肿瘤坏死因子α短期刺激促进骨髓间充质干细胞的成骨分化-东南大学学报医学版

肿瘤坏死因子α短期刺激促进骨髓间充质干细胞的成骨分化

单位：1. 西安市第九医院口腔科, 陕西西安 710054;
2. 军事口腔医学国家重点实验室, 国家口腔疾病临床医学研究中心, 陕西省口腔医学重点实验室, 第四军医大学口腔医院牙体牙髓病科, 陕西西安 710032;
3. 第四军医大学基础医学院, 陕西西安 710032

关键词：肿瘤坏死因子α 水凝胶骨髓间充质干细胞成骨分化

分类号：R329.2

出版年·卷·期（页码）：2021·40·第四期（500-505）

摘要：

目的：探究肿瘤坏死因子α（TNF-α）刺激3 d对骨髓间充质干细胞（BMMSCs）增殖和成骨分化的影响。方法：不同浓度（0、1、10、50 ng·ml^-1）的TNF-α刺激BMMSCs 3 d后，MTS细胞增殖检测试剂盒检测BMMSCs继续培养1、4、7 d后的体外增殖活性；细胞迁移实验检测其体外迁移能力；成骨诱导液继续诱导2周后检测其体外碱性磷酸酶（ALP）活性；水凝胶分别包裹1、50 ng·ml^-1的TNF-α在小鼠极限模型中植入，8周后取材通过微计算机断层扫描技术（micro-CT）和HE染色检测新骨形成能力。结果：与对照组比较，TNF-α处理组刺激3 d对细胞增殖无影响，但可显著促进BMMSCs的迁移，其中50 ng·ml^-1浓度迁移效果最好。ALP活性检测显示，在2周时1、10、50 ng·ml^-1 TNF-α均能显著促进BMMSCs的迁移（P<0.05），并呈浓度依赖性递增，50 ng·ml^-1的TNF-α效果最好。成功构建了水凝胶TNF-α短期缓释系统，小鼠体内micro-CT和HE染色显示50 ng·ml^-1 TNF-α短期缓释组成骨效果最好，能有效促进新骨形成。结论：本实验证实TNF-α成骨诱导液短时间刺激可有效促进BMMSCs在体内体外的成骨分化。

Objective: To explore the influence of tumor necrosis factor-α(TNF-α) on the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells(BMMSCs) after 3 days of short-term stimulation. Methods: BMMSCs were stimulated with different concentrations of TNF-α(0, 1, 10, 50 ng·ml^-1) for 3 days. The proliferation of BMMSCs in vitro was detected by MTS after 1, 4 and 7 days of continuous culture. Transwell migration assay was used to determine its migration ability in vitro. After 2 weeks of induction, the alkaline phosphatase activity(ALP) of the osteoblasts was detected. TNF-α coated with hydrogel of 1 and 50 ng·ml^-1 was implanted in the mouse limit model. After 8 weeks, micro-CT scanning and HE staining were employed to detect the new bone formation ability. Results: Compared with the control group, no effect on cell proliferation in the TNF-α treatment group was found after 3 days of short-term stimulation, but the stimulation could significantly promote the migration of BMMSCs, and the most effective migration ability was obtained with the concentration of 50 ng·ml^-1. ALP activity detection showed that the 1,10 and 50 ng·ml^-1 of TNF-α could significantly promote the migration of BMMSCs in a concentration dependent manner at 2 weeks(P<0.05), and the concentration of 50 ng·ml^-1 of TNF-α had the best effect. A short-term release system of hydrogel TNF-α was successfully constructed. The results of micro-CT and HE staining of mice in vitro showed that 50 ng·ml^-1 of TNF-α had the best effect on short-term release of bone formation, and could effectively promote new bone formation. Conclusion: Short-term stimulation of TNF-α can promote the osteogenic differentiation of BMMSCs both in vitro and in vivo.

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