waves.

In some embodiments of this invention, a reinforcing member made of welded seams on synthetic biodegradable homogenous layer frame, for example, welding seams provided in the form of a square and hexagonal patterns or along the edge of the implant.

In some valentinorossini of this invention, the synthetic biodegradable homogenous layer frame is a polymer with a molecular weight greater than about 1 kDa, for example, from about 1 kDa to about 1000000 kDa, for example from 25 kDa to 100 kDa.

In some embodiments, the implementation of the present invention, the implant further comprises in the frame of the one or more components that facilitate cell adhesion and/or germination to restore tissue, for example, a component selected from the group consisting of: estrogen, derived estrogen, thrombin, ECM powder, chondroitin sulfate, hyaluronan, hyaluronic acid (HA), heparansulfate, heparan sulfate, dermatosurgery, growth factors, fibrin, fibronectin, elastin, collagen, such as collagen type I and/or type II, gelatin and aggrecan or any other suitable component of the extracellular matrix.

In some embodiments, the implementation of the present invention, the implant includes a frame, one or more components selected from the group consisting of growth factors, for example, insulin-like growth factors (IGF), such as IGF-1 or IGF-2, or transforming growth factors (TGF) such as TGF-alpha or TGF-beta, or fibroblast growth factors (FGF) such as FGF-1 or FGF-2, or platelet-derived growth factors (PDGF), such as PDGF-AA, PDGF-BB or PDGF-AB, or nerve growth factor (NGF), or human growth hormone (hGH) and mechanical growth factor (MGF).

In some embodiments, the implementation of the present invention, the pulse is antat contains the specified frame the sample of cells or tissue explants.

In some embodiments of this invention, the implant is formed in the form of a tube and/or contains a valve and/or pocket, convenient for applying the suspension of the sample of cells or tissue explants on the implant.

In some embodiments, the implementation of the present invention, the implant includes two or more separate parts of the synthetic biodegradable homogenous layers of the framework, for example, 3, 4, 5 or 6 pieces of synthetic biodegradable homogenous layers of the frame attached to the reinforcing element, for example, the grid of the other polymer.

In some embodiments, the implementation of the present invention, the implant contains two or more, for example, 4 or 6 arms or lugs for attachment to structures in the implantation site, for example, in the pelvic region.

In some embodiments of the method according to this invention, the subject is suffering from a medical prolapse, for example, prolapse of pelvic organs or hernia.

In some embodiments of the method according to this invention the method includes the implantation of the specified biodegradable surgical implant together with a sample of cells or tissue explants specified subject at the site of implantation.

In some embodiments, the implementation of a sample of cells or tissue explants collected from a patient in the operating room and placed on impl ntat at the site of implantation during surgery. Alternatively, cells or tissue explants used in conjunction with the implant, were taken from the patient before surgery. In another embodiment, the implant and cells or tissue explants provide in the form of a set and used together during a surgical operation.

In some embodiments, the implementation of the methods according to this invention, the cells or tissue explants are autologous, homologous (allogeneic or xenogenic in origin relative to the cells specified the living tissue of the subject. In some embodiments of the method according to this invention, the cells or tissue explants are autologous to the subject, with the implant.

In some embodiments, the implementation of the methods according to this invention, the synthetic biodegradable frame is a homogeneous layer.

In some embodiments, the implementation of the biodegradable surgical implant according to this invention used in the methods of the present invention.

In some embodiments, the implementation of the methods according to this invention, the subject is suffering from a medical prolapse, for example, prolapse of pelvic organs, or hernia, or incontinence of urine.

In some embodiments, the implementation of the methods according to this invention, the number of cells specified in the applicable sample of cells or tissue Explant which is in the range from about 0.1×10 4cells to approximately 10×106cells / cm2the implant.

In some embodiments, the implementation of the methods according to this invention tissue explants obtained from muscle tissue, stem cells, e.g. stem cells, capable of differentiation in cultured myoblasts or fibroblasts; or combinations thereof.

In some embodiments, the implementation of the methods according to this invention, the cells or tissue explants obtained from a person.

In some embodiments, the implementation of the methods according to this invention, the cells or tissue explants before implantation cultured in vitro for a certain amount of time or within a specified synthetic biodegradable homogenous layer of the framework.

In some embodiments, the implementation of the methods according to this invention, the cells or tissue explants prior to implantation is not cultivated in vitro.

In some embodiments, the implementation of the methods according to this invention, the cells or tissue explants are collected and used according to the method in the operating room.

In some embodiments, the implementation of the methods according to this invention the method further includes applying to the specified biodegradable surgical implant composition comprising a component which facilitates the cell adhesion and/or sprouting of regenera the AI fabric, for example, a component selected from the group consisting of estrogen, derived estrogen, thrombin, ECM powder, chondroitin sulfate, hyaluronan, hyaluronic acid (HA), heparansulfate, heparan sulfate, dermatosurgery, growth factors, fibrin, fibronectin, elastin, collagen, such as collagen type I and/or type II, gelatin and aggrecan, or any suitable component of the extracellular matrix.

In some embodiments, the implementation of the methods according to this invention the method further includes applying to the specified biodegradable surgical implant composition comprising a component selected from the group consisting of growth factors, for example, insulin-like growth factors (IGF), such as IGF-1 or IGF-2, or transforming growth factors (TGF) such as TGF-alpha or TGF-beta, or fibroblast growth factors (FGF) such as FGF-1 or FGF-2, or platelet-derived growth factors (PDGF), such as PDGF-AA, PDGF-BB or PDGF-AB, or nerve growth factor (NGF), or human growth hormone (hGH) and mechanical growth factor (MGF).

In some embodiments, the realization sets in this invention include a component that is suitable for shredding or destruction, this component includes holes or mesh for crushing specified tissue sample by application of pressure, whereby the tissue image is C is forced through the specified mesh or holes.

In some embodiments, the realization sets in this invention include a component that is suitable for shredding or destruction, on the basis of the crusher, ultrasonic treatment, high pressure or physical impact of knives or other tools, one example is homogenizer with rotating knives.

Listed embodiments of the present invention

Biodegradable surgical implant for support, enhancement and regeneration of living tissue of a subject, including:

synthetic biodegradable homogenous layer frame;

one or more biodegradable reinforcing elements;

characterized in that said synthetic biodegradable homogenous layer frame is hydrophilic.

Biodegradable surgical implant according to a variant implementation 1, where the specified synthetic biodegradable homogenous layer frame has the ability within 5 minutes, for example, for 2 minutes at 30°C, to absorb water in an amount of at least 10%, for example at least 20%, e.g. at least 30%, such as at least 50%, of the volume of the framework.

Biodegradable surgical implant according to any of embodiments 1 or 2, where the % by volume of the specified reinforcing element is less than 40% of the implant.

Bioral the proposed surgical implant according to any of embodiments 1-3, where the specified synthetic biodegradable homogenous layer frame shows the percentage of elongation at break in the range of about 10-200%, for example, in the range of about 30-100%, for example, in the range of about 30-70%, for example, in the range of about 30-60%.

Biodegradable surgical implant according to any of embodiments 1-4, where the specified surgical implant shows the percentage of elongation at break in the range of about 20-1000%, for example, in the range of about 20-800%, for example, in the range of about 20-500%, for example, in the range of about 20-400%, for example, in the range of about 20-300%.

Biodegradable surgical implant according to any of embodiments 1-5, where specified synthetic biodegradable homogenous layer frame shows the tensile strength in the range of about 5-40 pounds per square inch, for example, in the range of about 8-30 pounds per square inch, for example, in the range of about 8 to 20 pounds per square inch, for example, in the range of about 8-16 pounds per square inch, for example, in the range of about 8-14 pounds per square inch.

