Stromal Cell-Derived Factor-1β Potentiates Bone Morphogenetic Protein-2-Stimulated Osteoinduction of Genetically Engineered Bone Marrow-Derived Mesenchymal Stem Cells In Vitro
Stromal Cell-Derived Factor-1β Potentiates Bone Morphogenetic Protein-2-Stimulated Osteoinduction of Genetically Engineered Bone Marrow-Derived Mesenchymal Stem Cells In Vitro
复制标题
DOI：
10.1089/ten.tea.2012.0085
复制
发表时间：
2013-01-01
影响因子：
4.1
通讯作者：
Hill, William D.
中科院分区：
文献类型：
--
作者：
Herberg, Samuel;Fulzele, Sadanand;Hill, William D.
骨骼损伤是最普遍的临床问题之一，骨髓来源的间充质干细胞/基质细胞（BMSC）已成功用于治疗骨骼损伤。基质细胞衍生因子 1 (SDF-1；CXCL12) 是 CXC 趋化因子家族的成员，具有多个剪接变体。两种最丰富的变体 SDF-1 α 和 SDF-1 β 具有相同的氨基酸序列，但 SDF-1 β C 末端有四个额外的氨基酸，这可能通过糖胺聚糖介导表面稳定并保护 SDF-1 β 免受蛋白水解裂解，使其效力是 SDF-1 α 的两倍。越来越多的证据表明，SDF-1 通过调节干/祖细胞的招募、植入、增殖和分化参与骨形成。然而，潜在的分子机制尚未完全阐明。在本研究中，我们测试了以下假设：SDF-1 beta 可以在体外增强骨形态发生蛋白 2 (BMP-2) 刺激的 BMSC 成骨分化和趋化性。利用逆转录病毒介导的基因转移生成新型Tet-Off-SDF-1βBMSC，我们发现条件性SDF-1β表达受到多西环素以剂量依赖性和时间性方式的严格调节，导致SDF-1βmRNA和蛋白质水平显着增加。此外，在体外基因工程 BMSC 中，发现 SDF-1 beta 可增强 BMP-2 刺激的矿化、关键成骨标志物的 mRNA 和蛋白表达，并通过细胞外信号调节激酶 1/2 (Erk1/2) 磷酸化调节 BMP-2 信号转导。我们还表明，SDF-1 beta 可在体外促进表达 CXC 趋化因子受体 4 (CXCR4) 的 BMSC 的迁移反应。总之，这些数据支持SDF-1β可以在BMP-2刺激的BMSC成骨分化中发挥重要作用，并且可能以自分泌和旁分泌的方式发挥其生物活性。
Skeletal injuries are among the most prevalent clinical problems and bone marrow-derived mesenchymal stem/stromal cells (BMSCs) have successfully been used for the treatment thereof. Stromal cell-derived factor-1 (SDF-1; CXCL12) is a member of the CXC chemokine family with multiple splice variants. The two most abundant variants, SDF-1 alpha and SDF-1 beta, share identical amino acid sequences, except for four additional amino acids at the C-terminus of SDF-1 beta, which may mediate surface stabilization via glycosaminoglycans and protect SDF-1 beta from proteolytic cleavage, rendering it twice as potent as SDF-1 alpha. Increasing evidence suggests that SDF- 1 is involved in bone formation through regulation of recruitment, engraftment, proliferation, and differentiation of stem/progenitor cells. The underlying molecular mechanisms, however, have not yet been fully elucidated. In this study, we tested the hypothesis that SDF-1 beta can potentiate bone morphogenetic protein-2 (BMP-2)-stimulated osteogenic differentiation and chemotaxis of BMSCs in vitro. Utilizing retrovirus-mediated gene transfer to generate novel Tet-Off- SDF-1 beta BMSCs, we found that conditional SDF-1 beta expression is tightly regulated by doxycycline in a dose-dependent and temporal fashion, leading to significantly increased SDF-1 beta mRNA and protein levels. In addition, SDF-1 beta was found to enhance BMP-2-stimulated mineralization, mRNA and protein expression of key osteogenic markers, and regulate BMP-2 signal transduction via extracellular signal-regulated kinases 1/2 (Erk1/2) phosphorylation in genetically engineered BMSCs in vitro. We also showed that SDF-1 beta promotes the migratory response of CXC chemokine receptor 4 (CXCR4)-expressing BMSCs in vitro. Taken together, these data support that SDF-1 beta can play an important role in BMP-2-stimulated osteogenic differentiation of BMSCs and may exert its biological activity in both an autocrine and paracrine fashion.