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目的 :研究不同浓度中药 Liu-OGN对体外成人成骨细胞增殖和分化的影响。方法 :采用胶原 -胰蛋白酶消化法获得成人松质骨中的成骨细胞 ,并进行纯化和培养 ,然后使用含不同浓度 Liu-OGN药物血清的培养液进行培养。通过倒置相差显微镜和电镜观察细胞生长情况 ,采用 MTT法检测细胞增殖情况 ,通过碱性磷酸酶和骨钙素等生化指标测定细胞分化情况。结果 :获得的成人成骨细胞生长情况良好 ,生化指标稳定可靠 ,纯化效果佳 ,可以满足相关研究的需要。进一步的研究结果表明 ,在成骨细胞密度为 1 0 0 0 0 /ml的条件下 ,与不加药对照组比较 ,1 0 %、2 0 % Liu-OGN中药组和地塞米松组的成骨细胞体积增大 ,上清液中碱性磷酸酶和骨钙素含量均明显增高 ;但各 Liu-OGN中药组成骨细胞的增殖情况无明显变化 ;结论 :中药 Liu-OGN对体外成人成骨细胞的分化有明显地促进作用 ,可以作为促进骨形成的药物使用。
Objective: To study the effects of different concentrations of Liu-OGN on the proliferation and differentiation of adult human osteoblasts in vitro. METHODS : Osteoblasts in adult cancellous bone were obtained by collagen-trypsin digestion, purified and cultured, and then cultured in culture medium containing different concentrations of Liu-OGN drug serum. Inverted phase contrast microscopy and electron microscopy were used to observe cell growth. MTT assay was used to detect cell proliferation, and cell differentiation was determined by biochemical markers such as alkaline phosphatase and osteocalcin. Results: The obtained adult osteoblasts grew well, the biochemical indicators were stable and reliable, and the purification effect was good. It can meet the needs of related research. Further research results showed that 10% and 20% of the Liu-OGN Chinese medicine group and dexamethasone group were compared with the non-dosing control group at the osteoblast density of 1 000/ml. The volume of bone cells increased, and the contents of alkaline phosphatase and osteocalcin in the supernatant increased significantly. However, the proliferation of osteoblasts in each Liu-OGN composition did not change significantly. Conclusion: Liu-OGN, a traditional Chinese medicine, on adult human osteoblasts in vitro The differentiation of cells has a significant promoting effect and can be used as a drug for promoting bone formation.