生长分化因子-5对间充质干细胞向肌腱细胞分化及迁移潜能影响的相关研究

Effects of GDF-5 on Differentiation into Tendon Cells and Migration of Bone Marrow Derived Mesenchymal Stem Cells

目的:研究生长分化因子-5(GDF-5)对间充质干细胞向肌腱细胞分化及迁移潜能的影响。方法:用GDF-5对骨髓间充质干细胞(MSCs)进行14 d诱导分化,取样对细胞外基质合成及向肌腱细胞分化相关基因表达进行测定分析;通过体外三维细胞迁移模型,评价GDF-5对间充质干细胞迁移能力的作用及影响。结果:20、100、500 ng/mL GDF-5诱导4 d后的细胞数均高于0 ng/mL GDF-5诱导(P<0.05),且随着GDF-5浓度增高增殖细胞数呈现增加的趋势。100 ng/mL GDF-5诱导4、12 d的细胞数均高于0 ng/mL GDF-5诱导(P<0.05),随着培养时间的延长,差异更加明显。100 ng/mL GDF-5诱导分化骨髓干细胞12 d后,Tenascin-C与Ⅰ型胶原蛋白的基因表达量均高于0 ng/mL GDF-5诱导(P<0.05)。100 ng/mL GDF-5诱导的细胞迁移能力大于0 ng/mL GDF-5诱导(P<0.05)。结论:GDF-5可以促进间充质干细胞向肌腱细胞分化,并诱导活化间充质干细胞的迁移,该机制可能在肌腱损伤的修复重建中具有重要意义。将GDF-5与间充质干细胞进行联合治疗可为肌腱损伤的修复提供新的治疗方法。

Objective:To investigate the effects of the growth and differentiation factor-5(GDF-5)on differentiation into tendon cells and migration of bone marrow derived mesenchymal stem cells(BMSCs).Method:BMSCs were treated with GDF-5 for 14 d,and the samples were collected.The extracellular matrix synthesis and the expression of genes related to differentiation into tendon cells were analyzed.The effects of GDF-5 on the migration ability of mesenchymal stem cells were evaluated by three-dimensional cell migration model in vitro.Result:The number of cells induced by 20,100 and 500 ng/mL GDF-5 for 4 d were higher than that induced by 0 ng/mL GDF-5(P<0.05),and the number of proliferating cells increased with the increase of GDF-5 concentration.The number of cells induced by 100 ng/mL GDF-5 on day 4 and 12 were higher than that induced by 0 ng/mL GDF-5(P<0.05),and the difference was more obvious with the extension of culture time.After inducing bone marrow stem cells with 100 ng/mL GDF-5 for 12 d,the gene expression levels of Tensascin-C andⅠtype collagen were both higher than those of 0 ng/mL GDF-5 induction(P<0.05).The cell migration ability induced by 100 ng/mL GDF-5 was higher than that induced by 0 ng/mL GDF-5(P<0.05).Conclusion:GDF-5 can promote differentiation into tendon cells of mesenchymal stem cells and induce migration of activated mesenchymal stem cells,which may play an important role in the repair and reconstruction of tendon injury.Combined treatment of GDF-5 with mesenchymal stem cells may provide a new therapeutic method for tendon injury repair.