Osteoblastic Differentiation of Stem Cells from Human Exfoliated Deciduous Teeth by Probiotic Hydroxyapatite

Abstract

Objective: Multipotent cells derived from human exfoliated deciduous teeth (SHED) possess the ability to differentiateinto various cell types, including osteoblasts. This study aims to simulate the growth induction and osteogenicdifferentiation of SHED cells using probiotics and their resultant biomaterials.Materials and Methods: This experimental study proceeded in two stages. Initially, we evaluated the effect ofautoclaved nutrient agar (NA) grown probiotic Bacillus coagulans (B. coagulans) on the SHED and MG-63 cell lines.Subsequently, probiotics grown on the Pikovskaya plus urea (PVKU) medium and their synthesised hydroxyapatite (HA)were identified using X-ray diffraction (XRD), scanning electron microscopy (SEM), energy-dispersive X-ray (EDX), andFourier transform infrared spectroscopy (FTIR), and then used to stimulate growth and osteogenic differentiation of theSHED cell line. Osteoblast cell differentiation was assessed by morphological changes, the alkaline phosphatase (ALP)assay, and alizarin red staining.Results: There was a substantial increase in SHED cell growth of about 14 and 33% due to probiotics grown on NAand PVKU medium, respectively. The PVKU grown probiotics enhanced growth and induced stem cell differentiationdue to HA content. Evidence of this differentiation was seen in the morphological shift from spindle to osteocyte-shapedcells after five days of incubation, an increase in ALP level over 21 days, and detection of intracellular calcium depositsthrough alizarin red staining-all indicative of osteoblast cell development.Conclusion: The osteogenic differentiation process in stem cells, improved by the nano-HA-containing byproducts ofprobiotic bacteria in the PVKU medium, represents a promising pathway for leveraging beneficial bacteria and theirsynthesised biomaterials in tissue engineering.

Keywords

• differentiation
• hydroxyapatite
• osteoblast
• probioticn