Effect of VEGF on Periodontal Ligament Derived Stem-Cell Differentiation

Objectives: Tissue engineering and regenerative medicine (TERM) is emerging as a potential therapeutic option for the treatment of congenital defects such as cleft palate. Recently the treatment modalities using TERM in pediatric patients is gaining attention due to the difficulties involved in auto-transplantation, such as obtaining huge quantity of bone and additional surgical procedures with donor site morbidity, in pediatric population. However, one of the major challenges in the craniofacial tissue engineering is repairing the bone with a proper vasculature within the newly formed bone in the critical size bone defect. It has been established that human periodontal ligament derived mesenchymal stem cells (PDLSCs) can be differentiated to be osteogenic precursors in bone regeneration. However, it is unknown whether PDLSCs have the capacity to differentiate into endothelial cells. The aim of this study is to investigate the capability of the PDLSCs to differentiate into endothelial cells.
Methods: Standard cell culturing procedures has been followed for PDLSCs. Then vascular endothelial growth factor (VEGF) was supplemented to PDLSCs to establish the endothelial differentiation of the cells. The genetic expression of the various endothelial markers was measured using qPCR technique with different markers. Tube formation assay was performed to investigate the differentiation progress. The surface marker protein CD31 was investigated using immunofluorescence technique.
Results: There is a dose dependent increase in the gene expression of the endothelial marker genes VCAM-1, KDR, PCDH-12, and FLT-1. One week of induction of 10 and 50mg/ml VGEF formed tubes in PDLSCs. Distinct presence of the CD31 protein was apparent with the data acquired from Immunofluorescence microscopy.
Conclusions: PDLSCs is found to have the potential to differentiate into endothelial cells. This study has potential to develop functional bone in craniofacial region.