Method for obtaining modified scleroplant m biomaterial usable in ophthalmology

FIELD: medicine.

SUBSTANCE: method involves carrying out mechanical treatment of biological connective tissue and following treatment with solutions. Animal or human connective tissue is hold in solution of collagen and/or its derivatives and/or collagen processing products at 25-65°C and exposed to ionizing radiation treatment.

EFFECT: improved biocompatibility and physicochemical properties; prolonged drug action.

6 cl

 

The invention relates to medicine, and more specifically to ophthalmology, and can be used for the manufacture of sclerocarya materials, materials for plastics connective tissue, filling materials, drainage systems, and devices to create a depot of drugs.

A method of obtaining a biomaterial for use in ophthalmology (RF Patent No. 2054283 dated February 20, 1996), which is in mechanical treatment biological connective tissue of animals with the following processing solutions. The disadvantage of this method is the lack of biocompatibility, low mechanical properties, in particular low elasticity of the biomaterial and long term engraftment.

The objective of the invention is to improve the biocompatibility, improved physico-mechanical properties of the biomaterial, increasing the adhesive surface properties.

The technical result of the invention is to improve the biocompatibility and improved physico-mechanical properties of biomaterials by surface modification and three-dimensional structure of collagen biomaterial and/or its derivatives. as well as the possibility of prolonged action of medicines in the area of operation and their deposition in the surrounding tissue. Improving the biocompatibility leads to increased e is the efficiency of surgical intervention for reducing the integration period.

The technical result is achieved in that in the method of obtaining modified biomaterial for use in ophthalmology, including mechanical biological treatment of connective tissue and subsequent processing solutions according to the invention the connective tissue in animals or humans stand in a solution of collagen, and/or its derivatives, and/or by-products of collagen at a temperature of 25-65°and irradiated with ionizing radiation.

In the proposed method is volumetric structuring of collagen biomaterial and/or modified collagen, and/or processing products of collagen with the formation of gel-like frame that supports the shape of the biomaterial. In addition, the material acquires the ability to retain a given shape, which greatly simplifies the implantation process and reduces the time of operation. In addition, in the process of the proposed method of obtaining a biomaterial surface is modified collagen hydrogel, which improves its biocompatibility and adhesive properties with respect to the sclera of the recipient. During implantation of the collagen surface layer stimulates fibroblastic reaction in the contact zone, which leads to a reduction of the integration period and the formation of mechanically stable complex sclera-implan is at.

The temperature range of 25-65°selected experiments. At temperatures below 25°With solutions of collagen and its derivatives heliroute and do not penetrate into the connective tissue. At temperatures above 65°there is a significant reduction and compaction of connective tissue, which also impedes the diffusion of collagen and its derivatives in connective tissue

One of the variants of this method is one in which before curing in a solution of collagen connective tissue is saturated with glycosaminoglycans in an amount of 0.01-10.0 wt.%.

Glycosaminoglycans are complex polysaccharides and used to strengthen the collagen matrix of the biomaterial, because they form strong complexes with collagen. These include hyaluronic acid and its salts, keratan and chondroitin sulfates, dermatooncology, heparin and heparansulfate. For saturation of the connective tissue use solutions of glycosaminoglycans in concentrations of 0.01-10.0 wt.%. At concentrations below 0.01% is not achieved, the effect of the structure of connective tissue. At concentrations above 10% there is no significant improvement of the mechanical properties of the biomaterial.

Before keeping in the solution of collagen connective tissue can be full of drugs in therapeutic doses, as well as Biol who cally active compounds. The result is a biomaterial capable prolongirovanne to allocate a drug or bioactive compound in the area of surgical intervention, i.e. to Deposit these funds. As biologically active compounds can be used vitamins, antioxidants, steroids, corticosteroids, hormones, peptides and polypeptides (e.g., carnosine, glutathione, and others)that affect the normalization of metabolic and reparative processes.

The solution of collagen and/or its derivatives, and/or by-products of collagen can be pre-filled with glycosaminoglycans and/or drugs in therapeutic doses and/or biologically active compounds. In the proposed method are the biomaterial-modified collagen and containing listed substances.

The method is as follows. From the mechanically cleaned and treated with solutions of connective tissue in animals or humans form fragments of a desired shape and placed in a solution of collagen and/or its derivatives (for example, chemically modified collagen), and/or in the solution of products of processing of collagen (e.g., partially hydrolyzed collagen or gelatin) at a temperature of 25-65°C. the Concentration of the solution may argirov shall be from 0.01 to 50 wt.%. Connective tissue is kept in a specified solution for some time. The duration of treatment depends on the size and shape of fragments of connective tissue and may be from 5 minutes to 24 hours. Then the biomaterial is removed from solution and irradiated with ionizing radiation. The biomaterial can also be irradiated directly in the specified solution of collagen and/or its derivatives.

