Carcasses and method for producing the carcasses for carrying out human bone tissue engineering

FIELD: biomedical engineering.

SUBSTANCE: device has composite materials containing biologically active microparticles stimulating human bone tissue regeneration. Silicon, calcium and phosphorus particles combination is used in given carcasses as biologically active substance stimulating human osteoblast proliferation and differentiation and promoting osteogenesis and new bone calcification. Beside that, organic polymer is used in given carcasses as carrier having three-dimensional structure and external anatomical shape. It shows several properties compatible with bone regeneration and blood vessel neogenesis.

EFFECT: enhanced effectiveness of treatment; improved safety and cost-effectiveness.

27 cl, 6 dwg, 1 tbl

 

The scope to which the invention relates.

The present invention relates to frames made of composite materials, engineering of human bone tissue, in particular to the frames, made from a new medical composite materials of the microparticles, which has the function of stimulating the regeneration of human bone tissue, methods for their manufacture and their application to engineering of human bone tissue.

Prerequisites to the creation of inventions

Engineering of human bone tissue is dealing with the process of using absorbable biological materials, such as frames, to stimulate the regeneration of bone. Physico-chemical properties and three-dimensional structure of these frameworks are the key factors affecting the regeneration of bone tissue. Based on molecular criteria of conformity of biological materials transplantation devices for the body or bodies for the engineering of tissues must be secure and possess the biological activity to stimulate regeneration of the relevant human tissue and recovery of associated physiological functions at the cellular and molecular level (Chou, et al. J. Cell Sci., 1995, 108: 1563-1573; Chou, et al. J. Biomed. Mater. Res., 1996, 31:209-217; Chou, et al., J. Biomed. Mater. Res., 1998, 39:437-445).

In PR destoyed the prior art combination of scaffold material, mainly extracted from the natural collagen, calcium phosphate, or organic polymers. Natural collagen has possible disadvantages such as high cost, the worst physiological quality, easier spread of disease and the induction of hypersensitivity in humans (Pachence and Kohn, Biodegradable polymers for tissue engineering in Principles in tissue engineering, 1997, pp.273-293). Calcium phosphate (Kukubo, et al. J. Mater. Science, 1985, 20:2001-2004; Feinberg, et al. Shanghai Journal of Stomatology, 2000, 9:34-38 and 88-93) have the disadvantage, such as poor ability to shrink, and exhibits biological activity, stimulates the regeneration of human bone tissue (Chou, et al. Biomaterials, 1999,20:977-985). Organic polymers, such as poly(lactic acid) (PLA), poly(glycolic acid) (PGA), or a mixture of PLA And ISC (PMGC), also have some disadvantages: acid degradation products released during the degradation of the specified polymer, can cause an inflammatory reaction and alien reaction in the tissues of the human body and, therefore, affect the regeneration of bone tissue. Moreover, these polymers do not possess biological activity, which stimulates the regeneration of human bone tissue (Hubel, Bio/Technology, 1995, 13(6):565-576; Thomson, et al. Polymer scaffolds processingin principles inTissueEngineering, 1997, pp.273-293; Cao, et al. Plast Reconstr. Surg. 1997, 100:297-304; Minuth, et al. Cell Tissue Research, 1998, 291(1): 1-11; Wong Yulai, et al. Shanghai Journal of Stomatology, June 2000, 9(2):94-96). In the previous ur the outside of equipment, there are attempts to transplant some of biologically active proteins such as cellular binding protein (cell binding protein) or bone inducing protein (bone inducing protein), inactive polymer frames (Barrea, et al, Macromolecules 1995, 28:425-432; Ugo and Reddi, Tissue engineering, morphogenesis, and regeneration of the periodontal tissue by bone morphogenetic proteins, 1997). But these methods are unlikely to be implemented in the clinical setting due to the high cost, instability and heterogeneity transplanted proteins and difficulties sterilization frames. U.S. patent 5977204 describes the frames, made from a composite material containing an organic polymer and biostable (bioceramics). The above-mentioned biostable was first described in U.S. Patent 4103002. There used a combination of silicon, calcium and phosphorus to improve the biocompatibility between the specified material and the bone tissue of a person, but not to stimulate bone regeneration. In fact, the U.S. Patent 5977204, and U.S. Patent 4103002 not describe with certainty the activity of silicon in relation to stimulate bone regeneration and do not mention a synergistic stimulatory effect of calcium and phosphorus. In addition, the materials disclosed in both patents include sodium. However, sodium has no stimulating activity against bone regeneration. Therefore, in accordance with the principles of molecular compatibility of biomaterials, Karka is s, as stated in U.S. Patent 5977204, do not have significant biological activity against stimulating bone regeneration. In addition, in the process described in this patent, the manufacture of frames from the specified composite material using organic solvents, which can lead to cytotoxicity in the human body. U.S. patent 6051247 discloses a composite material containing biostable from U.S. Patent 4103002 and polysaccharides (such as dextran), suitable for repair of bone defects. But the specified composite material is used only to create a paste or putty, which is not suitable for the manufacture of frames with the exact three-dimensional structure and a certain resistance to pressure for tissue engineering. In addition, the combination of biostable specified composite material is inactive in relation to stimulating bone regeneration. Biostable used in U.S. Patents 5977204, 4103002 and 6051247 has an average particle size (diameter) of more than 70 microns. Such large particles significantly affect the physical properties of composite materials, and inorganic elements cannot uniformly be released during the decomposition of the materials of the frames. U.S. patents 4192021 and 5017627 disclose composite material containing the second organic polymer and calcium phosphate, which can be used for the manufacture of frames for repair of bone defects. However, this composite material is inactive in relation to the stimulation of bone regeneration, and microporosity and the diameter of pores is provided for these frames, are not suitable for implantation and regeneration of bone cells. U.S. patent 5552454 discloses a composite material in which the calcium phosphate is applied to the surface of the organic polymer particles. This model does not have as a stimulating effect on bone tissue regeneration, and may not be used for precise three-dimensional structure of frames for tissue engineering.

Three-dimensional structure of frameworks for engineering of human bone tissue is important for the regeneration and bone, and blood vessels in new bone. In the prior art in U.S. Patent 5977204, 4192021, 5017627 and 5552454 design of all frames in the form of a homogeneous porous or non-porous model, where the configuration of the pores, the pore size and distribution of pores in porous frameworks uniform. However, such frames with similar and uniform pore distribution is not suitable for bone regeneration. In examples of these frameworks described in the prior art, the diameter of pores in the frames is in the range from 150 to 400 microns. is the diameter large enough to allow penetration of human cells in the Central part of the data frames. Thus, the regeneration of bone tissue occurs only 2-3 mm around these frames. In another aspect, a relatively large pore size (more than 400 microns) is not suitable for bone regeneration. (Cartner and Mhiatt, Textbook of Histology, 1997; Tsuruga et al, J. Biochem., 1997, 121:317-324; Gauthier et al, J Biomed. Mat. Res., 1998, 40:48-56). In accordance with the molecular compatibility of biological material, the regeneration of blood vessels in the Central part of the frames is key for the growth of new bone in the data frames, with blood vessels, mainly educated only in channels with a diameter of more than 400 microns. Therefore, the frames of the prior art having uniform pores, do not meet the various requirements of simultaneous bone regeneration and regeneration of blood vessels, and thus, the practical use of such frameworks for engineering bone tissue is limited.

Consequently, the prior art shows that there is a great need in frames that are suitable for the engineering of human tissues, which are biologically active to stimulate proliferation and differentiation of human osteoblasts, promote the formation and calcification of new bone and restore the appropriate physiological functions at the cellular and molecular level.

The object image is to be placed

The object of the present invention is to obtain frames, free from organic solvent and having a three-dimensional structure and the external anatomical structure, which was produced using the method of hot wheels without the use of organic solvent based on the principles of molecular compatibility of biomaterials using a composite material of the microparticles made from a combination of microparticles of silicon, calcium and phosphorus as biologically active substances frames, which could actively stimulate the proliferation and differentiation of human osteoblasts and to promote the formation and calcification of new bone. In combination with the organic polymer in a specific ratio, as the carrier specified composite material is biologically active to stimulate bone regeneration and has desirable physical properties. The resulting frames can be used in engineering of human bone tissue is safe, practical and effective to restore bone defects caused by tumor, inflammation or injury, or for orthopedic surgery on the bones of a man.

