猪去抗原半月板体外降解的实验研究

目的 研究猪去抗原半月板在体外的降解规律，以进一步调整其合适降解速度。方法选用4～6月龄健康小型猪5只，取其双膝关节半月板。一部分采用环氧化物进行去抗原处理，制备成去抗原半月板；另一部分不作任何处理，为新鲜半月板。将两种半月板冻干处理后，置于含胶原酶、木瓜蛋白酶、透明质酸酶和胰酶的PBS液中，于37C酶解3、7、15、30d。另取两种半月板在PBS液37℃下水解30d。采用失重法、紫外光谱分析及扫描电镜测定其降解特性，并测定两种半月板的亲水性。结果两种半月板于4种酶中的酶解效果显著，其中以胰酶最明显，但去抗原半月板酶解程度低于新鲜半月板。新鲜半月板及去抗原半月板水解率分别为0．76％、0．81％。两种半月板在4种酶的降解液中紫外吸收值范围大致相同，为200～300nm。扫描电镜观察，去抗原半月板的空间结构改变不及新鲜半月板明显。去抗原半月板的亲水性较新鲜半月板稍低：吸水率为157％、188％；保水率为152％、167％。结论去抗原半月板降解较新鲜半月板慢，为调整去抗原半月板在体内降解速度提供依据。

DEGRADATION OF ANTIGEN-EXTRACTED MENISCUS IN VITRO

OBJECTIVE: To study degradation of the antigen-extracted meniscus in PBS solution with no enzyme or with different enzymes. METHODS: Four types of enzymes (collagenase, hyaluronidase, trypsin, papain) were used to enzymolyze the antigen-extracted meniscus and the fresh meniscus for 3, 7, 15 and 30 days (37 degrees C). The antigen-extracted meniscus and the fresh meniscus were immersed in PBS solution (37 degrees C) for 30 days. Weight loss measurement, UV spectrophotometry, and scanning electron microscopy (SEM) were used to characterize the degraded materials. RESULTS: The two types of the materials were remarkably digested under the enzymes, especially under trypsin. The degradation curves showed that the antigen-extracted meniscus was enzymolyzed less than the fresh meniscus. The degradation products were grouped as amino, peptide, and polyose by the analysis. Both of the materials could hardly be hydrolyzed in PBS solution without the enzymes. The four different enzymes had different surface morphologies under the examination of SEM. CONCLUSION: The antigen-extracted meniscus is enzymolyzed more slowly than the fresh meniscus in vitro, and the result can be used as a guideline to the further research.