The crushed polymer hydrogels to prevent the formation of adhesions and methods for their production
The invention relates to medicine, namely to molecular-linked gel containing a variety of biological and non-biological polymers such as proteins, polysaccharides and synthetic polymers. Such molecular gels can be applied to the required areas in the patient's body by extruding the gel through the opening to the area of the target. Gels can mechanically destroy and use them in such implantable articles, such as a prosthesis for the breast. Use in a living body (in vivo) of such gels is useful to prevent the formation of post-operative spinal and other tissue adhesions, to fill tissue defects by tissue, body cavity, surgical defects, etc. 11 C. and 33 C.p. f-crystals, 5 Il., table 4. 1. The scope of the present invention the Present invention relates essentially to the cross-linked polymer compositions and, in particular, to the use of such compositions to prevent the formation of tissue adhesions, and for other purposes.Often after surgery adhesions are formed on the tissues, which can lead to the compromise of the results of surgical operations, as well as to the occurrence of postoperative complications. Spike Asociatia in different parts of the patient's body, including tissue of the pelvis, abdomen, spine, tendons, eyes, urinary system, cardiovascular system, when normal tissue is connected with the surfaces of internal organs of a patient that have been injured or have been damaged during surgery. Such adhesions can connect organs or other body tissues that are normally separate. To resolve these adhesions will require additional surgery and it involves additional cost, risk and/or discomfort for the patient.Of particular interest for this application are adhesions tissue, which often occur after surgery on the spine due to the formation of scar tissue between the spinal nerves and spaced below the adjacent tissues. The formation of this scar tissue can cause compression of the nerve root, thereby creating a neural complications, such as, for example, constant pain in the lower back and ischialgia (pain along the sciatic nerve). Currently, the epidural scar tissue should be removed for more surgery.To reduce or exclude postoperative adhesions have proposed various methods and materials. Such pic is on the site of the target. Currently, there are, for this purpose, a woven material of regenerated cellulose, supplied by the firm Johnson & Johnson under the trademark "Interceed". However, this material does not match well enough at the bottom of the fabric. Other polymeric materials that have been tried for this purpose, include nylon, cellophane, polytetrafluoroethylene (PTFE), polyethylene, siloxane, elastomers and films of copolymers of polylactic acid. Many of these materials are not capable of biodegradation and therefore they remain in the body with unpredictable and potentially adverse consequences.The reduction and elimination of postoperative adhesions on the spine are particularly problematic. It was proposed to apply different permanently implanted devices such as, for example, those described in U.S. patents 5437672 and 4013078. However, the use of permanent implants undesirable. They also offered to apply able to break down barriers and film. However, the use of such barriers and films is also problematic. The area between adjacent vertebrae are difficult to access and therefore very difficult to accurately place and FIXME such materials should be sufficiently fluid, so they could enter and adapt to the treated areas and they must be sufficiently viscous and resistant, so they can remain in the space until such time as the fabric does not go away. These goals must be balanced with the requirement of biocompatibility and rassasivaetsia compositions against the formation of adhesions.For these reasons, it is desirable to obtain improved compositions, methods and products to prevent the formation of tissue adhesions after surgery and other injuries. In particular, it is desirable to obtain compositions and methods of using the compositions in the living body (in vivo) to eliminate and prevent the formation of epidural adhesions after excision of an arc of a vertebra or other surgical operations on the spinal column. It is also desirable that such compositions are useful for preventing or exclusion of adhesions formation elsewhere in the body, as well as for other purposes in vivo, for example, as a filler of voids in the tissue, such as voids formed as a result of a biopsy, tissue, or other tissue injury with uneven edges, as filler prostheses such as breast prostheses, seal and/or to stop bleeding (hemostasia. Compositions and methods according to the present invention, should also be adapted for delivering drugs and other biologically active substances to the surfaces of the fabric adjacent to the areas where were implanted in these compositions. At least some of these objectives are achieved by variants of the present invention disclosed in this application, which will be described later.2. The known technical solutions of the Barrier film and the materials used to prevent or exclude the formation of adhesions on the spine or other adhesions are described in U.S. patents 5350173, 5140016; 5135751; 5134229; 5126141; 5080893; 5017916; in PCT publications WO 95/21354; WO 92/15747; WO 86/00912; and Boilers and others , (1988), "Fert. Ster., 49; 1066-1070, in U.S. patents 5437672 and 4013078, each of which describes a spinal protective device, which remain as permanent implants in the spinal cord.Collagen and other polymeric liners, intended to close the subcutaneous penetration, for example a tissue tract formed when accessing the femoral artery, is described in several patents, including U.S. patents 5540715; 5531759; 5478352; 5275616; 5192300; 5108421 and 5061274.Collagen compositions, which are mechanically destroy for ismanam and insoluble collagen. Injectable collagen composition is described in U.S. patent 4803075. Composition that can be injected into the bone/cartilage described in U.S. patent 5516532. Feed matrix-based collagen containing dry particle size in the range from 5 to 850 μm, which can be suspended in water and have a specific surface charge density, described in the patent WO 96/39159. Collagen preparation having a particle size of from 1 to 50 μm, which is useful as an aerosol for education dressings for wounds, described in U.S. patent 5196185.Polymer neurodynamic hydrogel, which can be made and which can be entered via a syringe described in WO 96/06883. Polyoxyalkylene polymer to prevent the formation of adhesions described in U.S. patent 5126141.Following the application under consideration, which are assigned to the assignee of this application, contain related subject matter: USSN 60/050437, filed June 18, 1997; 08/704852, filed August 27, 1996; 08/673710 filed June 19, 1996; USSN 60/011898, filed February 20, 1996; USSN 60/006321, filed November 7, 1996; USSN 60/006322, filed November 7, 1996; USSN 60/006324, filed November 7, 1996; and USSN 08/481712, filed June 7, 1995. Full disclosure of each of these applications is given here as a reference.The invention Sonia such compositions to areas of the target in the patient's body. These methods and compositions are particularly suitable for preventing or deterring the formation of tissue adhesions, such as adhesions tissue on the spine after surgery and traumatic injury, in addition, the present compositions and methods can also find application in the stop or stop bleeding (hemostasis), especially when used in combination with the corresponding hemostasis means, such as thrombin, fibrinogen, factors of the blood coagulation system, etc., These compositions are also useful for filling of the tissues, especially to fill in areas of soft and hard tissues, including channels, path, body cavities, etc., which are present in the muscles, skin, epithelial tissue, connective or supporting tissue, nervous tissue, eye and other tissues sensitive organs, vascular and cardiac tissues, gastrointestinal tissues and organs, pleura and other lung tissue, kidney, glands of internal secretion, male and female genital organs, adipose tissue, liver, pancreas, glands, lymph, cartilage, bone, intraoral tissue and gelatinous connective tissue. The compositions according to the present invention will also be useful for filling soft implan nicaenum for enzyme barrier or lining. These compositions are also useful in other operations where it is desirable to fill a limited cavity biocompatible and resorbable polymeric material. Moreover, these compositions can be combined with drugs and other biologically active means, where drugs can be released after a certain period of time on a given area of the target.The compositions according to the present invention, contain molecular cross-linked hydrogel, which is able to dissolve and contains a small derivative works from, with some size and other physical properties that improve the fluidity of the gel (for example, its ability to extrusion through a syringe) and the ability to flow and adapt to the areas on or within tissue, including tissue surface and formed cavity, such as the gaps between the vertebrae, cavities, holes, pockets, etc. in the tissues. In particular, derivative works (subgroups) have a size that allows them to flow, when the compositions are subjected to voltages above the threshold level, for example during extrusion through the opening or cannula or when they condense on the site feed using a spatula or similar Poro">105PA. However, these compositions will remain essentially stationary during the exposure of their stresses below the threshold level.These compositions can be dry, partially hydrated or fully hydrated, and they will demonstrate the degree of swelling of from 0 to 100% depending on the degree of hydration. Fully hydrated material will absorb about 400-1300% water or aqueous buffer solution by weight, which corresponds to a nominal increase in the diameter or width of the individual particles derived units in the range of from about 50% to about 500%, usually about 50-250%. Thus, the particle size of the initial material as a dry powder (to hydration) will determine the size of the partially or fully hydrated derived units (depending on the following factors). Typical and preferred size intervals for dry particles and fully hydrated derived units are given in table.1 the Compositions according to the present invention, will be present usually in the form of dry powder, partially hydrated gel or fully hydrated gel. The dry powder has a moisture content of below 20% by weight) is p is mounted gels, which usually 50-80% hydration, are useful for those applications in which you want the material is swollen after its application onto the wet area of the target, such as a hole in the fabric. Fully hydrated form will be useful for those applications where the swelling on site (in situ) is not required, for example in the spinal column and other areas in which there are nerves and other sensitive structures.Dimensions derived units (subgroups) can be obtained in various ways. For example, cross-linked hydrogel