JPS59500803A - Introducing polymers into implantable biological tissues to prevent calcification - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed description of the invention] Introducing polymers into implantable biological tissues to prevent calcification Uninvented technical field Glucuraldehyde preservation of 4F tissue has been shown to be particularly useful in porcine bovine prosthetic heart valves. a) The implanted tissue flap, which had been preserved in formaldehyde, 1〈1: Overcoming the good fortune of ability, b) Discontinuing the use of allograft valves, C) Thromboembolism prevention used in connection with the use of non-biological artificial (mechanical) heart valves. Avoid the use of anticoagulants, especially in children, when they are necessary or undesirable for It is now possible to However, like other important findings, glucuraldehyde Existing prosthetics have also created their own dilemmas.

The biologically relatively inert glucuraldehyde preservation of Carpentier et al. Although these valves have demonstrated excellent long-term stability in many cases, tissue fatigue It was not used continuously because it had serious drawbacks such as easy calcification. It was. This glucuraldehyde also eliminates the use of mechanical prostheses. The people who will benefit the most from a prosthetic heart valve are children and young children. It was thought of for the first time. However, recent clinical experiments have shown that fj'ii cannot be used for a relatively short period of time. There is an increasing number of reports that serious calcification of the underlying tissue is common in children and adolescents. did Therefore, despite its advantages of long-term stability and generally fewer complications, However, this glucuraldehyde-preserved heart valve is suitable for use in children. There are some people who think that there is no such thing.

Tissue calcification is still largely stigmatized.

However, calcium metabolic diseases, age, diet, and changes in tissue components (e.g. collagen) Until now, it has been proven that various factors such as quality and emotional disturbance are involved to some extent. It is. Recently created for transplantation of glucuraldehyde-preserved porcine xenografts The existence of specific calcium-binding amino acids has been demonstrated, which contributes to calcification. It is believed that there are. Collagen line of glucuraldehyde preservation of transplanted tissue In fibers, calcification is accompanied by degradative changes, or dystrophic calcification is accompanied by tissue alteration. It is unclear whether this is the cause or the result. However, a cure will soon be found. Efforts are continuing to investigate the causes of calcification in transplanted tissues, with the hope that ·reactor.

So far, no cause for calcification has been identified in biological transplants.

Calcification of the transplanted biological tissue is achieved by treatment with an anionic surfactant (C) and by treatment with an anionic surfactant (C) before transplantation. This can be reduced by avoiding contact with tissue phosphate ions. These things are said separately. These methods are effective in reducing calcification of bioprosthetic heart valve tissue. Therefore, it seems promising. This treatment not only reduces calcification but also The durability of the tissue after transplantation must also be maintained. In particular, this treatment Dynamic properties must be maintained and the rigid leaflet heads must not be compromised.

In the present invention, the present inventors effectively reduce calcification of transplanted living tissue, We developed a method to maintain the original hemodynamic properties of the leaflets of bioprosthetic heart valves.

It is effective in reducing the calcification tendency of transplanted living tissue, and xenotransplantation Several problems regarding the fillability of single-heart valves have been overcome.

Summary of the invention The present invention aims to prepare living tissue before transplantation so that calcification of the tissue is alleviated or reduced after transplantation. An improved method for processing is shown.

Illegal biocompatible polymers are effective in reducing tissue calcification after implantation. It is characterized by being introduced into body tissue. In one embodiment, the monomer is This method involves covalently fixing the compound to the compound and then polymerizing it.

Detailed description of the invention The present invention utilizes a variety of transplantable biological tissues obtained from various animals and their body parts. We believe that it is possible to make substances resistant to calcification (・ Ru. These tissues are particularly obtained from cows, pigs, horses or rabbits and include locks, ligaments, heart valves, or tissue used to make the heart (e.g. dura mater and 6 membranes); and Tissues used for augmentation (e.g. skin pieces, 6 membranes, aorta and tympanic membrane) may also The present invention (・τA preferred embodiment of the present invention is considered to be suitable for Porcine heart valve or 6-membrane tissue fixed with dehyde and treated with biocompatible polymeric material was Transferred under the skin of the heron. rIshita. This processed tissue makes it a surprising (but delicious) ingredient. In some cases, after tl σ) no sustained alleviation or reduction of calcification occurred. This calcification Continuation. Mitigation is a method to increase the durability of transplanted tissues, especially bioprosthetic heart valves. Rieru.