Biodegradable surgical implant according to any of embodiments 1-6, where the specified surgical implant is displayed on the t the tensile strength in the range of about 300-50000 pounds per square inch, for example, in the range of about 500-30000 pounds per square inch, for example, in the range of about 1000-20000 pounds per square inch, for example, in the range of about 1000-10000 pounds per square inch, for example, in the range of about 1000-8000 pounds per square inch.

Biodegradable surgical implant according to any of embodiments 1-7, where the specified synthetic biodegradable homogenous layer frame demonstrates flexibility when wetted to saturation with liquid.

Biodegradable surgical implant according to any of embodiments 1-8, where the specified synthetic biodegradable homogenous layer frame has otkrytostju structure with a size in the range of 30-200 μm.

Biodegradable surgical implant according to any of embodiments 1-9, where the specified synthetic biodegradable homogenous layer frame has a generally verticale.pamintul structure.

Biodegradable surgical implant according to any of embodiments 1-10, where specified synthetic biodegradable homogenous layer frame has otkrytostju structure with interconnected pores.

Biodegradable surgical implant according to any of embodiments 1-11, where the specified synthetic biodegradable homogenous layer frame recip is n using lyophilization.

Biodegradable surgical implant according to any of embodiments 1-12, where specified biodegradable reinforcing element is based fibers and/or filaments with a thickness of approximately 10 nm-1000 μm, for example, in the range of about 10 nm to 800 μm, for example, in the range of about 10 nm-500 μm.

Biodegradable surgical implant according to any of embodiments 1-13, where the specified biodegradable reinforcing element is a layer made of a woven material, knitted material, mesh, non-woven felt of filaments or fibers.

Biodegradable surgical implant according to a variant implementation of the 14, where the specified layer has a thickness of 30 μm-5 mm, for example, 3-5 mm, for example, 1-4 mm.

Biodegradable surgical implant according to any of embodiments 1-15, where the specified synthetic biodegradable homogenous layer frame is completely biodegradable within 1-12 monthsin situapplication.

Biodegradable surgical implant according to any of embodiments 1-16, where specified biodegradable reinforcing element promotes cell attachment and germination of cells derived from living tissue of a specified entity or from the application of cellular or tissue explants.

Biodegradable surgical implant according to any of the options on the westline 1-17, where specified biodegradable reinforcing element is completely biodegradable within 1-12 monthsin situapplication.

Biodegradable surgical implant according to any of embodiments 1-18, where specified biodegradable reinforcing element is made of polymer-based copolymer(lactide and glycolide) PLGA, e.g., polymer, where the molar ratio (i) of the elementary units of lactide and (ii) of the elementary parts of glycolide in the remainder of the copolymer(lactide and glycolide) is in the range from 90:10 to 10:90, for example, in the range from 80:20 to 10:90, for example, about 10:90.

Biodegradable surgical implant according to any of embodiments 1-19, where the specified synthetic biodegradable homogenous layer frame is a polymer with the General formula:

A-O-(CHR1CHR2O)n-B

where

A is a residue of a copolymer(lactide and glycolide) with a molecular weight of at least 4000 g/mol, the molar ratio (i) of the elementary units of lactide and (ii) of the elementary parts of glycolide in the remainder of the copolymer(lactide or glycolide) is in the range from 80:20 to 10:90;

B is either a remainder of the copolymer(lactide and glycolide), as defined for A, or selected from the group consisting of hydrogen, C1-6-alkyl and protective hydroxyl groups;

one of R1and R2 within each elementary group -(CHR1CHR2O)- is selected from hydrogen and methyl, and the other of R1and R2within the same elementary link -(CHR1CHR2O)- is hydrogen;

n represents the average number of elementary units -(CHR1CHR2O) in the polymer chain and is an integer in the range 10-1000; and where the

the molar ratio (iii) polyalkyleneglycol elementary links(CHR1CHR2O) to the combined amount of (i) the elementary units of lactide and (ii) of the elementary parts of glycolide in the residue(remainder) of the copolymer(lactide or glycolide) is at most 20:80;

and where the molecular weight of the copolymer is at least 10000 g/mol, preferably at least 15000 g/mol.

Biodegradable surgical implant according to a variant implementation of 20, where R1 and R2 each elementary element is hydrogen.

Biodegradable surgical implant according to a variant implementation of the 20 or 21, where B is the remainder of the copolymer(lactide or glycolide), as defined for A.

Biodegradable surgical implant according to any of embodiments 20-22, where B is C1-6-alkyl.

Biodegradable surgical implant according to any one of embodiments 20 to 23, where B is a protective group of hydroxyl.

Biodegradable hir is rychecky the implant according to any one of embodiments 20 to 23, where B is a hydroxy-group.

Biodegradable surgical implant according to any of embodiments 20-25, where the weight percentage (iii) polyalkyleneglycol elementary links(CHR1CHR2O) to the combined amount of (i) the elementary units of lactide and (ii) of the elementary parts of glycolide in the residue(remainder) of the copolymer(lactide or glycolide) is in the range of 4%-10% wt./weight.

Biodegradable surgical implant according to any of embodiments 1-26, where the specified synthetic biodegradable homogenous layer frame obtained by lyophilization of a solution comprising a biodegradable polymer in solution.

Biodegradable surgical implant according to any of embodiments 1-27, where the specified reinforcing element is made of a biodegradable suture materials and/or threads.

Biodegradable surgical implant according to a variant implementation 28, where the specified reinforcing element looks like a structure selected from the group consisting of: triangles, circles, connecting waves, mesoeconomics waves and overlapping waves.

Biodegradable surgical implant according to any of embodiments 1-29, where the specified reinforcing element is made of welded seams on synthetic biodegradable homogenous layer frame, for example, swaro the different seams, made in the form of square and hexagonal patterns or along the edge of the implant.

Biodegradable surgical implant according to any one of embodiments 1-30, where the specified synthetic biodegradable homogenous layer frame is a polymer with a molecular weight of more than about 1 kDa, for example, from about 1 kDa to about 1000000 kDa, for example from 25 kDa to 100 kDa.

Biodegradable surgical implant according to any one of embodiments 1-31, with the specified implant further comprises in the specified frame, one or more components that facilitate cell adhesion and/or germination for tissue regeneration, for example, a component selected from the group consisting of: estrogen, derived estrogen, thrombin, ECM powder, chondroitin sulfate, hyaluronan, hyaluronic acid (HA), heparansulfate, heparan sulfate, dermatosurgery, growth factors, fibrin, fibronectin, elastin, collagen, such as collagen type I and/or type II, gelatin and aggrecan or any other suitable component of the extracellular matrix.

Biodegradable surgical implant according to any of embodiments 1-32, and the implant further comprises in the specified frame, one or more components selected from the group consisting of growth factors, such as the er, insulin-like growth factors (IGF), such as IGF-1 or IGF-2, or transforming growth factors (TGF) such as TGF-alpha or TGF-beta, or fibroblast growth factors (FGF) such as FGF-1 or FGF-2, or platelet-derived growth factors (PDGF), such as PDGF-AA, PDGF-BB or PDGF-AB, or nerve growth factor (NGF), or human growth hormone (hGH) and mechanical growth factor (MGF).

Biodegradable surgical implant according to any one of embodiments 1-33, and the implant further comprises in the specified frame the sample of cells or tissue explants.

The way of support, enhancement and regeneration of living tissue in the subject with the medical prolapse, for example, prolapse of pelvic organs, or hernia, while this method involves the implantation of a biodegradable surgical implant comprising a synthetic biodegradable homogenous layer frame together with the sample of cells or tissue explants, a specified subject in the area specified prolapse or hernia.

