RESUMEN
OBJECTIVE: To investigate the in vitro effects of CCN2 on odontoblast-like cells proliferation and differentiation. DESIGN: MDPC-23 cells were cultured in DMEM supplemented with 5% FBS. CCN2 was either added to culture media or coated onto culture polystyrene, addition or coating of dH2O was served as control. In the addition group, CCN2 (100â¯ng/mL) was added into culture media. In the coating group, CCN2 at the concentration of 1000â¯ng/mL was employed. Cell proliferation was performed using CCK-8 assay. Cell differentiation and mineralization were analyzed by ALPase activity assay, real time RT-PCR and alizarin red staining. One-way ANOVA with post-hoc tukey HSD test was used for statistical analysis. RESULTS: MDPC-23 cells exhibited robust proliferative activity upon exposure to either soluble or immobilized CCN2. ALP activity of cells cultured on CCN2-modified surface was continuously strengthened from day six (0.831⯱â¯0.024 units/µg protein versus 0.563⯱â¯0.006 units/µg protein of control) till day eight (1.035⯱â¯0.139 units/µg protein versus 0.704⯱â¯0.061 units/µg protein of control). Gene expression of BSP, OCN and OPN were promoted by soluble CCN2 after 48â¯h exposure. Moreover, gene expression of BSP, OCN, OPN, ALP, COL1â¯A1, Runx-2, DSPP and DMP-1 was significantly enhanced by immobilized CCN2. Finally, mineralization of MDPC-23 cells was accelerated by both soluble and immobilized CCN2 to different extent. CONCLUSIONS: The findings indicate that CCN2 promoted proliferation, odontogenic gene expression and mineralization of MDPC-23 cells. It is proposed that CCN2 may be a promising adjunctive formula for dentin regeneration.