Evaluation of in vitro Efficacy of Vitamin D3 on the Osteogenic Differentiation and Mineralization Capabilities of Fetal and Adult Osteoblasts of Rabbit Reflects Therapeutic Potential
The ability of the osteoblasts to lay the extracellular matrix and mineralize determines the efficiency of bone formation. Vitamin D injections are used to enhance bone mineralization in clinical cases. The present study was designed to evaluate the in vitro efficacy of Vitamin D on the osteoblastic activity in both fetal and adult osteoblasts. The long bone was collected from fetal and adult rabbits and cultured in Dulbeccos Modified Eagles Medium (DMEM) containing 10% fetal bovine serum and antibiotics. The passaged cells, after attaining more than 80% confluence were induced for mineralization through osteogenic media with or without 1, 25-dihydroxyvitamin D3 (Vit D3). Fetal and adult cells were harvested after 7, 14, 21, 28 days for observation of osteoblastic differentiation and mineralization. Relative transcripts abundance of mRNA of alkaline phosphatase (ALP) and osteocalcin (OCN) were measured to assess the osteoblastic activity using quantitative real-time polymerase chain reaction (RT-qPCR) at different time intervals. The mineral accumulation was assessed by Von-Kossa staining. Fetal osteoblasts compared to adult osteoblasts showed a marked significant increase in mineralization on addition of Vit D. However, the adult osteoblasts reflected transcript abundance of ALP only after 14th day and OCN genes after 21 days post induction whereas fetal osteoblasts showed the same by 7th day post induction. This reflects a great therapeutic potential of fetal osteoblasts along with Vit D3 in bone regeneration. Further studies are required to correlate the influence of Vit D3 on ALP/OCN expression and also whether there is a comparative increase in the receptors of Vit D3 and osteocalcin protein in fetal cells as compared to adult osteoblasts.
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