The way of support, enhancement and regeneration of living tissue in the subject, while this method involves the implantation of a biodegradable surgical implant according to any one of embodiments 1-34 specified entity.

The method according to option exercise 36, where the specified subject is suffering from a medical prolapse, for example, prolap whom and organs of the pelvis, or hernia.

The method according to the options exercise 36 or 37, where the method includes implanting the specified biodegradable surgical implant together with a sample of cells or tissue explants specified subject at the site of implantation.

The method according to the options exercise of 35 or 38, where these cells or tissue explants are autologous, homologous (allogeneic or xenogenic in origin relative to the cells specified the living tissue of the subject.

Method according to any one of embodiments 35 or 38-39, where the number of cells in the specified applied the sample of cells or tissue explants is in the range from about 0.1×104cells to about 10×106cells / cm2the implant.

Method according to any one of embodiments 35 or 38-40, where tissue explants are those of muscle tissue, stem cells, e.g. stem cells, capable of differentiation in cultured myoblasts or fibroblasts; or combinations thereof.

Method according to any one of embodiments 35 or 38-41, where these cells or tissue explants obtained from a person.

Method according to any one of embodiments 35 or 38-42, where these cells or tissue explants cultivated in vitro in teeniefiles amount of time on or inside these synthetic biodegradable homogenous layers of the carcass prior to implantation.

Method according to any one of embodiments 35 to 43, the method further includes applying to the specified biodegradable surgical implant composition comprising a component which facilitates the cell adhesion and/or germination for tissue regeneration, for example, a component selected from the group consisting of: estrogen, derived estrogen, thrombin, ECM powder, chondroitin sulfate, hyaluronan, hyaluronic acid (HA), heparansulfate, heparan sulfate, dermatosurgery, growth factors, fibrin, fibronectin, elastin, collagen, such as collagen type I and/or type II, gelatin and aggrecan or any suitable component of the extracellular matrix.

Method according to any one of embodiments 35-44, the method further includes applying to the specified biodegradable surgical implant composition comprising a component selected from the group consisting of growth factors, for example, insulin-like growth factors (IGF), e.g., IGF-1 or IGF-2, or transforming growth factors (TGF) such as TGF-alpha or TGF-beta, or fibroblast growth factors (FGF) such as FGF-1 or FGF-2, or platelet-derived growth factors (PDGF), for example, PDGF-AA, PDGF-BB or PDGF-AB, or nerve growth factor (NGF), or human growth hormone (hGH) and mechanical growth factor (MGF).

A method of obtaining a biodegradable surgical implant according to any one of embodiments 1-34, the method simultaneously includes sequential steps:

get these synthetic biodegradable homogenous layers of the carcass;

obtain and activate the specified one or more biodegradable reinforcing details in these synthetic biodegradable homogenous layers of the carcass;

optional inclusion of one or more components, as defined in any one of embodiments 32 to 34.

The set includes:

biodegradable surgical implant according to any one of embodiments 1-34;

a sample of cells or tissue explants; and

optional instructions for use of the method of support, enhancement and regeneration of living tissue in the subject with the medical prolapse, for example, rectal prolapse or prolapse of pelvic organs, or hernia, while this method involves the implantation of a specified biodegradable surgical implant together with a sample of cells or tissue explants specified subject in the area specified prolapse or hernia.

The implant according to any one of embodiments 1-34 for use as a drug.

The implant according to any one of embodiments 1-34 for the use in the treatment of diseases, associated with prolapse of the pelvic organs and hernia.

EXAMPLES

Example 1

Welds

Frames are layers of lyophilised structures. They are made by freezing/lyophilization of a solution of the polymer. This results in otkrytostju porous structure in which pores are oriented in the direction of freezing. Such orientation can be seen in Fig.1.

This orientation means that the material has a very low tensile strength. To strengthen the material type welds. The material is compressed/melted, thus losing the structure described above, and acquiring strength. This means that the material can withstand tension when sewing and handling them. This welding may be performed using pulse welding, laser welding or similar thermal processing.

These welds can be added only at the edges or in the mesh structure for greater strength. Having a network structure, you can crop the frame size without loss of strength.

2-layer frame with different times of decomposition

It may be desirable to have a layer component, which supports cell growth and is rapidly degraded, and the layer which remains longer for support and strength. The frame may be 2-ply is a structure, a cast/lyophilization 2-layer structure and the subsequent joining her for strength, or joining layer, which decomposes more slowly in the mesh structure and re-attaching layer that decomposes faster to it (for example, by welding). The combination of polymers that will give fast/slow degradable composition, known to experts in this technology. It can also be made by welding of the frame, which decomposes faster to support grid, which decomposes more slowly.

Installation of the frame on the substrate layer

Another way to strengthen the frame is attaching the frame to a more durable substrate material.

The substrate material may be biodegradable nonwoven fibrous material, for example, a coating of electrospraying biodegradable complex polyester. Material the frame is made from a material that decomposes quickly in the body (8 weeks). It may be desirable to have a substrate material with a longer degradation time of, for example, 6 months or longer. Examples of materials with a longer degradation time of PLGA are content glycolide >50 mol. %, PLGA with a lactide content >50 mol. %, poly(D,L-lactide), poly(L-lactide), poly(caprolactone), poly(3-hydroxybutyrate). Other suitable materials, which is s can be applied, easily select and apply a specialist in this technology.

The frame can be fixed on the support by welding.

Strengthening threads

By incorporating mesh of biodegradable sutures can strengthen the frame structure.

EXPERIMENTAL PART

Frames

MPEG-PLGA 2-30 kDa with a molar ratio G:L 50:50 diluted to 4 wt%./volume of dioxane. The bottom 10×10 cm Alu-shaped cover with dioxane and the form is cooled to -5°C. When the dioxane freezes, 27 ml of the polymer solution is cast on top of the frozen layer, and form again cooled to -5°C. the Frozen polymer solution then lyophilizer and dried structure is kept for 5 days in a vacuum desiccator.

2-layer frame with 2 different times of decomposition

MPEG-PLGA 2-30 kDa with a molar ratio G:L 50:50 diluted to 4 wt%./volume of dioxane.

PDLLA is diluted to 4% wt./volume of dioxane.

The bottom 10×10 cm Alu-shaped cover with dioxane and the form is cooled to -5°C. When frozen dioxane, and 13.5 ml of a solution of PDLLA cast on top of the frozen layer, and form again cooled to -5°C. When frozen, 13,5 ml MPEG-PLGA cast on top of the frozen layer, and form again cooled to -5°C. the Frozen two-layer structure is then lyophilizer. The frame now consists of 2 layers with different times of Razlog the deposits. The layer of MPEG-PLGA will decompose at ~8 weeks (in-vivo) and layer PDLLA will decompose at ~12 months (in vivo).

Welding

HAWO hpl450 device for pulse welding set on the welding time 3, the cooling time 7. Welding seam width of 2 mm.

Electroprecizia

2.5 g of PLGA 10:90 (PURAC purasorb PLG®1017) diluted to 25 ml in hexafluoroisopropanol and electropages in layers.

6 g PDLLA (Phusis) was dissolved in 20 g of acetone and electropages (1 kV/cm) in the layers.

EXAMPLE 2

Elasticity

Regarding the elasticity of the frame, which is depicted in figure 6, when the frame is dry, it is hard. On the other hand, as soon as it becomes wet, it becomes very flexible. This was compared with polypropylene mesh, which is no less hard after exposed to water.

