原花青素在酸性条件下交联制备心脏瓣膜的性能研究  

Crosslinking of decellularized heart valve by procyanidins in the acidic buffer solution:Compliance,anti-calcification and biological compatibility

作　　者：王晓亚[1,2] 马旭兵[1,2] 翟万银[2] 张嘉敏[3] 朱自严[3] 张红锋[1] 常江[2] 

机构地区：[1]华东师范大学生命科学学院,上海200062 [2]中国科学院上海硅酸盐研究所生物材料与组织工程中心,上海200050 [3]上海市血液中心上海市输血研究所,上海200051

出　　处：《华东师范大学学报（自然科学版）》2014年第6期109-120,共12页Journal of East China Normal University(Natural Science)

基　　金：国家自然科学基金(31070870)

摘　　要：为了明确原花青素（procyanidins,PC）在酸性条件下交联脱细胞心脏瓣膜是否能够维持其柔顺度、抑制其钙化成核和具备良好的生物学特性,以1 mg/mL PC在酸性条件下（pH3.0～7.4）交联脱细胞心脏瓣膜,采用万能力学测试仪测试研究其柔顺度,并用体外钙化、体外酶降解、细胞黏附、抗凝血和血小板黏附实验研究其抗钙化和相关生物学特性.结果显示,弱酸性条件下交联的脱细胞瓣膜（PC1-pH6.0）呈现出近似于未交联脱细胞瓣膜的柔软程度,交联后拉伸强度显著提高（5.95士0.6） MPa vs （4.64士0.81） MPa,同时弹性模量与未交联对照组相当,显示出较好的柔顺度;PC1-pH6.0组体外可有效抑制瓣膜钙化;PC交联能保护脱细胞瓣膜不被Ⅱ型胶原酶降解,瓣膜间质细胞在交联脱细胞瓣膜表面黏附良好,交联后的脱细胞瓣膜具有更好的抗血栓潜能.因此,pH6.0条件下PC交联的脱细胞心脏瓣膜具有类似于未交联脱细胞瓣膜的柔顺性,能有效抑制钙化,具有抗酶降解、抗血栓和良好的细胞相容性,有望用作可再细胞化的生物瓣.The aim of this study was to explore whether procyanidins （PC）-crosslinked acellular heart valves in acidic buffer solution can obtain the identical matrix compliance similar to non-crosslinked acellular ones with anti-calcification and good biocompatibility properties.Based on the different pH acidic buffer solutions of 1 mg/mL PC,the crosslinked acellular valves were crosslinked.The compliance was determined by bending angle measurement,anti-calcification by soaking in simulated body fluid,resistance to enzymatic degradation by collagenase-Ⅱ hydrolysis.Under pH 6.0 condition,PC crosslinked valve （PC1-pH 6.0） group was as soft as its non-crosslinked counterpart.Compared with noncrosslinked group （4.64 MPa）,the ultimate tensile strength of PC1-pH 6.0 group reached up to 5.95 MPa while the elastic modulus maintained the same,showing higher tensile strength than and the same compliance as the decellularized valves.Calcification was completely inhibited after crosslinking at pH 6.0.Under this condition,the PC-crosslinked acellular valves can effectively resistant to proteolysis compatible to glutaraldehyde-crosslinked ones.Furthermore,acellular valves in PC1-pH 6.0 group provided a good substance for adhesion of heart valve interstitial cells and present anti-thrombosis potential.PC-crosslinked acellular valves in the Acidic Buffer Solution （pH 6.0） possess identical matrix compliance with non-crosslinked decellularized valves,potential of anti-calcification,resistance to enzymatic degradation,anti-thrombosis and good cytocompatibility,which may be used for preparing bioprosthetic heart valves.

关 键 词：原花青素 交联 柔顺度 酸性条件 抗钙化 生物相容性 

分 类 号：Q819[生物学—生物工程]