牛成牙骨质细胞中骨相关标志物的表达

Expression of Bone Associated Markers by Newborn Bovine Cementoblast.

目的 :探讨新生牛成牙骨质细胞 (cementoblast,CB)中几种矿化相关蛋白在不同水平的表达及CB在体外矿化的能力。方法 :体外分离培养CB ,免疫细胞化学及原位杂交检测几种矿化相关蛋白 :骨桥蛋白、骨涎蛋白、骨钙素、碱性磷酸酶及Ⅰ型胶原在CB中的表达 ;vonKossa染色观察它们在体外形成矿化结节的能力。均以大鼠成骨细胞 (osteoblasts,OB)UMR10 6为阳性对照。结果 :几种矿化相关蛋白在CB和OB中均呈阳性表达 ,在CB中的表达弱于OB ;OPNmRNA、BSPmRNA在CB与OB中阳性表达 ;CB在体外可形成矿化结节 ,CB形成的矿化结节较OB形成者少而且小。结论 :牛CB的生物学特性与OB相似 ,但其在体外形成硬组织的能力弱于OB。

Objective:To investigate the biological characteristics of CB and compare them with OB.Methods:Culture of primary bovine CB were established from new-born bovine teeth.Cementum was manually dissected, fragmented, and digested twice with collagenase. Immunohistochemistry staining showed that the CBs were positive to monoclonal Ab against cementum attachment protein (CAP),as well as polyclonal Ab against bone sialoprotein (BSP), osteopontin (OPN), osteocalcin (OCN), alkaline phosphatase (ALP), and collagen I. Mineral nodules were detected in vitro using van Kossa staining.Results:Cementoblasts and osteoblasts were both positive to polyclonal antibodies typeⅠcollagen, OPN, BSP, OCN and ALPase, although there were only weaker staining in CB;CB and OB were also positive to OPNmRNA and BSPmRNA; CB and OB all can form nodule in vitro, although the nodule formed by CB was lesser than OB.Conclusions:These results demonstrate that bovine cementoblasts and osteoblasts share many phenotypic characteristics, and the capacity of mineralization of CB is weaker than that of OB.