Centro/Departamento de Física, Universidade do Minho, 4710-057 Braga, Portugal.
Centre for Biomaterials and Tissue Engineering, CBIT, Universitat Politècnica de València, 46022 Valencia, Spain.
Int J Mol Sci. 2017 Nov 11;18(11):2391. doi: 10.3390/ijms18112391.
The aim of this work was to determine the influence of the biomaterial environment on human mesenchymal stem cell (hMSC) fate when cultured in supports with varying topography. Poly(vinylidene fluoride) (PVDF) culture supports were prepared with structures ranging between 2D and 3D, based on PVDF films on which PVDF microspheres were deposited with varying surface density. Maintenance of multipotentiality when cultured in expansion medium was studied by flow cytometry monitoring the expression of characteristic hMSCs markers, and revealed that cells were losing their characteristic surface markers on these supports. Cell morphology was assessed by scanning electron microscopy (SEM). Alkaline phosphatase activity was also assessed after seven days of culture on expansion medium. On the other hand, osteoblastic differentiation was monitored while culturing in osteogenic medium after cells reached confluence. Osteocalcin immunocytochemistry and alizarin red assays were performed. We show that flow cytometry is a suitable technique for the study of the differentiation of hMSC seeded onto biomaterials, giving a quantitative reliable analysis of hMSC-associated markers. We also show that electrosprayed piezoelectric poly(vinylidene fluoride) is a suitable support for tissue engineering purposes, as hMSCs can proliferate, be viable and undergo osteogenic differentiation when chemically stimulated.
这项工作的目的是确定在具有不同形貌的支撑物中培养时，生物材料环境对人骨髓间充质干细胞（hMSC）命运的影响。聚偏二氟乙烯（PVDF）培养支架的结构范围在 2D 和 3D 之间，是在沉积有不同表面密度的 PVDF 微球的 PVDF 膜上制备的。通过流式细胞术监测特征 hMSC 标志物的表达来研究在扩增培养基中培养时多能性的维持情况，结果表明细胞在这些支撑物上失去了其特征表面标志物。通过扫描电子显微镜（SEM）评估细胞形态。还在扩增培养基中培养七天后评估碱性磷酸酶活性。另一方面，当细胞达到汇合时，在成骨培养基中培养以监测成骨分化。进行骨钙素免疫细胞化学和茜素红测定。我们表明，流式细胞术是研究接种在生物材料上的 hMSC 分化的合适技术，可以对 hMSC 相关标志物进行定量可靠的分析。我们还表明，电喷压电聚偏二氟乙烯是组织工程的合适支撑物，因为 hMSCs 在化学刺激下可以增殖、存活并进行成骨分化。
