鱼源胶原蛋白海绵材料的构建及其生物学性能

Preparation and Biochemical Properties of Collagen Sponges from Fish

目的构建鱼源胶原海绵材料,评价其生物学性能并与哺乳动物来源的胶原海绵材料相比较。方法分别从草鱼鱼皮和猪皮中提取鱼皮酸溶性胶原蛋白(SKA)、鱼皮酶溶性胶原蛋白(SKP)和猪皮酶溶性胶原蛋白(PSP),采用氨基酸组成分析和SDS-PAGE凝胶电泳对胶原蛋白进行结构鉴定;以制备所得的3种胶原蛋白构建柱状胶原海绵材料,采用差式扫描量热仪(DSC)、体外模拟胶原蛋白酶降解、材料拉伸性能分析以及NIH/3T3细胞培养实验评价胶原海绵材料的热稳定性、降解性、生物相容性和材料机械力学性能。结果从鱼皮和猪皮中分离、纯化得到的SKA、SKP和PSP均为典型的Ⅰ型胶原,SKA和SKP的氨基酸组成基本一致,但它们与PSP在氨基酸组成上存在部分差异。SKA和SKP的热变性温度分别为(35.6±0.2)℃和(35.8±0.3)℃,明显低于PSP的热变性温度(41.6±0.3)℃(P<0.05)。与PSP海绵相比,SKA和SKP海绵材料具有更好的耐胶原酶降解能力和材料拉伸强度,但其材料的延展性能低于PSP海绵(P<0.05)。在NIH/3T3培养实验中,3种胶原海绵材料均没有出现抑制细胞生长现象(P>0.05),SKP和PSP在培养48h和96h时展现出促进细胞增殖的能力(P<0.05)。结论鱼源胶原海绵材料在生物学性能上与哺乳动物来源的胶原海绵材料各有优劣,具备应用于生物医学材料领域中的潜在价值。

Objective To prepare collagen sponges from fish and evaluate their biochemical properties by comparing with collagen sponges from mammal.Methods Acid-soluble collagen from grass carp skin（SKA）,pepsin-soluble collagen from grass carp skin（SKP）and pepsin-soluble collagen from porcine skin（PSP）were extracted from grass carp skin and porcine skin.The structure characterizations of collagens were investigated by amino acid composition and SDS-PAGE analysis.Collagen sponges were prepared from SKA,SKP and PSP.The thermal stability,enzymatic sensitivity,mechanical properties and cytotoxicity of those collagen sponges were tested by using the methods of DSC analysis,determination of collagen degraded products in collagenase solution,tensile strength analysis and cell culture of NIH/3T3.Results SKA,SKP and PSP comprised two different α chains（α1 and α2）and were classified as type collagen.SKA and SKP were similar in amino compositions but they were different from those of PSP.Both SKA and SKP had a lower denaturation temperature[（35.6±0.2）℃ and（35.8±0.3）℃]than PSP[（41.6±0.3）℃,P0.05].Both of SKA and SKP sponges showed more resistive to collagenase and higher tensile strength than PSP sponges（P0.05）,but the prolongation property of PSP was better than that of SKA and SKP（P0.05）.Both of SKA and SKP did not induce a significant cytotoxic effect according to the results of in vitro cytotoxicity test（P0.05）,and SKP exhibited higher cell viability than controls after 48 h and 96 h（P0.05）.Conclusion The biochemical properties of collagen sponges from fish and mammal have their own merits,and collagen sponges from fish could be used as an alternative source for further applications in medical biological materials.