Summary of the invention

To achieve these objectives, in one aspect of the present invention is predlagajutsja composite structures for engineering of human bone tissue, having a three-dimensional structure with micropores, and the connecting channels containing inorganic microparticles of silicon as the primary inducing substances for bone regeneration, calcium and/or phosphorus microparticles as a synergistic stimulatory substances and organic polymer as a carrier. In another aspect, the present invention proposes a method of manufacturing frames made of composite material for bone tissue engineering person, which includes a method of hot-casting without using any organic solvent. The following aspect of the present invention features the use of frameworks made of composite material for bone tissue engineering person when repair of the bone defect caused by the tumor, inflammation or injury, and orthopedic operations on the bones of a person, byin situimplantation of cells or implantation of osteoblasts, pre-proliferatingin vitroin the human body.

Brief description of drawings

Figure 1 - influence of microparticles of silicon, calcium and phosphorus on the proliferation of osteoblasts, the biological activity of alkaline phosphatase, the synthesis and secretion of osteocalcin and calcification of the bones in the body of healthy people.

Figure 2 - comparison of characteristics of the frames, made from the composition is traditional material, containing microparticles of silicon, calcium and phosphorus and organic polymer (PMGC), and frames, made of a material PMGC, the stimulation of proliferation of osteoblasts and biological activity of alkaline phosphatase in the body of a healthy person.

Figure 3 - spherical structures made of composite material containing nanoparticles of silicon, calcium and phosphorus, and organic polymer (PMHC).

4 is a SEM photograph (performed using scanning electron microscope) of micropores frames made of composite material containing nanoparticles of silicon, calcium, phosphorus, and organic polymer (PMHC).

Figure 5 - Photos of the cylindrical frame, having a three-dimensional structure, consisting of micropores and connecting channels, made of a composite material containing nanoparticles of silicon, calcium, phosphorus, and organic polymer (PMGC), using the method of hot wheels.

6 - experimental model reconstruction on the animal misakomoko process TMJ man, with the help of tissue engineering.

Detailed description of the invention

The present invention is based on a long and intensive study on the subject of search elements in the form of microparticles for use as chemical components in the La production of frameworks for engineering of human bone tissue, the specified element in the form of microparticles is a biologically active stimulator of regeneration of human bone tissue, samarasekera neutralizing acidic or alkaline substances around framesin vivo. Were originally developed frameworks for engineering bone tissue of the person in which microparticles of silicon used as the main active component for stimulating the proliferation and differentiation of human osteoblasts, bone formation and calcification; microparticles of calcium is used as the active component for synergistic stimulation of proliferation and differentiation of osteoblasts; microparticles of calcium and phosphorus are used as active ingredients for a synergistic stimulation of calcification of the regenerated bone. These combinations of elements of biologically non-toxic, actively stimulate the regeneration of bone tissue, destroyedin vivoand may be replaced by new bone. Thus it is possible to avoid the use of protein products in engineered tissue, reduce production costs and improve the safety and effectiveness of frameworks in clinical practice. Biologically active combination of frames according to the present invention differs from the frames of biostable disclosed in U.S. Patent 5977204, in which the biologically active combination for the present the invention includes only silicon or contains silicon as a main component and a specific ratio of calcium and/or phosphorus, but without sodium. In addition, the diameters of the particles of the elements of the present invention differ from those of the materials of the prior art. Therefore, the present invention relates to the selection of new chemicals and combinations of ratios for bone tissue regeneration. In the present invention is used, an organic polymer, such as PLA, ISC or PMGC, as a carrier for these microparticles of silicon, calcium, phosphorus, which connects these microparticles of silicon, calcium and phosphorus for forming, and provides resistance to pressure these frames. Microparticles of silicon, calcium and phosphorus in the composite material is used as biologically active components in these frames, and therefore, these frames serve as a reservoir for these bioactive components. These biologically active components slowly, continuously and uniformly released from the frame, when the organic polymer is destroyedin vivoto stimulate bone formation and neutralization of acidic degradation products of the specified organic polymer, providing a suitable environment for bone regeneration. Therefore, the present invention refers to the resolution of the issues underlying preaches the existing level of technology, consisting in the fact that the frames engineering of human bone tissue deprived of biological stimulation and may not be used for repair of bone defects of large size.

In the present invention are all inorganic elements are microparticles that are different from the elements of U.S. Patent 5977704 related to elements in the form of particles with diameter greater than 50 microns. Unless otherwise indicated, the "particles" in the present invention are defined as particles having a diameter equal to or less than 10 microns, preferably in the form of nanoparticles having a diameter less than 1000 nm, more preferably less than 100 nm, most preferably from 5 to 80 nm. In the scope of the present invention microparticles of silicon, calcium and phosphorus, having a diameter of 100 nm or less than 10 microns, can also be used to achieve the objective of the present invention, because they also have a biological stimulant. The only difference is that these microparticles have a weaker stimulant, as they slowly dissolve and diffuse. The diameter of the microparticles used in the present invention, is clearly less than the diameter of the microparticles used in the materials of the frames of the prior art for the engineering of bone tissue. In addition, the diameter of mikroC STIC, used in the present invention, promotes uniform distribution of chemical elements in the frame, even the release of these chemical elements of the frames and can improve the stability of the frames to the pressure.

In the present invention, unless otherwise indicated, a "biologically active stimulating substance" is defined as a substance that can actively stimulate normal cells to proliferation and specific differentiation, to perform their characteristic physiological functions. The elements silicon, calcium and phosphorus according to the present invention are biologically active stimulating substances, which can actively stimulate normal human osteoblasts to proliferation and to stimulate a number of specific physiological functions of osteoblasts (as, for example, the biological activity of alkaline phosphatase, the synthesis and secretion of osteocalcin and calcification of the bones). All combinations of inorganic elements in the frames of the prior art did not possess stimulating biological activity similar to the one which has the combination of the present invention.

In the present invention, unless otherwise indicated, "frameworks for engineering bone tissue of a person" is defined as frames, which have a characteristic three-dimensional structure is ru and configuration compatible with the anatomical morphology of the damaged area of the bone of a man, made of biological material, which is safe and biologically active, and can be absorbedin vivoduring a certain period. In those cases, when such frames are implantedin vivothey provide a favorable environment for proliferation and differentiation of osteoblasts and facilitate the gradual formation of new bone in the data frames, while frame material of these frames is gradually absorbed and, eventually, disappearsin vivothe location of these frames is replaced by new bone. All previous frames of the prior art do not have a specific three-dimensional structure similar to the structure of the frames of the present invention.

First tested and used inorganic element silicon as the main active substance in the frame, which has a biologically active stimulant in engineering of human bone tissue. Experimental data of normal human osteoblasts, shown in figure 1, confirm that the ions of silicon added to the cell culture medium, have considerable inducing and stimulating action (2-4 times) on key biological indicators when the creation is new bone, such as the proliferation of osteoblasts, the biological activity of alkaline phosphatase, the synthesis and secretion of osteocalcin and calcification of bones, etc. Data concerning the experimental animal models, shown in Figure 3, further acknowledge that after the inorganic particles of elemental silicon implantedin vivothe particles of inorganic silicon is diffused into the surrounding soft tissue and induce an increase in the ion concentration of sulfur that characterizes the early stage (two weeks) of new bone formation, and induce an increase in the concentration of ions of calcium and phosphorus, characteristic of late stage (8 weeks) formation of a Mature and dense bones. Based on these data, the present invention provides a technical breakthrough, i.e. confirms specific biological catalytic effect of inorganic elemental silicon and elemental silicon can be used in frames for engineering bone tissue. In addition, the data of Figure 1 show that the concentration of silicon is directly proportional to its biological stimulating action and maximum stimulating effect of silicon appears when saturated concentration of the silicon (100 ppm).

In addition, the experimental data shown in figure 1, confirm that the combination of inorganic alimentariomobile and inorganic elemental calcium and phosphorus obviously has a synergistic effect on the speed of proliferation of normal human osteoblasts and the synthesis and secretion of osteocalcin and calcification of bone. Therefore, the present invention uses an inorganic elements calcium and phosphorus as synergistic substances to promote biological activity of the ions of silicon.

In the present invention, unless otherwise indicated, all combinations of elements using ion silicon as the sole or primary biological stimulants, and calcium and/or phosphorus as a synergistically active substances, to actively and effectively stimulate the formation of new bone. Preferably, the percent content of the atoms in the combination of elements that are used as biological stimulants are 60-100% silica, 0-30% calcium and 0-20% of phosphorus, more preferably 60-90% of silica, 0-25% calcium and 0-15% phosphorus, and most preferably 60-70% silicon, 20-25% calcium and 10-15% of phosphorus.