having dimensions exceeding the range of the target (as hereinafter defined), one can mechanically destroy several points during the manufacturing process. The present composition can be destroyed, in particular, (1) before or after crosslinking of the original polymer material and (2) before or after hydration stitched or unstitched polymer starting material, for example, as a fully or partially hydrated material or dry powder particles. The term "dry" will mean that the moisture content is quite low, typically below 20% by weight of water, thus, the powder is free flowing, and that individual particles will not be aggregated. The term "hydrated" will mean, the shaft 80 to 95% of the equilibrium level of hydration, thus, the material will act as a hydrogel.Mechanical destruction of the polymer material in the dry state, preferably in cases where it is desirable to adjust the particle size and/or particle size distributions. It is easier to adjust the grinding of dry particles than hydrated hydrogenrich materials, thus, the more easily you can adjust the size of the resulting smaller particles. On the contrary, the mechanical destruction of the hydrated cross-linked hydrogels usually be achieved more simply and it involves fewer stages than the grinding of dry polymer starting material. Thus, the destruction of hydrated gels may be preferable in cases where the final size and/or size distribution of the derived units of the gel are not critical.In the first typical method of manufacturing a dry seamless polymeric starting material, for example dry gelatin powder, is subjected to mechanical failure during normal operation in Assembly, for example, by homogenization, grinding, koatservatsii, grinding, grinding in a jet mill, etc., the Powder will be enough fragmentation to achieve the size of the dry particles, which will be on the d gidratirovana. The relationship between the size of the dry particles and the size of the fully hydrated subgroup will depend on the swelling of the polymer material, as will be hereinafter defined.Or powdered polymer starting material can be obtained by spray drying. Methods of spray drying is based on the flow of the solution through a small opening such as a nozzle for the formation of droplets that are released into the gas stream in counter-current or single direction of flow, usually in a heated gas stream. Gas evaporates the solvent from the liquid source material, which may be a solution, dispersion or so on, the Use of spray drying for the formation of the source material dry powder is an alternative to the mechanical destruction of the source material. The operation of the spray drying will typically form a seamless product is a dry powder with a very uniform particle size. As will be described later, the particles can be made then.In many cases, the operation of mechanical destruction can be adjusted enough to get the size and distribution of particle size within the desired interval. However, in other cases, when you want a more accurate distribution of the size of h is raspredeleniya particle size, for example, by sieving, or aggregation, etc., Mechanically crushed polymer source material then sewn together, as will be described in more detail, and dried. The dried material may be desirable end product, when it can be re-gidratirovana and can swell immediately before applying. Either mechanically crushed crosslinked material can be re-hydration and then packaging re-hydrated material for storage and subsequent use. The following describes specific ways of packaging and application of these materials.Where the size of the derived units of the crushed hydrogel less important, the dried polymer starting material may be hydrated, to dissolve or to weigh in the appropriate buffer solution and sew to mechanical destruction. Mechanical destruction of the pre-formed hydrogel is typically achieved by passing the hydrogel through the hole, where the hole size and strength extruded together define the particle size and particle size distributions. Although this method is often simpler than the mechanical destruction of the dry polymer particles to hydration and crosslinking, however, the possibility reformirovania gels can be placed in a syringe or other applicator to mechanical destruction. The materials will then be mechanically destroyed during their flow through the syringe onto the area of the target tissue, as will be described in detail later. Or unground cross-linked polymer material can be stored in a dry form prior to its use. Dry material can then be loaded into a syringe or other suitable applicator, hydrate inside the applicator and mechanically grind at the time of submitting material to the site of the target, and again usually through a small hole or cavity of a tubular organ.The polymer can be made and be hydrated to form a hydrogel, as will be described in more detail later. Typical polymers include proteins selected from gelatin, collagen (e.g., soluble collagen, albumin, hemoglobin, fibrinogen, fibrin, fibronectin, elastin, keratin, laminin, casein and derivatives thereof, and combinations. Or the polymer may contain a polysaccharide, such as glycosaminoglycan, a derivative of starch, a derivative of cellulose, derived hemicellulose, xylan, agarose, alginate, chitosan, and combinations thereof. As another alternative, the polymer may contain non-biological polymer that forms a hydrogel, such as polyacrylates, polymethacrylates, poly is f and the combination.The binding polymer can be achieved by any known method. For example, in the case of protein binding can be achieved using an appropriate cross-linking agent such as aldehyde, periodate sodium, epoxy compounds, etc., Or alternatively, the binding may be caused by exposure to radiation, for example,-irradiation or electron beam irradiation. Applying appropriate cross-linking agents and radiation, can also sew polysaccharides and non-biological polymers. Non-biological polymers can also be synthesized as cross-linked polymers and copolymers. For example, the reaction between mono - and polyunsaturated monomers can lead to the formation of synthetic polymers, having an adjustable degree of crosslinking. Typically, the polymer molecule has each molecular weight in the range from 20 to 200 KD and has at least one relationship with another polymer molecule in the structural grid, and often she has from 1 to 5 links where the true level of crosslinking chosen in part to ensure the required rate of biodegradation in the intervals, which will be mentioned later.The degree of crosslinking of the polymer effect on some functional properties of the hydrogel, including the ability to fill the cavity, the ability to swell and the ability to adhesion to the tissue. The degree of crosslinking of the polymeric gel composition can be adjusted by adjusting the concentration of the crosslinking agent, exposure to radiation for crosslinking, a change in the relative amounts of mono - and polyunsaturated monomers, changes in the reaction conditions, etc., Usually the degree of crosslinking regulate by adjusting the concentration of the crosslinking agent.Exposure to radiation, for example,the irradiation can also be used for sterilizing compositions before or after packaging. In that case, if the composition consists of a sensitive radiation materials, the composition must be protected from exposure to sterilization. For example, in some cases it will be necessary to add ascorbic acid to the exclusion of further crosslinking of the material as a result of free-radical mechanisms.Hydrohalide compositions according to the present invention have a solids content in the range of from 1 mass% to 70 mass%, preferably 5-20% by weight, better from 5 to 16% by weight. For gels having a higher solids content, usually more than 16% by weight, preferably incorporated into the composition dtweedie plasticizers include polyethylene glycols, sorbitol, glycerol, etc.,The equilibrium swelling of the crosslinked polymers according to the present invention, will be usually in the range of 400%-1300%, preferably 500-1100% depending on their intended applications. This equilibrium swelling can be adjusted by changing the degree of crosslinking, which is achieved, in turn, by changing the conditions of the crosslinking, for example fashion style staple, the duration of exposure to the action of a crosslinking agent, the concentration of crosslinking agent, the temperature cross-linking agent, etc.In the section "Experiments" described and prepared materials having values of equilibrium swelling about 400-1300%. Found that materials having different values of equilibrium swelling, act differently in different applications. For example, the ability to stop bleeding in the model divot liver was easily achieved with crosslinked gelatin materials having a degree of swelling in the range of 700-950%. For the femoral artery, the more successful the values of equilibrium swelling of the tampon in the range of 500-600%. Thus, the ability to regulate the binding and the equilibrium swelling allows you to optimize the compositions according to the present invention, for nesporedstvenno to its application to the area of the target. Of course, hydration and equilibrium swelling are closely linked. Material with 0% hydration will not swell. Material with a 100% hydration will be at its equilibrium water content. Hydration between 0% and 100% will correspond to swelling between the minimum and maximum amounts. In practice, many dry kinabuhi materials according to the present invention, will have some residual moisture is usually less than 20% by weight, often from 8 to 15% by weight. When used herein, the term "dry", it refers to materials having a low moisture content, where the individual particles are free flowing and usually anabuki.Hydration can be easily adjusted by regulating the amount of aqueous buffer solution added to the dry or partially hydrated cross-linked material before applying it. Usually, as a minimum, it is desirable to introduce a sufficient amount of aqueous buffer solution, so that the material can be squeezed through a syringe or other feeding device. However, in other cases it may be desirable to use a spatula or other applicator to supply less flowable materials. The required degree of hydration pomogaet, it is generally preferable to use partially hydrated gel, which can swell and fill the cavity due to the absorption of moisture from the area of the target. On the contrary, a fully or substantially fully hydrated gels are preferred for use in the brain, near the spine and on the sites of a target near nerves and other sensitive body structures that can be damaged during swelling after applying the gel. You can also make use of the gel composition according to the present invention, excessive contents of buffer solution, having a two-phase composition having a fully hydrated gel and free phase buffer solution.The preferred hydrogel material according to the present invention is gelatin, which is made to achieve 700-950% of swelling at equilibrium hydration. The material will be cut to have a size of the gel particles in the range of from 0.01 to 1.5 mm, preferably 