Department of Biology, Faculty of Science, University of Mohaghegh Ardabili, Ardabil, Iran.
Department of Engineering Sciences, Faculty of Advanced Technologies, University of Mohaghegh Ardabili, Namin, Iran.
Cell Tissue Bank. 2020 Jun;21(2):181-188. doi: 10.1007/s10561-020-09812-9. Epub 2020 Feb 3.
The hair follicle is a dynamic structure which contains different niches for stem cells, therefore; it has been considered as valuable and rich sources of stem cells, due to easy access, multipotency and non-oncogenic properties. In the present study, the differentiation capacities of hair follicle stem cells into bone cells on the natural collagen scaffolds were investigated. The stem cells were extracted from the hair follicle bulge area of male Wistar rats' whisker and cultured until 3rd passage, then osteogenic differentiations were induced by culturing the cells in the specific osteogenic medium. After 3 weeks, the differentiation parameters, including morphological changes, levels of calcification and expression of the bone specific genes were detected. The hydrogel preparation and scaffold fabrication was carried out using the extracted collagen and was studied by scanning electron microscope. Comparison of the stem cells' growth and changes on the scaffold and non-scaffold conditions showed that, in the both situation, the cells revealed differentiation signs of osteocytes, including large and cubic morphology with a star-shaped nucleus. Staining by Alizarin-red and Von-Kossa methods showed the presence of red and black calcium mass on the scaffold. Expression of the osteopontin and alkaline phosphatase genes confirmed the differentiation. Considerable porosity in the surface of the scaffold was recorded by scanning electron microscopy, which made it convenient for cells' attachment and growth. The data showed that the bulge stem cells possess significant capacity for osteoblastic differentiation and collagen scaffolds were found to be an appropriate matrix for growth and differentiation of the cell.
毛囊是一种动态结构，包含干细胞的不同龛位，因此，由于其易于获取、多能性和非致瘤性，被认为是有价值的和丰富的干细胞来源。本研究探讨了毛囊干细胞在天然胶原支架上向骨细胞分化的能力。从雄性 Wistar 大鼠胡须的毛囊隆突区提取干细胞，并培养至第 3 代，然后在特定的成骨培养基中培养诱导成骨分化。3 周后，检测分化参数，包括形态变化、钙化水平和骨特异性基因的表达。使用提取的胶原蛋白进行水凝胶制备和支架制造，并通过扫描电子显微镜进行研究。比较支架和非支架条件下干细胞的生长和变化，结果表明，在这两种情况下，细胞都表现出成骨细胞的分化特征，包括大而立方的形态和星形核。茜素红和 Von-Kossa 染色显示支架上存在红色和黑色钙团块。骨桥蛋白和碱性磷酸酶基因的表达证实了分化。扫描电子显微镜记录到支架表面有相当大的孔隙率，这使得细胞附着和生长变得更加方便。数据表明，隆突干细胞具有显著的成骨分化能力，胶原支架是细胞生长和分化的合适基质。