Microparticles silicon/calcium/phosphorus in the specified biologically active composite material are in the form of a mixture of various types of individual elementary particles or produced by the mixing of various types of items and dry grinding conventional physical or chemical methods. In accordance with Figure 1 corresponding to the number of atomic elements in the mixture of particles or microparticles complex elements is not an essential factor for the achievement of the objectives of the present invention, because of the different atomic or mass ratio of lead in the result only to different levels of stimulating activity. Therefore, all combinations with arbitrary atomic or mass ratios of these three elements, where silicon is used as the primary biologically active stimulating substances, and calcium and phosphorus are used as synergistically biologically active stimulating substances, can be used for frames of the present invention.

Inorganic elements silicon, calcium and phosphorus are defined as biologically active elements, which have the ability to stimulate the proliferation of human bone tissue, differentiation of osteoblasts and bone calcification. It is also an important scientific achievement in the field of biomaterials. In the prior art it was considered that the synthesized or extracted exogenous osteogenic, auxin or connexin and so on have a biological stimulant, but these biological products are not safe, have low biological stability and high cost, and thus can hardly be used in bioengineering. In addition to the above, the stimulating activity of combinations of inorganic elements according to the present invention additional is but confirmed a close relationship between bone regeneration and distribution of the released ion silicon on the surface of the partition between the implanted material and cloth as shown in animal models of Figure 3, the bone regeneration - stimulating effect of a composite material containing microparticles silicon/calcium/phosphorus and PMHC on the model of normal human osteoblasts, as shown in figure 2, and data, animal models, as shown in Fig.7. For the above reasons, it is shown that inorganic elements silicon, calcium and phosphorus can be used to replace a biologically active proteins and to achieve significant biological stimulating action. Additionally, these inorganic elements can be used as biologically active materials in the frames for the engineering of human bone tissue, to obtain safe and stable frames, the material which can be easily prepared with lower cost and greater security and stability, and to expand the practical application of data frames.

In prior art organic polymers (PLA, ISC and PMGC) is usually used as the sole material for frames. However, these polymers do not possess stimulating biological activity, and acid products of their decomposition in the body impede the regeneration of bone tissuein vivo. In the present invention organic p is limer used only as a carrier for a specific combination of microparticles of silicon, calcium and phosphorus. According to the results of the test frames having other media properties, when the content of inorganic elements in combination of more than 80%, the stability of the frames to the pressure will be relatively weaker, consequently, the specific spatial structure cannot be maintained in the animal body, and when the content of the combination of inorganic elements less than 20%, stimulating biological activity is insufficient to accelerate the final formation of new bone within 8 weeks. To achieve compromise between the resistance to pressure and biological activity data of the frames of the present invention determines the volume ratio of the combination of the silicon/calcium/phosphorus to organic polymer in the range from 80:20 to 20:80, preferably from 70:30 to 30:70, in accordance with the biological testing of examples 2, 4 and 6. In this range can be adjusted solubility of composite material frames. With the increasing content of the combination of the silicon/calcium/phosphorus increases the biological activity of stimulating bone regeneration. The amounts of these two materials can be installed in this range to suit the various requirements for the recovery of human bone tissue. In this subramaniyapuram combination of biologically active substances and organic polymer, forming a framework that can serve as a reservoir for such biologically active substances. By dissolving the organic polymer (PLA, ISC, PMGC) in vivo (from 1 to 8 weeks) microparticles of silicon/calcium/phosphorus continuously and steadily released, stimulating the proliferation and differentiation of osteoblasts and the formation and calcification of the bone during the process of bone regeneration. In addition, the release of nanoparticles of silicon/calcium/phosphorus can neutralize the acidic products of destruction of organic polymer, resulting in the local environment around the frame is favorable for the regeneration of bone tissue.

In the prior art any calcium phosphate or biostable for repair of bone defects are large particles having a diameter greater than 50 microns. When using such large particles in the composite material will be affected by the physical properties of the specified composite material. In addition, the release of large particles embedded in an organic polymer frames is not uniform. Therefore, in the present invention used microparticles of silicon/calcium/phosphorus, which has a diameter less than or equal to 10 microns, preferably less than 1000 nm, more preferably less than 100 nm and most preferably in the range of 5-nm, so that these microparticles are uniformly embedded in an organic polymer and slowly and evenly released during the destruction of the specified organic polymer. These microparticles obtained by mixing microparticles of each of the three elements according to atomic content, as defined in the present invention.

In prior art three-dimensional structure of different frames is microporous with the same pore size and uniform distribution. The disadvantage of these frames is that relatively small micropores (which has a diameter less than 300 microns) adverse for the passage of osteoblasts and blood vessels, and relatively large micropores (having a diameter of about 400 microns) is unfavorable for the regeneration of bone tissue. Therefore, the practical use of these frameworks in bioengineering bone is clearly limited. In the present invention is used, the frames having a three-dimensional structure, containing both micropores and connecting channels, as shown in Fig.6. According to the results of the tests of the other diameters of the pores with diameter less than 100 microns are not suitable for the passage of the cells and pores of diameter greater than 300 microns are not suitable for the formation of new bone. Therefore, all frames used in the Examples of manufacturing and biological tests is s, as shown in Figure 4-7 have micropores with diameters in the range from 100 to 300 microns. Micropores with a diameter defined in the present invention, suitable for proliferation of osteoblasts and regeneration of new bone. Employment of micropores according to the present invention is from 50% to 90%. For example, the employment of micropores of frames used in the Examples, as shown in Fig.4, Fig.6 and Fig.7, is 80%, 50% and 50%, respectively.

In accordance with the results of tests for other employment micropores, physical resilience frameworks to pressure the employment of micropores more than 90% is clearly weaker and insufficient to resist the pressure of the surrounding tissues, whereas osteoblasts are unlikely to penetrate the frames for the formation of new bone in the employment of micropores less than 50%. In accordance with the test results of different diameters of the connecting channels, with a diameter greater than 500 microns, the resistance of the frames to the pressure is clearly weaker, and difficult neogenes large volume of bone tissue, while the penetration of cells and bone formation is difficult in the case where a diameter of less than 350 microns. Therefore, the diameter of the connecting channels of the present invention is in the range from 350 to 500 microns, for securing the entry of cells into the depth of the frames and the supply of nutrients and sour is one of new bone through new blood vessels, that grow along these connecting channels inside the frames. In accordance with the results of the tests of the distances between the connecting channels, the stability of frames to pressure weaker in those cases when the distance is less than 3 mm, while the occurrence of cells in all the micropores of frames for the formation of new bone is difficult in cases where the distance is more than 6 mm, therefore, it is not suitable for the formation of new bone. Therefore, the gap between the connecting channels of the present invention is preferably in the range from 3 to 6 mm to ensure uniform entry of cells inside all of micropores through the connecting channels. The present invention uses a combination of elements with three-dimensional structure containing as the connecting channels and concentrically arranged micropores, which can be re-grouped (like bricks) to form a variety of frames larger for repair of large bone defects. This new three-dimensional structure contains micropores, which is useful for the regeneration of bone and connective channels, favorable for uniform distribution of cells, transport of nutrients to the tissues and regeneration of blood vessels in new bone, and followed the Sabbath.) can be used to repair bone defects is large in size, which could not be restored in the prior art. In respect of the skeletons of small size, having a size less than 5 mm, or various bone defects remains sclerotic, in accordance with the present invention can be used frames with only micropores and a variety of forms, such as spherical shape, a cylindrical shape or a square shape, as shown in Fig.4 and Fig.6.

The anatomical shape of the frames according to the present invention for the engineering of human bone tissue can be divided into two groups, namely pre-made form and custom, customize the form, depending on the location and size of the bone defect. Made in advance frames can be of various shapes, such as spherical shape, a cylindrical shape or a square shape, etc. In cases where the diameter of pre-made frames is less than 5 mm, the frames will only micropores without connecting channels. These skeletons of small size can be of various diameters in the range from 0.5 mm to 5 mm, pre-Made frames that have a size greater than 5 mm, constructed in the form of a set of combined elements containing both micropores and connector the s channels of different sizes and shapes, so as to fill the space of the bone defect. Made of pre-frames are used to fill the damaged areas of the bones of various sizes and shapes and in different locations in the body for bone tissue regeneration. In custom, customize frames using human bone scan as a template for designing the shape of the frames corresponding to the anatomical morphology of human bone, and data frames are combined structure, containing both micropores and connecting channels that can be used to repair large bone defects in orthopedic operations on human bone and to treat in the absence of the surviving wall of the bone to maintain the shape.

