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为了研究缺锌对体外培养大鼠成骨细胞增殖和分化的影响，从大鼠颅骨分离出成骨细胞，并于ＤＭＥＭ培养基中进行传代培养。锌特异性络合剂ＴＰＥＮ络合掉培养基中的锌建立细胞体外培养的缺锌模型。采用３Ｈ－ＴｄＲ掺入法确定成骨细胞在不同时点ＤＮＡ的合成状况，同时应用流式细胞仪进行细胞周期的分析；采用组织学方法观察细胞骨架的变化、３Ｈ－脯氨酸掺入法确定胶原蛋白的合成；采用酶动力学和放免方法分别测定碱性磷酸酶活性和骨钙素含量。结果表明，缺锌组成骨细胞３Ｈ－ＴｄＲ掺入量在各个时点明显低于对照组，细胞被阻滞于细胞周期中Ｇ２／Ｍ期，这与细胞骨架受损密切相关。缺锌组胶原蛋白合成、骨钙素含量及碱性磷酸酶活性明显低于对照组。但缺锌补锌组的各个指标与对照组相比无统计学差异。因此，缺锌能够抑制体外培养大鼠成骨细胞的增殖和分化。
In order to study the effect of zinc deficiency on the proliferation and differentiation of cultured rat osteoblasts, osteoblasts were isolated from the rat skull and subcultured in DMEM medium. Zinc-specific complexing agent TPEN complexed zinc in culture medium to establish a zinc-deficient model of cultured cells in vitro. 3H-TdR incorporation method was used to determine the DNA synthesis of osteoblasts at different time points. At the same time, flow cytometry was used to analyze the cell cycle. The changes of cytoskeleton were observed by histological method. 3H-proline incorporation Determine the synthesis of collagen; enzyme kinetics and radioimmunoassay method were measured alkaline phosphatase activity and osteocalcin content. The results showed that incorporation of 3H-TdR into zinc-deficient osteoblasts was significantly lower than that of the control at each time point, and the cells were arrested in the G2 / M phase of the cell cycle, which was closely related to the cytoskeleton damage. Zinc deficiency group collagen synthesis, osteocalcin content and alkaline phosphatase activity was significantly lower than the control group. However, zinc-deficient zinc group showed no significant difference compared with the control group. Therefore, zinc deficiency can inhibit the proliferation and differentiation of cultured rat osteoblasts in vitro.