Khader Ateka, Sherman Lauren S, Rameshwar Pranela, Arinzeh Treena L
1 Department of Biomedical Engineering, New Jersey Institute of Technology , Newark, New Jersey.
2 Department of Medicine-Hematology/Oncology, Rutgers-New Jersey Medical School , Newark, New Jersey.
Stem Cells Dev. 2016 Dec 15;25(24):1909-1918. doi: 10.1089/scd.2016.0158. Epub 2016 Oct 17.
Articular cartilage has a limited ability to heal. Mesenchymal stem cells (MSCs) derived from the bone marrow have shown promise as a cell type for cartilage regeneration strategies. In this study, sodium tungstate (NaWO), which is an insulin mimetic, was evaluated for the first time as an inductive factor to enhance human MSC chondrogenesis. MSCs were seeded onto three-dimensional electrospun scaffolds in growth medium (GM), complete chondrogenic induction medium (CCM) containing insulin, and CCM without insulin. NaWO was added to the media leading to final concentrations of 0, 0.01, 0.1, and 1 mM. Chondrogenic differentiation was assessed by biochemical analyses, immunostaining, and gene expression. Cytotoxicity using human peripheral blood mononuclear cells (PBMCS) was also investigated. The chondrogenic differentiation of MSCs was enhanced in the presence of low concentrations of NaWO compared to control, without NaWO. In the induction medium containing insulin, cells in 0.01 mM NaWO produced significantly higher sulfated glycosaminoglycans, collagen type II, and chondrogenic gene expression than all other groups at day 28. Cells in 0.1 mM NaWO had significantly higher collagen II production and significantly higher sox-9 and aggrecan gene expression compared to control at day 28. Cells in GM and induction medium without insulin containing low concentrations of NaWO also expressed chondrogenic markers. NaWO did not stimulate PBMC proliferation or apoptosis. The results demonstrate that NaWO enhances chondrogenic differentiation of MSCs, does not have a toxic effect, and may be useful for MSC-based approaches for cartilage repair.
关节软骨的愈合能力有限。源自骨髓的间充质干细胞（MSCs）作为软骨再生策略的一种细胞类型已显示出前景。在本研究中，首次评估了作为胰岛素模拟物的钨酸钠（NaWO）作为增强人MSCs软骨生成的诱导因子。将MSCs接种到生长培养基（GM）、含胰岛素的完全软骨生成诱导培养基（CCM）以及不含胰岛素的CCM中的三维静电纺丝支架上。将NaWO添加到培养基中，最终浓度分别为0、0.01、0.1和1 mM。通过生化分析、免疫染色和基因表达评估软骨生成分化。还研究了使用人外周血单核细胞（PBMCS）的细胞毒性。与不含NaWO的对照相比，低浓度NaWO存在时MSCs的软骨生成分化增强。在含胰岛素的诱导培养基中，在第28天时，0.01 mM NaWO中的细胞产生的硫酸化糖胺聚糖、II型胶原蛋白和软骨生成基因表达明显高于所有其他组。在第28天时, 0.1 mM NaWO中的细胞与对照相比，II型胶原蛋白产量明显更高，sox-9和聚集蛋白聚糖基因表达也明显更高。GM和不含胰岛素且含有低浓度NaWO的诱导培养基中的细胞也表达软骨生成标记物。NaWO不刺激PBMC增殖或凋亡。结果表明，NaWO增强了MSCs的软骨生成分化，没有毒性作用，可能对基于MSCs的软骨修复方法有用。
