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背景:骨组织在特殊物理环境如失重环境下,代谢活动会发生显著的变化,而成骨细胞是骨代谢和骨形成的核心部分,其对重力环境的变化敏感。目的:观察模拟失重条件对大鼠股骨骨髓基质细胞数量体外成骨能力的影响,揭示骨丢失的机制。设计:随机配对,对照实验。单位:解放军第四军医大学航空航天医学系和口腔医学院病理科。材料:选用20只成年健康雄性SD大鼠。实验开始当日按体质量随机分为对照组和悬吊组,每组10只。碱性磷酸酶试剂盒由北京中生生物工程高技术公司生产。方法:实验于1999-11/2000-07在解放军第四军医大学口腔医学院病理科完成。将SD大鼠随机配对分为鼠尾悬吊组和对照组,每组10只。悬吊组大鼠做尾部悬吊28d,大鼠始终保持30°头低位及后肢自由悬垂不负重状态。对照组正常饲养。实验期满,取股骨,将股骨骨髓基质细胞进行原代和传代细胞培养。主要观察指标:采用细胞计数法和噻唑蓝法绘制原代和传代培养细胞的生长曲线,进行碱性磷酸酶活性及体外矿化小结形成量的检测。结果:①碱性磷酸酶活性:原代和传代培养悬吊组低于对照组,差异有显著性意义(P<0.05)。②钙化小结形成数:悬吊组少于对照组,差异有显著性意义(P<0.05)。③细胞生长:原代和传代股骨间充质细胞的生长曲线呈“S”形,悬吊组和对照组细胞倍增时间相近。④股骨骨髓基质细胞数:原代细胞培养系中,悬吊组比对照组约少50%(P<0.05)。结论:模拟失重条件下,大鼠骨髓基质细胞数明显减少,后肢承重骨成骨细胞数减少,体外成骨能力降低。
BACKGROUND: Bone tissue undergoes significant changes in metabolic activity under special physical conditions such as weightlessness. Osteoblasts, the core of bone metabolism and bone formation, are sensitive to changes in the gravitational environment. OBJECTIVE: To observe the effects of simulated weightlessness on in vitro osteogenic capacity of rat femur marrow stromal cells and to reveal the mechanism of bone loss. Design: Random pair, control experiment. Unit: Department of Aeronautics and Astronautics, Fourth Military Medical University of PLA and Department of Pathology, School of Stomatology. MATERIALS: Twenty healthy male SD rats were selected. On the day of the experiment, the rats were randomly divided into control group and suspension group according to the body weight, with 10 in each group. Alkaline phosphatase kit by Beijing Zhongsheng bio-engineering high-tech companies. METHODS: The experiment was performed at the Department of Pathology, School of Stomatology, Fourth Military Medical University of Chinese PLA from November 1999 to July 2000. The SD rats were randomly paired into tail suspension group and control group, 10 rats in each group. Suspension rats tail suspension 28d, rats always maintain a 30-degree low head and hind limbs free hanging weightless state. The control group was reared normally. After the expiration of the experiment, the femur was taken and the primary and subculture cells of femur marrow stromal cells were cultured. MAIN OUTCOME MEASURES: Growth curves of primary and subcultured cells were drawn by cell counting and thiazolyl blue assay, alkaline phosphatase activity and the formation of mineralized nodules in vitro were measured. Results: ① Alkaline phosphatase activity was lower in the primary and subculture suspension groups than in the control group, with significant difference (P <0.05). ② The number of calcified nodules: suspended group less than the control group, the difference was significant (P <0.05). ③ Cell growth: The growth curve of primary and passage femoral mesenchymal cells was “S” shape, the cell doubling time was similar in suspension group and control group. ④ Femoral bone marrow stromal cells: primary cell culture line, the suspension group about 50% less than the control group (P <0.05). CONCLUSION: Under the condition of simulated weightlessness, the number of bone marrow stromal cells in rats decreased significantly, the number of osteoblasts in hindlimb-bearing bone decreased and the osteogenic capacity in vitro decreased.