Influence Of Bone Marrow Mesenchymal Stem Cells On Proliferation And Osteogenic Differentiation Of Dental Follicle Cells

ObjectiveThis study was undertaken to investigate the effect of bone marrowmesenchymal stem cells on proliferation and osteogenic differentiation ofdental follicle cells under co-culture in vitro.Methods1. Isolation, culture and identification of dental follicle cells and bonemarrow mesenchymal stem cells of SD rats.2. A co-culture system of rat dental follicle cells and bone marrowmesenchymal stem cells in vitro was established, and including thefollowing controls: a co-culture of dental follicle cells in the lower chamberand bone marrow mesenchymal stem cells in the upper chamber(a pore sizeof0.4μm in diameter) using a Transwell system, dental follicle cells withno cells in the upper chamber.3. Dental follicle cells proliferation assay with Cell Counting Kit-8.Changes in dental follicle cells growth viability were analysed using theCell Counting Kit-8on days1,3,5and7after co-culturing dental follicle cells with bone marrow mesenchymal stem cells.4. Measurement of ALP activity. After7and14days of co-culture, ALPactivities in the experimental and control dental follicle cells wereanalyzed.5. Real-time fluorescent quantitative PCR.10days after co-culturing dentalfollicle cells with bone marrow mesenchymal stem cells, mRNA levels ofRunx2, COL-I, OCN, BSP in dental follicle cells were assessed.Results1. Rat dental follicle cells and bone marrow mesenchymal stem cellswere obtained and identified.2. The dental follicle cells showed a higher level of proliferation in theexperimental group after co-culturing dental follicle cells with bonemarrow mesenchymal stem cells. After5and7days of co-culture, the cellproliferation level in dental follicle cells was significantly (P <0.05) higherthan that in the single culture cells.3. After7and14days of co-culture with bone marrow mesenchymalstem cells, ALP activity in dental follicle cells was significantly (P <0.05)higher than that in the single culture cells.4. After dental follicle cells were co-cultured for10days, Runx2, COL-I,OCN, BSP mRNA expression levels in the experimental group wereincreased relative to levels in the control groups(P <0.05). ConclusionAfter dental follicle cells were co-cultured with bone marrowmesenchymal stem cells, the cell proliferation level, ALP activity and Runx2, COL-I, OCN, BSP mRNA expression levels were promoted. Aco-culture of dental follicle cells and bone marrow mesenchymal stem cellsis potentially used for periodontal tissue engineering.