0.05-0.5 mm, and in order to achieve 70-100% equilibrium swelling before applying it on the site, it will preferably be subjected to hydration to a sufficient level.In some cases, the hydrogel composition according to the present invention, can and/or semipolymer, and combinations of two or more separate materials of each type of polymer, such as two or more proteins, polysaccharides, etc.,Polymer compositions according to the present invention may contain a combination of powdered cross-linked polymer hydrogels, which have been described, and seamless polymeric materials. The crushed crosslinked polymer hydrogels consist of sets of subgroups, with the amount determined by the method of preparation. Choose the size that it was suitable for sealing a limited volume, had the fluidity and described hereinafter, the rate of biodegradation. The discrete nature of the crosslinked derivatives of units, however, leaves the area of voids that can be filled in combination with unstitched polymeric material. Seamless polymeric or other material of the filler may contain any of the above polymeric materials, and it may, but not necessarily, be the same polymeric material which is sewed to obtain custom made mechanically destroying the gel. Choose the relative amount of cross-linked polymer and unstitched polymer to obtain a relatively continuous (free of voids) of the composition after an optional mechanical destruction ichitomo the polymer), usually in the range from 10:1 to 2:1, preferably from 5:1 to 2:1.The hydrogels according to the present invention can be applied using a syringe, spatula, brush, spray by hand by force, or by any other known means. Gels are usually applied using a syringe or similar applicator, able to squeeze out the gel through a nozzle hole, a needle, tube or other channel for the formation of beads, layer or similar piece of material. Mechanical destruction of the gels may occur during the extrusion of the gel through the opening of the syringe or other applicator, typically having a size in the range from 0.01 to 5.0 mm, preferably 0.5 to 2.5 mm, Preferably, however, first prepare the polymeric hydrogel from a powder having the desired particle size (which after hydration gives derivative works of the hydrogel with the desired size), or it can be partially or fully mechanically grind to a specified size to the final extrusion or other method of application.The composition can be applied at different degrees of hydration, typically, but not necessarily, at least partial hydration. If they are UN-hydrated form, the composition will be nabuk put at their equilibrium levels of hydration of the composition will exhibit essentially the equilibrium hydration and slight, or no swelling, swelling while applying them to the fabric. Swelling of the non hydrated and partially hydrated compositions is due to the absorption of moisture from the fabric and the environment in which the composition is applied.According to the present invention is also suggested kits containing any of the above hydrated or non hydrated materials gel in combination with written instructions for use that specify any of the above methods of application of the gel on the target tissue. Composition and written instructions will be placed together in a conventional container such as a box, jar, bag, tray, or similar Written instructions can be printed on a separate sheet of paper or other material and placed on/or within the container, or they can be printed on the container. Composition (compositions) usually placed in a separate sterile bottle, vessel, vial, or so on, When the gel material is not gidratirovana, the kit may include a separate container with an appropriate aqueous buffer solution for hydration. It may include other elements of the system, for example the applicator, i.e. the syringe.A brief description of h is, on formed during the surgery, the defect on the vertebral body to prevent the formation of adhesions after surgery on the spine.Figures 2A and 2B show the application of the compositions of the molecular cross-linked polymer gel according to the present invention, a defect in soft tissue, where the treated area may cover a protective cover after filling defect of the polymer composition.Figures 3A and 3B show the use of molecular-crosslinked polymer compositions according to the present invention for filling a hypodermic penetration of tissue into a blood vessel, for example a fabric path, formed as part of the commissioning procedure of the catheter into a blood vessel.Figure 4 shows a kit containing sterile packaging for applicator comprising molecular crosslinked polymer composition according to the present invention.Figure 5 shows the relationship between the swelling percentage and the percentage solids content in the polymer gel.Description of the preferred variants of the Composition according to the present invention, contain absorbable biocompatible molecular cross-linked hydrogels. The term "biocompatible" osni Standardization (NAS, Northwood, Ohio). The term "biodegradable" means that the composition will decompose or dissolve when they are placed directly on the target area on the patient's body (not protected within the implantable device, such as a prosthesis for the breast) for some period of time, for example, in one year or less, usually from 1 day to 1 year, and more often from 1 day to 120 days. Specific Protocol for measuring recable and destruction are presented in the section "Experiments". The term "molecular crosslinked" means that the materials contain in the polymer molecule (that is, a single chain), which are connected by bridges composed of an element, group, or connection, where the atoms of the main chain of the polymer molecules are connected by primary chemical bonds. As will be described in detail hereinafter, the crosslinking can be achieved in different ways.The term "hydrogel" means that the composition contains a single-phase aqueous colloid, in which the biopolymer or nabopolassar, as will be defined in more detail further absorb water or aqueous buffer solution. The hydrogel contains multiple links, where each link represents a molecular cross-linked hydrogel having dimensions that depend on the degree gidratnoi water, that is, the water cannot be removed from the hydrogel by simple filtering.The term "percentage of swelling" means the dry weight (mass) is subtracted from the mass in wet state, divided by the dry weight and multiplied by 100, where "wet weight" is measured after removal of the wetting agent, as fully as possible, outside of the material, for example, by filtration, and dry weight was measured after exposure to elevated temperatures for a time sufficient to evaporate the wetting agent, for example 2 hours at 120oC."Equilibrium swelling" is defined as the percentage of swelling at equilibrium after immersing the polymeric material of the wetting agent over a period of time sufficient so that the water content became constant, usually 18-24 hours."Target area" is a place that must be made of a gel material. Typically, the target area is the site of the tissue of interest, however, in some cases, the gel can be entered or to hand out at a point near a location of interest, for example, when the material swells on site (in situ), to close places of interest.The hydrogels according to the present invention, can be obtained from biopolymer llagen, albumin, hemoglobin, casein, fibrinogen, fibrin, fibronectin, elastin, keratin, laminin, and their derivatives and combinations. Especially preferred is the use of gelatin or instant defibrillator collagen, and preferably gelatin; typical compositions of gelatin follows. Other relevant biopolymers include polysaccharides, such as glycosaminoglycans, derivatives of starch, xylan, cellulose derivatives, derivatives, hemicellulose, agarose, alginate chitosan and their derivatives and combinations. Appropriate abipolar choose those which are capable of destruction either of two mechanisms: that is, (1) break the main chain of the polymer or (2) degradation of the side chains, resulting in solubility in water. Typical abipolar for the formation of hydrogels include synthetic materials, such as polyacrylates, polymethacrylates, polyacrylamides, polyvinyl resins, polylactide-Lida, polycaprolactones, polyoxyethylene and their derivatives and combinations.The polymer molecule can be made by any method suitable for formation of a water of the hydrogel according to the present invention. For example, the polymer molecules can be made using bi - or polyfun the ranks of bifunctional cross-linking agents include aldehydes, epoxides, suktinimida, carbodiimide, maleinimide, azides, carbonates, isocyanates, diphenylsulfone, alcohols, amines, imidate, anhydrides, halides, silanes, diazoacetate, aziridine, etc., Crosslinking can be achieved using the oxidative or other agents, such as periodates that activate the side chain or part of the polymer, thus they can react with other side chains or parts for cross-linking. An additional way of stitching is the exposure of polymers to radiation, e.g. gamma radiation with the aim to activate the polymer to the reaction was carried out stitching. You can also apply dihydroceramides ways of stitching. Dehydrothermal crosslinking of gelatin can be achieved by maintaining it at an elevated temperature, typically 120oSince, during the period of time of at least 8 hours. With increasing degree of crosslinking, as reduced the percentage of swelling at equilibrium can be achieved by increasing the temperature, increasing the exposure time or a combination of both. The crosslinking reaction can be accelerated work under reduced pressure. The following describes preferred methods of crosslinking of the gelatin molecules.Moley degradation of the gel. As the plasticizer may be an alcohol, such as glycol, sorbitol or glycerol, preferably a polyethylene glycol having a molecular weight in the range of from about 200 to 1000 L, preferably about 400 D. the Plasticizer will be present in the compositions in amounts of about 0.1% by weight to 30% by weight, preferably from 1% by mass to 5% by weight of the composition. Plasticizers are particularly useful for use with gels having a high solids content, usually more than 10% by weight of the composition (without plasticizer).What follows is a description of typical methods for obtaining molecular-stitched gelatino. Get gelatin and placed in aqueous buffer solution for education unstitched gel, typically having a solids content of 1 to 70% by weight, often from 3 to 10% by weight. Gelatin sew typically by exposure to the action of glutaraldehyde (e.g., 0.01 to 0.05% by weight, throughout the night, at a temperature 0-8oWith in aqueous buffer solution), periodate sodium (for example, 0.05 M, at a temperature 0-8oC for 48 hours) or 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide ("EDC") (for example, 0.5 to 1.5% by weight, in the over-night at room temperature, or under what should be suspended in alcohol, preferably methyl or ethyl alcohol when the solids content 1-70% by weight, typically 3-10% by weight, and sew by exposure to the action of a crosslinking agent, usually glutaric aldehyde (e.g., 0,01-0,1% weight by weight, overnight at room temperature). Cross-linking with glutaraldehyde formed cross-links (bridges) through tiffanie reason that you can stabilize, followed by treatment with sodium borohydride. In the case of aldehydes, you should maintain the pH at a value of about 6 to 11, preferably 7-10. After blending the resulting granules may be washed in distilled water and can be rinsed in alcohol, dried and re-suspended to a given degree of hydration in an aqueous medium having the desired buffer solution and pH. The obtained hydrogels can be downloaded then the applicators according to the present invention, as will be described in more detail later. Or hydrogels can be mechanically grind to/or after stitching, as will be described in detail.This is followed by typical methods for obtaining molecular-linked gelatin compositions, with the percentage of equilibrium swelling in the range of about 400-1300%, preferably 600-950%. Get gelatin, a first crosslinking agent in solution (usually glutaric aldehyde, preferably in a concentration of 0.01-0.1% by weight) for gel formation, typically having a solids content of 1 to 70% by weight, typically 3-10% by weight. The gel mix well and maintain at a temperature of 0-8oWith all night during stitching. It then washed with deionized water three times, twice with alcohol (preferably methanol, ethanol or isopropyl alcohol) and allow it to dry at room temperature. To further stabilize the stitching gel can be treated with sodium borohydride.The compositions according to the present invention can also be combined with other materials and components, for example, the bioactive component (components), enter the patient with a viscosity modifier, such as carbohydrates and alcohols, and other materials designed for other purposes, for example, to regulate resorbtive. Typical bioactive components include, but are not limited to, proteins, carbohydrates, nucleic acids, and inorganic and organic biologically active molecules, such as enzymes, antibiotics, antineoplastic agents, bacteriostatic agents, bactericidal agents, antiviral the substances, antibodies, neurotransmitters, psychoactive drugs, drugs affecting reproductive organs, and the oligonucleotides, for example protivocesterne oligonucleotides. These bioactive components are normally present in relatively low concentrations, typically less than 10% by weight of the compositions, essentially less than 5 mass%, and often below 1% by mass.Typical hemostatic agents include thrombin, fibrinogen and factors of the blood coagulation system. Hemostatic agents such as thrombin, can be added in a concentration of from 50 to 10,000 units of thrombin per ml of gel, preferably from about 100 units of thrombin per ml of the gel to about 1000 units of thrombin per ml of gel.Molecular cross-linked hydrogels according to the present invention, can be subjected to mechanical failure during their submission to the site of the target by extrusion through an orifice or other restriction of flow, or they can mechanically destroy during periodic process prior to their submission to the site of the target. The main purpose of this stage is mechanical damage is the formation of multiple sub-groups of the hydrogel, having a size, which increases their ability to fill and seal the floor through the tube, cannula and/or other applicators small diameter surface of the target. Without mechanical destruction of the molecular cross-link hydrogels will be difficult to adjust and fill uneven cavity of the processed target, for example, the intervertebral spaces in the spinal column, tissue cavity, subcutaneous tissue path, etc., by shredding the gel into particles of a smaller size such gaps can be filled more efficiently, while maintaining mechanical integrity and durability of cross-linked gel, which is essential for him, allowing him to act as a tool against the formation of adhesions, filler for tissue or etc., it has been Unexpectedly discovered that a simple extrusion manually composition, usually with the use of the syringe, having an aperture size in the range from 0.01 to 5.0 mm, preferably from 0.1 to 2.5 mm, provides the appropriate amount of mechanical destruction to improve properties of the gel as described above.Gel compositions according to the present invention, can be subjected to mechanical failure are also up to their end use or supply. Molecular crosslinking of polymer chains of the gel can be made before or after its mechanical , as, for example, mixing, because the composition of the gel is crushed into subgroups that have a size in the above range of 0.01 to 5.0 mm When the composition of the gel is subjected to destruction before it is applied, the gel can be applied or to introduce other ways, not by extrusion, for example, using a spatula, spoon or similar Other periodic mechanical methods of destruction include pumped through a homogenizer, a mixer or a pump, which compresses, stretches or poses its shearing force to a level that exceeds the yield strength of the hydrogel at destruction. In some cases, the extrusion of the polymer composition causes the gel to become, essentially, a continuous structural grid, i.e. grid, which increases the size of the initial weight of the gel, in the aggregate branch grids or subgroups, which have dimensions in the above intervals. In other cases, you may need to partially destroy the gel composition before loading it into a syringe or other applicator. In such cases, the gel material will achieve the desired size of the subgroup to the final extrusion.In the now preferred embodiment, it is possible first to prepare a polymer (e.g., drying, spraying and/or megawide finely ground or powdered dry solid material, which can be destroyed by further grinding to form particles having the desired size, which is usually limited to narrowly outside of a small interval. You can also, in addition, a stage of a choice of size and modification, such as screening, cyclone classification, etc. described below For typical gelatinous materials the size of the dry particles is preferably in the range from 0.01 to 1.5 mm, more preferably from 0.05 to 1.0 mm, a Typical grain size distribution is such that more than 95% by weight of the particles will be in the range of 0.05 to 0.7 mm, the Methods of grinding the polymer starting material include homogenization, grinding, coacervation, crushing, the grinding in a jet mill, etc. Powdered polymer starting materials can also be obtained by spray drying. The distribution of particle size can also be adjusted to improve the known methods, for example by sieving, aggregation, additional grinding and etc.Dry powdered solid material can then be suspended in an aqueous buffer solution, as described here, and sew. In other cases, the polymer can be suspended in an aqueous buffer solution, to sew, and ZAT is SQL in aqueous buffer solution. In all cases, the resulting material contains cross-linked hydrogel having discrete illumination with the above dimensions.The compositions according to the present invention, after mechanical grinding, will be able to resorption, i.e. they are capable of biodegradation in the body of the patient over a period of time less than one year, usually from 1 to 120 days, preferably from 1 to 90 days, and more preferably from 2 to 30 days after their initial application. This is particularly true, when these materials are used to prevent the formation of postoperative and other adhesions, when necessary, the only barrier between the treated surfaces of the fabric, while there will be a healing of tissue. Methods of measuring the length of time required for resorbtive, shown in Example 11 under "Experiments". In other cases, for example, when the present compositions are contained within an implantable device, such as a prosthesis for the breast, the resorption of the material will be prevented by the membrane or other mechanical barrier surrounding the song (unless damaged).Refer now to Figure 1, which describes a method of preventing the formation of adhesions after oblasna the present invention, for applying the gel so that it covers the open solid membrane of the brain. Typically, the gel will dissolve over a period of time in the range from 7 to 60 days.Refer now to Figures 2A and 2B, showing that the molecular cross-linked hydrogels according to the present invention can also be used for filling cavities D in the soft tissue So the Syringe 50 includes a cylinder 52, a piston 54 and the cannula 56, contains a molecular cross-linked hydrogel inside the cylinder 52. The hydrogel caught With the cannula 56 by pressing on the piston 54 in a known manner. As shown in Figure 2B, is squeezed out enough gel to fill this cavity. It is preferable to use partially hydrated hydrogel, which will continue to swell during its exposure to the action of moist surrounding tissue environment. It may be desirable to place the pad P over the outer surface of the gel, as shown in Figure 2B. As the pads can be adhesive or other conventional self-adhesive pad. However, preferably, the overlay contains collagen, gelatin or other film which can be fixed by application thereto of energy, such as optical or radio and 3B, which show that the compositions and methods according to the present invention can also be applied to fill the subcutaneous tissue tract TT, which form via the button on top of the fabric to gain access to the blood vessel BV. At the distal end of the channel in the tissue of the TT can be placed on the inner wall of the blood vessel, the barrier element 70. To hold the barrier element 70 in place, you can use the thread 72. For extrusion of molecular crosslinked hydrogel material according to the present invention, in a channel fabric on the barrier element 70 is used syringe 74 containing the cylinder 76, the piston 78 and the cannula 80. As shown in Figure 3B, to fill the entire internal volume of the channel in the tissue used hydrogel, which is preferably partially gidratirovana for swelling after placement, as described above. On the exposed surface of the channel fabric (not shown) can place the pad or other coverage.Refer now to Figure 4, which shows kits, according to the present invention containing hydrated, partially hydrated and/or UN-hydrated described polymer composition, packaged in an appropriate container is ATOR 90, which contains a pre-extruded molecular crosslinked hydrogel according to the present invention. The applicator may take a variety of forms, including syringes, as described above. The Figure 4 shows the applicator 90, containing a tube 92 having a neck 94, which forms the opening for extruding the hydrogel. The gel contained inside the tube and it can squeeze through the neck 94 by pushing the tube. The applicator 90 is contained preferably in a sterile package 96. Sterile packaging may take various forms, but it is shown in the form of a shell containing a base sheet and a transparent plastic cover. Such packages can be sterilized in a known manner. The radiation used for the crosslinking of the hydrogel can also be used for sterilization of the entire package. Instructions for use may be printed on the packaging or on a