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摘要
Background aims. Suspension mononuclear cells (MNCs) can be differentiated into osteoblasts with the induction of ascorbic acid and beta-glycerophosphate. The aim of this study was to determine the ability of suspension MNCs to differentiate into osteoblasts using ascorbic acid only. Methods. Suspension MNCs were obtained by a combination of gradient centrifugation and culture selection. Suspension MNCs were subjected to differentiation assay by culturing them inside proliferation medium supplemented with 10 mu g/mL, 30 mu g/mL, 50 mu g/mL, 60 mu g/mL, 90 mu g/mL and 500 mu g/mL of ascorbic acid. Proliferation medium supplemented with 50 mu g/mL ascorbic acid and 10 mmol/L beta-glycerophosphate was used as a positive control for osteoblast induction, and proliferation medium without ascorbic acid was used as a negative control. Differentiation analysis was performed using alkaline phosphatase (ALP) assay, von Kossa staining and expression of osteoblastrelated genes. Results. With all concentrations of ascorbic acid used, there was a significant increase (P < 0.05) in ALP-specific activity and mineralized nodule formation throughout the differentiation course compared with negative control. Ascorbic acid was also able to activate the expression of osteopontin (SPP1), osteonectin (SPARC) and runt-related transcription factor 2 (RUNX2) messenger RNA in positive control and ascorbic acid-induced MNCs (30 mu g/mL and 90 mu g/mL) but not in negative control. Conclusions. Ascorbic acid alone was sufficient to induce osteoblast differentiation from suspension MNCs; 30-90 mu g/mL of ascorbic acid was found to be the optimal concentration. Ascorbic acid can be used as a nutritional supplement for cellular therapy of bone-related disease.
- 出版日期2014-5
