الفهرس 
Title : The Differentiation Enhancing Potential of Platelet Rich Fibrin Scaffold Prepared Under Different Centrifugation Protocols on Stem Cells from The Apical Papilla (In Vitro Study)
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Abstract
Introduction: Platelet rich fibrin was widely used in dentistry for its biocompatibility and enhanced tissue regeneration ability. Since the low-speed concept (LSC) like advanced platelet rich fibrin (A-PRF), shown an increase in total leucocyte numbers and growth factors release within PRF matrix scaffolds compared to the standard protocol for PRF preparation/ leucocyte PRF (L-PRF). Aim: The present work was designed to evaluate the proliferation and differentiation potential of the stem cells of apical papilla (SCAP) seeded along with platelet rich fibrin (PRF) scaffolds prepared under two different centrifugation protocols.
Methodology: SCAP were obtained from impacted third molar teeth with immature roots. PRF scaffold was prepared under two different previously published centrifugation protocols. Standard protocol, Leucocyte PRF (LPRF) (10 ml; 2700 rpm for 12 minutes) and LSC protocol, advanced PRF (A-PRF) (10 ml; 1500 rpm for 14 minutes). The PRF scaffolds were freshly used in culture. After characterization the SCAP were divided according to the study design into four groups, 1. negative control (NC), 2. positive control (PC), 3. L-PRF and 4. A-PRF groups. Then seeded for seven days for osteogenic differentiation. Cell count (trypan blue stain) and cell viability (MTT assay) were carried out to assess the proliferation capacity. Alizarin red S (ARS) stain, Alkaline phosphatase (ALP) activity and Receptor activator of nuclear factor-kappa B ligand (RANKL) immunofluorescence staining were used to evaluate the osteogenic potential in the differentiated SCAP.
Results:L-PRFshowed statistically no significant increase in the cell count compared to both control groups, while the A-PRF showed a highly significant one. Regarding the viable cell count in the A-PRF was higher and statistically significant than the L-PRF. ARS mineralization nodule formation, ALP activity and RANKL protein expression were significantly higher in A-PRF and L-PRF, respectively when compared to the control groups. Conclusion: Both types ofPRF increased the cell count, cell viability with no cytotoxicity that reflected on increased proliferation and differentiation in terms of increased mineralization formation, ALP activity and RANKL protein expression that enhanced the osteogenic differentiation, compared to the control groups. A-PRF showed significant increase in proliferation and differentiation potentials when seeded with SCAP compared to L-PRF.