separate sheet enclosed in the package.The present invention can also be used to stop bleeding (causing hemostasis) on the injured tissue surface, for example on the surface of any organ, including the liver, spleen, heart, kidneys, intestines, blood vessels, vascular organs, etc. For applying gel on inosensu the hemostatic agent. The gel is to be applied so that the actively bleeding worn or damaged area is completely covered with absorbable molecular-linked gel. Appropriate hemostatic tools include thrombin, fibrinogen and other factors of the blood coagulation system, as described, for example, in U.S. patents 5411885; 4627879; 4265233; 4298598; 4362567; 4377572 and 4442655 listed here for reference. The catalytic components of the hemostatic tool, such as thrombin, can be connected essentially in the syringe immediately before use, so their combined activity will persist until such time as they will be printed on the fabric.When they are used in the areas surrounding nerves and other sensitive structures of the body, is used preferably fully hydrated hydrogels (i.e., > 95% hydration at the equilibrium swelling) to avoid damage to the nerves because of swelling in a closed environment.To illustrate and not to limit offered the following examples.Experimental EXAMPLE 1: MATERIALS AND METHODS for producing POWDERED POLYMER PRODUCT Powdered polymer compositions have, in General, as follows.Applying loose the PTA), having a solids content of 10%, allowed to swell in 0.1 N aqueous sodium hydroxide and 0.05 periodate sodium and kept at a temperature of 0-8oWith in 2-3 days. The swollen granules were washed in distilled water until he received a pH of 8. Neutralized swollen granules were dried in a fume hood with laminated flow and again suspended them in 0.05 M phosphate, 0.15 M sodium chloride at pH of 7.2 +/- 0.2 and 10% solids content. The composition was then downloaded into the syringe a volume of 3.0 cm3and was irradiated with electron beam radiation value of 3.0 megarad for her sterilization.EXAMPLE 2: MATERIALS AND METHODS for producing POWDERED POLYMER PRODUCT
Gelatine (Woburn) was allowed to swell in an aqueous buffer solution (for example, 0.05 M phosphate, 0.15 M sodium chloride at pH of 7.2 +/- 0,2) when the solids content of 1-10%, and then subjected to crosslinking using glutaraldehyde (0.01 to 0.05% weight by weight, during the whole night at room temperature), periodate sodium (0.05 M, 0-8oWith, within 48 hours) or by irradiation 3,0 megarad gamma or electron beam radiation. The gels were then squeezed out of the syringe through the normal application of manual effort.EXAMPLE 3: MATERIALS AND Spooge when the solids content of 1-10% (weight/weight), cooled to 5oC. the resulting gel to grind by stirring with a mixer, motor-driven. Then added periodate sodium and sodium hydroxide, and mixed to obtain a 0.05 M of periodate sodium and 0.10 M sodium hydroxide. The cooled mixture was maintained at a temperature 0-8oWith in 2-3 days. Particles of cross-linked gel was then washed with water at a temperature of 5oC to achieve a pH of 8. Finally, the gel particles were washed aqueous buffer solution (for example, 0.05 sodium phosphate and 0.15 sodium chloride at a pH of 7.2+/-0,2) and left at a temperature of 0-8oTo reach the equilibrium state buffer solution. Free buffer solution was decanted from the fragmented mass of the gel, and the gel particles was loaded into a syringe and subjected to electron beam or gamma irradiation at 3,0 megarad for their sterilization. Such sterilized particles of gel squeezed directly from the syringe, forcing them further fragmentation.EXAMPLE 4: MATERIALS AND METHODS of preventing FORMATION of ADHESIONS AFTER SURGERY
This study has demonstrated the efficacy of powdered polymer composition of the gel to eliminate or reduce scarring after excision of the arc of the vertebra. The purpose is Noah injection of ketamine hydrochloride in combination with xylazine made anesthesia. Each rabbit was injected intramuscularly with 5 mg/kg dose Baytril. From the middle of the chest (approximately T-10) to tail shaved back of each rabbit. For proper preparation of the epidermis expanded far enough shaved area on the abdomen. The rabbit was placed on a pad, circulating water in the supine position under sterile conditions. To obtain a small trough in the lumbar region around it placed a small towel. The epidermis on the lumbosacral region was prepared by cleaning with iodoform and washing with 70% alcohol. In the middle of the epidermis made an incision from L-1 to L-5 and inflicted upon him the armband designed for the lumbar-sacral area. Hemostasia was achieved by a combination of mechanical compression and electrocautery. The bandage made the cut for placing the tips of the prongs of the spinal cord. The muscles around the spinal cord was separated from the processes of the spinal cord through the opening and the plate L-4 using peristernia Elevator. Muscles kept at a certain scattering using the retractor with cremalleras. General dorsal Lamin-ectomy L-4 was carried out by removing the spinous process cost the s measures to prevent damage to the spinal cord or roots of the lumbar and sacral plexuses in the spinal cord. The laminectomy defect was washed with sterile saline and removed any remaining bone fragments. Removed yellow fat and epidural fat ligaments, leaving a clean Dura, open to some degree of laminectomy.After the operation is completed at site L-4 was carried out identical to the laminectomy at the level L-2. During this operation, the plot L-4 was protected from drying with a swab soaked in sterile saline. The cavity L-3 has not been subjected to processing, to provide between the two treated plots barrier of soft tissue. After cooking both plots these plots were subjected to the treatment experienced by the material or leave them as control samples according to a randomly selected code.Rabbits were determined in the experimental group and after performing a laminectomy was carried out the following operations. Unprotected Dura at the site of lumbar handled 0.5 to 0.9 ml fragmented gelatin composition of Example 1. The material was placed in such a way that all exposed Dura was closed test material. At another part of the waist unprotected Dura mo is a new bandage (fascia) closed absorbing seam of the appropriate size (for example, 4-0) with a simple interrupted pattern. Subcutaneous tissue was closed by imposing absorbing seam with a simple continuous pattern, and the skin was closed with appropriate suture material, or surgical staples.During the first five days after surgery, animals were injected twice daily intramuscular injection of 5 mg/kg Baytril (Baytril).At 7 or 28 days after surgery the animals killed and subjected to autopsy. Did free surgical incision and explored the area excision arc vertebra.The spikes on the meninges were sorted and marked according to the degree and severity and their expense were as follows:
The spikes on the meninges: the adhesion of connective tissue between bone or a deep scar and Dura mater inside the spinal canal. It was determined by entering the probe between the bone and Dura mater to separate the two structures (see tab.2).In any case, which applied the fragmented gelatin composition was not observed in the formation of adhesions after surgery. However, adhesions (adhesion) was formed in 71% of the control plots. The results of the test sites on sehato study has demonstrated the effectiveness of fragmented resin composition for sealing (sealing) of openings in the vessel as a result of the injection. Investigated the femoral artery at the crossed white pigs Gempshirsky/Yorkshire breeds, which were grown on the farm (pork power the farms, Tart, California), the femoral artery was identified and was Coulibaly with a needle (SmartelTMCardiovascular Dynamics, Irvine, California). For the formation of a passage to the vessel and increasing the hole of the femoral artery was used by the expander (dilator) 9 French once posted a guide wire. Next, the expander is removed and introduced the shell 7 French in the femoral artery. After this has removed a guide wire. Check the installation by diversion of blood into the side branch of the shell. Pulsating arterial bleeding was also noted at the insertion point of the membrane at the site of the incision on the skin. After removal of the sheath was introduced into the canal fragmented gelatin composition of Example 1 using the Teflon tip of the catheter (18 gauge) attached to a syringe for subcutaneous injection. The exit point of the catheter tip bleeding is not found, and it demonstrated the effectiveness of fragmented gelatin composition for sealing of the injection site and surrounding vessel tissue.EXAMPLE 6: FRAGMENTED Primararily 1 as a carrier for filling and sealing holes in liver tissue was demonstrated by this study. In the liver of the white hybrid pigs Gempshirsky/Yorkshire breeds (pork power the farms, Turlock, California) caused the formation of the three RAS (2 holes in fabric and 1 shot in the fabric).After the surgical formation of a wound in the tissue of the #1 liver she was actively bleeding. For the complete filling of the defect in the tissue on it pushed the fragmented gelatin composition is contained in an amount of 1 ml syringe, includes approximately 500 units of thrombin - 1000 units/ml After 2-3 minutes of the formed clot that caused immediate cessation of bleeding. When the applied composition is captured by the tweezers, it turned out that it is fairly well stuck to the fabric and has good integrity. Against manual use of filling material objected, and additional bleeding was not observed.After surgical education hole #2 in liver tissue, it was actively bleeding. Approximately 1 ml of fragmented gelatin composition containing thrombin (approximately 500 units/ml) was dispensed from a syringe and caused to completely fill the defect in the tissue. Using an argon beam coagulator (Valleylab, boulder, Colorado or Birtcher Medical systems, Irvine, California), put the pad Raple surgical education blunt puncture holes in liver #1 was actively bleeding. For the complete filling of the defect tissue from a syringe extruded and applied to the defect approximately 0.8 ml fragmented gelatin composition containing thrombin (approximately 500 units/ml). After the filing of the fragmented gelatin composition all the bleeding stopped after about 2 minutes.After surgical education blunt holes noted active bleeding from holes #1 in the spleen. For the complete filling of the defect in the tissue inflicted on him by syringe approximately 0.8 ml fragmented gelatin composition containing thrombin (approximately 500 units/ml). Approximately 2 minutes after applying the fragmented gelatin composition all the bleeding stopped.In all four of the above examples used by the delivery system was a syringe with a volume of 3 cm3(Bacton Dickinson, Frank-Lin on the lake, new Jersey). It contained the fragmented gelatin composition of Example 1.The material according to the present invention, for filling holes and other tissue defects can be prepared as follows. A solution of thrombin (0.5 ml; 4000-10000 units/ml) added to 4.5 ml of fluid gel in order to obtain 5 ml of gel containing 400-1000 units/ml thrombin. Prominately composition AS a FILLER FOR TISSUE AND ANATOMICAL FILLING MATERIAL
