Sundaramurthi Dhakshinamoorthy, Krishnan Uma Maheswari, Sethuraman Swaminathan
School of Chemical & Biotechnology, Centre for Nanotechnology & Advanced Biomaterials (CeNTAB), SASTRA University, Thanjavur, 613401, India.
Ann Biomed Eng. 2014 Dec;42(12):2589-99. doi: 10.1007/s10439-014-1124-3. Epub 2014 Sep 25.
Nanomaterials with stem cells have evolved as a promising therapeutic strategy to regenerate various tissues. Tissue engineered grafts with bone marrow derived mesenchymal stem cells (BM-MSCs) can offer a cell-based therapeutic strategy for deep wounds like burns and traumatic ulcers. In this study, we have fabricated poly(3-hydroxybutyrate-co-3-hydroxyvalerate (PHBV) nanofibers through electrospinning. The adhesion, proliferation and epidermal differentiation of BM-MSCs on PHBV nanofibers were investigated. Epidermal differentiation media containing epidermal growth factor (EGF), insulin, 3,3',5-triiodo-L-thyronine (T3), Hydrocortisone and 1α, 25-dihydroxyvitamin (D3) were used to trigger differentiation of BM-MSCs on PHBV. The proliferation of BM-MSCs on PHBV was significantly higher than the tissue culture polystyrene (TCPS) control (p < 0.05). Live/dead staining of BM-MSCs on PHBV nanofibers confirmed the change in morphology of BM-MSCs from spindle to polygonal shape indicating their differentiation into keratinocytes. The expression levels of the genes keratin (early), filaggrin (intermediate) and involucrin (late) that are involved in epidermal differentiation were upregulated in a stage-specific manner. Our results demonstrate the potential of PHBV nanofibers in promoting adhesion and differentiation of mesenchymal stem cells. This novel cellular nanofiber construct can be a better alternative to the existing therapies for skin tissue engineering.
纳米材料与干细胞相结合已发展成为一种很有前景的组织再生治疗策略。含有骨髓间充质干细胞(BM-MSCs)的组织工程移植物可为烧伤和创伤性溃疡等深度伤口提供基于细胞的治疗策略。在本研究中,我们通过静电纺丝制备了聚(3-羟基丁酸酯-共-3-羟基戊酸酯)(PHBV)纳米纤维。研究了BM-MSCs在PHBV纳米纤维上的黏附、增殖及表皮分化情况。使用含有表皮生长因子(EGF)、胰岛素、3,3',5-三碘-L-甲状腺原氨酸(T3)、氢化可的松和1α,25-二羟基维生素(D3)的表皮分化培养基来诱导BM-MSCs在PHBV上分化。BM-MSCs在PHBV上的增殖明显高于组织培养聚苯乙烯(TCPS)对照组(p < 0.05)。对PHBV纳米纤维上的BM-MSCs进行活/死染色,证实BM-MSCs的形态从纺锤形变为多边形,表明它们已分化为角质形成细胞。参与表皮分化的角蛋白(早期)、丝聚蛋白(中期)和兜甲蛋白(晚期)基因的表达水平呈阶段特异性上调。我们的结果证明了PHBV纳米纤维在促进间充质干细胞黏附和分化方面的潜力。这种新型的细胞纳米纤维构建体可能是皮肤组织工程现有疗法的更好替代方案。