Differentiation of human mesenchymal stem cells on niobium-doped fluorapatite glass-ceramics

Abstract

ObjectiveOur goal was to characterize the response of human mesenchymal stem cells (hMSCs) to a niobium-doped fluorapatite-based glass-ceramic (FAp). MethodsThe glass was prepared by twice melting at 1525°C for 3h, and cast into cylindrical ingots later sectioned into discs and heat-treated to promote crystallization of fluorapatite submicrometer crystals. Tissue culture polystyrene (TCP) was used as control. The surface of the FAp discs was either left as-heat treated, ground or etched. Initial cell attachment was assessed at 3h. Proliferation and alkaline phosphatase (ALP) expression data were collected at days 1, 4, and 8. Cell morphology was examined using SEM, at days 2 and 4. Mineralization was evaluated by Alizarin Red staining and SEM. ResultsInitial cell attachment on as heat-treated, etched, or ground surfaces was similar to that of the positive control group (p>0.05). The percentage of area covered by living cells increased significantly on as heat-treated, etched, or ground surfaces between days 1 and 8 (p<0.05). There was no significant difference among groups in cell coverage at day 8, compared to TCP control. SEM revealed well spread polygonal cells with numerous filopodia, either attached to the ceramic surface or connected to neighboring cells. ALP expression at day 8 was significantly higher in osteogenic media compared to growth media on both FAp and control. FAp discs stained positively with Alizarin Red and calcium-rich mineralized granules associated with fibrils were observed by SEM at day 35. SignificancehMSCs displayed excellent attachment, proliferation, and differentiation on niobium-doped FAp glass-ceramic.