This study demonstrated the effectiveness of fragmented gelatin composition as a system for closing a wound, which fills and seals the tissue defects. Formed surgically four holes in the fabric: 1 - easy, 2 - in the liver and 1 - spleen white crossed Gempshirsky/Yorkshire breeds of pigs grown on the farm (pork Pour the farms, Turlock, California).After the formation of the puncture in the tissue surgically observed air leakage from the lung. To completely fill the tissue defect caused approximately 1 ml fragmented gelatin composition from Example 1 using a syringe. Struck the pad "EpisilTM(Fujin Medical Technologies, Inc., Mountain view, CA) using an argon beam coagulator (Valleylab, boulder, Colorado or Birtcher Medical System-Thames, Irvine, California). Immediately stopped the air leak. When superimposed overlay captured with tweezers, it turned out that she quite firmly stuck to the fabric and had good integrity. The issue of fragmented gelatin composition was obtained by aeration of the lung to a pressure of 28 cm of water column.After surgical images syringe is dispensed approximately 1 ml fragmented gelatin composition and struck her on the tissue defect. Fragmented composition swollen and respectively stopped the bleeding, but still there was some leakage bleeding.The liver was noted to be bleeding after surgical education the holes in the fabric. Of the syringe is dispensed approximately 1 ml fragmented gelatin composition and caused her to completely fill the tissue defect. Using an argon beam coagulator (Valleylab, boulder, Colorado or Birtcher Medical systems, Irvine, California, struck the pad "EpisilTM(Fujin Medical Technologies, Inc., Mountain View, California). Happened the immediate cessation of bleeding. When the pad seized by the forceps, it turned out that she quite firmly stuck to the fabric and has good integrity.Puncture 1 in the spleen was actively bleeding after surgical education blunt puncture. To completely fill the defect tissue from a syringe is dispensed approximately 0.8 ml fragmented gelatin composition and caused her to defect. Approximately 2 minutes after applying the fragmented gelatin composition all the bleeding stopped.With appropriate anesthesia used a young female goats fermerskoi breed (Covertop Dairy, Mlno tore a blood vessel. The vessel caught using atravmaticheskikh clips to blood vessels, separated by a distance of about 2-3 cm Vessel was cut using the blade of a standard scalpel to reveal the two free ends of the blood vessels. Using 6-0 Prolene continuous suture, made the anastomosis end to end. After completion of the anastomosis clamps were removed. At the site conducting anastomotic bleeding was noted. Around the anastomosis is dispensed from a syringe about 2 cm3the fragmented gelatin composition containing thrombin (approximately 500 units/ml). The song put a gauze bandage. Approximately 3 minutes after applying the fragmented gelatin composition all bleeding has stopped. The incision was closed, respectively, and the animal allowed to recover for his further review.EXAMPLE 8: MATERIALS AND METHODS to PREVENT the formation of POSTOPERATIVE ADHESIONS IN the ABDOMEN
This study demonstrated the effectiveness of fragmented gelatin composition to eliminate and/or reduce the formation of adhesions in the abdominal cavity, when it is used alone or in combination with an overlay "RemisilTM(Fujin Medical Technologies, Inc., Mesovarium rat Sprague Dawley (Harris, E. C.(1995) "Analysis of the kinetics of education peritonitic adhesions in rats and to evaluate the potential anti-adhesions, Surgery, 117: 663-669). This model can measure objectively the only concrete bond between them.For this study used 15 rats Sprague Dawley. The anesthesia made by intramuscular injection of cetomimidae in combination with xylazine. After anesthesia in appropriate training for the operation was conducted by the middle line. The defect in the abdominal wall formed approximately at a distance of 1 cm transverse incision in the midline. The defect is formed by a cut segment 1x2 cm parietal peritoneum, including the superficial layer of muscles. The defect size 1x2 cm then formed on the serosal surface of the cecum. Blind intestine was subjected to the scraping blade of the scalpel so that scraped the surface formed a homogeneous surface the point of bleeding. The blind gut is then lifted and placed so that when closing the cecum will be in contact with a defect in the wall of the peritoneum. The defect in the wall of the peritoneum has been scraped in the same way. Both scraped area exposed to the air for 10 minutes.Has established the following 3 experimental groups. Each group consisted of 5 alive is latinova composition of Example 1, placed between the defect in the wall of the peritoneum and the defect in the cecum before closing.Group 3: Fragmented gelatin composition (Example 1) + overlay "EpisilTM"posted on defect in the cecum before closing.The incision on the midline of the closed overlapping absorbent 4-0 suture, and the skin was closed with 4-0 suture of silk. All animals were subjected to recovery from surgery and observed for 7 days.On the 7th day after surgery, rats were subjected to Athanasii, and the peritoneum was opened for the evaluation of surgically created defect. Adhesions between the defect in the wall of the peritoneum and the defect in the cecum, if they were present, were subjected to evaluation of strength by pulling the two tissues. For measuring the force required to break adhesions, used a strain gauge.Both processing fragmentirovannoj gelatinous compositions alone or in combination with an overlay Episil resulted in reducing the number of animals with adhesions, compared with the control group. The percentage of animals in each group who had adhesions, are presented in Table 4.EXAMPLE 9: MATERIALS AND METHODS (INPUT) SUPPLEMENTATION of ASCORBATE IN the GEL BEFORE IRRADIATION
Particles gelatinous, Wisconsin), containing 0.01 to 0.1% by mass glutaraldehyde (Sigma, street Louis, Missouri), and stirred overnight at ambient temperature. Alternatively, particles of gelatin derived from an extract of calf skin (Spier Co., Pennsylvania), suspended in the amount of 5-15% by weight in aqueous buffer solution at pH 9, containing from 0.01 to 0.1% by weight glutaraldehyde (Sigma) for gel formation, which is well mixed and cooled overnight. Particles of crosslinked gelatin then washed three times with ethanol and dried at ambient temperature. Then he measured the equilibrium swelling washed crosslinked gelatin and 0.5-1.0 g of this material was Packed in a 5 syringes with a capacity of 5 cm3, 3.0 ml to 4.5 ml of aqueous buffer solution containing ascorbic acid or a salt of ascorbic acid, for example, 0.02 M phosphate (J. T. Baker, Phillipsburg, new Jersey), 0.15 M sodium chloride (WR, West Chester, PA), 0.005 M of sodium ascorbate (Aldrich) at pH 7.0, was added to the syringe containing crosslinked gelatin, using the second syringe and three-way passing crane, while steps were taken against the penetration of outdoor air in the syringes for the hydrogel within a few syringes. Nearbynow acid or a salt of ascorbic acid, but, however, had a similar composition and pH value, for the formation of a gel inside them. The syringes containing the hydrogel was then subjected to gamma irradiation in the frozen state, when 3,00,3 megarad. After irradiation was measured by equilibrium swelling of the hydrogel contained within the syringes. The hydrogels obtained by the use of buffer solutions, which contained ascorbic acid or a salt of ascorbic acid, typically had values of equilibrium swelling after exposure, within the20% and typically10% of the value before irradiation, whereas gels formed using buffer solutions not containing ascorbic acid or a salt of ascorbic acid, showed a decrease in the value of the equilibrium swelling up to 25-30% of its value before irradiation.EXAMPLE 10: MATERIALS AND METHODS STITCHING AND measuring the PERCENT SWELLING
Particles of gelatin was allowed to swell in an aqueous buffer solution (for example, 0.2 M phosphate, pH of 9.2) containing a crosslinking agent (for example, from 0.005 to 0.5% by weight glutaraldehyde). The reaction mixture is kept overnight in a refrigerated, and then its three times washed with deionized water and in again suspended in aqueous buffer solution at a low concentration of solid particles (2-3%) at ambient temperature for a fixed period of time. Buffer solution was greater than the concentration required for equilibrium swelling, and attended two phases (phase hydrogel and the phase buffer solution). Then an aliquot of the suspension containing the wet hydrogel was subjected to filtering by application of vacuum to 0.8 μm nominal on the membrane filter with a limited bandwidth (Millipore, Bedford, mA). After removal of extraneous buffer solution recorded the total weight withheld wet hydrogel, wet membrane filter. The hydrogel and the membrane is then dried at a temperature of approximately 120oC for at least two hours and recorded the total weight of the dried residue and dried hydrogel membrane filter. Spent several sample wet membrane filter residue and dried hydrogel membrane filter without hydrogel and used them to derive the formula of net weight of wet and dry hydrogel hydrogel. Then figure out the Percentage of swelling as follows:
Measurement of swelling was performed three times and peregnyli for this sample of gelatin. Is swelling, expressed in percent, for the samples, re-suspended in the buffer is.The obtained crosslinked gelatin materials have demonstrated the values of equilibrium swelling in the interval from 400 to 1300%. The equilibrium degree of swelling depends on the specific method and degree of crosslinking.EXAMPLE 11: DESTRUCTION
