骨脱细胞后的有机成分分析

COMPONENT ANALYSIS OF BONE MATRIX VIA CELL EXTRACTION

目的 探讨脱细胞骨细胞外基质 (AECM)的制备 ,并对其进行有机成分分析。 方法 采用低渗和除垢剂脱细胞方法制备 AECM。使用 HE、Mallory- Heidenhain氏快速一步法染色和阿利新蓝染色观察形态学改变 ;免疫组织化学检测纤维连接蛋白 (FN)和层连接蛋白 (L N)。 结果 HE、Mallory- Heidenhain氏快速一步法染色后镜下可见AECM胶原纤维排列规则 ,骨陷窝空虚 ;阿利新蓝染色可见 AECM骨陷窝周边染色呈不同程度蓝绿色 ;免疫组织化学染色可见 FN和 L N的抗体染色阳性部位在细胞外基质 (ECM)。 结论 低渗和除垢剂二步法脱细胞是 AECM制备的有效方法。脱细胞过程中去除了细胞成分 ,保留了天然的胶原纤维分布及 ECM主要支架成分蛋白多糖、FN和 L N。

Objective To investigate the preparation of bone acellular extra cell matrix(AECM) and to analyze its component. Methods With low osmosis theory and method of cell extraction by detergent, bone acellular extra cell matrix was prepared. We observed morphologic changes with HE, Mallory Heidenhain rapid one step dyeing and Alcian blue dyeing and examined fibronectin(FN) and laminin(LN) with immunohistochemistry. Results Light microscope showed that the collagen fibers arranged regularly in AECM with blankness of bone lacunas by HE, Mallory Heidenhain rapid one step dyeing and that the region around bone lacunas was stained different degrees of blue green by Alcian blue dyeing. The result of immunohistochemistry showed there are positive markers of FN and LN in ECM. Conclusion This method for preparation of bone acellular extra cell matrix is effective, and it can keep natural structure of collagen fibers and maintain components of ECM, such as proteoglycan, FN and LN.