In addition, in the prior art for the manufacture of organic polymer frames for bioengineering usually use organic solvents. Because organic solvents practically can not be completely removed from the frames, it's bad for the regeneration of human bone tissue. In contrast to the method of manufacture of bodies for the engineering of human bone tissue, known in the prior art, the method according to the present invention uses a conventional method of hot wheels for the manufacture of the manufactured item is evritania or non-standard, driven frameworks for bioengineering. The method according to the present invention can avoid the cytotoxicity caused by residual organic solvent in the frames of the prior art, and can reduce the cost of mass production frames.

Clinical use of frames according to the present invention for the engineering of human bone tissue includesin situimplantation of cellsin vivoor implantation of the cells, proliferatingin vitro.In situimplantation of cellsin vivoincludes direct implantation of pre-made skeletons of small size in the cavity of the bone defect of a person during surgical intervention, directly using undifferentiated interstitial cells, abundant in blood and tissue bleed and have exudate, trapped in the cavity of the bone defect during surgery, for infiltration into the space between the frames and then stimulate bone regeneration under the influence of the material of the frames. Therefore, this method can be used to repair bone defects in unstressed plot with residual outer wall of the bone. Implanted pre-made frames are a combination of frames with size of more than 0.5 mm, for Example, spherical and cylindrical frames, as shown is as Fig.4 and Fig.6, use in the above methods. Implantation of cells, proliferatingin vitroused to repair bone defects in the impact area or in an area with no residual outer wall of the bone. The source of normal autologous osteoblasts person required in large quantities for repair of bone defects in large size by implanting them inside the frames, is always a serious problem in the field of medicine. Implantation proliferating in vitro autologous osteoblasts healthy person, used in the present invention can solve the problem. The present invention uses autologous superficial skeletal fragment, obtained from the patient as the source of osteoblasts. 0.2 cm3superficial skeletal fragment can proliferatein vitrowith the formation of 6-10 million autologous osteoblastic cells with normal activity osteogenesis, as shown in Fig.7. Moreover, it leaves no scars, there is no functional or physical impact on the site collection. 55 million proliferating cells osteoblasts sufficient to supply frames for regeneration 2 cm3normal autologous bone. In clinical practice, custom customized frames are investing in the region of the bone defect after esmerine the osteoblastic cells implanted inside these frames in vitro, and bone fixed ligiously supporting tires with conventional surgery on the bone. When the regeneration of new bone in the frame, a supporting force of these tires gradually weaken, the load on the new bone is gradually increased, and eventually restored the physiological function of the regenerated bone (see below "Experimental model animal reconstruction misakomoko process of the temporomandibular joint of the human engineered tissue").

Compared with the prior art, the positive properties of the present invention are that the use of microparticles of silicon/calcium/phosphorus, having a stimulating effect on bone regeneration of man, as a biologically active material provides biological efficacy of frames according to the present invention is significantly higher than in those frames, which are known in the prior art without biological stimulating action; the combined elements containing both micropores and connecting channels in the data frames that promotes uniform distribution of human cells and regeneration of blood vessels in these frameworks, solving the problem that the regenerated bone only limited in the local area surrounding frames predshestvuyuschego technology. Moreover, the repair and regeneration of large bone defects of the person, unattainable in the prior art, at the present time can be achieved in the present invention by re-grouping combinations of elements frames matched with three-dimensional structure for the formation of sufficient volume.

The present invention is further illustrated by the following non-limiting Examples in combination with the drawings.

Example 1

Microparticles of silicon, calcium, phosphorus biologically significantly stimulate the proliferation of osteoblasts, the biological activity of alkaline phosphatase, the synthesis and secretion of osteocalcin and calcification in the body of a healthy person.

Osteoblast human cells used in the test were obtained from healthy donors aged 20 to 25 years. Each group of cells obtained from 0.2 cm3surface fragments of the skeleton of a donor. Just test used 5 groups of cells. Average values and standard deviations of survey data for 5 groups shown in figure 1. It is seen that 0.2 cm3superficial skeletal fragment donors can proliferate with the formation of 6-10 million autologous osteoblastic cells having osteogenic activity in vitro. In the cell culture medium used in this is pytanie, pre-add particles of silicon, calcium, phosphorus with diameter less than 10 microns in certain concentrations or proportions as shown in tables 1, with the saturation concentration of the silicon component 100 ppm. During the cultivation, the culture medium with defined concentrations of particles replaced with fresh medium containing particles of the same concentration every 3 days. On the 12th day and 20-day conduct the following studies: 1) study of the proliferation of osteoblasts: counting the total number of cells grown in culture medium with different concentrations or quantitative ratios of chemical additives using a conventional flow cytometer, and the counting multiplicity proliferation of osteoblasts, as shown in figure 1, based on the number of cells adhering to the culture of the Cup in the first 24 hours, indicating that the inorganic elemental silicon has a clear stimulating effect on the proliferation of osteoblasts and stimulating effect is directly proportional to the concentration of silicon. Moreover, inorganic elements calcium and phosphorus synergistically enhance biological stimulatory effect of silicon; 2) determining the biological activity of alkaline phosphatase. An important characteristic of the normal osteoblasts is the secretion of alkaline phosphatase is normal functional activity. The investigated cells, cultured under the conditions shown in table 1 for 12 days and 20 days; these cells are isolated using plasmase destroyed a conventional ultrasonic generator and the cell suspension analyze conventional chromatography; then count microequivalents the amount of substrate cleaved under the action of alkaline phosphatase produced 10 million cells per hour. These results confirm that the inorganic elemental silicon can enhance the biological activity and the incentive effects of alkaline phosphatase is proportional to the concentration of silicon; 3) determination of the synthesis and secretion of osteocalcin. The synthesis and secretion of osteocalcin are specific and important indicators of activity of normal human osteoblasts. The content of osteocalcin, secreted into the culture medium, determine the normal histochemical method using monoclonal antibodies against osteocalcin person. The results are expressed as values in femtograms of osteocalcin secreted 10 million cells on day 12 and day 20. The results showed that inorganic elemental silicon could significantly stimulate increased secretion of osteocalcin normal human osteoblasts. This stimulating effect right about ortional concentration of silicon. Moreover, inorganic elements calcium and phosphorus acted synergistically, contributing to biological stimulating effect of silicon; 4) study of the calcification of bone. The deposition of calcium in the span of osteoblasts is one of the important indicators during the final stage of the formation of new bone. Cells of each group were stained for the presence of calcium conventional methods on the 12th and 20th, and the intensity of staining was determined by conventional chromatographic device. The results showed that higher concentrations of silicon, calcium and phosphorus significantly and synergistically acted by stimulating and increasing calcification of normal human osteoblasts.

Example 2

Composite material containing microparticles of silicon, calcium, phosphorus and organic polymer (PMGC), has the advantage, compared with only one PMGC material in stimulating the proliferation of normal human osteoblasts and biological activity of alkaline phosphatase

This biological study illustrates the catalytic effect of one group millimicron composite material according to the present invention in the cell culturein vitroand performs a comparison with only one organic polymer PMHC and conventional polystyrene cups for cell culture. The content of the atoms reorgan the economic elements in the combinations of elements of composite material comprise 67% of silicon, 22% calcium and 11% of the phosphorus and the volumetric ratio of the combination of inorganic elements to PMHC is 50:50. This composite material and only one organic polymer PMGC separately processed to obtain a disk with a diameter of 2 cm and a thickness of 1.5 mm by hot casting pattern at 200°C for 8 hours (detailed stage see also Example 4). Received the CDs individually placed on a standard polystyrene cups for cell culture, 2 cm in diameter, cells seeded on various molded disks or directly to conventional polystyrene cups for cell culture without molded disk, and then determine the effects of various materials on the cultured cells. Three groups of test cells obtained from three healthy donors. Cell proliferation and biological activity of alkaline phosphatase is determined by represented in figure 1, then the cells are cultured for 7 days. The data shown in figure 2 represent the average values and the average deviations of the three groups of cells. The results show that a disc made of a composite material of the present invention, has a strong biological stimulating effect than the disk only one PMNC and than conventional polystyrene Cup for cell culture.

