1. Advanced-platelet-rich fibrin extract promotes adipogenic and osteogenic differentiation of human adipose-derived stem cells in a dose-dependent manner in vitro
- Author
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Dan-Dan Zhu, Maojian Chen, Meng-Xin Wang, Donglin Huang, Hong-Mian Li, Jinping Mo, Zhijie Liang, Changyuan Wei, and Wenhai Nong
- Subjects
0301 basic medicine ,Adipose tissue ,Complex Mixtures ,Biology ,03 medical and health sciences ,Paracrine signalling ,chemistry.chemical_compound ,0302 clinical medicine ,Osteogenesis ,Platelet-Rich Fibrin ,Humans ,Oil Red O ,Adipogenesis ,Dose-Response Relationship, Drug ,Cell growth ,Cell Differentiation ,Mesenchymal Stem Cells ,030206 dentistry ,Cell Biology ,General Medicine ,Cell biology ,Reverse transcription polymerase chain reaction ,RUNX2 ,030104 developmental biology ,chemistry ,Stem cell ,Developmental Biology - Abstract
Advanced platelet-rich fibrin (A-PRF) is an autogenous biological material obtained from peripheral blood. A-PRF extract (A-PRFe) contains a high concentration of various cytokines that are increasingly appreciated for their roles in improving stem cell repairing function during tissue regeneration. However, the optimal A-PRFe concentration to stimulate stem cells is unknown. This study aimed to identify the optimal concentrations of A-PRFe to promote adipogenic and osteogenic differentiation of human adipose-derived stem cells (ASCs). We produced A-PRFe from A-PRF clots by centrifuging fresh peripheral blood samples and isolated and identified ASCs using surface CD markers and multilineage differentiation potential. Enzyme-linked immunosorbent assay (ELISA) showed the concentrations of several cytokines, including b-FGF, PDGF-BB, and others, increased gradually, peaked on day 7 and then decreased. Cell proliferation assays showed A-PRFe significantly stimulated ASC proliferation, and proliferation significantly increased at higher A-PRFe doses. The degree of adipogenic and osteogenic differentiation increased at higher A-PRFe concentrations in the culture medium, as determined by oil red O and alizarin red staining. Reverse transcription polymerase chain reaction (RT-PCR) showed that expression levels of genes related to adipogenic/osteogenic differentiation (PPARγ2, C/EBPα, FABP4, Adiponectin, and ALP, OPN, OCN, RUNX2), paracrine (HIF-1α, VEGF, IGF-2) and immunoregulation (HSP70, IL-8) function were higher in groups with a higher concentration of A-PRFe than in lower concentration groups. This study demonstrates that A-PRFe is ideal for use in ASC applications in regenerative medicine because it improves biological functions, including proliferation, adipogenic/osteogenic differentiation, and paracrine function in a dose-dependent manner.
- Published
- 2021
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