Comparison of the expression level of osteogenic genes of DPSC transferred to scaffolds containing nano hydroxyapatite in presence of A-PRF and L-PRF

Abstract

Background: Due to the Importance of osteoblast differentiation using autologus blood derivatives in the reconstruction of bone defects in reconstructive medicine, the aim of this study was the extent of expression of genes related to extraction in human pulp stem cells transferred to existing nanohydroxyapatite scaffolds in the presence of L-PRF, A -PRF. Materials and Methods: In this in vitro study, human dental pulp stem cells from permanent teeth extracted for orthodontic or periodontal purposes were purchased from Bonyakhteh research center. Polymer scaffolds made of PCL-PEG-PCL and containing nanohydroxyapatite particles were placed in 9 houses out of 12-house plates. Within 12 cells, 500,000 cells were added (3 cells were considered as a control group without scaffolding). CC10 venous blood was taken from the ears of rabbits and their A-PRF was obtained according to L-PRF and A protocol. And clots with approximate dimensions of 0.5 × 0.5 cm were placed in 6 cells (3 wells containing L-PRF and 3 cells containing A-PRF). Three wells were considered for the cell and the scaffold alone. After 14 days, the expression of four genes RUNX2, COL1A1, OSTEOCALSIN, BSP, GAPDH (refrence gene) were examined by real time PCR. Results: Both A-PRF and L-PRF protocols significantly increased the expression of ossification genes in dental pulp stem cells over 14 days. This increase was greater in the A-PRF group than in the L-PRF group. Conclusion: Both the A-PRF and L-PRF protocols can be used in bone remodeling for more detailed animal studies.