The synergistic effects of IL-6/IL-17A promote osteogenic differentiation by improving OPG/RANKL ratio and adhesion of MC3T3-E1 cells on hydroxyapatite.

OBJECTIVE: In this study, we evaluated the potential role of IL-6 and/or IL-17A in regulating the OPG/RANKL (osteoprotegerin/receptor activator of nuclear factor kappa b ligand) system of murine osteoblast cell line (MC3T3-E1) cultured on hydroxyapatite (HA). METHODS: MC3T3-E1 cells were seeded on HA and treated with recombinant IL-6 or rIL-17A or combination of the two cytokines. Cell proliferation and differentiation activity were measured by MTS and alkaline phosphatase assays respectively. Observation of cell adhesion and proliferation was examined by scanning electron microscopy. Gene and protein expressions were performed on RANKL and OPG using qPCR, Western blot and ELISA. RESULTS: We demonstrated that treatment with recombinant IL-17A (rIL-17A) and the combination rIL-6/rIL-17A promoted better adhesion and higher proliferation of cells on HA. Cells treated with rIL-17A and the combination cytokines showed a significant increase in differentiation activity on day 7, 10 and 14 as indicated by ALP activity (p < 0.001). Gene and protein expressions showed significant up-regulation of OPG and ALP (p < 0.001) and down-regulation of RANKL (p < 0.001) expression by all the treated groups. Interestingly, the combination of the two cytokines resulted in a significant increase of OPG/RANKL ratio (p < 0.001). CONCLUSION: These findings indicated that treatment with the combination of the two cytokines (IL-6/IL-17A) has synergistic effects to promote osteoblastic differentiation but suppress osteoclastogenesis by altering the OPG/RANKL ratio.

Gypsum-based biomaterials: Evaluation of physical and mechanical properties, cellular effects and its potential as a pulp liner.

This in vitro study aimed to evaluate setting time and compressive strength of gypsum-based chitosan biomaterials and its effect on proliferation of stem cells from human exfoliated deciduous teeth (SHED) and alkaline phosphatase (ALP) activity. Pure-GYP was mixed with water (2.5 g: 1.9 mL); Gyp-CHT was prepared with gypsum, chitosan, and water (2.5 g: 0.285 g: 1.9 mL). Cell viability and ALP activity were assessed at different periods. Data were analyzed using SPSS (p<0.05). The setting times were 2.7 min and 2.8 min for pure-GYP and Gyp-CHT, respectively. Significantly higher compressive strength was observed with Gyp-CHT. SHED treatments with both materials were not cytotoxic. ALP was consistently higher in the treated groups compared with the control. Cellular attachments were evident with SEM. Excellent cellular viability with pure-GYP and Gyp-CHT, as well as increased ALP activities, suggested the possibility of tertiary dentin formation. Further studies are necessary to evaluate the biomaterials for its pulp protective potentialities.