If necessary, saturation connective tissue drugs or biologically active compounds fragments of connective tissue are placed in solutions of these substances, and then incubated in a solution of collagen.

For comparison studied the response of fibroblasts to the biomaterial of the pericardium bovine performed according to the method described in the prototype, and the modified biomaterial treated with a solution of collagen in the proposed method. These biomaterials were used as substrates for cultivation of keratinocytes. Compared the number of cells in culture by the end of the experiment, investigated the shape and morphology of the cells. As a result of the experiment was that in the population of cell culture on modified biomaterial shape and morphology of the cells was correct, undisturbed, and the number was above 30%.

On offer is built the way it was made 15 sets of implants, which were used for the surgical treatment of progressive myopia, macular degeneration, plastic scleral layer of the eye, as cerclage. In all cases noted areactive the postoperative period, which indicates a high biocompatibility obtained by the proposed method biomaterial. In addition, the implants were easy to implant, because they did not twisted and kept straightened shape during implantation due to its elasticity.

The proposed method allows to obtain a wide range of implants made of modified biomaterial for various areas of ophthalmic surgery.

The invention is illustrated by examples.

Example 1. From the pre-cleaned and washed by the solution of pericardium bovine form implants rectangular shape with rounded corners size 2×1 cm and incubated in 0.01% solution of collagen for 8 hours at a temperature of 25°C. Then the implants are packaged and subjected to irradiation with ionizing radiation dose of 3.0 Mrad. The material used to perform operations meridional scleroplasty for the treatment of progressive myopia.

Example 2. Mechanically cleaned and treated with solutions of the pericardium bovine cut into strips with a length of 8 cm and a pulse is 1 cm with rounded corners and placed in a 0.1% solution of biologically active compounds carnosine to saturate for 2 hours. Then the biomaterial is placed in a 30% solution of gelatin rich hyaluronic acid at a concentration of 0.01%, and incubated for 5 minutes at a temperature of 45°C. thereafter, the strips of biomaterial straighten, hermetically sealed and irradiated with ionizing radiation dose of 1 Mrad. The resulting biomaterial used for plastics sclera after remote silicone cerclage.

Example 3. Amniotically membrane person was purified mechanical, process solutions and kept in a solution of gelatin and a derivative of collagen, a molecule which introduced double bond, at a concentration of 5 and 0.1%, respectively, at a temperature of 37°C for 30 minutes. Then formed fragments of approximately 3×3) cm2and irradiated with ionizing radiation. The resulting biomaterial used in the plastic of the cornea and eyelids.

Example 4. The pericardium bovine purified mechanical, process solutions and placed in a solution of the following composition: collagen - 1%, derived collagen with double bonds in the molecule - 1%, hydrolyzed collagen - 3%. In this solution, the pericardium was kept at a temperature of 55°C for 4 hours, then cut into slices (6×25) mm2and irradiated with ionizing radiation. The resulting biomaterial used as fillings for local breaks in the retina.

Example 5. Per the card of cattle were cleaned mechanically, worked solutions, and cut in slices (2×2) cm2. The fragments were placed in a 10% solution of glycosaminoglycans to saturation, and then in a solution of 2% collagen and 0.1% solution derived collagen with double bonds in the molecule. In the latter solution, the tissue fragments kept at a temperature of 45°C for 1 hour, then corked and irradiated with ionizing radiation. The resulting biomaterial used in plastic scleral layer of the eye.

Example 6. Scleral shell eyes pigs were mechanically cleaned, worked solutions, cut into pieces in the form of a disc with a diameter of 0.5 cm and filled with gentamicin. Then the fragments were placed in a solution of biologically active substances methyluracil in a concentration of 0.2%. After that, the fragments of the sclera was kept in 3% solution of collagen at a temperature of 25°C for 15 minutes and subjected to ionizing irradiation. The resulting biomaterial used as episcleral implant in the treatment of uveitis.

Example 7. The pericardium bovine purified mechanical, process, solutions, cut into pieces (0,5×1.5) cm2and saturated chloramphenicol. Then the fragments were kept in 1% solution of collagen containing in its molecule a double bond within 15 minutes at a temperature of 37°C. the thus Treated fragment the tissue was irradiated with ionizing radiation. The resulting biomaterial used for the surgical treatment of uveitis.

Example 8. Scleral shell of the eyes of cattle was purified mechanical, process, solutions, cut into pieces (2×1) cm2with rounded corners and put to saturation in a solution of glycosaminoglycans in concentrations of 2%, and then in a solution of gentamicin therapeutic concentrations. After that, the fragments of the sclera were placed in a solution containing 0.1% of chemically modified collagen, a molecule which added positively charged group, and 5% gelatin. In the specified solution fragments of the sclera was kept for 15 minutes at a temperature of 40°C, then placed in sealed containers and irradiated with ionizing radiation. The resulting biomaterial used in the surgical treatment of uveitis.

