# Role of Extracellular Vesicles of Stem Cells from Human Exfoliated Deciduous Teeth in Osteogenesis

## TL;DR

This study shows that extracellular vesicles from stem cells in baby teeth can boost bone cell growth and regeneration, offering potential for new therapies.

## Contribution

The study identifies extracellular vesicles from SHED-CM as key drivers of osteogenesis, offering a novel cell-free therapeutic approach.

## Key findings

- SHED-EVs enhance the proliferation and migration of human calvarial osteoblasts.
- SHED-EVs increase alkaline phosphatase expression and promote bone differentiation.
- SHED-EVs show potential for use in bone regeneration therapies.

## Abstract

The tissue regenerative potential of the liquid component of mesenchymal stem cells has gained significant attention. Stem cells from human exfoliated deciduous teeth-conditioned medium (SHED-CM), which is often extracted during orthodontic treatment, promotes bone regeneration. However, further investigation is warranted to determine which component of SHED-CM affects bone regeneration. Therefore, we focused on the extracellular vesicles contained in SHED-CM (SHED-EVs) and aimed to study their effects on osteoblasts. SHED-EVs were isolated using a pellet-down EV extraction kit and identified using transmission electron microscopy and NanoSight. SHED-EVs were added to human calvarial osteoblasts (HCOs), and cell proliferation and migration ability were examined with Incucyte® and BrdU. Alkaline phosphatase (ALP) expression was confirmed using real-time PCR and ALP quantification. The bone differentiation potential was examined using Alizarin Red S (ARS) staining. SHED-EVs promoted proliferation and migration of HCOs. Real-time PCR and ALP quantification results demonstrated that HCOs cultured with SHED-EVs exhibited increased ALP expression. ARS staining revealed that SHED-EVs promoted bone differentiation of HCOs. These results suggest that SHED-EVs promote cell proliferation and migration and bone regeneration of osteoblasts, highlighting their potential in the development of bone regeneration therapies.