EXAMPLE 3

Determination of the strength of the frames with and without welds

Apparatus: the apparatus for testing tensile Lloyd with a torque sensor 50 N. Speed: 100 mm/min, the spacing of the mounting bracket 20 mm.

Cages (40×40×2 mm) is cut into strips of a width of 5 mm On some 3 mm weld bead perform over the entire length of the strip (this weld seam has a thickness of approximately 0.1 mm). The maximum force and elongation at break is measured for both unmodified and welded strips.

Maximum load (N)Deflection at break (mm)% elongation at breakN/m2Pounds per square inchUnmodified0,9110,2851,399,13 E+0413Unmodified0,92or 10.6053,029,16 E+0413Unmodified0,70a total of 8.7443,716,99 E+0410

Unmodified0,8411,0955,46of 8.37 E+0412
Weld15,0537,81189,075,02 E+07 7275
Weld13,5159,49297,444,50 E+076533
Weldat 9.5341,31206,543,18 E+074607
Weld11,1944,28221,38to 3.73 E+075409

EXAMPLE 4

The use of muscle biopsies in the implant, comprising a frame

Muscle sampling is placed in the container with an appropriate buffer, for example, the environment for the cells, PBS, etc. Cells and muscle fibers isolated from the biopsies using a tissue grinder (for example, Sigma-Aldrich). Then muscle suspension applied to the surface of the frame prior to implantation.

In one series of experiments, the muscle fibers were isolated from biopsy or by dissection of the muscles by using, for example, scalpels, or by dissolving muscles using enzymatic treatment, for example, collagenase, to obtain the individual fibers with the satellite cells. These fibers are applied to the surface of the frame before implantation.

In another series of experiments, tissue explants from the muscle tissue are those of the muscles, dissected into muscle puree using, for example, scalpels, or where muscle fibers isolated from the remaining tissue by mechanical or enzymatic means, or where the muscle shredded muscle suspension, all of which include a population of fibroblasts, muscle fibers, and muscle precursor cells as satellite cells, and cultured myoblasts.

EXAMPLE 5

How crushing tissue

In some embodiments, the implementation of this invention are grown in vitro, cells can be seeded on a component, comprising a frame, prior to implantation. Cells for this purpose can be achieved by taking biopsies and retrieval and expansion of cells in vitro prior to implantation. However, this procedure is expensive and may have regulatory problems.

Instead, tissue puree (containing cells), obtained directly in the operating room, may be applied to the component, as described below.

The principle is that the tissue from the biopsy passes through a sorting grid under pressure. It crushes the sample into a puree, which can be applied to the frame before implantation

The grid may be round and may be increased at the edges, as shown in Fig.7.

The grid can be loaded into a plastic syringe, which is then loaded cloth before note the application of pressure.

However, the component, which uses a higher pressure can be applied with advantage. Accordingly, it is possible to apply metal piston in a metal cylinder, as shown in figure 8.

Alternative sampling can be crushed using a commercial grinder tissues or homogenizer with rotating knives.

EXAMPLE 6

Biodegradable surgical implant comprising a frame, and, in particular, the implant used to repair pelvic applied according to this invention, designed to ensure the application of the cells prior to implantation. In addition to the forms shown in the following example, the implant can give a definite shape to be inserted into the pelvic region.

All models can be reinforced by additional welds on the edges as reinforcement and attachment points for sutures for attaching the component to the pelvic bottom.

1. Valve

A rectangular layer of nonwoven material with the attached valve:

The valve may be any of:

a) The same non-woven material of the same thickness. In this case, the component can be carved from a single layer of material. The bend line may need partial cutting or stamping.

b) the same material of a different thickness. In this case, the valve must be attached to Pramogu is to these lamps suitable means (preferably by welding).

c) Other material, but still non-woven, any thickness. As before, the valve must be attached.

d) Other material and other process (e.g., lyophilized). The valve is attached by welding.

Cell is applied, the flap is folded over and the design does not necessarily closed using, for example, welds, as shown below.

The valve may be partial (Fig.9) full size (Fig.11 left) or segmented (Fig.11 right).

2. Tube

Tube, either seamless or with seams.

Options for tubes similar to those for valve:

a) a Single material, a single thickness. Can be seamless or made with a single welded seam on a rectangular formation.

b) a Single material, one or two thicknesses. Two rectangles of different or the same thickness, United the two welds.

c) Two materials, one or two thicknesses. Two rectangles of different materials with different or the same thickness, United the two welds.

Cells are inserted into the tube and the tube tapers and implanted. The closure phase is not needed.

3. Pocket

Option tube and valve. The rectangle are welded to the component using 3 stitches. All possible options for the valve and tubing used for the pockets.

4. Absorbent 3D frame

Hydrophilic 3D frame are welded to the grid. Cells on oat on the frame, and they are absorbed by the frame. This model can be viewed as a subgroup of the other models. The model may include a single frame (as in Fig.14) fixed on the grid, or 2 or more frames recorded on the grid.

5. Simple layer

Simple rectangular formation. Special techniques may be required to facilitate wetting and/or attachment of cells to layer:

1) Glue (e.g., fibrin) for adhesion of cells.

2) modification of the fibers to facilitate wetting.

a) Coaxial spinning with an outer layer of a hydrophilic polymer.

b) Coating a hydrophilic polymer.

(c) Joint spinning 2 different fibers, one of which is hydrophilic:

i) a Mixture of fibers.

ii) Layers, one hydrophilic, one hydrophobic.

iii) a Gradient starting with hydrophobic and ending hydrophilic.

iv) any combination of i, ii and iii.

3) Any combination of the signs of 1 and/or 2.

An example of the layer modified to facilitate wetting shown in Fig.15. Poly(ε-caprolacton) is a hydrophobic polymer, but by coating the fibers with a small amount (~3%) hydrophilic polymer (MPEG-PLGA 2-30 50DL) wetting the blood is accelerated.

All models in the examples 1-5 can optionally have shoulders/tabs to attach the frame to the structures in the pelvic region, as can be seen in Fig.19.

EXAMPLE 7

The use of the implant for reconstructive surgery of the pelvis

Used absorbable implant, consisting of MPEG-PLGA (methoxypolyethyleneglycol copolymer of lactic and glycolic acids). It liofilizirovanny and made more hydrophilic to facilitate the germination of cells and improve the recovery process. Figure 16 illustrates the structure.

The aim of the research was to study the biocompatibility and durability of the three MPEG-PLGA implants: a simple, enriched extracellular matrix (ECM, ACell, Inc.) or estrogen (estradiol, Sigma-Aldrich, Inc.).

Study design, materials and methods

Twenty implants each drug a 1×2 cm were implanted subcutaneously on the abdomen of rats, two in each. As control was used plot simulation with a blunt compartment and one stitch suture Vicryl. Explantation was performed at week 3 (15 rats) and 8 week (15 rats). The explants were fixed in a 10% solution of formalin buffer and processed in the usual way for pathological histology and stained with hematoxylin and eosin and by the Institute.

Inflammation, vascularization and organization of connective tissue was assessed semi-quantitatively on a scale of 0-4 (not-much/hard). On week 3 was assessed in the implant. 8 week where the implant has disappeared, the evaluation was made in the remaining granulocytecolony on the site.

The thickness of the scar tissue was measured at 100x magnification. Every 10 units was equal to this increase of 1.28 mm

Two 3-week sample (both of implants, enriched with estrogen) and one 8-week sample simulation were excluded because of errors encountered during the processing for pathological histology.

Data are presented as mean and standard error (SE) and analyzed using non-parametric analysis of variance the Kruskal-Wallis test with subsequent U-test of Mann-Whitney for pairwise comparisons between groups.

