Proliferation and Osteogenic Differentiation of Dental Stem Cells in a Nanofibrous Hydrogel

Background: Cleft palate is the second most common congenital defect in the US. This genetic disability can be extremely debilitating affecting individual’s speech and facial growth. Bone tissue engineering emerged as a potential option for the repair and regeneration of the bone in the defect area. Objective: The objective of the study was to investigate proliferation and osteogenic differentiation of periodontal ligament derived stem cells (PDLSCs) encapsulated in a self-assembling, nano-fiber hydrogel scaffold, PuramatrixTM. Methodology: PDLSCs were cultured in PuramatrixTM. The cell proliferation was measured by a colorimetric WST assay at 1, 2, 3 and 5 day intervals. Cell viability was assessed using live dead cell assay. PDLSCs encapsulated in PuramatrixTM were either grown in the complete medium (CM), which served as the control, or osteogenic medium (OM). The expression of osteogenic marker genes alkaline phosphatase (ALP) and collagen type I (COLI) were assessed by quantitative PCR. The results were analyzed by one-way ANOVA. Results: Our results revealed that PDLSCs were viable at all concentrations observed. There was significant increase (P<0.05) in the proliferation in the cells seeded with (0.1 and 3) x105/ml from day 1 to Day 5. The viability of the cells seeded with (30x105)/ml were comparable with control. The results of live dead cell assay demonstrated PDLSCs encapsulated in PuramatrixTM survived at all concentrations. There was a twofold upregulation of ALP, COL I in PuramatrixTM encapsulated cells compared to control. Conclusion: PuramtrixTMsupported PDLSCs growth and differentiation. This tissue engineered construct has potential applications for bone regeneration.