人脱细胞半月板的制备

Preparation of human meniscus acellular matrix

目的：研究人脱细胞半月板的制备及其形态结构和生物力学性能。方法：切取人半月板制备人脱细胞半月板，用HE、甲苯胺蓝、天狼星红、番红花O、AB染色和Hoechst-33258染色等方法进行定性检测，扫描电镜进行形态学观察，用生物力学机测定其在不同压缩比率下的瞬时形变恢复率，最大形变恢复率及最大受压强度。结果：各种染色均显示细胞陷窝内已无细胞结构，Hoechst-33258染色显示无细胞残留，天狼星红染色为阳性，电镜观察示半月板表面及内部不规则存在空虚的陷窝。人脱细胞半月板的孔隙较大，孔隙直径为80～760μm，孔隙率在67％以上。生物力学测定其压缩30％时，最大瞬时形变恢复率为（89．62±1．04）％，最大形变恢复率为100％，最大强度是（3．04±0．13）N。结论：人半月板经去污剂等处理后，去除了细胞成分，保留了三维立体结构的细胞外基质，具有一定的生物力学性能；成功制备的人脱细胞半月板可作为半月板组织工程的支架载体。

Objective：To investigate development of a cell extraction process for preparing human meniscus acellular ma-trix,and morphology and biomechanical properties. Methods：Human meniscus were subjected to modified eight step deter-gent,then,the specimens were assessed by staining with haematoxylin eosin,toluidine blue,sirius red,saffron O,alcain blue and hoechst-33258,et al. The ultrastructure of the specimens was observed with scanning electron microscope. Transient re-covery rate of deformation,maximal recovery rate of deformation and maximal compressive strength were tested to determine the biomechanical properties of the scaffold. Results：Every stain confirmed that the celluar constituents of the specimens were removed. The specimens stained positively by staining with sirius red. Lacuna were found irregularly not only on the surface of the meniscus,but also in the meniscus with scanning electron microscope. Pores in the specinmens were large ,the diameter of pores was 80 to 760μm,porosity was over 67%. The transient recovery rate of deformation was （89.62±1.04）%,the maximal recovery rate of deformation was 100%and the maximal compressive strength was （3.04±0.13）N,when the specimens were compressed 30%. Conclusion：The modified eight step detergent can remove the immunogenic cell components from human meniscus,in addition,3D extracellular matrix can be retained. The scaffold has good biomechanical properties. This scaffold stands a good chance to be an implant for future tissue engineering of the human meniscus.