In the present invention, various polymeric substances can be introduced into the bovine body. used in the present invention 911, which is a high-molecular substance, includes acrylamide, acrylic acid, and acrylic acid ester. There are ster, tsucrylic acid, and tsucrylamide.

The present inventors have shown that acrylamide is effective in reducing calcification of transplanted living tissues, Acrylamide has been found to be a suitable polymeric material for the present invention.

In the present invention, this polymer substance is contained in the interstitium of the tissue to form a physical or mechanical bond. can be formed and impregnated or chemically bonded. Covalent bond This method prevents the polymer material from separating from the tissue after implantation, and prevents layers from forming within the tissue. The covalent bonding method is a preferred method because it is less susceptible to the effects of chemical reactions. protein There are several groups suitable for covalent bonding, including α- and and ε-amino groups: α-, β-, and γ-carboxyl groups, stein and serine Sulfhydryl group, hydroxyl group, imidaal group of histidine.

5 and the phenolic ring of tyrosine. Chemical groups are chemically bonded to these residues. Methods to do this are well known. Furthermore, mucopolysaccharides in biological tissues have free carboxyl groups. In particular, various monomers can be combined. The present inventors have discovered that monomers in biological tissue The method of covalently bonding the lime and then polymerizing it with a biocompatible polymer is found that this is a preferred method of the present invention. Ta. This bond can be created by direct compulsion or by using a cassilling agent such as the one described below. It is carried out using

Introducing macromolecules into tissues (() results in the spaces within the tissue being filled, thus allowing the host's plasma to It is thought that the entry of proteins and cellular materials into the transplanted tissue is prevented. This approach is the heart It is thought that this causes deterioration of the valve tissue and the valve becomes unusable sooner. Share more It is believed that the bound polymeric material also increases the mechanical strength of the tissue.

In one preferred embodiment of the invention, a species is used to bind the polymeric substance to tissue. Various coupling agents can be used. In some embodiments, the carbodiimide Free carboxylic acid on tissue proteins and mucopolysaccharides with the help of activators such as Diamines are used to attach various monomers to the residues. This kind of cup An example of pulling is JournalofP by Llogd and Burns. polymer 5science: Polymer Chemistry Edi tion.

vol,,17. Described in PP3459-3483 (1979) 6 Special table BH59 -500803(3). Diamines suitable for the present invention include formulas) (- (NH2)2 (wherein R is an aliphatic functional group having a straight chain, branched chain, or cyclic chain, or an aromatic functional group). The length or size of the chain is The diamine must be able to freely diffuse through the protein network of the tissue. child Preferably, the diamine is water-soluble. The most preferred diamine of the present invention is ethyl It is diamine.

In a preferred embodiment of the present invention, further polymerizable monomers are mentioned above. It is attached to tissue via an amine spacer. In one embodiment, acrylic acid or derivatives thereof are preferred. In a further preferred embodiment of the invention, on the tissue: Only covalently bonded macromolecules are preferred. Therefore, after the introduction of acrylic acid, the tissue is completely Wash to flush away acrylic acid monomers that are non-covalently bound and trapped within the tissue. leave. This unbound monomer self-polymerizes when more macromolecules are introduced into the tissue. promote. Self-polymerization, the formation of macromolecules that are not covalently bonded to tissues, by reducing available monomers and thus altering the intended properties of the treated tissue. -Mari.

As described above, various polymeric substances are suitable for introduction into living tissues in the present invention. Takamin Give various properties (charge, hydrophobicity, and hydrophilicity) by changing the properties of the child. be able to. To give the property of JU1 by changing the degree of polymerization and the degree of crosslinking You can also do it. In a preferred embodiment of the invention, the tissue-bound monomer is about 0.5- By suspending it in a solution of about 6 layers of acrylamide or similar monomers. Further polymerization can be carried out in effective amounts. More preferably the solution is about 0. 5 - about 2 parts by weight, most preferably 1 part by weight. Bi of approximately 0.25 weight salary This acrylamide is crosslinked using sacrylamide.

In the present invention, the pH range that has a tissue stabilizing effect (that is, the pH that is harmless to tissue components) Preferably, the tissue is preserved and fixed within the confines. The preferred pH range is about 7.0 to about 7. ．． 6, more preferably about 7.1 to about 7.4.

The most preferred pH of the present invention is 7.3.

