Polyvinyl alcohol improves resistance of epoxy-treated bovine pericardium to calcification in vitro

A. Kostyunin, M. Rezvova, T. Glushkova, D. Shishkova, A. Kutikhin, T. Akentieva, E. Ovcharenko
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Abstract

Background. Around half of bioprosthetic heart valves become dysfunctional 15 years postimplantation because of structural valve deterioration notable for the degradation and calcification of the prosthetic tissue. Protection of bioprosthetic heart valves from structural valve deterioration requires innovative materials, science approaches including enveloping of the bioprosthetic heart valves into the polymer sheath.Aim. To develop a polyvinyl alcohol sheath for improving resistance of bioprosthetic heart valves to calcification.Material and methods. Bovine pericardium fixed with ethylene glycol diglycidyl ether was incubated with distinct concentrations of polyvinyl alcohol (5, 10, 12, or 15%) with the following freezing and thawing to perform cryotropic gelation. Surface and structure of unmodified and polyvinyl alcohol-modified bovine pericardium have been investigated by fluorescence microscopy and scanning electron microscopy, whilst tensile testing was carried out by uniaxial tension test. Haemocompatibility was assessed through the measurements of haemolysis and platelet aggregation/adhesion upon the contact of donor blood with the samples. Resistance to calcification was tested by incubation of the samples in calcium and phosphate supersaturated (10 µmol/L) cell culture medium for 3 and 6 weeks with the following tissue lysis and colorimetric measurement of Ca2+ ions.Results. Using cryotropic gelation, we obtained a polyvinyl alcohol-coated and filled bovine pericardium matrix. Out of all polyvinyl alcohol concentrations, 12% polyvinyl alcohol solution sealed pores and hollows within the bovine pericardium (what was not achieved using 5% or 10% polyvinyl alcohol solutions) and demonstrated the best processability as compared to 15% polyvinyl alcohol solution. Cryotropic gelation did not deteriorate durability, elasticity, or haemocompatibility of bovine pericardium. After 3 and 6 weeks of the incubation in calcium-supersaturated solution, polyvinyl alcoholmodified bovine pericardium contained 5- and 3-fold reduced amount of calcium compared to unmodified bovine pericardium.Conclusions. Enveloping of bovine pericardium into polyvinyl alcohol increases its calcification resistance, retains its tensile properties and haemocompatibility, and can be considered as a promising approach for the modification of bovine pericardium during the manufacturing of bioprosthetic heart valves.
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聚乙烯醇提高环氧处理牛心包体外抗钙化的能力
背景。大约一半的生物心脏瓣膜在植入15年后会出现功能障碍,原因是瓣膜的结构性退化,尤其是假体组织的退化和钙化。保护生物人工心脏瓣膜不受结构瓣膜恶化的影响需要创新的材料和科学的方法,包括将生物人工心脏瓣膜包裹在聚合物鞘中。目的:研制聚乙烯醇护套,提高生物人工心脏瓣膜的抗钙化能力。材料和方法。用乙二醇二缩水甘油醚固定牛心包,用不同浓度的聚乙烯醇(5%、10%、12%或15%)孵育,然后冷冻和解冻,以进行低温凝胶化。用荧光显微镜和扫描电镜研究了未改性和聚乙烯醇改性牛心包的表面和结构,并用单轴拉伸试验进行了拉伸试验。血液相容性是通过测量供体血液与样品接触时的溶血和血小板聚集/粘附来评估的。将样品在钙和磷酸盐过饱和(10µmol/L)的细胞培养基中孵育3周和6周,然后进行组织裂解和Ca2+离子比色测定,以测试样品对钙化的抗性。采用低温凝胶法,我们获得了一种聚乙烯醇包被和填充的牛心包基质。在所有聚乙烯醇浓度中,12%聚乙烯醇溶液密封了牛心包内的孔隙和空洞(使用5%或10%聚乙烯醇溶液无法实现这一点),并且与15%聚乙烯醇溶液相比,表现出最佳的加工性。低温凝胶化不会降低牛心包的耐久性、弹性或血液相容性。在钙过饱和溶液中孵育3周和6周后,聚乙烯醇改性的牛心包膜的钙含量比未改性的牛心包膜减少了5倍和3倍。聚乙烯醇包裹牛心包膜,可提高其抗钙化性能,保持其拉伸性能和血液相容性,可被认为是在生物人工心脏瓣膜制造过程中对牛心包膜进行改性的一种有前途的方法。
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