Osteogenic stimulation of human dental pulp stem cells with a novel gelatin-hydroxyapatite-tricalcium phosphate scaffold

Abstract

The aim of the present study was to construct and compare gelatin-HA-TCP scaffolds with a gelatin-only scaffold and to investigate the effect of the scaffold on osteogenic differentiation of human dental pulp stem cells. We developed a novel scaffold for bone tissue engineering via a solution casting/particle washing method, and the physical and mechanical properties of the scaffolds were examined using scanning electron microscopy and a universal testing machine, respectively. Scaffold cytotoxicity toward human dental pulp stem cells (hDPSCs) was evaluated with the CCK8 method, and hDPSC differentiation was evaluated with an alkaline phosphatase activity assay, alizarin red S staining, and reverse transcription-polymerase chain reaction (RT-PCR). Our results indicate that the gelatin-HA-TCP scaffolds exhibited good homogeneity, interconnected pores, and relatively high mechanical strength and water absorption rates. A significant increase in hDPSC proliferation and ALP activity that stimulated mineralization of the hDPSC-generated matrix was also seen on gelatin-HA-TCP scaffolds compared with the gelatin-only scaffolds. In addition, RT-PCR revealed that the gelatin-HA-TCP scaffold upregulated gene expression of the osteogenic markers Runx2, bone sialoprotein, and OSX. In conclusion, gelatin-HA-TCP scaffolds presented better mechanical properties, cytocompatibility and differentiation-inducing characteristics than gelatin scaffolds. These results indicate that the novel hydrogel gelatin-HA-TCP scaffolds may be a promising biomaterial for bone tissue engineering. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 1851-1861, 2018.

Keywords: dental pulp stem cell; gelatin; hydroxyapatite; scaffold; tissue engineering.