The buffer used in one of the preferred embodiments of the invention is stable and suitable for stabilization steps. It is non-reactive and useful for maintaining a favorable pH, especially during tissue fixation. It is preferable that it has sufficient buffering capacity. Types of buffers used and their The concentration varies depending on the preparation conditions of each individual tissue. Several manufacturing companies with various manufacturing conditions This is being carried out by Buffer solution is conventional 0.01M-0.02M phosphate Buffered saline (PBS) or phosphate-deficient solutions, such as lysate, may be used. A solution with a phosphate salt content of 0.01-〇, 02M may be less than that of the PBS solution, and is preferred. Preferably, it is about 0.001 to about 0.002 M or less. suitable for the present invention Liquid solutions include borates, carbonates, bicarbonates, and cacodylates (non-toxic to animals). ), and other synthetic, artificial, or organic buffers (even PES (N-2-hydroxyethyl 2perazine-N'-2-ethalsulfone acid), MOPS (morpholinepropanesulfonic acid), and PIPES (1 , 4-piperazine diethanesulfonic acid)). The inventors have developed a HEPES buffer. We found that tissue prepared with liquid is effective in reducing calcification after transplantation, Therefore, it is optimal for the present invention.

The buffered or non-buffered solution used in the present invention may be a fixative such as glucuraldehyde. It is preferable not to interfere with the tissue stabilization process. i.e. these solutions are fixed It must not react with the fixative or interfere with proper fixation of the tissue by the fixative. child Examples include buffers containing primary and secondary amines, e.g. (hydroxymethyl)aminomethane (Tris) is glucuraldehyde is known to react with the aldehyde groups of interfere with

In the present invention, the tissue is fixed with glucuraldehyde of 0.625 weight coefficient (tanned). ).

The invention is further illustrated in the following examples, but the invention is limited thereto. It's not a thing.

Example 1 Extracted porcine arterial heart valve tissue was treated with 0.54 g/liter HE. Sodium salt of PES and 0.885 2% by weight zurtrium chloride (PH7,6) Thoroughly wash and transport at 4°C with an isotonic (285 ± 15 mm IJ osmoles) solution containing did. This was added to 5.69 g/liter of T (sodium salt of EPES, 0.0 g/liter). 440% by weight sodium chloride and 2.6 grams/liter MgCl26H2 Solidify at room temperature with 0.625 wt 9% glucuraldehyde in an isotonic solution containing 0. Established.

Example 2 The excised tissue of Example 1 was treated in a solution containing about 4% formaldehyde. further sterilized and rinsed with sterile saline immediately before transplantation to remove any remaining Glucool alcohol. The dehyde was removed and the cells were implanted subcutaneously into growing rabbits.

The heart valve tissue was collected at weekly intervals until 6 weeks later.

After recovery, the degree of calcification of the tissue was quantitatively evaluated using atomic absorption spectrometry. It is performed by tracking the weight of calcium in the tissue, and histologically, Von Kossa This was done by observing the degree of calcification in the stained tissue.

Example 2 Approximately 5 f/(iB weight fa) of the excised tissue prepared by the method of Example 1 was , 40111/! containing about 1 g of ethylenediamine. in a solution of ())H4,75) immersed in. After approximately 30 minutes, the pH was maintained at 4.75 during the 30 minute apparatus time at room temperature. aqueous) raw 1-ethyl-3(ro-dimethylaminozorosol)carbodiimide- g of HRI was added stepwise.

The pH is 4.75 to ensure maximum reactivity between this diamine and the carboxyl group. It is preferable to control io to 1. Next, HEPEr; buffered saline (, p) I 7.4. Wash the tissue thoroughly with water containing 0.2M acrylic acid. The sample was immersed in solution (pH 4,75) K for about 60 minutes. Then HEPES buffered saline The tissue was thoroughly washed to remove unbound acrylic acid from the tissue. Then this The tissue bound with acrylic acid was further suspended in approximately 4Qme of distilled water for approximately 60 minutes. 1. Bubble nitrogen for 30 minutes. 06. %(V/V)N.

2% ammonium persulfate containing N, N', N'-tetramethylene diamine After 30 minutes, the free radical initiation step was completed and 0.25 0 containing % bisacrylamide (N,N'-methylbisacrylamide) , transferred to 4Qml of 5% by weight acrylamide solution for 30 minutes and replaced with persulfate solution. let All tissue transfers were performed in a nitrogen atmosphere. Incubate the reaction mixture for approximately 60 minutes. After combining, the tissue was washed with distilled water and soaked in a solution containing 4 parts formaldehyde. The medium was sterilized, washed again with sterile physiological saline, and implanted subcutaneously into a growing rabbit. child Heart valve tissue was collected every 1Y1 for 6 weeks, and the degree of tissue calcification was quantified. Specifically, atomic absorption spectrometry is used to track and evaluate the degree of calcium content in dry tissue. Histologically, it is used to observe the degree of calcification of Van Kossa-stained tissues. I evaluated it more. The results of this histological and quantitative measurement are both 1/i: The transplanted heart valve tissue introduced with Mido was compared with the heart valve tissue treated by the method of Example 2. It has been shown to significantly reduce calcification.