Results

On week 3, all implants had satisfactory germination cells. Germinated cells were distributed throughout the implant. Indicators of inflammation were significantly different between the different implants. Levels were higher in enriched ECM than in simple implants (table 1). Indicators of vascularization, the organization of the connective tissue and thickness of scar tissue differed slightly.

No trace of the implant is not left on the 8th week. There was no reaction to the foreign body and lingering signs of a chronic inflammatory reaction. Possible effects of enrichment implant disappeared for 8 week (table 2). No significant differences were found in the thickness of the connective tissue after implant compared with the slice and simulation.

Table 1
nInflammationThe presence of blood vesselsConnective tissueThickness
A: Simple implant 103,3 (0,15)1,9 (0,18)0,7 (0,26)12,8 (2,3)
B: Implant w/ ECM 103,9 (0,10)*1,5 (0,17)1,5 (0,27)11.8 in (1,2)
C: Implant w/ estrogen 83,8 (0,16)**1,6 (0,26)1,3 (0,37)14,9 (2,0)
A compared to B compared with C p=0,02p=0,33p=0,12p=0,52
Assessment of inflammation, presence of blood vessels and organization of connective tissue after 3 weeks 0-4 (not-much/hard). Thickness in absolute values. The mean (standard error).
*A compared with B: p=0.02, ** A. is in C: p=0,08

Table 2
nInflammationThe presence of blood vesselsConnective tissueThickness
A: Simple implant 101,4 (0,16)1,5 (0,17)3,0 (0,0)8,7 (1,3)
B: Implant w/ ECM 101,6 (0,16)1,6 (0,16)3,0 (0,0)9.1 (0.9)
C: Implant w/ estrogen 101,4 (0,16)1,6 (0,22)3,1 (0,1)11,1 (2,2)
D: Simulation 91,0 (0,0)0,8 (0,20)3,0 (0,0)11,6 (2,3)
A compared to B compared with C p=0.72p=1,0p=1,0p=0,79
A compared to B compared to C than the s with D p=1,0p=0,87
Indicators of inflammation, presence of blood vessels and organization of connective tissue after 8 weeks 0-4 (not-much/hard). Thickness in absolute values. The mean (standard error).

Interpretation of results

Results at week 3 showed a more progressive stage of the healing process from implant enriched ECM. The initial effects of enrichment ECM disappeared for 8 week. Implants MPEG-PLGA were fully biocompatible, disappearing at 8 week and not leaving a trace. Qualitatively tissue response at 8 weeks after implant was the same as after surgical simulation.

Durability less than 8 weeks was unexpected and is too short for use as such in reconstructive surgery of the pelvis. However, due to the characteristics presented here, the implant may play a role in the future as a carrier for cells, for example, stem cells or cells crushed muscle, contributing to their growth and without affecting the tissue of the host.

Conclusion

MPEG-PLGA in all three preparations had excellent biocompatibility. However, the durability was unexpectedly less than 8 weeks, which makes the implant is best suited for use in the gas reconstructive surgery when combined with cells, for example, stem cells or crushed muscle tissue, including cultured myoblasts and fibroblasts.

EXAMPLE 8

The use of autologous muscle cells and fragments of muscle fibers along with the implant for reconstructive surgery of the pelvis

Used absorbable implant, consisting of MPEG-PLGA (methoxypolyethyleneglycol copolymer of lactic and glycolic acids). It was dried and made more hydrophilic to facilitate the germination of cells and improve the recovery process. Figure 16 illustrates the structure.

The aim of the research was to study the biocompatibility and durability derived from muscle cells and fragments of muscle fibers together with implants MPEG-PLGA to support the regeneration of muscles.

Study design, materials and methods

Experiments on animals were carried out in the vivarium at the Panum Institute, Copenhagen, and approved by the Danish Inspectorate of animal experiments by resolution No. 2009/561-1585.

Experimental animals were 30 excluded from breeding female rats Sprague Dawley weighing 300-420 g (Taconic, Denmark). Animals and their care were provided by the Panum Institute in accordance with national guidelines.

The implants were made of MPEG-PLGA. Three different preparations were used: (A) clean the implant, B) implant with a fragment of the mi autogenic muscle fibers (MFF), C) the implant enriched autologous muscle cells-precursors (MPC).

Each implant was tested in 10 rats for 3 weeks and 10 rats for 8 weeks. Rat abdominal subcutaneous model allowed testing of the two parts of the implant in the rat.

Rats were shot using Hypnorm/Dormicum. Was made an incision of 4 cm in the midline on the abdomen. After subcutaneous stupid detachment of the implant having dimensions of 10×20×1 mm, were placed superficially on the abdominal muscle fascia and attached at one stitch 4-0 Vicryl (Ethicon). The implants were placed longitudinally to the midline. The skin was closed with 4-0 Vicryl. Antibiotic and pain medication was administered in accordance with veterinary recommendations. Rats were euthanized at 3 and 8 weeks after implantation.

For implants with MFF was made 2 cm incision in the hind limb of rats, and was taken muscle sampling with a diameter of 4 mm immediately prior to abdominal surgery. MFF were prepared in sterile Petri dishes with two scalpels by israsena biopsies in small gruel in a drop of saline solution, and the skin was closed, as described above. The implant was placed on the MFF, which immediately was attached to the implant. When the implant is covered MFF side of the implant facing the fascia./p>

MPC were grown in Interface Biotech A/S, Denmark, from biopsy specimens, obtained as described above, but 2 weeks before surgery.

Isolation and cultivation derived from muscle cells

Two weeks before implantation muscle biopsies were obtained as described above. Biopsies were transferred to sterile transport medium and left overnight at 4°C. the Selection was carried out according to the modified Protocol “Gene Delivery to Muscle” {Springer, 2002 250 /id}. Briefly: sampling carefully crushed; 0.5 ml collagenase/dispute/CaCl2added and grinding was continued; the mixture is incubated at 37°C for 1 hour, centrifuged for 5 minutes at 350×g at room temperature, the supernatant was removed, the cells again resuspendable in 10 ml of medium cultivation on the basis of the environment F-10 and placed in covered with collagen bulb. Cells were planted in 25 cm2the flask.

After 7 days of cultivation, the cells were trypsinization and brought in covered with collagen bulb. Even after 7 days of cultivation, the cells were trypsinization, conducted a count of the cells was inoculated at a concentration of 2×106cells on the implant. Cells were sown in implants for 24 hours prior to implantation, the implants with cells were incubated overnight and was sent to the vivarium.

The implants with the surrounding through the entire thickness of the tissue of the host were collected, fixed in 10% buffered Rast is the PR formalin and routinely processed for pathological histology and immunohistochemistry. The slice thickness of 5 microns.

Character growth and survival MPC and MFF assessed by immunohistochemical staining. To identify the cells of skeletal muscles in contrast to smooth muscle cells applied two different primary antibodies: mouse monoclonal antibody to desmin man (1:100, clone D33, DAKO, Denmark) and mouse monoclonal antibody to α-smooth muscle actin person (SMA) (1:100, clone 1A4, DAKO). Known cross-specificity of antibodies to the equivalent proteins in rats was confirmed by the positive and negative controls. Desmin stained the cytoplasm of skeletal and smooth muscle cells, whereas SMA stained the cytoplasm of smooth muscle cells, but not cells of skeletal muscles. As a secondary antibody was applied set Histostain ®-Plus (InVitrogen). AEC or DAB was used as Chromogen for determination of peroxidase activity. Cell nuclei were clearly stained with hematoxylin.

If 8-week sections stained with desmina were negative relative to the MPC and MFF, an additional 6 slices at intervals of 30 μm from this sample were stained in order to ensure that no residue MPC or MFF in the nearby areas of the sample.

Results

Surgery was well tolerated by all animals. Erosion or signs of infection were not observed, and there were no signs of encapsulation of impla the Tata.

After explantation, all implants were visible at the macroscopic study on week 3, and none on the 8th week. In the latter case, a tiny granuloma, representing suture material, was the only indicator of the site of implantation.

The growth pattern established immunohistochemically with desmina and SMA.

3 week growth model MPC and MFF was qualitatively different when painting desmina, therefore, quantitative assessment of desmin+cells were not carried out. Negative staining for SMA desmin+structures in the relevant sections have established that they were musculoskeletal type.

Desmin+cells, as was observed, were thinly distributed inside the implants, which sowed the MPC. MFF identified as fragmented muscle tissue with striation, unevenly distributed under the implants (Fig.17). MPEG-PLGA had nonspecific staining desmina in varying degrees, therefore, the morphology was a key factor in the interpretation of the painting.

In one of the pure implants desmin+cells were found in the configuration characteristic of the implants seeded MPC, but to a lesser extent.

8 week MFF survived and were identified as fragmented striated muscle in 6 of 10 samples (Fig.18). Further two were doubtful, since mo is folage and location desmin +structures differed: it may be artifacts representing distorted/twisted skin muscles. One specimen was found homogeneous weak positive region, probably representing the dead MFF eaten by macrophages. One sample was completely negative.

MPEG-PLGA themselves and MPC disappeared for 8 week.

In conclusion, MPEG-PLGA in all three preparations had excellent biocompatibility and disappeared within 8 weeks in this abdominal rat model. When autologous muscle precursor cells were combined with MPEG-PLGA, skeletal muscle cells were identified in 3 weeks, but not after 8 weeks. In contrast, skeletal muscle identified after 3 and 8 weeks, when the MPEG-PLGA combined with fragmented muscle fibers. This shows a high level of survival for the muscle fibers in combination with MPEG-PLGA.

1. Biodegradable surgical implant for support, enhancement and regeneration of living tissue of a subject suffering from prolapse of pelvic organs, including
a) synthetic biodegradable homogenous layer frame,
b) one or more biodegradable reinforcing element made of welded seams on synthetic biodegradable homogenous layer frame.
c) a sample of tissue explants in the specified frame, and specified tissue Explant is ispolnen from the muscle tissue, when the implant is formed in the form of a tube and/or includes a valve and/or pocket, suitable for applying suspension of the sample of cells or tissue explants on the specified implant
while specified synthetic biodegradable homogenous layer frame is a polymer of the General formula:
A-O-(CHR1CHR2O)n-B
where
A represents a residue of a copolymer of lactide and glycolide with a molecular weight of at least 4000 g/mol, with the molar ratio (i) lactide elementary units and (ii) glycolide elementary units in the residue of a copolymer of lactide and glycolide is in the range from 80:20 to 10:90,
In represents a residue of a copolymer of lactide and glycolide as defined for A, or selected from the group consisting of hydrogen, C1-6-alkyl and protective groups of hydroxyl,
one of R1and R2within each elementary group -(CHR1CHR2O)- is selected from hydrogen and methyl, and the other of R1and R2within the same elementary link -(CHR1CHR2O)- represents hydrogen,
n represents the average number of elementary units -(CHR1CHR2O) in the polymer chain and is an integer in the range 10-1000; and where the
the molar ratio (iii) of the elementary parts of polyalkyleneglycol - (CHR1CHR2O)- total number of (i) lactide e the elemental units and (ii) glycolide elementary units of residue(s) of a copolymer of lactide and glycolide is at most 20:80;
and where the molecular weight of the copolymer is at least 10000 g/mol, preferably at least 15000 g/mol.

2. Biodegradable surgical implant according to p. 1, where the specified synthetic biodegradable homogenous layer frame is hydrophilic.

3. Biodegradable surgical implant according to p. 1, where the specified synthetic biodegradable homogenous layer frame has the ability within 5 minutes, for example, for 2 minutes at 30°C, to absorb water in an amount of at least 10%, for example at least 20%, e.g. at least 30%, such as at least 50% of the frame.

4. Biodegradable surgical implant according to p. 1, where the volume % of the specified reinforcing parts is less than 40% of the implant.

5. Biodegradable surgical implant according to p. 1, where the specified synthetic biodegradable homogenous layer frame obtained through lyophilization.

6. Biodegradable surgical implant according to p. 1, where the specified synthetic biodegradable homogenous layer frame is completely biodegradable within 1-48 months of use in situ.

7. Biodegradable surgical implant according to p. 1, where the specified biodegradable reinforcing element promotes cell attachment and germination of cells derived from tissue explants.

8. Borislava the text of the surgical implant according to p. 1, where specified biodegradable reinforcing element is made of a polymer of a copolymer of lactide and glycolide PLGA, e.g., polymer, where the molar ratio (i) lactide elementary units and (ii) glycolide elementary units in the residue of a copolymer of lactide and glycolide is in the range from 90:10 to 10:90.

9. Biodegradable surgical implant according to p. 1, where the weight percentage (iii) of the elementary parts of polyalkyleneglycol -(CHR1CHR2O)- total number of (i) lactide elementary units and (ii) glycolide elementary units of residue(s) of a copolymer of lactide and glycolide is in the range of 4%-10% wt./weight.

10. Biodegradable surgical implant according to p. 1, where the specified synthetic biodegradable homogenous layer frame obtained through lyophilization solution comprising a biodegradable polymer in solution.

11. Biodegradable surgical implant according to p. 1, and the implant further includes in the specified frame, one or more components that facilitate cell adhesion and/or germination for tissue regeneration, for example, a component selected from the group consisting of: estrogen, derived estrogen, thrombin, powder ECM, chondroitin sulfate, hyaluronan, hyaluronic acid (ON), heparansulfate, heparan sulfate, dermatosurgery, growth factors, fibre is a, fibronectin, elastin, collagen, such as collagen type I and/or type II, gelatin and aggrecan or any suitable component of the extracellular matrix.

12. Biodegradable surgical implant according to p. 1, and the implant further includes in the specified frame, one or more components selected from the group consisting of growth factors, for example, insulin-like growth factors (IGF), e.g., IGF-1 or IGF-2, or transforming growth factors (TGF) such as TGF-alpha or TGF-beta, or fibroblast growth factors (FGF) such as FGF-1 or FGF-2, or platelet-derived growth factors (PDGF), for example, PDGF-AA, PDGF-BB or PDGF-AB, or nerve growth factor (NGF), or human growth hormone (hGH) and mechanical growth factor (MGF).

13. Biodegradable surgical implant according to p. 1, the implant comprises two or more separate parts of the synthetic biodegradable homogenous layers of the frame attached to the reinforcing element, for example, a grid of different polymer.

14. Biodegradable surgical implant according to p. 1, the implant comprises two or more shoulders or lugs for attachment to structures in the implantation site, for example, in the pelvic region.



 

Same patents:

FIELD: medicine.

SUBSTANCE: invention refers to medicine and tissue engineering, and may be used in cardiovascular surgery for small-vessel bypasses. A vascular graft is made by two-phase electrospinning with the staged introduction of the ingredients into the polymer composition.

EFFECT: making the bioresorbed small-diameter vascular graft possessing the improved biocompatibility ensured by using the polymer composition of polyhydroxybutyrate (PHBV) with oxyvalerate, and epsilon-polycaprolactone with type IV collagen, human fibronectin and human fibroblast growth factor (hFGF) additionally introduced into the composition.

2 cl, 1 ex

FIELD: medicine.

