致敏淋巴细胞对兔颅骨成骨细胞功能的影响

Effect of sensitized lymphocytes on rabbit calvarial osteoblasts

目的 研究致敏淋巴细胞对兔颅骨成骨细胞增殖、分化的作用。方法 经酶消化法从新生兔颅骨中分离成骨细胞 ,进行成骨细胞的碱性磷酸酶染色及成骨能力的检测。从经铬 (Cr6+)致敏的新西兰兔外周血分离淋巴细胞并提取其培养上清液 (LCM) ,并分别观察在无LCM、加未经抗原(Cr6+)刺激的LCM和加经Cr6+刺激的LCM等 3种情况下成骨细胞增殖、碱性磷酸酶和骨钙素分泌的情况。结果 从兔颅骨分离的细胞碱性磷酸酶染色阳性 ,在长期培养中能够形成钙化结节。致敏淋巴细胞培养介质抑制成骨细胞的增殖、促进碱性磷酸酶的分泌 ,抑制骨钙素的产生。经方差分析检验均具有明显的统计学差异。结论 致敏淋巴细胞能够抑制成骨细胞的功能 ,可导致骨形成障碍。

Objective To study the effects of sensitized T lymphocytes on osteoblastic proliferation and differentiation. Methods Sensitized lymphocytes were obtained from a rabbit sensitized by chromium, and lymphocyte cultured mediums (LCM) were collected. Osteoblasts were obtained from new born rabbit calvaria. The osteoblasts were identified with alkaline phosphatase staining and mineralization in vitro. By osteoblast cultured in vitro, osteoblastic proliferation, and production of alkaline phosphatase and osteocalcin were evaluated in three conditions: without LCM, with LCM not stimulated by Cr 6+ and with LCM stimulated by Cr 6+ . Results The cells from rabbit calvaria were positive to ALP staining and could form calcium node in vitro. The sensitized LCM could inhibit proliferation of osteoblasts, increase alkaline phosphatase production and decrease osteocalcin production. These results all had statistically significant difference by ANOVA. Conclusion Sensitized T lymphocytes could decrease the number of mature osteoblasts and impede the formation of mature mineralized bone.