中国组织工程研究

中国组织工程研究 ›› 2019, Vol. 23 ›› Issue (1): 35-40.doi: 10.3969/j.issn.2095-4344.1523

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

骨髓干细胞在羟基磷灰石涂层钛片上向软骨的分化

覃李玲，严 妍，余 卓，覃巧玲，黎 伟，刘 莉   

  1. 三峡大学第三临床医学院·葛洲坝集团中心医院口腔科，湖北省宜昌市 443002
  • 修回日期:2018-09-04 出版日期:2019-01-08 发布日期:2018-11-28
  • 通讯作者: 刘莉，主任医师，副教授，三峡大学第三临床医学院·葛洲坝集团中心医院口腔科，湖北省宜昌市 443002
  • 作者简介:覃李玲，女，1985年生，湖北省宜昌市人，土家族，2011年咸宁学院毕业，主治医师，主要从事口腔修复、口腔正畸研究。
  • 基金资助:

    宜昌市医疗卫生科研项目(A18-301-27)，项目负责人：刘莉；三峡大学青年科学基金项目(KJ2017A009)，项目负责人：黎伟

Chondrogenic differentiation of bone marrow stem cells on a hydroxyapatite coated titanium plate

Qin Liling, Yan Yan, Yu Zhuo, Qin Qiaoling, Li Wei, Liu Li   

  1. Department of Stomatology, Third Clinical College of China Three Gorges University, Central Hospital of China Gezhouba (Group) Corporation, Yichang 443002, Hubei Province, China
  • Revised:2018-09-04 Online:2019-01-08 Published:2018-11-28
  • Contact: Liu Li, Chief physician, Associate professor, Department of Stomatology, Third Clinical College of China Three Gorges University, Central Hospital of China Gezhouba (Group) Corporation, Yichang 443002, Hubei Province, China
  • About author:Qin Liling, Attending physician, Department of Stomatology, Third Clinical College of China Three Gorges University, Central Hospital of China Gezhouba (Group) Corporation, Yichang 443002, Hubei Province, China
  • Supported by:

    the Medical and Health Science Research Project of Yichang City, No. A18-301-27 (to LL); the Youth Science Fund Project of Three Gorges University, No. KJ2017A009 (to LW)

PDF

222

摘要：

文章快速阅读：

文题释义：
羟基磷灰石涂层：采用等离子体喷涂、电化学沉积或仿生沉积等技术将羟基磷灰石沉积在基体表面的涂层。羟基磷灰石已被大量研究证实具有促进干细胞黏附、增殖和骨诱导分化的作用，并可增加复合支架的强度及延长降解时间。
电沉积：是指金属从非化学水溶液中电化学沉积的过程，在外电场的作用下电流通过电解质溶液在电极上发生氧化还原反应。

 

