Evaluation of Osteogenic Differentiation of PDLSCs Encapsulated in Self-Assembling Hydrogel Scaffold in the Presence of BMP-2

Nova Southeastern University, Davie, Florida, USA

Objective. To investigate optimal BMP-2 concentration to enhance osteogenic differentiation of periodontal ligament derived stem cells (PDLSCs) embedded in PuraMatrix™ hydrogel. Background. Bone Morphogenic Protein-2 (BMP-2) provides the primary signal specifically targeting progenitor cells to stimulate production of osteoblasts and promote mineral deposition. PuraMatrix™, a nanofiber hydrogel scaffold, promotes cell growth and provides sufficient porosity for adequate nutrient diffusion within the hydrogel suspension, mimicking in vivo conditions. It is hypothesized that PuraMatrix™ encapsulated PDLSCs stimulated with BMP-2 accelerates osteogenic differentiation. Methods. Cell proliferation was measured by WST and live-dead cell assays. Osteogenic differentiation of PDLSCs in hydrogel supplemented with 50, 100, and 200ng/ml of BMP-2 for one week was measured through a pNPP assay and quantitative PCR analysis for ALP and RUNX2 gene expression. Results. Live dead cell assay of PDLSCs encapsulated in PuraMatrix™ induced with BMP-2 demonstrated that cells were viable at all concentrations observed. A dose dependent increase in the ALP activity was observed in treated cells. PCR analysis demonstrated a significant increase in the ALP and RUNX2 gene expressions at all concentrations. ALP levels significantly increased in cells treated with 50ng/ml compared to 200ng/ml BMP-2 and no significant difference between 100ng/ml and 200ng/ml BMP-2 groups. These results suggest accelerated osteogenic differentiation of PDLSC’s with increased BMP-2 dosages. Conclusions. These findings can be implicated towards use of this BMP-2 mediated cell-scaffold-system in craniofacial bone tissue engineering. Grants. This study was funded by a grant from FloridaBlue.