兔肋软骨脱细胞基质的制备

Preparation of rabbit costal cartilage acellular matrix

背景:研究表明新西兰兔软骨组织可作为组织工程支架材料,其中关节软骨及耳软骨的脱细胞基质的研究较多,但采用肋软骨作为组织工程软骨支架的研究较少。目的:制备新西兰兔肋软骨脱细胞基质,探讨天然软骨支架作为组织工程支架的可行性。方法:用联合去垢剂-酶法获得软骨支架,根据脱细胞过程中TritonX-100第2次处理时间0,24,48,96h分为4组。脱细胞完毕后各组支架固定行扫描电镜采集图像观察计算支架孔隙率、孔径长度,并对支架进行苏木精-伊红染色、甲苯胺蓝及Ⅱ型胶原免疫组织化学染色,并将脱细胞支架植入异体新西兰兔皮下观察其相容性。结果与结论:兔肋软骨脱细胞基质呈乳白色,大小均一,染色示支架结构完整,仍保存大量酸性黏多糖及Ⅱ型胶原成分,扫描电镜观察经一定时间的脱细胞处理后可得到结构完整,孔隙均匀的天然软骨支架,其孔隙率为(61.31±8.45)%;孔径长度为(32.80±5.15)μm,符合正态性分布,各组脱细胞支架植入异体新西兰兔皮下7d后生物相容性良好,周围软组织无明显充血、化脓等炎症排斥反应出现。结果显示,兔肋软骨脱细胞支架具有良好的基质组成,有较完整、均匀的孔隙结构及孔径分布,可作为组织工程支架材料。

BACKGROUND： Several studies have suggested that New Zealand rabbits cartilage tissue can be used as a kind of tissue engineering scaffold material and is widely used in scientific research, including articular cartilage and ear cartilage cells of the matrix of the emergence of studies comparing fully, but adopting costal cartilages as tissue engineering cartilaginous framework of research little, yet few reports. OBJECTIVE： Based on the preparation of costal cartilage acellular matrix from New Zealand rabbits, to discuss the feasibility of natural cartilage matrix scaffold for tissue engineering scaffolds. METHODS： By using detergent-enzyme method, cartilaginous framework was obtained, and acellular matrix was divided into four groups of 0, 24, 48 and 96 hours according to Triton X-100 second processing time. Scanning electron microscope was used for image acquisition to calculate stents porosity, pore diameter, and then the scaffolds underwent hematoxylin-eosin, toluidine blue and Ⅱ collagen type immunohistochemical staining. The compatibility of the acellular matrix scaffolds transplanted subcutaneously was observed. RESULTS AND CONCLUSION： The acellular matrix presented with milk-white, size uniformity with intact stained structure, and there were a lot of acid mucopolysaccharide and collagen type Ⅱ. The porosity was （61.31± 8.45）%, and the pore diameter was （32.80±5.15） μm. The biocompatibility of the transplanted acellular matrix was good at 7 days after transplantation. No obvious hyperemia and purulent occurred. These findings show that there is a good substrate composition of rabbit coastal cartilage acellular matrix scaffolds with intact and even pore structure, which can be used as tissue engineering scaffold materials.