IADR Abstract Archives
Accelerated Osteogenesis of BMP-2 Induced PDLSCs Encapsulated in a Hydrogel
Objectives: Bone Morphogenic Protein-2 (BMP-2) is an osteogenic growth factor, provides the primary signal, which specifically targets progenitor cells to stimulate production of osteoblasts and promotes 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 adding BMP-2 to cells embedded within a PuraMatrix™, accelerates the osteogenic differentiation of mesenchymal stem cells (MSCs). Thus the objective of the study was to investigate the optimal concentration of BMP-2 to enhance osteogenic differentiation of periodontal ligament derived stem cells (PDLSCs) embedded in PuraMatrix™ hydrogel.
Methods: The cell proliferation was measured by WST and live-dead cell assay. The osteogenic differentiation of PDLSCs embedded in hydrogel supplemented with 50, 100, and 200ng/ml of BMP-2 was measured through pNPP assay, quantitative PCR analysis for the genes Alkaline Phosphatase (ALP) and RUNX2, and Alizarin Red and Von Kossa staining techniques to measure mineral deposition.
Results: The results of our study demonstrated that cells were viable at all concentrations observed. Live dead cell assay demonstrated PDLSCs encapsulated in PuraMatrix™ induced with BMP-2 survived throughout the experimental period. There was a dose dependent increase in the ALP activity in the cells treated with BMP-2 for one week. The quantitative PCR study results demonstrated a significant increase in the ALP and RUNX2 and gene expressions at all concentrations. While there was significant increase in the ALP levels in the cells treated with 50ng/ml compared to 200ng/ml BMP-2, there was no significant difference between 100ng/ml and 200ng/ml BMP-2 treated group. Our study results suggest accelerated osteogenic differentiation of PDLSC’s with increased BMP-2 dosages.
Conclusions: The findings can be implicated towards the use of this BMP-2 mediated cell-scaffold- system in craniofacial bone tissue engineering.
Methods: The cell proliferation was measured by WST and live-dead cell assay. The osteogenic differentiation of PDLSCs embedded in hydrogel supplemented with 50, 100, and 200ng/ml of BMP-2 was measured through pNPP assay, quantitative PCR analysis for the genes Alkaline Phosphatase (ALP) and RUNX2, and Alizarin Red and Von Kossa staining techniques to measure mineral deposition.
Results: The results of our study demonstrated that cells were viable at all concentrations observed. Live dead cell assay demonstrated PDLSCs encapsulated in PuraMatrix™ induced with BMP-2 survived throughout the experimental period. There was a dose dependent increase in the ALP activity in the cells treated with BMP-2 for one week. The quantitative PCR study results demonstrated a significant increase in the ALP and RUNX2 and gene expressions at all concentrations. While there was significant increase in the ALP levels in the cells treated with 50ng/ml compared to 200ng/ml BMP-2, there was no significant difference between 100ng/ml and 200ng/ml BMP-2 treated group. Our study results suggest accelerated osteogenic differentiation of PDLSC’s with increased BMP-2 dosages.
Conclusions: The findings can be implicated towards the use of this BMP-2 mediated cell-scaffold- system in craniofacial bone tissue engineering.