Odontogenic responses of human dental pulp cells to collagenanobioactive glass nanocomposites

摘要

Objectives. Collagen-based nanocomposite incorporating nanobioactive glass (ColBG) was developed as a scaffolding matrix for dentin-pulp regeneration. The effects of the novel matrix on the proliferation of human dental pulp cells (hDPCs) and their differentiation into odontoblastic lineage were investigated. Methods. Nanocomposite scaffold was prepared by incorporating nBG within the Col solution and then reconstituting them into a membrane form. Cell growth by MTS assay, adhesion by scanning electron microscopy (SEM), and odontoblastic differentiation by alkaline phos-phatase (ALP) activity, mineralization, and the mRNA expression of differentiation-related genes of DPCs on each scaffold were evaluated. Results. The introduction of nBG significantly improved the bone mineral-like apatite formation in the simulated body fluid, suggesting excellent acellular bone-bioactivity. The hDPCs cultured on the ColBG nanocomposite have shown active growth behavior during culture for 14 days. The mRNA levels of major organic extracellular matrix of dentin, collagen type I and III were highly expressed in the ColBG matrix. Moreover, the alkaline phosphatase (ALP) activity and the mineralized nodule formation were increased in the ColBG nanocomposite compared to those in Col. Odontoblatic differentiation genes, including dentin sialophosphoprotein, dentin matrix protein I, ALP, osteopontin and osteo-calcin were significantly stimulated in the Col containing nBG. Moreover, the key adhesion receptor integrin components α2 and βi, specifically binding to collagen molecule sequence, were upregulated in ColBG compared to Col, suggesting that odontogenic stimulation was closely related to the integrin-mediated process. Significance. In our study, the nanocomposite ColBG matrix induced the growth and odontogenic differentiation more effectively than Col alone, providing a promising scaffold condition for regeneration of dentin-pulp complex tissue.