摘要
目的:对比分析不同方法制备的猪肝脱细胞基质中蛋白的保留情况,进一步优化制备工艺。方法:分别采用聚乙二醇辛基苯基醚(Triton X-100)、十二烷基硫酸钠(SDS)两种表面活性剂并结合胰酶酶解等手段制备猪肝ECM。再以H&E染色、免疫组化、生物质谱分析等检测方法对比分析两种ECM的成分及含量,进而优化ECM的制备工艺并探讨其中的机制。结果:Triton X-100和SDS两组都可以制备符合国际标准的ECM,其中Triton X-100组对胶原蛋白和生长因子的保留优于SDS组;但GAG流失较多。结论:综合运用胰蛋白酶+氨水+Triton X-100的脱细胞策略可以制备符合国际标准的ECM,同时减少了ECM蛋白的流失。
Objective To compare and analyze the protein retention in decellularized porcine liver extracellular matrice(ECMs)prepared via two methods,so as to further optimize the preparation process.Methods Decellularized porcine liver ECM was prepared respectively by using either one of two surfactants,polyethylene glycol octyl phenyl ether(Triton X-100)and sodium dodecyl sulfate(SDS),combined with enzymolysis by trypsin.Hematoxylin and eosin(HE)staining,immunohistochemistry,and biomass spectrometry were used to compare and analyze the components and their contents in the two ECMs.Thereby,we optimized the preparation process of ECM and discussed the underlying mechanism.Results Either Triton X-100 or SDS was eligible for preparation of ECM that satisfies international standards.The ECM prepared using Triton X-100 showed better retention of collagens and growth factors,but more loss of glycosaminoglycan(GAG)compared with that prepared using SDS.Conclusion Combination use of trypsin,aqueous ammonia and Triton X-100 for decellularization may lead to preparation of ECM that satisfies international standards,meanwhile with less loss of ECM proteins.
作者
赵超尘
邬杏连
周俊晶
Zhao Chaochen;Wu Xinglian;Zhou Junjing(Department of Hepatobiliary Surgery,First Affiliated Hospital of Guangzhou Medical University,Guangzhou 510120,China;Central Pharmacy,First Affiliated Hospital of Guangzhou Medical University,Guangzhou 510120,China;Department of Hepatobiliary and Pancreatic Surgery,Affiliated Hospital of Jiangnan University,Wuxi 214062,Jiangsu,China)
出处
《中华生物医学工程杂志》
CAS
2022年第5期528-533,共6页
Chinese Journal of Biomedical Engineering
关键词
脱细胞基质
聚乙二醇辛基苯基醚
十二烷基硫酸钠
Decellularized matrice
Polyethylene glycol octyl phenyl ether
Sodium dodecyl sulfate
