天津医药

天津医药

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羟基磷灰石诱导脂肪间充质干细胞成骨分化的实验研究

程胜承，刘义，景亚青，鞠明艳，李光△   

  1. 天津医科大学遗传学系（邮编300070）
  • 出版日期:2018-07-15 发布日期:2018-07-15
  • 作者简介:程胜承（1990），男，硕士在读，主要从事肺部细菌菌群的鉴定、成骨不全患者的诊断及干细胞治疗研究

Experimental study on osteogenic differentiation of adipose-derived mesenchymal stem cells induced by hydroxyapatite

CHENG Sheng-cheng, LIU Yi, Jing Ya-qing, JU Ming-yan, LI Guang△   

  1. Department of Genetics, Tianjin Medical University, Tianjin 300070, China △Corresponding Author E-mail: lig@tmu.edu.cn
  • Published:2018-07-15 Online:2018-07-15

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摘要： 目的 探究羟基磷灰石（HA）对脂肪来源的间充质干细胞（ADSCs）向成骨分化的影响。方法 分离、纯化 并鉴定C57BL/6小鼠的ADSCs，将HA与ADSCs共培养，CCK-8法检测不同浓度（0、5、10、20、50、100、500 mg/L）的HA 对ADSCs增殖的影响；碱性磷酸酶（ALP）染色检测不同浓度HA对ADSCs成骨分化的影响；实时荧光定量逆转录-聚 合酶链反应（qRT-PCR）检测ADSCs成骨相关基因骨钙素（BGLAP）、碱性磷酸酶（ALP）、Ⅰ型胶原（COL1A1）、骨桥蛋 白（OPN）、Runt相关转录因子2（Runx2）的mRNA表达情况。结果 低浓度HA（≤20 mg/L）对ADSCs增殖的影响较 小，随着HA浓度增加，细胞的增殖活性下降。HA与ADSCs共培养可显著增加其ALP活性，并促进成骨相关基因的 表达（P＜0.01），且HA为20 mg/L时诱导效果较好。结论 HA具有诱导ADSCs向成骨细胞分化的能力，为两者混合 制作成新的骨支架修复材料提供了理论基础。

关键词: 羟基磷灰石类, 间质干细胞, 碱性磷酸酶, 逆转录聚合酶链反应, 脂肪间充质干细胞, 成骨分化

Abstract: Objective To investigate the effect of hydroxyapatite (HA) on the osteogenic differentiation potency of adipose-derived mesenchymal stem cells (ADSCs). Methods ADSCs were isolated and purified from C57BL/6 mice. Toxic effects of hydroxyapatite on ADSCs were detected by cell proliferation assay. Alkaline phosphatase (ALP) assay was used to detect the effects of different concentrations of HA on osteogenic differentiation of ADSCs. Relative mRNA expression levels of osteogenic genes (BGLAP, ALP, COL1A1, OPN and Runx2) were measured by RT-PCR. Results The low concentration of HA (≤20 mg/L) showed less effect on proliferation of ADSCs. With the increase of HA concentration, the cell proliferation decreased. The co-culture of 20 mg/L HA with ADSCs significantly increased the ALP activity, and promoted the expression of osteoblast-related genes (P＜0.01). Conclusion Hydroxyapatite have the ability to induce the osteogenic differentiation of ADSCs, which provide a theoretical basis for combinating HA and ADSCs into a new bone engineering scaffold.

Key words: hydroxyapatites, mesenchymal stem cells, alkaline phosphatase, reverse transcriptase polymerase chain reaction, adipose-derived mesenchymal stem cells, osteogenic differentiation