ABSTRACT
BACKGROUND:Synovial fluid mesenchymal stem cells can be amplified rapidly in vitro and collected by a minimally invasive method. Recent studies have suggested that synovial fluid mesenchymal stem cells have become an important kind of seed cells for bone tissue engineering. Osteogenic differentiation is required to be optimized prior to the application of synovial fluid mesenchymal stem cells in the bone regeneration. OBJECTIVE: To describe a protocol to generate osteoblast-lineage cells from human synovial fluid mesenchymal stem cells of the temporomandibular joint using a cocktail that includes glutamax, dexamethasone, β-glycerophosphate, vitamin C, 1,25-dihydroxyvitamin D3, and to investigate the effect of 1,25-dihydroxyvitamin D3 and dexamethasone on the osteogenic capacity of synovial fluid mesenchymal stem cells. METHODS:Synovial fluid mesenchymal stem cells from the human temporomandibular joint were expanded in vitro and cultured in different osteogenic induction media. The mineralization capacity of osteogenic differentiation was evaluated by alizarin red and Von kossa staining. And the osteogenic markers, including ALP, RUNX2 and OCN, were assessed by reverse transcription-PCR. RESULTS AND CONCLUSION: The mineralization formation increased greatly in the medium with 100 nmol/L dexamethasone and 10 nmol/L 1,25-dihydroxyvitamin D3. The expression of ALP, RUNX2 and OCN was enhanced distinctly in the 1,25-dihydroxyvitamin D3-induced differentiated cells. These findings reveal that appropriate concentration of 1,25-dihydroxyvitamin D3 and dexamethasone can be ideal ingredients to induce the osteogenic differentiation of human synovial fluid mesenchymal stem cells of the temporomandibular joint. Thus, this effective condition can be used to induce the osteogenic differentiation of synovial fluid mesenchymal stem cells for the bone regeneration in the temporomandibular joint.