Example 9. Amniotically membrane person, mechanically cleaned and processed solutions, cut into pieces (1×1) cm2fed in a solution containing glycosaminoglycans 5%, and hydrocortisone in therapeutic concentrations, carnosine - 0.1%, and then placed in a solution of collagen at a concentration of 3%. In the specified solution fragments of tissue was kept for 30 minutes at a temperature of 25°and irradiated With ionizing radiation. The resulting biomaterial used for plastics is effektov of the cornea.

Example 10. From the mechanically cleaned and treated with solutions of tracheal cartilage of cattle formed fragments of length 4 cm and a thickness of approximately 3 mm. These fragments kept at a temperature of 65°C for 15 minutes in a solution of the following composition, wt.%: collagen - 0,5%, chemically modified collagen containing in the molecule the double bond is 0.1%gelatin - 5%. This solution was pre-filled with glycosaminoglycans at a concentration of 10%, the local anesthetic action of lidocaine in therapeutic concentrations and in a biologically active compound glycine in a concentration of 1%. Then fragments of cartilage tightly corked and irradiated with ionizing radiation. The resulting biomaterial used for plastics orbit eyeball injuries.

Example 11. The pericardium bovine, pre-cleaned and treated with solutions, cut into disks with a diameter of 12 mm Solution containing 2% collagen and 5% of hydrolyzed collagen, was saturated by glycosaminoglycans at a concentration of 1% and cytokines (biologically active polypeptides) in a concentration of 0.01%. To this solution was placed disks from the pericardium and kept at a temperature of 30°C for 4 hours. Then the processed disks of the pericardium in the specified solution was placed in a flat container and irradiated with IO is siraudeau radiation. The resulting biomaterial used in surgical treatment of chorioretinal dystrophy.

Example 12. The pericardium bovine, pre-cleaned and treated with solutions, cut into pieces (1×1) cm2. A solution containing 0.1% of chemically modified collagen, a molecule which is introduced groups with double bonds, and 1% processing product of collagen, hydrolyzed collagen, pre-saturated hydrocortisone in therapeutic concentrations. Fragments of the pericardium was placed in the resulting solution and kept at a temperature of 25°C for 24 hours. Then the fragments were placed in sealed containers and irradiated with ionizing radiation. The resulting biomaterial used for episcleral implantation for the treatment of uveitis.

Example 13. Dura person, mechanically cleaned and treated solutions were cut into disks with a diameter of 1 cm and placed in rubbed 3% collagen-rich glycosaminoglycans at a concentration of 1%hydrocortisone in therapeutic concentrations and glutathione in a concentration of 0.5%. Fragments kept in the specified solution at a temperature of 30°C for 2 hours, and then irradiated with ionizing radiation. The resulting biomaterial used for episcleral filling in uveitis.

When is EP 14. Scleral shell eyes were mechanically cleaned, worked solutions and formed fragments (2×10) mm2. Pre-1% solution derived collagen - containing covalently linked cytostatic agent mitomycin C, saturated carnosine concentration of 1%. To this solution was placed fragments of the sclera and kept at a temperature of 45°C for 30 minutes. Then the fragments were irradiated with ionizing radiation. The resulting biomaterial used in the surgical treatment of glaucoma.

Example 15. The bone tissue of cattle were mechanically cleaned, treated by solutions of molded plates (2×1) cm2a thickness of approximately 2-3 mm and kept in a 20% solution of gelatin, pre-saturated with lincomycin at therapeutic concentrations, at a temperature of 65°C for 1 hour. Then the plates were placed in a sealed package and irradiated with ionizing radiation. The resulting biomaterials used for bone grafting.

1. The method of obtaining modified biomaterial for use in ophthalmology, including mechanical biological treatment of connective tissue processing solutions and radiation, characterized in that the use of connective tissue in animals or humans, which after machining and processing solutions : ribaut in a solution of collagen, and/or its derivatives, and/or by-products of collagen at a temperature of 25-65°and irradiated with ionizing radiation.

2. The method according to claim 1, wherein before curing in a solution of collagen connective tissue in animals or humans injected with glycosaminoglycans in an amount of 0.01-10.0 wt.%.

3. The method according to claim 1, wherein before curing in a solution of collagen connective tissue in animals or humans injected drugs in therapeutic doses.

4. The method according to claim 2, wherein before curing in a solution of collagen connective tissue in animals or humans injected drugs in therapeutic doses.

5. The method according to claims 1 to 4, characterized in that before curing in a solution of collagen connective tissue in animals or humans saturate biologically active compounds.

6. The method according to claim 1, characterized in that the solution of collagen and/or its derivatives, and/or processing products of collagen pre-saturate glycosaminoglycans in an amount of 0.01-10.0% and/or drugs in therapeutic doses and/or biologically active compounds.



 

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