摘要
背景：羟基磷灰石具有良好的生物相容性和生物活性，具有促进干细胞黏附、增殖和骨诱导分化的作用。
目的：观察羟基磷灰石涂层钛片对大鼠骨髓间充质干细胞增殖、软骨性分化的影响。
方法：采用电沉积法在钛金属基底表面沉积羟基磷灰石涂层(电沉积时间分别为30 min，1.5 h)，用场发射扫描电子显微镜观察涂层形态，X射线衍射分析其化学组成。取4只3周龄SD雄性大鼠(购于湖北省实验动物研究中心)双侧股骨的骨髓，采用全骨髓贴壁法培养骨髓间充质干细胞，将第3代细胞接种于有涂层钛片及没有涂层纯钛片上，并进行4周的成软骨诱导分化。通过CCK-8试剂盒分析羟基磷灰石涂层对大鼠骨髓间充质干细胞增殖的影响，二甲基亚甲基蓝比色法检测各组细胞糖胺多糖含量，RT-PCR法检测各组细胞软骨相关基因的表达，评价各组骨髓间充质干细胞的成软骨能力。
结果与结论：①羟基磷灰石涂层均匀分布在钛基底上，大小结构较一致，X射线衍射图谱与羟基磷灰石标准峰基本吻合；②羟基磷灰石涂层促进骨髓间充质干细胞增殖，羟基磷灰石涂层亦表现了比空白钛片组具有更好的生物活性；③羟基磷灰石涂层组较空白钛片组产生了更多的糖胺多糖，差异有显著性意义(P < 0.01)；④羟基磷灰石涂层组软骨相关基因(Sox9、多聚蛋白聚糖、Ⅱ型胶原、Ⅹ型胶原)的相对表达量显著高于空白钛片组(P < 0.05)，电沉积时间为1.5 h的羟基磷灰石涂层组的Ⅱ型胶原表达量明显高于电沉积时间30 min羟基磷灰石涂层组和空白钛片组(P < 0.01)；⑤结果表明，应用电化学沉积技术可以在钛片上制备均匀的羟基磷灰石涂层，且该涂层可以促进大鼠骨髓间充质干细胞增殖，具有良好的生物相容性，有利于骨髓间充质干细胞向软骨方向分化。

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程
ORCID: 0000-0001-5188-1838(覃李玲)

关键词: 钛片, 羟基磷灰石, 骨髓间充质干细胞, 糖胺多糖, Ⅱ型胶原, 软骨分化, 电化学沉积

Abstract:

BACKGROUND: Hydroxyapatite has good biocompatibility and bioactivity and can promote adhesion, proliferation and chondrogenic differentiation of stem cells.
OBJECTIVE: To investigate the effect of hydroxyapatite coated titanium plate on chondrogenic differentiation of bone marrow mesenchymal stem cells.
METHODS: Hydroxyapatite coating was fabricated on the titanium surface by electrochemical deposition technique (deposition time: 3 minutes and 1.5 hours) and characterized by field emission scanning electron microscopy for coating morphology and by X-ray diffraction for the chemical composition of the coating. Bone marrow samples from four Sprague-Dawley rats of 3 weeks old (purchased from Hubei Provincial Laboratory Animal Center, Wuhan, China) were taken to culture bone marrow mesenchymal stem cells using whole bone marrow adherent method. Passage 3 cells were inoculated onto the titanium plate with or without coating followed by 4 weeks chondrogenic induction. For assessing the ability of chondrogenesis, cell counting kit-8 assay was applied to test the cell proliferation; dimethylmethylene blue colorimetric assay was used to detect the content of glycosaminoglycans; and RT-PCR was used to test the expression of chondrocyte specific genes.
RESULTS AND CONCLUSION: (1) The hydroxyapatite coating was evenly distributed on the titanium plate and its structure was uniform. The X-ray diffraction peaks of hydroxyapatite powder generally matched the characteristic X-ray diffraction pattern of hydroxyapatite. (2) The hydroxyapatite-coated titanium plate improved the proliferation of bone marrow mesenchymal stem cells and showed better bioactivity compared to the non-coated titanium plate. (3) The glycosaminoglycan content was significantly higher in the hydroxyapatite coating group than the non-coated group (P < 0.01). (4) The RT-PCR results showed higher expression levels of chondrocyte specific genes (SOX9, type II collagen, type X collagen, aggrecan) in the hydroxyapatite coating group than the non-coated group (P < 0.05). The expression of type II collagen was significantly higher in the 1.5-hour group than the 0.5-hour group and non-coated group (P < 0.05). To conclude, the hydroxyapatite coating could evenly distribute on the titanium plate via electrophoretic deposition. Hydroxyapatite coated titanium surface has good biocompatibility and can promote proliferation and chondrogenic differentiation of rat bone marrow mesenchymal stem cells in vitro. 

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Bone Marrow, Mesenchymal Stem Cells, Titanium, Hydroxyapatites, Electrochemical Techniques, Tissue Engineering

中图分类号: