HPD Research Day | February 16, 2018

9 Marder Auditorium Marder Auditorium 10:45 – 11:15 a.m. Evaluation of Osteogenic Differentiation of PDLSCs Encapsulated in Self-Assembling Hydrogel Scaffold in the Presence of BMP-2 Karen Ben-Elazar, BA, D3, College of Dental Medicine, Nova Southeastern University Alireza Heidari, Ph.D., College of Dental Medicine, Nova Southeastern University Nicole DeLorenzo, BS, College of Dental Medicine, Nova Southeastern University Jerry T. Ennolikara, Halmos College of Natural Sciences and Oceanography, Nova Southeastern University Umadevi Kandalam, Ph. D., Associate Professor, College of Dental Medicine, Nova Southeastern University 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. Marder Auditorium 11:15 – 11:45 a.m. RGD-Modified Alginate Scaffold Supplemented with BMP2 Supports Osteogenic Differentiation of Periodontal Ligament Derived Stem Cells (PDLSCs) Alireza Heidari, Ph.D., Research Lab Assistant II, College of Dental Medicine, Nova Southeastern University Nicole DeLorenzo, BS, Research Lab Technician I, College of Dental Medicine, Nova Southeastern University Umadevi Kandalam, Ph.D., Associate Professor, College of Dental Medicine, Nova Southeastern University Objective. To develop in vitro injectable cell-growth factor-scaffold system and to assess osteogenic differentiation of (PDLSCs). Background. Multipotent stem cells derived from human periodontal ligament stem cells (PDLSCs) are able to promote the formation of bone tissue making this cells interesting candidate for craniofacial bone tissue engineering. RGD-coupled alginate is considered as promising scaffold for the capacity to mimic many functions of the extracellular matrices. Bone Morphogenic Protein-2 (BMP-2) is a potent growth factor, plays an essential role in osteogenic induction. Our hypothesis is that, the combination of BMP-2 to cells embedded within alginate beads scaffold, accelerates the osteogenic differentiation. Methods. Human PDLSCs was isolated and cultured using standard culture conditions. The cells were encapsulated in the alginate gels and their proliferation was measured. The PDLSCs in alginate beads were supplemented with 50, 100, and 200ng/ml of BMP-2. The activity of alkaline phosphatase was measured through a pNPP assay. The osteogenic differentiation of ALP, RUNX2 and COL1 gene expression was measured using quantitative PCR. Results. The WST demonstrated that PDLSCs were viable at all concentrations. Furthermore, the cells were viable until day7. The quantitative PCR results reported upregulation expressions in the ALP, and COL1 genes at all BMP2 concentrations with peak at 200ng/ml. Conclusions: BMP2 enhances the osteogenic differentiation. RGD modified alginate- BMP2- PDLSCs- system represents a promising source for bone tissue engineering. Grants. This study was funded by Florida Blue foundation grant # 333248.