Example 3

Most the of (diffusion) and the distribution of ions of silicon after implantation of silicon millimicrons material in an experimental model on the animal and the distribution of ions in stimulating the regeneration of new bone

In this biological test as an experimental animal model using white adult rabbits. On the fibula experimental animal using sewed doing the bone cavity with a diameter of 0.5 cm, then the specified cavity is filled with particles of the composite silicon/calcium/phosphorus (atomic ratio of Si:Ca:P=67:22:11) with a diameter of 50-80 nm, in conclusion, the injured area is sutured. The test animals are fed for 2 or 8 weeks, and then plot the filled composite material and the surrounding tissue are removed repeated surgical intervention, fixed with 10% formaldehyde, dipped in resin, cut in 1 mm layers in longitudinal section and finally determine the distribution of ion concentrations on both sides of the interface between land filled with composite material and the surrounding tissue of the animal using the radiation of the ion analyzer. The data shown in Table 1 represent average values for 5 groups of animals in atomic percent.

Table 1< / br>
The distribution of the concentrations of silicon, calcium, phosphorus, sulfur and chlorine at the interface between the body and implanted material
Two weeks +1 mm ← side materialThe boundary between the material and the body of the animalSide of the body of the animal → +1 mm → +2 mm
Silicon14,784,128,7913,922,09
Calcium28,378,709,0114,47for 9.47
Phosphorus7,317,88of 8.4711,6418,51
Sulfur7,8124,2611,8815,4731.90 beef
Chlorine24,990000
Eight weeks← side materialSide of the body of the animal →
Silicon12,7221,220,410,580,29
Calcium56,6437,9664,6359,9359,44
Phosphorus17,7634,4532,4035,9537,96
Sulfur00000
Chlorine00000

The results show that silicon ions are released from the nanoparticles of silicon and diffused into the body of the animal after implantation of the composite material in the body of the animal for 2 weeks, resulting in a dramatic increase in the local concentration of ions of silicon, and also increases the concentration of sulfide ions, which indicates that the active regeneration of new bone at an early stage. 8 weeks silicon ions disappear in the body of the animal and significantly increases the concentration of calcium and phosphorus, which indicates the formation of Mature bone. This model biological tests was also tested histologically, and the results confirm that the image fabric with two sides of a surface of the partition correspond to the dynamic changes of new bone formation, as described above, changes in the ion distribution. The results of this biological tests confirm that ion silicon has a major biological effect on the stimulation of new bone formation.

the example 4

Spherical frames, made of a composite material containing nanoparticles of silicon, calcium, phosphorus and organic polymer (PMHC) by hot casting

This is an example of making spherical frames of a composite material. The source materials are silicon dioxide (SiO2), calcium oxide (CaO) and triphosphate calcium (Ca5HO13P3), where the concentration of atoms make up 67% of silicon, 22% calcium and 11% phosphorus, respectively, and the mass ratio of initial substances are 40% silica, 6% of calcium oxide and 54% triphosphate calcium, respectively. This preparative process comprises mixing the above-mentioned silicon-, calcium - and phosphorus-containing starting materials in accordance with these mass ratios, grinding using a self-rotating "Retsch track mill for 3 days until the diameter of the microparticles is from 5 to 80 nm. The diameter of the microparticles is confirmed using a scanning electron microscope. Organic polymer PMGC pulverized using an electric grinder stainless steel and sift through a fine sieve to obtain particles PMGC diameter from 25 to 50 microns. Spherical frames, shown in Figure 3, is made with a combination of inorganic elements and PMHC in the ratio of 70:30. The template is made from politer is floretina, then fill in the template particles of the original silicon, calcium, phosphorous-containing inorganic substance and an organic polymer in the above ratio. After filling in the template aglomerados at 200°C for 8 hours in a ceramic furnace is gradually cooled (10°C / minute) and finally take the form of obtaining spherical frames, as shown in Figure 3 (employment of micropores is 80% and the diameter of the micropores is from 100 to 300 microns).

Example 5

Image scanning electron microscope show the micropores in the frames, made from a composite material containing nanoparticles of silicon, calcium, phosphorus and organic polymer (PMHC).

Received frames made of composite material containing nanoparticles of silicon, calcium, phosphorus and organic polymer (PMGC), cut in longitudinal section and their internal micropores studied using conventional scanning electron microscope. The results, shown in figure 4, confirm that the data frames produced by hot casting, have a structure with micropores, following each other (specified diameter of micropores is in the range from 100 to 300 microns).

Example 6

Cylindrical frames produced by hot molding of a composite material, terasawa silicon nanoparticles, calcium and phosphorus and organic polymer (PMGC), have a three-dimensional structure, containing both micropores and the connecting channels connecting these micropores

This is an example of cylindrical frames of a composite material. The source materials are silicon dioxide (SiO2), calcium oxide (CaO) and triphosphate calcium (Ca5HO13P3), where the concentration of atoms make up 67% of silicon, 22% calcium and 11% phosphorus, respectively, and the mass ratio of initial substances are 40% silica, 6% of calcium oxide and 54% triphosphate calcium, respectively. This preparative process comprises mixing the above-mentioned silicon-, calcium - and phosphorus-containing inorganic starting materials in the above mass ratio, grinding using a self-rotating "Retsch track mill for 3 days until the diameter of the microparticles is from 5 to 80 nm. The diameter of the microparticles is confirmed using a scanning electron microscope. Organic polymer PMGC pulverized using an electric grinder stainless steel and sift through a fine sieve to obtain particles PMHC with a diameter from 25 to 50 microns. Standard cylindrical frames, shown in Fig.6, made from a combination of inorganic elements and PMHC in the ratio of 50:50, as described below. Shab what he is made of polytetrafluoroethylene, and then specified in the template set several wires of stainless steel with a diameter of 350-500 microns and with an interval of 4 nm in the corresponding direction. Above the original substances fill the template, aglomerated at 200°C for 8 hours in a ceramic furnace is gradually cooled (10°C / minute) and then the form is removed. After removal of the cylindrical frame, wire, stainless steel extract, the resulting frames are shown in Figure 5 (employment of micropores is 50%; the diameter of the micropores is from 100 to 300 μm; the diameter of the connecting channels is 500 microns).

Example 7

Experimental model animal reconstruction misakomoko process of the temporomandibular joint of the person by means of tissue engineering

As shown in Figa, the pattern of polytetrafluoroethylene prepared in accordance with the anatomical shape misakomoko process TMJ human and non-standard, customized frames are prepared in accordance with the method of manufacture of frames, shown in figure 5. As shown in Fig.6b, superficial skeletal fragments are obtained from the surface segment of the patient through the following steps: incision of soft tissue in the hidden place of the body surface under local anaesthesia, excoriation about 0.2 cm3on ernstig skeletal fragments and cultivation in cell culture medium. The incision is sutured. He heals from 3 to 5 days and does not affect the activities or views of the patient. The obtained skeletal fragment placed on a standard polystyrene culture plate in the chamber for cell culture and cultivated at 37°C. After 2 weeks from 0.2 cm3superficial skeletal fragments proliferate from 6 to 10 million autologous osteoblasts from normal osteogenic activity, as shown in Fig.6b. Fig.6b indicates positive test results calcium deposits using a conventional method Vancusa" method, where brown particles in accumulation of bone cells, painted pink, are evidence of calcification of bone. For the above reasons, these proliferance cells can be used in frames in medical practice. According to medical practice, for enough frames 5 million cells for regeneration and education 2 cm3normal autologous bone. Critical stages include the Department proliferating cells using plasmase from the culture Cup, dip the frames in the cell solution so that the cells penetrated into the side frames through the connecting channels and the connecting micropores frames. Frames with cells in them implanted in the body of an animal model for the study (see Figs) through conventional surgery. In the Kli of the systematic practice frameworks, implanted in the body, fixed with the help of ligious tire by means of normal bone surgery. When the regeneration of new bone in the frames supporting force of the clamping rail is gradually weakening, the load on the new bone is gradually increased and, ultimately, restore the physiological function of the regenerated bone. As shown in Fig.6d, new bone tissue is formed within 6 weeks after implantation of cells and frames in the body. For example, a new bone of the animal model studied by extracting implanted frames surgically, commit 10% formaldehyde solution for 24 hours, immersion in paraffin wax, making slices and tissue staining the conventional method. Newly formed normal bone man can be seen under an ordinary optical microscope, and the emergence of Garanovich tubules (Harvard tubule) confirms the formation of high bone density. For the mean time, the newly formed blood vessels found in the initial place of the connecting channels in the frames. These histological data show that neogenes normal bone tissue is sufficient.

