Regeneration of dental–pulp complex-like tissue using phytic acid derived bioactive glasses

Abstract

Phytic acid derived bioactive calcium phosphosilicate (PSC) glasses with a high phosphate content were synthesised by using non-toxic phytic acid as a phosphorus precursor. This study aimed to verify the effects of PSC on the odontogenic differentiation and dentin–pulp complex-like tissue regeneration of dental pulp cells (DPCs). Nitrogen adsorption, field-emission scanning electron microscopy, Fourier transform infrared spectroscopy, pH measurement, and inductively coupled plasma optical emission spectroscopy analyses were performed to characterise PSC. Classical 45S5 bioactive glasses (45S5) were used as positive control. Cell proliferation (1, 3, 5, 7 and 9 days), odontogenic-related gene expression levels (3 and 7 days) and mineralisation ability (21 days) of human DPCs (hDPCs) were evaluated with methylthiazol tetrazolium assay, real-time polymerase chain reaction and alizarin red staining after hDPCs from third molars were treated with PSC or 45S5 extractions. Rat molar crowns with pulp tissues covered by PSC or 45S5 were transplanted subcutaneously into nude mice for 2 and 6 weeks to demonstrate their biological effects in vivo. Results revealed that the specific surface area of PSC was larger than that of 45S5. The PSC also induced hydroxycarbonate apatite precipitation earlier than 45S5. pH was slightly increased when the amount and dissolution time of PSCs were increased. By comparison, pH was remarkably increased by 45S5. The amounts of Si and P ions released by PSC (0.1 mg mL⁻¹) were larger than those released by 45S5. Cell proliferation, mRNA expression levels of dentin sialophosphoprotein, dentin matrix protein 1 and osteocalcin and mineralisation of hDPCs were also more strongly promoted by PSC than by 45S5. In vivo, the amount of induced typical dentin-like tissues with odontoblast-like cells generated on the interface between materials and pulp tissues was higher in PSC than in 45S5. Only collagen-like tissues were observed in groups without bioactive glasses. These findings suggested that PSC enhanced the odontogenic differentiation of DPCs and dentin–pulp complex-like tissue regeneration. The PSC might be a potential candidate for vital pulp preservation and regeneration of the dentin–pulp complex.