Influence of calcium and phosphorus release from bioactive glasses on viability and differentiation of dental pulp stem cells

Abstract

The release of ions that can significantly contribute toward cellular response is an important characteristic of bioactive glasses (BG). Here, ionic extracts of three different compositions of BG powders in 60 mol% SiO2, x mol% CaO (x = 28, 32 and 36), x mol% P2O5 (x = 12, 8 and 4) compositional system were utilized to study their effect on the viability, differentiation and mineralization of dental pulp stem cells (DPSCs) in vitro. ICP was applied to detect the exact ionic concentrations released from different composition of BG. DPSCs treated with conditioned media from the glass with 4 mol% P2O5 (BGCM1, media containing 44.01 ± 0.6 mg/L Si, 61.72 ± 0.1 mg/L Ca and 7.57 ± 0.01 mg/L P) were more metabolically active compared to conditioned media from the glass with 8 mol% P2O5 (BGCM2, media with 47.36 ± 0.7 mg/L Si, 57.4 ± 0.1 mg/L Ca and 14.54 ± 0.2 mg/L P), at all times tested, but in all cases the process was slower than the control. Cells exposed to media conditioned by the glass with 12 mol% P2O5 (BGCM3, 40.46 ± 0.5 mg/L Si, 61. 85 ± 0.3 mg/L Ca and 28.43 ± 0.3 mg/L P) responded differently, such that cells showed to be more metabolically active than control at day 3, but then similar to or lower than control at higher time points. Differentiation of DPSCs toward osteogenic lineage in the presence of BGCM was assessed by Alizarin red staining. Cells treated with high phosphate BGCM3 displayed a higher density of red mineralized nodules than cells treated with BGCM1 and BGCM2 after 21 days of culture in non-osteogenic medium. BGCM3 was therefore chosen for gene expression studies. Osteogenic differentiation of DPSCs in the presence and/or absence of BGCM3 or osteogenic supplements were studied by RT-PCR. Overall, the results demonstrated that, in the absence of osteogenic supplements, BGCM3 group showed a significantly higher mRNA expression levels for alkaline phosphatase at day 7, osteopontin and osteonectin at days 7 and 14, and a high level of collagen I at day 14, compared to negative control group (BM−). Overall, the results obtained from BGCM3 group are beneficial for the design and manufacture of scaffolds or particulates with tailored ion release for a range of bone repair applications.