Reference documents

1. Barrera DA, Zylstra E, Lansbury PT, Langer R. (1995), Copolymerization and degradation of poly(lactic acid-co-lysine), Macromolecules, 28:425-432.

2. Cao Y, Vacanti JP, Paige KT, Upto J, Vacanti CA. (1997), Transplantation of chonrocytes utilizing a polymer-cell construct to produce tissue-engineered cartilage in the shape of a human ear, Plast. Reconstr. Surg. 100:297-304.

3. Cartner LP, Brian Harvey JL. (1997), Textbook of Histology, V.B. Saunders Company, Philadephia.

4. Chou L, Firth JD, Uitto V-J, Brunette DM. (1995), Substratum surface topograph alters cell shape and regulates fibronection, mRNA level, mRNA stability, secretion and assembly in human fibroblasts, J. Cell Sci. 108:1563-1573.

5. Chou L, Firth JD, Nathanson D, Uitto V-J, Brunette DM (1996) Effects of titanium on transcriptional and post-transcriptional regulation of fibronectin in human fibroblasts. J. Biomed. Mater. Res. 31:209-217.

6. Chou L, Firth JD, Uitto V-J, Brunette DM. (1998a), Effects of titanium substratum and grooved surface topography on metalloproteinase-2 expression in human fibroblasts. J. Biomed. Mater. Res. 39:437-445.

7. Chou L, Marek B, Wagner WR. (1999) Effects of hydroxyapatite coating crystallinity on biosolubility, cell attachment efficiency, and proliferation in vitro. Biomaterials, 20:977-985.

8. Feinberg SE, Holloster SJ, Halloran JW, Chu TMG, Krebsbach PH. (2000) A tissue engineering approach to site-specific reconstruction of skeletal structures of the maxillofacial regions. Shanghai Journal of Stomatology. 9:34-38; 88-93.

9. Gauthier AJ, Ducheyne P. Bettiger D. (1999) Effect of surface reaction stage on fibronectin-mediated adhesion of osteoblast-like cells to bioactive glass. J. Biomed. Mat. Res. 40:48-56.

10. Hubbell JA. (1995) Biomaterials in tissue engineering. Bio/Technology. 13(6):565-576.

11. KuKubo T. ITO S, Shigematsu M, Sakka S, Yamamuro T. (1985) Mechanical properties of a new type of apatite-containing glass ceramics for prosthetic application. J. Mater. Science, 20:2001-2004.

12. Minuth WW, Sittinger M, Kloth, S. (1998) Tissue engineering: generation of differentiated artificial tissues for biomedical applications. Cell Tissue Research. 291(1):1-11.

13. Pachence JM, Kohn J. (1997) Biodegradable polymers for tissue engineering, in Principles in Tissue Engineering. By Lanz RP, Langer R, Chick WL. Academic Press, London. P.273-293.

14. Thomson RC, Yaszemski MJ, Mikos AG. (1997) Polymer scaffolds processing. In Principles in Tissue Engineering. By Lanze RP, Langer R, Chick WL. Academic Press, London. P.263-272.

15. Tsuruga E, Hiroko T, Hideaki I, Yuichi W, Yoshinori K. (1997) Pore size of prous hydroxyapatite as the cell-substratum controls BMP-induced osteogenesis. J. Biochem. 121:317-324.

16. Ugo R, Reddi AH. (1997) Tissue engineering, morphogenesis, and regeneration of the periodontal tissue by bone morphogenetic proteins. 8:154-163.

17. Wong Yulai, Yilin Cao, Vacanti, (2000) Experimental studies on condyle of human temporomandibular joint by tissue engineering, Shanghai Journal of Stomatology. Vol.9(2):94-96.

18. U.S. patent No. 4103002, July 1978.

19. U.S. patent No. 4192021, March 1980.

20. U.S. patent No. 5017627, may 1991.

21. U.S. patent No. 5552454, September 1996.

22. U.S. patent No. 5977204, November 1999.

23. U.S. patent No. 6051247, April 2000.

1. Frameworks for engineering bone tissue of a human, comprising silicon-containing inorganic microparticles as the main biologically active stimulating substances, inorganic particles of calcium and phosphorus as auxiliary substances for synergistic amplification of biological stimulating action of silicon and an organic polymer as a carrier, and a combination of microparticles of silicon/calcium/phosphorus is used as biologically active substances in the frameworks to stimulate bone regeneration, these frames have a three-dimensional structure, containing both micropores and connecting channels, where the diameter of the micropores is from 100 to 300 μm, and the diameter of the connecting channels is from 350 to 500 microns.

2. Frameworks for engineering of human bone tissue according to claim 1, where the particles are a mixture of microparticles of silicon, microcast the calcium and particulate phosphorus, or are microparticles of a mixture of the elements silicon, calcium and phosphorus.

3. Frameworks for engineering of human bone tissue according to claim 1 or 2, where the diameter of the microparticles is less than or equal to 10 microns.

4. Frameworks for engineering of human bone tissue according to claim 3, where the diameter of the microparticles is less than 1000 nm.

5. Frameworks for engineering of human bone tissue according to claim 4, where the diameter of the microparticles is less than 100 nm.

6. Frameworks for engineering of human bone tissue according to claim 5, where the diameter of the microparticles is from 5 to 80 nm.

7. Frameworks for engineering of human bone tissue according to claim 1, where the content of the atoms of the elements in inorganic microparticles comprise 60-100% silicon, 1-30% calcium and 1-20% of phosphorus.

8. Frameworks for engineering of human bone tissue according to claim 7, where the content of the atoms of the elements in inorganic microparticles comprise 60-90% silicon, 1-25% calcium and 1-15% of phosphorus.

9. Frameworks for engineering of human bone tissue of claim 8, where the content of the atoms of the elements in inorganic microparticles is 60-70% silicon, 20-25% calcium and 10-15% of phosphorus.

10. Frameworks for engineering of human bone tissue according to claim 1, where the organic polymer as a carrier selected from the group consisting of polylactic acid (PLA), a polymer of glycolic acid (PGA) or composite material (PMHC) from the PMK and the ISC.

11. Frames for ingen the history of human bone tissue of claim 10, where the volume ratio of the particles of active ingredients to the organic polymer as the carrier ranges from 80%:20% up to 20%:80%.

12. Frameworks for engineering bone man in claim 11, where the volume ratio of the particles of active ingredients to the organic polymer as the carrier is 70%:30% to 30%:70%.

13. Frameworks for engineering of human bone tissue according to claim 1, where the employment of micropores ranges from 50% to 90%.

14. Frameworks for engineering of human bone tissue according to claim 1, where the distance between the connecting channels is 3 to 6 mm

15. Frameworks for engineering of human bone tissue according to claim 1, where the connecting channels and concentric micropores form a combined structural elements.

16. Frameworks for engineering of human bone tissue according to claim 1, where the data frames are pre-made or specially manufactured driven in accordance with the anatomical morphology.

17. Frameworks for engineering bone man in clause 16, where pre-made frames have the form selected from the group consisting of spherical, cylindrical and square shape.

18. Frameworks for engineering bone man in clause 16, where non-standard customized frame is shaped, constructed in accordance with the anatomical morphology of the defect in the human bone in the form of a template, and represents a frame connected with the teams of structural elements, including connecting channels, and concentric micropores.

19. The method of manufacture of bodies for the engineering of human bone tissue according to any one of claims 1 to 18, characterized in that use a method of hot wheels without the use of an organic solvent.

20. The use of frameworks for engineering of human bone tissue according to any one of paragraphs 1-18 for regenerative repair of bone damage caused by the tumor, inflammation or injury, or orthopedic surgery.

21. The application of claim 20, where the application is achieved by in situ implantation of cells in the human body, or by implantation of cells, proliferating in vitro.

22. Use item 21, where in situ implantation of cells in the human body includes the direct implantation of pre-made frames in different sizes and forms in the damaged area of bone defect containing undifferentiated interstitial cells.

23. Use item 21, where the implantation of the cells, proliferating in vitro, involves implanting these proliferating in vitro autologous osteoblasts in large specially made customized frames, and subsequent implantation of these frames in the human body.

