Int J Stem Cells.  2017 May;10(1):38-47. 10.15283/ijsc16067.

Efficacy of Human Platelet Rich Fibrin Exudate vs Fetal Bovine Serum on Proliferation and Differentiation of Dental Pulp Stem Cells

• Saeed MA
• El-Rahman
• Helal ME
• Zaher AR
• Grawish ME

• KMID: 2384600
• DOI: http://doi.org/10.15283/ijsc16067

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.

Keyword

Human platelet rich fibrin exudate; Fetal bovine serum; Dental pulp stem cells; MTS cell proliferation assay; Alizarin Red S staining assay

MeSH Terms

Blood Platelets*
Cell Proliferation
Cell Survival
Culture Media, Serum-Free
Dental Pulp*
Enzyme-Linked Immunosorbent Assay
Exudates and Transudates*
Fibrin*
Humans*
Methods
Molar, Third
Stem Cells*
Tissue Donors
Culture Media, Serum-Free
Fibrin