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

Document Type : Original Article

Authors

1 Department of Cell, Molecular Biology and Microbiology, Faculty of Biological Science and Technology, University of Isfahan, Isfahan, Iran

2 Center for Nanotechnology and Biomaterials Application and Research at Marmara University, Goztepe Campus, Istanbul, Turkey

Abstract

Objective: Multipotent cells derived from human exfoliated deciduous teeth (SHED) possess the ability to differentiate
into various cell types, including osteoblasts. This study aims to simulate the growth induction and osteogenic
differentiation of SHED cells using probiotics and their resultant biomaterials.
Materials and Methods: This experimental study proceeded in two stages. Initially, we evaluated the effect of
autoclaved 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), and
Fourier transform infrared spectroscopy (FTIR), and then used to stimulate growth and osteogenic differentiation of the
SHED 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 NA
and PVKU medium, respectively. The PVKU grown probiotics enhanced growth and induced stem cell differentiation
due to HA content. Evidence of this differentiation was seen in the morphological shift from spindle to osteocyte-shaped
cells after five days of incubation, an increase in ALP level over 21 days, and detection of intracellular calcium deposits
through alizarin red staining-all indicative of osteoblast cell development.
Conclusion: The osteogenic differentiation process in stem cells, improved by the nano-HA-containing byproducts of
probiotic bacteria in the PVKU medium, represents a promising pathway for leveraging beneficial bacteria and their
synthesised biomaterials in tissue engineering.

Keywords

Main Subjects

References

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Cell Journal (Yakhteh)
Volume 25, Issue 11 - Serial Number 118
November 2023
Pages 753-763

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