Furthermore, (when attaching C tissue to a bioprosthetic heart valve, perform regular constant flow and pulse flow tests. In cases where acrylamide-loaded heart valve tissue has a detrimental effect on the stiffness of the valve leaflets, I didn't give it.

% 4 out of 1 The collected tissues of Examples 2 and 3 were further analyzed for the presence or absence of changes after transplantation. . As a result, no significant differences were observed in condensation temperature, water content, or amino group analysis results. does not contain HEPES sodium salt and 0.885% by weight of sodium chloride. Isotonic (285i 15 mm IJ osmoles) solution (pr-+ 7.3 > medium 4 Thoroughly washed and transported at °C. and 5.39 g/13 HEPES sodium salt, [1,440% by weight of thorium chloride and 2.6 g/l! MgC42 ・6) (0.625% by weight of geltaraldehyde in an isotonic solution containing 20 Fixed at room temperature.

Example 6 The excised tissue of Example 5 was soaked in a solution containing about 4 parts formaldehyde. Further sterilize with i+ solution and wash with sterile physiological saline immediately before transplantation to remove any remaining glucose. Raldehyde was removed and the cells were implanted subcutaneously into growing rabbits. Ask yourself questions about this organization for a week. The animals were collected up to 6 weeks later.

After collection, the degree of tissue calcification can be quantitatively determined by atomic absorption spectrometry. It was evaluated by tracking the weight percent of Si, and histologically, VOII Koss Evaluation was made by observing the degree of calcification of the a-stained tissue piece.

Implementation: 5 g (wet weight) of the excised tissue prepared by the method of Example 5 was weighed approximately 2.5 Immersed in a solution (pH 4,75) 4Q+++J containing g of ethylenediamine/ Ta. After about 3θ minutes, pJ-7 was maintained at 4.75 for 30 minutes at room temperature. Water-soluble 1-ethyl-3(ro-dinothylaminopropyl)carbodiimide HC42! 9 was added in stages.

In order to obtain the maximum σ) reactivity between this diamine and carboxyl group, the pH was adjusted to 4.75. It is preferable to set ti3i1 to io, 1.

Next, the tissue was thoroughly washed with HEPES grade buffered saline (T) (17.4) ．． Aqueous solution containing 2M acrylic acid (p.

It remained patent for about 30 minutes at 4.75). The tissue is then rinsed with HEPES-buffered saline. Unbound acrylic acid was removed from the tissue by washing with The connected tissue was suspended in approximately 40 ml of distilled water and aerated with nitrogen bubbles for approximately 60 minutes. 0.6% (v/v) of nitrogen was bubbled through the filter beforehand. 2% ammonium persulfate containing N, N', N'-tetramethylene diamine The solution was replaced with 43 ml of Ni solution. After 30 minutes, the free radical initiation phase is over and the tissue is 0.25% bisacrylamide (N.

1% by weight acrylamide solution containing N'-methylbisacrylamide) 4 01111 for 30 minutes and replaced with persulfuric acid solution. All tissue transfers are performed in a nitrogen atmosphere. I did it in the atmosphere. After allowing the reaction mixture to polymerize for approximately 60 minutes, the tissue was washed with distilled water. Sterilize with a 4% formaldehyde solution and rinse again with sterile saline. , implanted subcutaneously in rabbits at the stage of development. this organization every other week until 6 weeks later Recovered. The degree of tissue calcification can be quantitatively determined by atomic absorption spectrometry. Evaluated by tracking the weight percent of Lucium and histologically VanKossa Evaluation was made by observing the degree of calcification of stained tissue pieces. histological and Both quantitative evaluation results showed that the transplanted tissue introduced with acrylamide was that of Example 6. It has been shown that calcification is significantly reduced compared to tissue treated with this method. difference Furthermore, when attaching tissue to the bioprosthetic heart valve, regular constant flow and pulse flow tests were performed. In some cases, the acrylamide-infused tissue may have a detrimental effect on the stiffness of the valve leaflets, causing won.