Thirty rabbits (15 untreated control animals and 15 animals treated with fragmented gelatin composition was subjected to a surgical operation to simulate damage to the spleen and bleeding. The defeat of the spleen created by education regulated wound through a 6-mm syringe biopsy. In the "Treated" group formed by experimenting damage immediately processed fragmented gelatin composition to cause hemostasis of the wound. "Control" group of animals was subjected to treatment within the first 7.5 minutes to demonstrate the amount of bleeding due to damage to the body. After 7.5 minutes after the formation of the damage applied fragmented gelatin composition to stop bleeding from a wound to prevent spontaneous bleeding and death of the animal. All animals were allowed to recover. Each of the ten animals were subjected to Athanasii at 14 and 28 day animals on the 28th day. Animals collected on the 28th day, it was difficult to determine by simple tests is present or not test material, so part of the remaining animals were taken on a study on the 42nd day, and the other part is on the 56th day. During the dissection of the corpse of the affected area of the spleen and the peritoneal cavity was examined macroscopically. It was noted and appreciated the presence of the fragmented gelatin composition in the cavity of the peritoneum at some distance from its host, and the presence or absence of lesions of the spleen. Noted and evaluated the presence or absence of postoperative adhesions in the area of the lesion of the spleen. The spleen was carefully cut and processed for histological evaluation of biocompatibility and biodegradation.The application of the fragmented gelatin composition formed on surgical wounds in the spleen was possible to obtain a good means of plugging to stop bleeding. After applying the fragmented gelatin composition during the surgery, the rabbits remained alive for 14, 28, 42 and 56 days after surgery. One rabbit died from non-associated pneumonia on the 5th day after the operation and his selecte the abdominal cavity have been investigated fully. Appreciated the presence of the fragmented gelatin composition in the abdominal cavity at a distance from its host, and the presence or absence of the fragmented gelatin composition on the defeat of the spleen. It was noted and appreciated the presence or absence of adhesions on the lesion of the spleen. The spleen was carefully cut and processed for histological analysis.During all time points was fully visible to the affected area of the spleen in all animals. Macroscopically, the fragmented gelatin composition was absent in two of the ten animals, taken on the 14th day. At all other time points it was impossible to determine macroscopically fragmented gelatin composition. The absence of a hydrogel material, measured macroscopically in this rabbit model, determines the degradation of the hydrogel according to the term used here and in the claims.In three out of ten animals, those that were slain on the 14th day after surgery, found a small amount of fragmented gelatin composition, free-floating in the abdominal cavity. In all probability it represents the excess material which maurelli at some distance from the area of lesion of spleen, it was not any evidence of reaction tissue from the intestines or omentum. Animals taken at any other time, the material was not detected at some distance from the area of the lesion of the spleen.On the lesion of the liver from any animal were not detected postoperative adhesions associated with the use of fragmented gelatinous material. As expected, all animals had a seal attached to the defeat of the spleen. Other adhesions affecting the spleen, were rare, and when they were discovered, they were random and usually associated with cutting the wall of the body.Two of the ten animals taken from the 14-day time period, the fragmented gelatin composition was no macroscopically and microscopically. On the 28th day after the implantation of the fragmented gelatin composition was not visible upon gross observation, and microscopically it was entirely absent in five of the ten rabbits exposed to research in other animals it was present in very small amount and it was indicated that the fragmented gelatin was biodestruction essentially to 28 days. Fragmented gelatin to which it was discovered in a minimum quantity of only one of the four rabbits, studied at day 56 after it is entered. Wound healing in the spleen proceeded normally on the 42nd day, and more rapidly on day 56.Although the present invention has been described in detail by way of illustration and example for purpose of clarity of understanding, however, it is clear that the scope of the applied claims possible, certain changes and modifications.
1. Powdered polymer composition comprising a biocompatible cross-linked hydrogel having a particle size when it is fully gidratirovana, 0.05 to 5 mm; the equilibrium swelling of the 400 - 1300% and the time degradation in the living body (in vivo) in a wet environment tissue from 1 day to 1 year.2. The composition according to p. 1, characterized in that it contains a dry powder having a particle size of 0.01 to 1.5 mm, and the moisture content below 20% by mass.3. The composition according to p. 1, characterized in that it contains partially hydrated hydrogel having a degree of hydration of 50 - 95% hydration at equilibrium swelling.4. The composition according to p. 1, characterized in that it contains fully gidratirovannyi hydrogel having a degree of hydration of over 95%.5. Composition according to any one of paragraphs.1-4, characterized in that the hydrogel to change the project.7. Composition according to any one of paragraphs.1-4, characterized in that it additionally contains a plasticizer selected from the group consisting of polyethylene glycol, sorbitol and glycerin.8. The composition according to p. 7, characterized in that the plasticizer is present in an amount of 0.1 - 30% by weight of the polymeric component.9. Composition according to any one of paragraphs.1-4, characterized in that it includes the additional active agent.10. The composition according to p. 9, characterized in that the active agent is a hemostatic agent.11. The composition according to p. 10, characterized in that the active agent is thrombin.12. Composition according to any one of paragraphs.1-4, characterized in that the molecular cross-linked gel containing crosslinked protein hydrogel.13. The composition according to p. 12, wherein the protein is selected from the group consisting of gelatin, soluble collagen, albumin, hemoglobin, fibrovera, fibrin, casein, fibronectin, elastin, keratin, laminin, and their derivatives and combinations.14. Composition according to any one of paragraphs.1-4, characterized in that the molecular cross-linked gel containing crosslinked polysaccharide.15. The composition according to p. 14, characterized in that the polysaccharide is selected from the group consisting of glycosaminoglycans, production and combinations.16. Composition according to any one of paragraphs.1-4, characterized in that the molecular cross-linked gel containing crosslinked nabopolassar.17. The composition according to p. 16, wherein the polymer is selected from the group consisting of polyacrylates, polymethacrylates, polyacrylamides, polyvinyl resins, polylactic-glycolide, polycaprolactones, polyoxyethylene and their derivatives and combinations.18. Composition according to any one of paragraphs.1-4, characterized in that it is capable of resorbtive molecular cross-linked hydrogel contains at least two components selected from the group consisting of crosslinked proteins, cross-linked polysaccharides and stitched semipolymer.19. Composition according to any one of paragraphs.1-4, characterized in that the molecular cross-linked hydrogel contains a polymer and a crosslinking agent, the polymer and crosslinking agent react under conditions which provide crosslinking of polymer molecules.20. Composition according to any one of paragraphs.1-4, characterized in that the molecular cross-linked hydrogel is obtained by irradiation of the polymer under conditions which provide crosslinking of polymer molecules.21. Composition according to any one of paragraphs.1-4, characterized in that the molecular cross-linked hydrogel obtained by the reaction of monounsaturated and polyunsaturated monomers under conditions, which is, the fact he is in the manufacture of biocompatible, capable of resorbtive polymer; the polymer compound with the aqueous buffer solution for gel formation, the crosslinking of the gel and the grinding crosslinked gel to obtain compositions under item 1.23. A method of obtaining a polymer composition, characterized in that it consists in the manufacture of biocompatible, capable of resorbtive polymer, the dispersion of the polymer, cross-linking of powdered polymer and the stitched connection powdered polymer with an aqueous buffer solution to obtain a composition under item 1.24. The method according to p. 22 or 23, characterized in that stage stitching consists of exposure of the polymer to radiation.25. The method according to p. 24, characterized in that it further includes the connection crosslinked compositions with a certain amount of stabilizer that is effective for inhibiting the modification of the polymer when it is subjected to irradiation for sterilization.26. The method according to p. 25, characterized in that the stabilizer is ascorbic acid, sodium ascorbate, other salts of ascorbic acid or antioxidant.27. The method of multiplexing the path of the fabric, characterized in that it consists of at least partial filling of the way fabric composition with the n is in the feed composition under item 1 on the plot target number, enough to stop the bleeding.29. The method of delivery of bioactive substances to the site of the target in the patient's body, characterized in that it consists in applying the composition under item 1 in combination with a bioactive substance to the site of the target.30. The method of delivery to the site of the target tissue Nabakevi composition, characterized in that it consists in preparing the compositions of p. 1, while the composition hydratious less than its equilibrium swelling and cause the composition to the target area, where it swells to the values of equilibrium swelling.31. The kit contains a composition comprising a sterile biocompatible, capable of resorbtive molecular cross-linked gel under item 1, written instructions for applying gel to the area of the target on the fabric and the container containing the composition, and written instructions.32. Kit p. 31, characterized in that the gel dehydration.33. Kit p. 31, characterized in that the gel gidratirovana.34. The method of applying a polymer composition to the target area in the patient's body, characterized in that it consists in the preparation of biocompatible molecular cross-linked hydrogel under item 1 and the extrusion of the gel through the opening at the site of the target.35. The way p is s 0,01 - 5,0 mm and36. The method according to p. 34, characterized in that the hydrogel is crushed to the stage of extrusion.37. The method according to p. 34, characterized in that the area of the target tissue selected from the group consisting of muscle, skin, epitheliale tissue, connective or supporting tissue, nervous tissue, eye tissue, and other tissue sensitive organs, vascular and cardiac tissues, gastrointestinal tissues and organs, pleura and other pulmonary tissues, the tissues of the kidneys, glands of internal secretion, male and female reproductive organs, adipose tissue, liver, pancreas, lymph, cartilage, bone tissue, oral tissue and gelatinous connective tissue, spleen and other abdominal organs.38. The method according to p. 37, wherein the target area is the area of the cavity within the selected tissue.39. The method according to p. 38, characterized in that the area of the cavity is selected from the group consisting of channels, paths in the tissues of the intervertebral spaces and body cavities.40. The method according to p. 38, characterized in that it includes a fixed barrier layer on the cavity after extrusion of the gel.41. The method according to p. 34, characterized in that the stage of extrusion causes the hydrogel to break down into subgroups that have a size of 0.05 to 3.0 mm
31.07.1997 on PP.1-21; 23-33, 35-41; 43-44;
27.08.1996 on PP.22, 34 and 42.
