# PTEN Inhibition Suppresses Differentiation in Periodontal Ligament Stem Cells

## TL;DR

This study shows that inhibiting PTEN in periodontal ligament stem cells reduces their ability to form colonies, migrate, and differentiate into bone or fat cells.

## Contribution

The study reveals PTEN's role in regulating stem cell differentiation and tissue homeostasis in periodontal ligament cells.

## Key findings

- PTEN inhibition with VO-OHpic reduced colony formation and cell migration in PDLSCs.
- PTEN inhibition impaired osteogenic and adipogenic differentiation in PDLSCs.
- RNA sequencing showed upregulated TGF-β and calcium signaling pathways with PTEN inhibition.

## Abstract

Phosphatase and Tensin Homolog (PTEN) functions in numerous biological processes, encompassing cell proliferation, growth, self-renewal, and differentiation. This study examined the modulatory function of the PTEN inhibitor in periodontal ligament stem cells (PDLSCs). PDLSCs were treated with VO-OHpic at a concentration range from 0.625 to 5 μM. MTT assay and Coomassie Blue staining were conducted to determine cell viability and colony-forming unit ability, respectively. The scratch assay was employed to examine cell migration. Mineral deposition and intracellular lipid accumulation were assessed. The qRT-PCR and immunofluorescence were used to evaluate mRNA and protein expression, respectively. RNA sequencing was employed for transcriptomic analysis. VO-OHpic exposure showed no cytotoxic effects in PDLSCs; however, at 5 μM, it markedly decreased colony-forming efficiency and impaired cell migration. Under osteogenic induction conditions, 5 μM VO-OHpic markedly attenuated mineralisation and downregulated the osteogenic marker gene expression partly through ERK signalling. Indeed, VO-OHpic impaired intracellular lipid accumulation during adipogenic differentiation, as evidenced by reduced expression of adipogenic marker genes. RNA sequencing analysis revealed that VO-OHpic treatment upregulated genes in the TGF-β and calcium signalling pathways, suggesting a regulatory role in PDLSC differentiation. In conclusion, PTEN regulates PDLSC colony formation, migration, and differentiation, suggesting a pivotal role for PTEN in maintaining periodontal tissue homeostasis.