24. Application neorg the organic microparticles, containing silicon, calcium and phosphorus, for making cages for engineering of human bone tissue for regenerative repair of bone defects in orthopedic operations.

25. The application of paragraph 24, where the diameter of the inorganic microparticles is less than or equal to 10 microns.

26. Use A.25, where the diameter of the inorganic particles is less than 1000 nm.

27. Use p, where the diameter of the inorganic microparticles is less than 100 nm.

28. The application of item 27, where the diameter of the inorganic particles is from 5 to 80 nm.

29. The application of paragraph 24, where the content of the atoms of the elements in inorganic microparticles comprise 60-100% silicon, 1-30% calcium and 1-20% of phosphorus.

30. The application of clause 29, where the content of the atoms of the elements in inorganic microparticles comprise 60-90% silicon, 1-25% calcium and 1-15% of phosphorus.

31. The application of article 30, where the content of the atoms of the elements in inorganic microparticles comprise 60-70% of silicon, 20-25% calcium and 10-15% of phosphorus.



 

Same patents:

FIELD: medicine, chemical-pharmaceutical industry, pharmacy.

SUBSTANCE: invention relates to creature of agents used in prophylaxis and treatment of diseases and pathological states associated with the locomotor system. Agent represents the complex preparation including the composition № 1 and № 2. The composition № 1 on hydrophobic base comprises the following active substances: phytocomponents of peppermint, common wormwood, mountain arnica, grape seeds, matricary, pine cones, pot-marigold, bur-marigold, sea-buckthorn oil, red pepper oil, red palm oil, mixture of essential oils of peppermint, lavender and clove in the ratio 1:1:1, bee venom, menthol, synthetic camphor and the following accessory substances: form-forming thickening agent - bee wax; preserving agent - propylparaben; emolents - cetiol CC, DC 345 and vaseline oil; antioxidant - kovi-ox. The composition № 2 on hydrophilic base comprises the following active substances: aqueous extracts of lilac flowers, mountain arnica, walnut peel; propocyanides of grape berries, propolis alcoholic extract, Kova-B-trox, and accessory substances: gel-forming agent - natrosol 250; moistening agents - propylene glycol and urea; chelating agent - disodium-EDTU; emolent - PLN phospholipids; preserving agents - methylparaben and caton CG; pH regulating component - triethanolamine and deionized water. Proposed agent possesses the expressed and prolonged curative effect that allows its using both for prophylaxis and treatment of different diseases and damages of structures in locomotor system and for improving skin state and adjacent tissues. Agent shows high therapeutic activity, namely its rapid and deep penetration into tissues, heating and relaxing effect, improving circulation in tissues and enlargement of blood vessels. Agent shows analgesic, anti-inflammatory, anti-edematous effect, accelerates reparative processes and improves tissue trophism. Agent shows the complex directivity on all mechanisms in base of diseases and pathological states of the locomotor system and their clinical symptoms and possesses the broad spectrum of pharmacological effect and both recovers the damaged functions of organ and shows the systemic effect on all body.

EFFECT: valuable medicinal properties of agent.

2 tbl

FIELD: organic chemistry, pharmaceuticals.

SUBSTANCE: invention relates to new chemical compound of general formula I , or salts, or hydrates thereof. In formula I R1 represents group of formula -G1-R1a (wherein G1 represents single bond, oxygen, sulfur; R1a represents C1-C10-alkyl optionally substituted with halogen or C3-C8-cycloalkyl); R2, R3 and R4 are independently hydrogen or -G20-R20 (wherein G20 represents single bond, oxygen, sulfur, sulfinyl or sulfonyl; R20 represents C1-C6-alkyl optionally substituted with 1-3 halogen atoms or C3-C8-cycloalkyl); R5 and R6 are independently -X5-X6-X7 group (meanings of X5, X6 and X7 are as defined in specification) or R3 and R4 may together form pyrrol ring optionally substituted with C1-C6-alkyl; Ar represents phenyl, 1,3-benzodioxolyl, naphthyl, pyridyl, optionally substituted with 1-3 substituents (as defined in specification) Compounds of formula I has antagonistic activity in relates to receptors of corticotrophin releasing factor (CRF). Also disclosed are compounds, characterized by preferable structures, pharmaceutical compositions, using such compounds, intermediates for production thereof and method for treatment of various diseases mediated by CRF.

EFFECT: new compounds as antagonists of CRF receptors.

33 cl, 1 tbl, 316 ex

FIELD: medicine.

SUBSTANCE: material comprises deproteinized allobone granules as filler, the granules being at least 1 mm large, and hydrophylic dimethylacrylate as monomer. Bone cement, filler and monomer are taken in 1:1:0.5 proportion, respectively.

EFFECT: improved biocompatibility and osteointegration properties.

FIELD: organic chemistry, pharmaceuticals.

SUBSTANCE: Described are derivatives of general formula I (all symbols are as described in specification), pharmaceutically acceptable salts thereof or cyclodextrin clathrates. Such compounds hardly bind of EP2 subtype of PGE receptor and are useful in prophylaxis of immune diseases, allergy, death of neuronal cells, liver or kidney insufficiency, etc.

EFFECT: new agent for prophylaxis of various diseases.

18 cl, 388 ex, 68 tbl, 3 dwg

FIELD: organic chemistry, medicine, gynecology.

SUBSTANCE: invention relates to novel tetracyclic heterocompounds of the formula (I): wherein X, Y, Z, R1 - R4, n and m has values given in the invention description and used as selective modulating agents for estrogen receptors. Also, invention relates to a method for synthesis of these compounds and pharmaceutical compositions comprising thereof, and their using in treatment and/or prophylaxis of disorders mediated by one or more estrogen receptors. Proposed compounds are useful in treatment and/or prophylaxis of disorders associated with depleting estrogen and comprising such disorders as rush of blood, vaginal dryness, osteopenia and osteoporosis, hormone-dependent cancer and hyperplasia of breast, endometrium, uterus cervix and prostate, endometriosis, uterus fibroma, osteoarthritis that can be used as contraceptive agents both separately and in combination with progestogen or progestogenous antagonist.

EFFECT: valuable medicinal properties of compounds and pharmaceutical compositions.

25 cl, 7 tbl, 171 ex

FIELD: medicine, pharmacy.

SUBSTANCE: invention relates to creature of ointment for external using. The proposed ointment used for treatment of degenerative-dystrophic diseases of locomotor system comprises vegetable as a fatty product, wax as a bee product, heparin as anticoagulant of direct effect, and glycerol, hydrocortisone and dimethylsulfoxide also. After applying ointment the method involves effect by ultrasound with frequency 88 MHz, power 0.4 Wt/cm2 for 10-15 min. Treatment course is 10-15 procedures. Ointment has no adverse complications, it activates metabolic processes in joint region, improved microcirculation and recovers cartilage surface.

EFFECT: improved and valuable properties of ointment.

1 tbl, 2 ex

FIELD: pharmaceutical industry, pharmacy.

SUBSTANCE: invention relates to producing combined preparations of a definite composition and ratio of components, namely, to a glucosamine-containing composition and possessing the enhanced anti-inflammatory activity. Also, invention relates to development of formulation as hydrogel based on this composition that provides enhancing bioavailability of the proposed composition. Invention proposes addition of citric acid and willow extract to the composition. The composition comprises the following components, g: glucosamine, 0.5; willow extract, 0.1; citric acid, 0.02; pectin, 0.25, and sucrose, 0.25. Invention provides the synergistic effect of the known components.

EFFECT: improved and valuable medicinal properties of composition.

2 cl, 1 tbl, 2 ex

FIELD: organic chemistry, medicine, pharmacology, pharmacy.

SUBSTANCE: invention relates to novel compounds possessing properties of EP4 agonist and their using as EP4 agonist for preparing a pharmaceutical composition used in treatment of disorders associated with reducing the osseous mass. Invention provides the enhanced effectiveness of treatment.

EFFECT: valuable medicinal properties of pharmaceutical compositions.

13 cl, 125 tbl, 32 ex

FIELD: medicine.

SUBSTANCE: method involves introducing drug of formula (I) where variables have known values shown in invention application.

EFFECT: increased treatment efficiency; reversed or prevented articular cartilage or subchondral bone degeneration.

10 cl, 10 dwg

FIELD: medicinal industry.

