NaOH消蚀法制备胎儿脱细胞真皮基质的研究

The study on fetus' acellure dermal matrix prepared by NaOH-maceration

目的:探索NaOH消蚀法对胎儿真皮基质的成分及其形态结构的影响。方法:使用NaOH消蚀法制备胎儿皮肤的脱细胞真皮基质(ADM),应用组织化学和免疫组织化学染色方法检测细胞的余留及细胞外基质成分的破坏情况,应用扫描电镜观察ADM的构筑。结果:NaOH消蚀处理16h即可去除真皮细胞成分。制备的ADM韧性好,勿需戊二醛交联。弹性纤维的含量有所减少而胶原纤维形态完整,排列正常,保留了基本的真皮构筑。HLA-DR呈阴性,Fibronectin(FN)呈弱阳性,Laminin(LN)和Ⅳ型胶原均呈阴性。结论:NaOH消蚀法简易经济,去除细胞成分较彻底,所制备的ADM韧性好,胶原纤维形态完整,排列正常,但对基膜的成分及结构破坏较重。

Objective: To study the effects of NaOH-maceration on the composition and structure of acelluar dermal matrix(ADM). Methods: The retention of cellular components and composition of ADM prepared by NaOH-maceration were detected using histochemical and immunohistochemical staining methods. The three-dimensional structures of ADMs were observed under scanning electron microscope (SEM). Results: The method of NaOH-maceration could completely remove the cellular components from the dermal matrix within 16h. The ADM was tough, so can be easily used in operation without cross-linking with glutaraldehyde. The morphological character of collagen bundles and the basic dermal architecture remained intactly, however the elastic fibers, fibronectin, laminin and collagen IV were injured duing the process of preparation. Conclusions:The procedure of NaOH-maceration is simple and easy to perform. The morphological character of collagen bundles is normal but the structure of the basement membrane is injured severely.