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

Second Advisor

Romer Ocanto

Third Advisor

Toshihisa Kawai

Publication Date / Copyright Date

2019

Publisher

Nova Southeastern University

Abstract

Background: Birth defects that result in structural malformation of major organs are one of the major causes of mortality in children. Tissue engineering and regenerative medicine (TERM) strategies are particularly beneficial for the pediatric patients to correct these defects owing to their greater regenerative capacity compared to the adult population. Limited availability of pediatric patient’s auto-transplantation of organs makes TERM strategies as a promising alternative toconventional clinical procedures that utilize the grafts. Recent developments in stem cells technologies, there is an enormous potential for TERM in correcting birth defects and enhancing the life span in young population. However, the major challenge to the craniofacial bone tissue engineering is the repair and regeneration of critical size bone defects accompanied by functional vascular network. Insufficient vasculature in regenerated bone is the main causes of large graft failure, leading to inner graft necrosis and lack of integration with the host tissue. While, it is well established that human gingiva derived mesenchymal stem cells (GMSCS) are robust source of osteogenic precursors in bone regenerative TERM, it is unknown if GMSCS also can differentiate into endothelial cells. The aim of this study was toexplore the potential of GMSCS to differentiate into endothelial cells.

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Methods: GMSCs were cultured under standard conditions. The angiogenic differentiation was induced administering Vascular Endothelial Growth Factor (VEGF). The expression of endothelial marker gene expression was measured by using quantitative PCR method. A functional tube formation assay was performed to assess the endothelialdifferentiation. Immunofluorescence was conducted to detect the surface marker protein CD31.

Results: Our results showed a dose dependent increase in the expression of the endothelial marker genes, PECAM-1, VCAM-1, KDR, FLT-1 and PCDH12. The tube formation assay revealed the ability to form capillary structures in the cellsinduced with 10 and 50ng/ml VEGF. The data on Immunofluorescence demonstrated the distinct presence of CD31 surface marker.

Conclusions: Our results showed that GMSCS have the potential to differentiate into endothelial cells. Thus, they serve as sources for programmed angiogenic and osteogenic cells to contribute for the regeneration of the vascularized bone tomake a paradigm-shift in stem cell therapy.

Disciplines

Dentistry

Keywords

VEGF induced differentation, Gingival stem cells, Endothelial cells

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