SUBSTANCE: invention refers to porous microsphere granules with the adjusted particle size for bone tissue regeneration. The above microspheres have a size within the range of 1-1000 mcm, have through pores of the size of 1-100 mcm and total porosity 40-75%. The declared microsphere granules are prepared by granulation by electrospinning, and heat-treated. A mixture used to form the granules by electrospinning contains a mixture of magnesium orthophosphate and biological hydroxyapatite of bovine demineralised bones in ratio 0.5:1.0, as well as 1-3% sodium alginate in distilled water and a hardener representing saturated calcium chloride.

EFFECT: invention provides preparing the microsphere granules possessing biocompatibility, biodegradation, osteoinduction and osteoconduction properties and able to be substituted by the bone tissue.

2 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine and tissue engineering, namely to cardiovascular surgery and may be used in coronary artery bypass surgery, as well as in surgical reconstruction of peripheral vessels. What is described is a method for making a porous tubular matrix of a vascular graft of a biodegradable polymer by two-phase electric spinning, with biologically active molecules stimulating the vascular regeneration being incorporated into a matrix wall matrix incorporated biologically active molecules.

EFFECT: creating the tissue-engineered high-patency and durability small-diameter vascular graft for biological re-modelling of the damaged vessels in vivo.

2 cl, 1 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine. What is described is an implanted multilayer chondral reparation flap showing biological compatibility and physiological resorption, and what is also described is a method providing surgical management in situ for intra-articular regeneration of cartilaginous tissue in joint damages and/or defects. The chondral reparation flap comprises a first external cell-impermeable layer and a second external cell-permeable layer adapted for placement in an immediate proximity from a subchondral bone on a wound portion, and also a cartilage-forming matrix located between the first and second layers. The cartilage-forming matrix represents an accepting medium for diffusion of autologous stem cells and contains chemical components promoting formation of a hyaline-like cartilage in the presence of said autologous stem cells. The method prevents a fibrous cartilaginous replacement tissue from forming within the injury region.

EFFECT: method provides autologous compositions which when used in a combination with the reparation flap form the medical system for formation of the replacement hyaline-like intra-articular cartilage.

17 cl, 7 dwg, 1 ex

FIELD: chemistry.

SUBSTANCE: porous matrix based on biocompatible polymer or polymer mix for tissue engineering is obtained by compression of polymer and sodium chloride particle mix with defined particle size, and further removal of sodium chloride by dissolution. Porosity grade of matrix lies within 93 to 98%, its pores fall into different sizes, with definite pore distribution by size within certain limits.

EFFECT: obtained matrices are free-shaped yet pertain stability and hardness characteristics required to withstand surgical implantation methods and counteract mechanical forces applied at the implantation point.

40 cl, 2 tbl, 8 ex

FIELD: medicine.

SUBSTANCE: present group of inventions concerns medicine, more specifically coated implants and devices. There is offered ceramic composition-precursor for making high-strength bio-elements used as an absorbable or partially absorbable biomaterial where the composition contains at least one silicate with Ca as a base cation with the absorption rate less or equal to the bone growth rate, and this at least one silicate acts as a base binding phase in a biomaterial, and this at least one silicate Ca is present in amount 50 wt % or more, and all other components if any are presented by additives, such as an inert phase, and/or additives which make a biomaterial to be radiopaque. There is offered hardened ceramic material which is based on the ceramic composition-precursor and is in the hydrated form. There is offered a medical implant, application of the medical implant, and also a device or a substrate coated with the uncured ceramic composition-precursor and/or hardened ceramic material.

EFFECT: invention provides a biomaterial having initial and constant durability which is dissolved in due time and reacts with an organism to generate a new tissue.

29 cl, 1 ex, 3 tbl

FIELD: medicine.

SUBSTANCE: invention relates to field of medicine. Claimed is composition with hyaluronic acid (HA), which includes gel particles of bound water-insoluble hydrated HA. HA includes bindings, represented with the following structural formula: HK'-U-R2-U-TK'. Where each group HA' represents the same or other molecule of bound HA'; each U independently represents optionally substituted 0-acylisourea or N-acylurea; and R2 represents optionally substituted alkyl, alkenyl, alkinyl, alkoxy, cycloalkyl, cycloalkenyl, cycloalkinyl, aryl, heteroaryl, heterocyclic radical, cycloaliphatic alkyl, aralkyl, heteroaralkyl or heterocyclolalkyl. Also claimed is method of developing tissues in individual, including introduction of needle into individual in place where development of tissues is necessary, needle is connected to syringe filled with composition with HA, and applying force to syringe in order to supply composition with HA to individual. Method of obtaining composition with HA includes formation of water-insoluble dehydrated particles of bound HA, separating insoluble in water particles by their average diameter, selection of subset of particles by average diameter and hydration of subset of dehydrated particles by means of physiologically compatible water solution. Other method of obtaining composition with bound HA includes binding precursor of bound HA by means of bis-carbodiimide in presence of pH buffer and dehydration of bound HA. Also included is method of developing tissues in individual that needs tissue development. Method of stabilisation of bound HA includes hydration of water-insoluble dehydrated bound HA by means of physiologically compatible water solution which includes local anesthetic, so that value of elasticity module G' for stabilised composition constitutes not less than approximately 110% from value G' for non-stabilised composition.

EFFECT: claimed composition of hyaluronic acid and method of preparation and application of HA composition are efficient for development of tissue and/or drug delivery.

27 cl, 22 ex, 2 tbl, 7 dwg

FIELD: chemistry.

SUBSTANCE: effect is achieved by using compositions based on different stereoregular amorphous biodegradable polymers - polylactides and copolymers of lactides with glycolides (18-72 mass ratio) as the second component of biocompatible mineral filler - hydroxyapatite with particle size of the main fraction of 1-12 mcm (8-41 mass ratio), as well as an organic solvent with boiling temperature equal to or higher than softening temperature by 3-20°C (20-41 mass ratio). After preparation of a homogenous mixture, the composition is undergoes thermal treatment at 80-130°C in a vacuum in a shaping vessel with the required shape. A porous product is obtained due to removal of solvent. Density of the obtained porous product is about 0.4-0.8 g/cm3.

EFFECT: design of a method of obtaining porous biodegradable composite polymer products based on polylactides or copolymers of lactides and gylcolides.

3 cl, 3 ex

FIELD: medicine.

SUBSTANCE: described are implants based on biodegradable thixotropic compound with pseudo-plastic properties and implant injected under skin or into skin in fibrous tissue. Containing microparticles of at least one biocompatible ceramic compound in suspension, in at least one liquid carrier containing at least one compound based hyaluronic acid and at least one biodegradable thixotropic compound with pseudo-plastic properties. Also disclosed is kit for preparation such implants directly before application, as well as implant production and using for filling of crinkles, and/or skin cavity, and/or cicatrices.

EFFECT: implants of simplified injection.

14 cl, 4 ex

The invention relates to medicine, namely to a restorative or cosmetic surgery and aesthetic dermatology

FIELD: medicine.

SUBSTANCE: invention can be used in the surgical management of non-inflammatory maxillary sinus diseases (MSD). An endonasal opening and sanitation of the maxillary sinus are performed. A plate of a synthetic polymer, porous polytetrafluoroethylene (ePTFE) 1 mm thick and with an open porosity of 70% is used to model a transplant of an adequate shape and by 5% more than the area of a bone postoperative defect of a posterior wall of the maxillary sinus. The transplant is laid on the residual anterior bone wall of the maxillary sinus. The soft tissues are closed completely rigidly fixing the transplant to the bone.