Although the present invention has been described in detail with reference to preferred embodiments, it will be apparent to those skilled in the art that improvements and changes can be made to the present invention. However, this does not depart from the spirit and scope of the invention.

Submission of a translation of the written amendment (Special I Ugly Article 184-7, No. 1, 59-21゜128 Commissioner of the Patent Office 1. Indication of patent application: POT/US83100671, title of invention: Portable Introducing polymers into biological tissues to prevent calcification 3. Patent applicant Name (First Name B) American Foszotal Sasilai Corporation 4, Agent Man Address: Shin-Otemachi Building 3, 2-2-1 Otemachi, Chiyoguchi 1-ku, Tokyo 100 31 5. Date of submission of written amendment: September 60, 1981; scope of claims corrected; (Received by the International Bureau on September 6.0, Ci9 B5.

There is no change in the scope of the original 9 Seikyu, Section 1, Paragraph 1 to Paragraph 1A.

New claims 15 to 48 are as follows. ] 15. A method for treating naturally occurring living tissue before transplanting it into an animal, (a,) The tissue is fixed under tissue fixation conditions, and (b) the tissue is polymerized into two parts. a first monomer that can be covalently attached to the tissue under monomer binding conditions, and (c) Adding a second monomer bath under polymerization conditions to reduce tissue calcification after implantation. contact the tissue in an amount effective to cause the second monomer to bind to the tissue. A method for treating the above-mentioned tissue, which is characterized by polymerizing with a monomer.

16. Between the steps (b) and (C), the monomers 1 to 16. The method of claim 15, further comprising the step of removing from the tissue.

17 Claim 15, wherein the first monomer is acrylic acid or methacrylic acid The method described in section.

18 The second monomer solution is acrylamide, Aku’J”k, 7cryl Claim 15 consisting of an acid ester, methacrylic acid, or methacrylamide The method described in section.

19 The first monomer is acrylic acid or methacrylic acid, and the second monomer is Mer solution contains acrylamide, acrylic acid, acrylic ester, methacrylic acid, or tutacrylamide.

20. Non-covalently bonded monomers are assembled between steps (b) and (c). 20. The method of claim 19, further comprising the step of removing from the fabric.

21. Claim Part 15, wherein the first monomer is covalently bound directly to the tissue. The method described in section.

22, the first monomer covalently bound to the tissue is acrylic acid and the second monomer is Claim 15, wherein the nomer solution comprises acrylamide and bisacrylamide. The method described in section.

26, the first monomer covalently bound to the tissue is acrylic acid and the second monomer is Claim 16, wherein the nomer solution comprises acrylamide and bisacrylamide. The method described in section.

24 The second monomer solution contains about 0.5 to about 6 parts by weight of acrylamide. 27. The method according to claim 22 or 26.

25 The second monomer solution further contains about 0.25 wt. bisacrylamide. 25. The method of claim 24, comprising:

26 Whether the biological tissue is an elbow, ligament, heart valve, dura mater, or epithelium, Claim 15 or the method described in paragraph 16.

27 The living tissue according to claim 26 is fixed with geltaraldehyde. How to put it on.

28 In the method of treating naturally occurring living tissue before transplanting it into an animal, hand, (a) Fix the tissue under tissue fixation conditions, and (1) apply a camping agent to the tissue. The first monomer that can be further polymerized is made into a 1,000-mer bond. covalently attached to the camping agent under conditions, and ((1) The second monomer solution under polymerization conditions reduces the calcification of the tissue after implantation. contact the tissue in an amount effective to induce characterized by polymerization with the first seven strands bonded to the fabric, -1- Method for processing the tissue described above.

29, (c) upper o? and (d), the non-covalently bonded monomers are 29. The method of claim 28, further comprising removing from the tissue.

60 Claim 2, wherein the first monomer is acrylic acid or methacrylic acid The method described in Section 8.

31 The person according to claim 28, wherein the camping agent is a diamine compound. Law.

32. Between the steps (c) and ((''), contacting the tissue with the activation factor is required. Scope of Claim 28. The method according to item 28.

33. The method of claim 32, wherein the Roro activator is a carbosomide.

34 The second monomer solution is acrylamide, acrylic acid, acrylic acid ester as claimed in claim 28. the method of.

35 The first monomer is acrylic acid or methacrylic acid and the second monomer If the solution is acrylamide, acrylic acid, Nisder acrylate, tutaacrylic acid, or 29. The method of claim 28, comprising methacrylamide.