- when the endoprosthesis through targeted injections hydrogel for plastics soft tissues of the face, breast, penis, calf muscles, vocal cords and other tissues, the density of which corresponds to the density of the hydrogel;
- as a filler in the manufacture of implants, including implants breast;
- as a depot for drugs with long-term medication, such as tumors or abscesses;
- as a carrier for culturing human cells and animals with subsequent implantation of hydrogel containing cells in the mammalian organism
FIELD: medicine-destination polymers.
SUBSTANCE: invention relates to biologically stable hydrogels to be employed as endoprosthesis consisting essentially of following components: polyacrylamide including acrylamide, crosslinked methylene-bis-acrylamide, wherein acrylamide and methylene-bis-acrylamide are linked at molar ratio from 150:1 to 1000:1. Hydrogel is rinsed with water or physiologic solution so that it contains about 0.5-3.5% polyacrylamide and less than 50 ppm acrylamide and methylene-bis-acrylamide monomers, while modulus of elasticity of hydrogel is approximately 10 to 700 Pa and its complex viscosity about 2 to 90 Pa*sec. Rinsing stage allows removal of nearly all amounts (even trace amounts) of above-indicated monomers resulting in lower toxicity and higher stability of hydrogel. Biologically stable hydrogel is used as injectable prosthesis to fill soft tissues and also to treat or prevent urinary incontinence or anal incontinence. Hydrogel, obtained in a few stages including combining acrylamide and methylene-bis-acrylamide, initiating radical polymerization, and rinsing with apyrogenic water or physiologic solution, is also useful in treatment or prevention of bladder-ureter reflux in mammalians. In all these cases biologically stable hydrogels contain between 0.5 and 25% polyacrylamide.
EFFECT: enlarged resource for manufacturing endoprostheses.
10 cl, 3 dwg, 7 tbl
SUBSTANCE: method of antibiotics fixation within porous implants is described. Result of method application lies in possibility of reliable fixation of antibiotic solution within porous implant and arrangement of favourable conditions for haemostasis in operative wound due to application of 10% gelatine solution as antibiotic carrier. Specified result is achieved by filling porous implants with antibiotic solution in liquid gel. For this purpose implant is dipped in solution by 3/4. Filling occurs under the influence of capillary forces. After solution cooled to form dense gel, antibiotic is fixed in implant pores and gradually released after installation to bone defect area.
EFFECT: reliable fixation of antibiotic solution within porous implant and arrangement of favourable conditions for haemostasis in operative wound.
3 cl, 1 ex
SUBSTANCE: invention concerns medicine, namely to reconstructive surgery, traumatology-orthopedy, maxillofacial surgery, stomatology and can be applied at osteo-plastic operations. For delivery of medical products immediately in a zone of defect and their prolonged influence in the centre of a lesion medicinal preparations are dissolved in a normal saline solution in a dose providing local effect, collagen-containing component is added to a solution in the ratio 9-20 g: 100 ml of a solution also admix with the carrier from dispersed allotransplants in the ratio of 1:1-3.
EFFECT: method allows lowering a dose necessary for reception of medical effect in 10 times, and also allows accelerating reparative processes in a defect zone.
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
SUBSTANCE: invention concerns medicine. Particles of the viscoelastic material chosen from group, consisting of polysaccharides and their derivatives which are suitable for injection with gel particles having the size in a range from 1 to 5 mm at action of a physiological saline solution are described. An implant for increase of volume of the soft tissues, containing particles of the viscoelastic material chosen from group, consisting of polysaccharides and their derivatives where the basic volume of the specified particles represents the gel particles, suitable for injection and having the size in a range from 1 to 5 mm at action of a physiological saline solution, is described. The way of increase of volume of soft tissues at a mammal, including a human being, including subepidermal introduction in a place of a body of the specified mammal in which it is desirable to enlarge volume of soft tissues is described.
EFFECT: augmentation of volume of soft tissues at a mammal.
24 cl, 4 ex
SUBSTANCE: invention relates to a method of producing biologically compatible gel which is thickened with cross-linked polymer by cross-linking a given amount of at least one biologically compatible natural polymer in a solution by adding a defined amount of cross-linking agent, an additional amount of polymer with molecular weight over 500000 dalton in a solution, in which the reaction mixture is diluted to reduce concentration of polymer in the solution, and the cross-linking reaction is stopped by removing the cross-linking agent.
EFFECT: gel and its use for separating, replacing or filling biological tissue or for increasing volume of such tissue, or supplementing or replacing biological fluid.
11 cl, 1 tbl, 4 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: group of inventions refers to medicine, more specifically to biocompatible alginate systems with the delayed gelatinisation process. There are offered sets and compositions for making a self-gelatinised alginate gel containing sterile water-soluble alginate and particles of sterile water insoluble alginate with a gelling ion. There are offered methods for dosing self-gelatinised alginate dispersion for making the self-gelatinised alginate gel. The methods can include dosing the dispersion in an individual. There is offered the self-gelatinised alginate gel of the thickness more 5 mm and not containing one or more sulphates, citrates, phosphates, lactates, EDTA or lipids. There are offered implanted devices coated with the homogeneous alginate gel. There are offered methods for improving viability of pancreatic islets or other cell aggregate or tissue, after recovery and while stored and transported.
EFFECT: group of inventions provides creation of the alginate gelling system which contains alginate and the gelling ions with high biological compatibility; enables the gelatinisation process without pH variations, connected with the other systems, and requires minimum ingredients, thus provides variation of gelatinisation time and gel strength depending on the specific requirements.
62 cl, 11 dwg, 2 tbl, 27 ex
SUBSTANCE: invention relates to field of medicine, in particular to method of obtaining form-preserving aggregates of gel particles, in which aggregates are held together by physical forces of non-covalent bonds, such as hydrophobic-hydrophilic interactions and hydrogen bonds. Method of obtaining form-preserving aggregates of gel particles includes introduction of preliminarily obtained suspension of gel particles in polar liquid, where gel particles have absolute electrochemical potential, into receiving medium, in which absolute electrochemical potential of gel particles decreases, which results in fusion of gel particles into form-preserving aggregate.
EFFECT: invention allows to obtain form-preserving gel aggregates in situ so that form of aggregate is determined by place of application.
49 cl, 35 ex, 11 tbl, 33 dwg
SUBSTANCE: invention relates to medicine. Described is implant, which can be injected in subcutaneous or intracutaneous way in form of monophase hydrogel, which contains gel, obtained from cross-linked hyaluronic acid and one of its physiologically acceptable salts.
EFFECT: obtaining subcutaneous implant used for filling wrinkles and stimulation of epidermal cells and/or supporting mechanical properties of skin density and elasticity.
15 cl, 2 ex
SUBSTANCE: claimed invention is aimed at manufacturing intraocular lens (IOL), for introduction of posterior eye chamber in form of PC Phakic lens. IOL is formed from hydrogel material, formed by cross-linked polymer and copolymer component. Lens includes UV chromophore, which is benzotriazole.
EFFECT: IOL hydrogel material usually has relatively high index of refraction and/or possesses desirable degree of protection against irradiation.
12 cl, 3 tbl