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Efficacy of Human Platelet Rich Fibrin Exudate vs Fetal Bovine Serum on Proliferation and Differentiation of Dental Pulp Stem Cells
International Journal of Stem Cells
Published online February 28, 2017;  
© 2017 Korean Society for Stem Cell Research.

Mahmoud A. Saeed1, Mohamed Abd El-Rahman2, Mohamed E. Helal2, Ahmed R. Zaher2, Mohammed E. Grawish1,2

1Department of Oral Biology, Faculty of Oral and Dental Medicine, Delta University for Science and Technology, Gamasa, Mansoura, Egypt,
2Department of Oral Biology, Faculty of Dentistry, Mansoura University, Mansoura, Egypt
Correspondence to: Mohammed El-Awady Grawish
Department of Oral Biology, Faculty of Dentistry, Mansoura University, Mansoura 35516, Egypt
Tel: +02-50-6328744, Fax: +02-50-2200706
E-mail: grawish2005@yahoo.com
; Accepted January 27, 2017.
This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
Background and Objectives: The imperative role of dental pulp stem cells (DPSCs) in regenerative therapy demands an in-vitro expansion which must deal with the safety and ethical problems associated with fetal bovine serum (FBS). The primary aim of this study was to compare the effects of human platelet rich fibrin (hPRF) exudate Vs FBS on proliferation and osteodifferentiation of human dental pulp stem cells (hDPSCs). The secondary one was to determine the optimum concentration of hPRF exudate inducing hDPSCs proliferation and osteodifferentiation.
Methods: The direct method was used to prepare hPRF exudate. hDPSCs were isolated from impacted mandibular third molars of twelve donors by the outgrowth method. For cell viability and proliferation rate testing, 96 well plates were used and the assay was done in duplicate and the trial repeated four times under the same conditions. Six wells were used to contain 10% FBS, serum free media, 1%, 5%, 10% and 20% concentrations of hPRF exudates, respectively. The proliferation assay was carried out by MTS tetrazolium cell proliferation assay kit and Elisa reader. The study design for osteodifferentiation protocol was exactly as the proliferation one and instead the assay was carried out by alizarin red with Elisa reader.
Results: Compared to 10% FBS, 10% hPRF exudate was the optimum concentration for hDPSCs proliferation, while 1% hPRF exudate was the optimum concentration for osteodifferentiation of hDPSCs.
Conclusions: Avoiding the risk of zoonosis which may be occurred with FBS, it is recommended to use 10% hPRF exudate for proliferation and 1% for osteodifferentiation.
Keywords : Human platelet rich fibrin exudate, Fetal bovine serum, Dental pulp stem cells, MTS cell proliferation assay, Alizarin Red S staining assay


November 2016, 9 (2)