Student Theses, Dissertations and Capstones

Document Type

Thesis

Degree Name

Master of Science (M.S.) in Dentistry

Copyright Statement

All rights reserved. This publication is intended for use solely by faculty, students, and staff of Nova Southeastern University. No part of this publication may be reproduced, distributed, or transmitted in any form or by any means, now known or later developed, including but not limited to photocopying, recording, or other electronic or mechanical methods, without the prior written permission of the author or the publisher.

Department

College of Dental Medicine

First Advisor

Umadevi Kandalam

Publication Date / Copyright Date

2017

Publisher

Nova Southeastern University

Abstract

Purpose: The aim of the study was to investigate effect of the growth factor, Bone Morphogenetic Protein 2 (BMP2), on the differentiation of Human gingiva derived stem cells (HGMSCs) in a self-assembled three-dimensional (3D) peptide hydrogel. The study has two parts; part 1 comprised of optimizing the dose of BMP2 and Part II was investigating the effective delivery method of BMP2 in enhancing the osteogenic differentiation of HGMSCs Methods: Human gingiva derived mesenchymal stem cells (HGMSCs) cultured in the peptide hydrogel (3-D cultures) were treated with 50, 100 and 200 ng/ml BMP2. The cells in the osteogenic differentiation of HGMSCs in the peptide gel was evaluated at one week. The expressions of osteogenic marker genes Alkaline-Phosphatase (ALP), Runt related transcription factor 2 (RUNX2), Collagen Type I (COL1) were measured using quantitative PCR. The results were compared with monolayer cells treated with BMP2 (2-D culture). Furthermore, to evaluate the effective delivery method, cells were encapsulated in Puramtrix and BMP2 was administered either in culture medium or encapsulated in Puramatrix. The cells were treated for one week and early osteogenic markers genes were measured. ANOVA was used to evaluate the results and P < .05 was considered statistically significant. Results: The results of the study demonstrated a dose dependent upregulation (P<.05) of the genes including ALP, RUNX2 and COL1 at all concentrations (50,100 and 200ng/ml). There was significant up regulation of gene expression in 200ng/ml compared to 50ng/ml (P<.05). BMP2 treatment accelerated the mineral deposition in HGMSCs. Overall results of our study demonstrated that the application of osteoconductive agent rhBMP2, stimulated the osteogenic differentiation regardless of the delivery method that was used in this study. However, BMP2 entrapped in PuramatrixTM showed significantly high ALP expression at 100ng/ml compared to 200ng/ml concentration Conclusions: The PuramatrixTM hydrogel in combination with BMP2 supported osteogenic differentiation of HGMSCs. This novel tissue engineered cell-scaffold system with growth factor has potential for the regeneration of bone in craniofacial defects.

Disciplines

Dentistry

Keywords

Bone morphogenetic protein, Craniofacial defects, Gingiva derived stem cells, Osteogenic differentiation, Peptide hydrogel

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