SUBSTANCE: invention relates to a method for isolation of biologically active substance from mammalian cartilage and to preparing a medicinal formulation for parenteral administration. Agent is made as a medicinal formulation for parenteral administration and represents peptide complex with the content of low-molecular fraction from 70% to 90%, molecular mass of its peptide components in the range 75-846 Da and concentration of polypeptides 2.5-2.9 mg/ml. Agent is prepared from cartilage of calves (age is 12 months, not above) or pigs by extraction with acetic acid in the presence of zinc chloride. Proposed method for preparing agent from cartilage of calves (age is 12 months, not above) or pigs involves tissue freezing at temperature -40°C (not less), keeping at temperature -20-22°C for 2 months (not less) followed by milling and addition of 3% acetic acid solution in the volume ratio = 1:5 at temperature 20 ± 5°C. Extraction is carried out at constant stirring up to preparation homogenous suspension and then 1% zinc chloride solution is added to suspension in the volume ratio = 50:1. Mixture is cooled at constant stirring up to temperature 7-16°C, stirred for 1 h in each 4 h settling for 48 h. Extract is separated from inert substances by separating and acetone is added to extract in the volume ratio = 1:5 and kept at temperature 3-5°C for 4 h. Formed homogenized deposit is precipitated with acetone repeatedly twice (not less) and deposit containing active substance is washed out on Nutch filter with two-fold volumes of acetone cooled to temperature 7-16°C up to preparing light-gray deposit. Deposit is rubbed through metallic sieve, dried, dissolved in distilled water at room temperature and constant stirring up to the concentration of polypeptides 2.5-2.9 mg/ml. Solution is centrifuged, filtered and subjected for ultrafiltration treatment in device at anti-pressure 1.0 kgf/cm2 (not above) through materials with retaining capacity 15000 Da. Glycocol is added to ultrafiltrate up to its final concentration 10-20 mg/ml at pH = 5.6-6.6, solution is subjected for sterilizing filtration under pressure 2.0 kgf/cm2 (not above), poured into ampoules in volume 2 ml and subjected for autoclaving at temperature 120°C for 8 min under atmosphere pressure 1.1 kgf/cm2. Invention provides optimal technology in isolation of peptide complex with the content of low-molecular fraction from 70% to 90%, molecular mass of peptide components in the range from 75 to 846 Da and preparing aqueous extract with the concentration of polypeptides 2.5-2.9 mg/ml. Invention provides both purification of the end product from impurities and to enhance its yield. The isolated substance differs from the known substances prepared early from mammalian raw by molecular mass of peptide components, absence of toxicity and apyretic properties based on the complete removing impurities. Proposed substance can be used in medicine as an agent normalizing functions of cartilage tissue.

EFFECT: improved preparing method, valuable medicinal properties of agent.

3 cl, 1 tbl, 1 dwg, 4 ex

FIELD: medicine.

SUBSTANCE: device has at least two porous polytetrafluoroethylene layers of different structure. The layer adjacent to parenchymal organ is composed of at least ten 30-120 mcm thick porous polytetrafluoroethylene layers having volume share of hollow space equal to 79-93%, specific surface of hollow space of 0.65-0.80 mcm2/mcm3, mean volume chord length of 25-30 mcm. The external layer has at least one 40-90 mcm thick porous polytetrafluoroethylene layer having 43-50% share of hollow space, 0.35-0.45 mcm2/mcm3 large specific surface of hollow space, mean distance between the bubbles in the volume being equal to 9,0-15,0 mcm, mean volume chord length of 8-11 mcm. General implant porosity is not less than 80%, implant thickness is equal to 1.0-2.0 mm.

EFFECT: producing implants allowing vasoselective parenchymatous sutures application.

6 cl, 5 dwg, 1 tbl

FIELD: medicine; traumatology; orthopedics.

SUBSTANCE: vertebra's body implant for front spondylodesis is made of porous powder bio-compatible matter in form of rod with support edge surfaces. Substance edge surfaces are made rough. Size of macroscopic roughness equals to 0,3-0,6 average size of particles of powder. Side surface of rod is made smooth and rod is made anisotropic-porous in total. Maximal value of porosity close to and onto support surfaces equals to 0,6-0,8, close to central part it equals to 0,2-0,3 and minimal value - at side surface and close to it - belongs to 0,10-0,15 range. Cross-section of rod is made symmetrical to central plane. Front part of cross-section has shape of circle and rear part has shape of square with rounded angles which has side to equal to diameter of circle.

EFFECT: higher stability of primary fixation; reduced traumatism; shorter operation time; simplified design of implant; simplified procedure of implantation.

12 cl, 5 dwg

Keratoprosthesis // 2270643

FIELD: medical engineering.

SUBSTANCE: device has optical member and supporting plate attached to it from porous poly tetrafluoroethylene having structure composed of polymer elements and empty space elements with elements joined into three-dimensional network possessing empty space element volume share of 15-40%, specific space element surface of 0.25-0.55 mcm2/mcm3, mean distance between empty spaces in a volume of 25-50 mcm and mean spatial chord of 8-25 mcm. The supporting plate is manufactured as convexo-concave lens having curvature radius of 7-10 mm. The optical member is manufactured from polymethyl methacrylate or polycarbonate and is optionally collapsible.

EFFECT: improved implantability characteristics; reduced rejection risk.

6 cl, 7 dwg

FIELD: medical engineering.

SUBSTANCE: implant is manufactured from porous polytetrafluoroethylene having structure composed of polymer elements and empty space elements with elements joined into three-dimensional network possessing empty space element volume share of 15-40%, specific space element surface of 0.25-0.55 mcm2/mcm3, mean distance between empty spaces in a volume of 25-50 mcm and mean spatial chord of 8-25 mcm. The supporting plate is manufactured as convexo-concave lens having curvature radius of 7,0-10,0 mm. Cylindrical hole is optionally available in lens center.

EFFECT: low costs and improved implant production; improved implantability characteristics.

3 cl, 7 dwg

FIELD: medical engineering.

SUBSTANCE: implant is manufactured from porous polytetrafluoroethylene having structure composed of polymer elements and empty space elements with elements joined into three-dimensional network possessing empty space element volume share of 15-40%, specific space element surface of 0.1-0.3 mcm2/mcm3, mean distance between empty spaces in a volume of 50-100 mcm and mean spatial chord of 12-38 mcm. The orbital implant is optionally shaped as ball or rounded pyramid.

EFFECT: reliable integration into connective eye tissue.

3 cl, 8 dwg

FIELD: medical engineering.

SUBSTANCE: implant is manufactured from porous polytetrafluoroethylene produced by pressing thermostatically controlled polytetrafluoroethylene powder fraction of 0.25-1.60 mm at 30±10°C and having structure composed of polymer elements and empty space elements with elements joined into three-dimensional network possessing empty space element volume share of 15-40%, specific space element surface of 0.25-0.55 mcm2/mcm3, mean distance between empty spaces in a volume of 25-50 mcm and mean spatial chord of 8-25 mcm.

EFFECT: high heat resistance properties; reliable integration into connective eye tissue.

6 dwg

FIELD: stomatologic techniques and materials.

SUBSTANCE: proposed membrane comprises at least two porous polytetrafluoroethylene layers: one adjusting parodentium tissues and the other being outside layer, the two having different porous structure. Layer adjusting parodentium tissues is characterized by volume portion of hollow space equal to 78-94%, specific surface of hollow space 0.5-0.9 mcm2/mcm3, average distance between hollows 20.0-50.0 mcm, and average volumetric chord 20.0-30.0 mcm. Outside layer is characterized by volume portion of hollow space equal to 30-60%, specific surface of hollow space 0.1-0.5 mcm2/mcm3, average distance between hollows 1.8-15.0 mcm, and average volumetric chord 1.0-15.0 mcm.

EFFECT: increased regenerative activity and simplified use.

7 cl, 7 dwg, 1 tbl, 6 ex

The invention relates to ophthalmology and is intended for operations as retinal detachment and progressive myopia
The invention relates to medicine, namely to materials for reconstructive bone-reconstructive surgery in traumatology-orthopedics

FIELD: medicine, gel composition.

SUBSTANCE: claimed composition contains calcium sulfate and viscous biopolymers and can be easily introduced into defect part of damaged bone. Composition of present invention is capable of penetration into adjacent organs and is useful as physiologically acceptable filling material for bone concretion.

EFFECT: composition to prevent growth of undesired conjunctive tissues and to induce blood vessel growth and bone osteoblast development in earlier stage.

7 ex, 9 dwg, 1 tbl, 6 cl

Up!