36. Between step (c) and step (1), the non-covalently bonded monomers are organized. 65. The method of claim 64, further comprising the step of removing.

37 It binds to tissues (the first 1,000 polymers are acrylic acid, and the second 1,000 polymers are acrylic acid). Claim 29, wherein the mer bath liquid comprises acrylamide and bisacrylamide. The method described in.

38. The first monomer bath solution contains about 0.5 to about 6 Mfiiφ of acrylamide. 67. The method according to item 66, wherein the range of B blue water is 66.

39 Diamino compound formula %formula%) (In the formula, 5 is an aliphatic functional group having a straight chain, a branched chain, or a cyclic chain, or an aromatic functional group. 62. The method according to claim 61, wherein the method has the structure (which is a functional group).

40 The second monomer solution further contains about 0.25 weight percent of bisacrylamide. The method according to claim 28, comprising:

41 Claim 28, wherein the biological tissue is an elbow, a ligament, a cardiopulmonary valve, a dura mater, or an epithelium. or the method described in paragraph 29.

42 The fourth range of H'ljf where the bovine body tissue is fixed with geltaraldehyde The method described in Section 1.

46 In a method for treating naturally occurring living tissue before transplanting it into an animal, (g)) Fix the tissue under tissue fixation conditions, and (b) further polymerize. The initial monomer was filtered for a sufficient period of time to allow the monomer to penetrate the tissue ( contact with the fabric, (c) Adding a second monomer bath under polymerization conditions to reduce tissue calcification after implantation. The first monomer is applied to the tissue using a falcon, which is effective for A method for treating the above-mentioned tissue, characterized by polymerizing it with a monomer.

44 Claim 46, wherein the first monomer is acrylic acid or methacrylic acid The method described in section.

45 Second monomer bath liquid is Acrylic IJ) Reamide, acrylic acid, acrylic acid Claim 43, wherein the ester, tutaacrylic acid, is tutaacrylic acid amide. or the method described in paragraph 44.

46 Furthermore, polymerization of the polymer was immobilized in living tissue, and the immobilization in the tissue was The first monomer is polymerized with the twelfth monomer. A method to induce calcification after transplantation of living tissue into an animal Law.

47 The first monomer is acrylic acid or methacrylic acid bound to the tissue; The second monomer is acrylamide, acrylic acid, acrylic ester, ivy acid, or methacrylamide, and the tissue contains glucuraldehyde [8j 46. The method of claim 45.

484F The body tissue is IR, 6F, heart valve, dura mater, or epithelium, Claim 4 The method described in Section 7.

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Claims (1)

[Claims] 1. Apply an effective amount of polymeric substance to the biological tissue to reduce calcification of the biological tissue after transplantation. A method for processing biological tissue before transplantation, the method comprising introducing the tissue into a tissue. 2. The method according to claim 1, wherein the polymer substance is infiltrated into living tissue. 6. The method according to claim 1, wherein the polymeric substance is covalently bonded to biological tissue. . 4 Claims in which a polymeric substance is bonded to biological tissue through a camping agent. The method described in section 6. 5. The method according to claim 4, wherein the camping agent is Diami/. 6. After bonding the acrylic acid monomer to diamino, add the polymeric substance to this acrylic acid. The method according to claim 5, wherein the method is polymerized with: Z　Polymer substances include acrylamide, acrylic acid, acrylic ester, and methacrylate. and methylacrylamide. Method described. 8. The method according to claim 6, wherein the polymeric substance is acrylamide. 9. The method according to claim 8, wherein the camping agent is ethylenediamine. . 10 The polymeric substance is bonded to the acrylic acid monomer at a concentration of about 0.5 to about 6% by weight. The method according to claim 9. 11. The method according to claim 10, wherein the living tissue is used to create a bioartificial heart valve. Method. 12a, covalently bonding the diamine to the tissue with a carbodiimide; b. covalently bonding an acrylic acid monomer to the diamine-bound tissue, and A method consisting of polymerizing a C0 biocompatible polymer onto acrylic connective tissue. Living human beings with less tendency to calcify, containing tissues in which more biocompatible polymers have been introduced. Engineered heart valve. 13. The valve according to claim 12, wherein the diamino is ethylene diamino. 14 Completely remove unbonded acrylic acid monomer from the tissue between Step and Step C. 13. The valve of claim 12, further comprising the step of flushing.