以牛心包为来源的天然衍生引导骨组织再生膜制备方法的实验研究

目的:探索天然牛心包脱细胞和交联处理的理想方法。方法:用胰酶去垢剂法(I)、胰酶核酸酶法(II)、胰酶去垢剂+核酸酶联合法(III)、 冻融去污剂法(IV)、冻融核酸酶法(V)、冻融去污剂+核酸酶联合法(VI)进行脱细胞和交联处理,采用光镜和SEM、细胞毒性实验、机械性能测定、细胞趋化性实验评估处理效果,筛选出理想的脱细胞和交联方法。结果:冻融去污剂+核酸酶联合法是理想的脱细胞方法。京尼平交联法是理想的交联方法。结论:冻融去污剂+核酸酶联合法制备的脱细胞牛心包再采用京尼平交联法可制备出理想的GBR膜材料。

Preparation of Naturally Derived Acellular Bovine Pericardium for Guiding Bone Regeneration.

Objective: To find out the best way for decellularization and cross-linking of naturally derived bovine pericardiumto prepare GBR membrane which can meet the clinical demand. Methods: Natural bovine pericardium were decellularized using trypsin+ detergent method (I), trypsin +enzymatic nucleic acid method (II), trypsin+ detergent +enzymatic nucleic acid method (III), freeze- thaw+detergent method (IV), freeze-thaw + enzymatic nucleic acid method (V), or freeze- thaw+detergent + enzymatic nucleic acid method (VI), and then were cross-linked using glutaraldehyde or genipin. The best way of decellularization and cross-linked method were screened by means of optical microscope, scanning electron microscope, cell toxicity test, determination of the mechanical properties, and measurement of the chemotactic properties. Results: Freeze- thaw+detergent + enzymatic nucleic acid method with bovine pericardium was the best method of decellularization. Genipin crosslinking method by which acellular bovine pericardium was crosslinked was better. Conclusion: Freeze- thaw+detergent + enzymatic nucleic acid method and genipin crosslinking method were ideal method for preparation of natural bovine pericardium.