EFFECT: method enables preventing early postoperative complications related to an in-growth of the cicatrical tissue into the sinus lumen by forming a fibrous frame of the connective tissue closing the postoperative defect.

1 ex

FIELD: medicine.

SUBSTANCE: bioactive porous 3D-matrix for tissue engineering involves a resorbed partially crystalline polymer having a porosity of 60-80% and a pore size of 2 to 100 mcm. A biopolymer gel having a particle size of 30-100 mcm is incorporated into a portion of the pores. A polymer/gel ratio makes 99:1 to 50:50 wt %. The matrix is prepared by grinding a mixture of gel and polymer powder having an average particle size of 100 mcm, and the prepared mixture fills prepared moulds to be placed in a high-pressure chamber wherein the temperature is increased to 25-40°C first, and then the CO2 pressure is increased to 4.0-25.0 MPa. The system is kept in the above environment for 1 hour, and then the chamber pressure is discharged to an atmospheric one for 30-120 minutes; thereafter the temperature is decreased to a room value, and the patterns are removed.

EFFECT: ensuring flexibility of using the matrix in various organs and systems, no toxicity, higher ability to tissue regeneration stimulation, prolonged effect of biostimulation.

6 cl, 5 ex, 1 tbl, 4 dwg

FIELD: medicine.

SUBSTANCE: tissue regeneration or healing is stimulated when using a structure comprising a multilayer plate of a collagen membrane material, which contains a lamellated barrier material of pure collagen prepared of a natural collagen tissue; the lamellated barrier material containing a barrier layer with an outer smooth barrier surface and a fibre surface, which is opposite the outer smooth barrier surface. The structure additionally contains a matrix layer of a collagen sponge material adjoining the fibre surface.

EFFECT: matrix layer of the collagen sponge material is absorbed by an individual's body at a higher rate, than the lamellated barrier material.

20 cl, 3 dwg, 5 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine. There are described methods for making implantable medical devices, preferentially of PEEK, having antimicrobial properties. The antimicrobial action is ensured by implantation of ceramic particles containing antimicrobial metal cations into the molten PEEK resin to be cooled and finally shaped by injection moulding, cutting and mechanical treatment or by other processing methods.

EFFECT: implants possess effective antimicrobial action for reducing a bacterial growth and a risk of infection.

12 cl, 1 dwg, 3 tbl

FIELD: medicine.

SUBSTANCE: invention refers to medicine. What is described is a method for preparing a cell-free organic tissue of a human or animal origin for the vitality recovery, particularly for introducing living cells, involving a stage of making a number of holes (4; 14) in the cell-free organic tissue (2; 12) through its surface (8; 18) and setting in the tissue (2; 12); wherein the said number of holes (4; 14) is formed using a needle or a kit of needles. The holes (4; 14) are partially intersected thereby forming partially connected holes (4; 14).

EFFECT: invention also refers to a respective cell-free organic tissue (2; 12) of the human or animal origin.

17 cl, 3 dwg

FIELD: process engineering.

SUBSTANCE: invention relates to medicine. Proposed method can be used in stomatology and orthopedics for production of medical materials stimulating recovery of bone tissue defects, for making dental stopping and dental pastes. It comprises preparation of mix containing compounds of calcium, phosphorus, silicon and sodium, impregnation of bioinertial incombustible porous matrix with made mix, matrix is composed of ceramics from aluminium or zirconium oxides followed by calcination. Note here that silicon compound represents tetraethoxysilane. Note also that phosphoric acid ether is used as phosphorus compound. Calcium and sodium compounds are represented by their carboxylates in polar organic solvent. This method includes making the thin layers on more strong bioinertial porous ceramics. Note also that said process involves no special complicated equipment and expensive reagents.

EFFECT: production of glass ceramics directly from solution omitting sol preparation stage, simplified and accelerated process.

7 cl, 5 ex

FIELD: medicine.

SUBSTANCE: invention refers to porous microsphere granules with the adjusted particle size for bone tissue regeneration. The above microspheres have a size within the range of 1-1000 mcm, have through pores of the size of 1-100 mcm and total porosity 40-75%. The declared microsphere granules are prepared by granulation by electrospinning, and heat-treated. A mixture used to form the granules by electrospinning contains a mixture of magnesium orthophosphate and biological hydroxyapatite of bovine demineralised bones in ratio 0.5:1.0, as well as 1-3% sodium alginate in distilled water and a hardener representing saturated calcium chloride.

EFFECT: invention provides preparing the microsphere granules possessing biocompatibility, biodegradation, osteoinduction and osteoconduction properties and able to be substituted by the bone tissue.

2 ex

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to ophthalmosurgery, and in particular to scleroplasty. Transplant for scleroplasty has polymeric base, covered with porous layer of the same polymer. As polymer base, transplant includes layer, made from porous stretched polytetrafluoroethylene, which has nodular-fibrillar structure. As porous layer, it includes layer of porous polytetrafluoroethylene, which has volume fraction of void space 15-40%, specific surface of void space 0.25-0.55 mcm2/mcm3, average distance between voids in volume 25-30 mcm and average chord volume 8-25 mcm, with the total width of transplant constituting 0.15-0.35 mm (first version). Transplant for scleroplasty can also include porous layer of polymer, whose surface is processed to add compatibility with sclera tissue. Transplant surface is processed by application of allogenic dermal fibroblasts of 3-5 passages of culturing, with the total width of transplant being 0.15-0.35 mm (second version).

EFFECT: chemically and biologically inert transplant, which ensures effect of invasion of sclera tissues, is obtained.

2 cl

FIELD: medicine.

SUBSTANCE: invention relates to chemical-pharmaceutical industry and represents artificial dura mater, produced from electrospinning layers by technology of electorspinning, with electrospinning layer, consisting of, at least, hydrophobic electrospining layer, which is produced from one or several hydrophobic polymers, selected from polylatic acid and polycaprolactone.

EFFECT: invention ensures creation of artificial dura mater, which has good tissue compatibility, anti-adhesiveness and possibility of introducing medications, preventing cerebrospinal fluid outflow during regeneration of person's own dura mater.

30 cl, 7 ex, 11 dwg

FIELD: medicine.

SUBSTANCE: invention refers to medicine, particularly to ophthalmic and maxillofacial surgery, and aims at repairing post-traumatic defects and deformations of the eye-pit bottom and walls. What is described is an implant in the form of a solid perforated plate which is formed by photocuring of a light curing composition; when heated the implant (a cure temperature 70-90°C) keeps the shape after self-cooling that provides high strength and biocompatible properties of the material. The cure temperature of the material is much higher than a temperature of a human body that ensures maintaining the physical and mechanical properties of the implant inserted in the human body, namely the material remains strong, when fixed it shows no cutting and breaking. Before the implantation, the plate is heated to the required temperature and bent so that it is congruently repeats the eye-pit shape in a place of the defect, overlaps the defect and supports the eye in the right anatomical position.

EFFECT: reducing the likelihood of graft rejection and reducing a quantity of various complications by improving the biocompatible properties of the material.

1 tbl, 5 ex

FIELD: medicine.

SUBSTANCE: tissue regeneration or healing is stimulated when using a structure comprising a multilayer plate of a collagen membrane material, which contains a lamellated barrier material of pure collagen prepared of a natural collagen tissue; the lamellated barrier material containing a barrier layer with an outer smooth barrier surface and a fibre surface, which is opposite the outer smooth barrier surface. The structure additionally contains a matrix layer of a collagen sponge material adjoining the fibre surface.

EFFECT: matrix layer of the collagen sponge material is absorbed by an individual's body at a higher rate, than the lamellated barrier material.

20 cl, 3 dwg, 5 ex

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