Advanced-platelet-rich fibrin extract promotes adipogenic and osteogenic differentiation of human adipose-derived stem cells in a dose-dependent manner in vitro

• Published in: Tissue & cell Vol. 71; p. 101506
• Main Authors: Liang, Zhijie, Huang, Donglin, Nong, Wenhai, Mo, Jinping, Zhu, Dandan, Wang, Mengxin, Chen, Maojian, Wei, Changyuan, Li, Hongmian
• Format: Journal Article
• Language: English
• Published: Scotland Elsevier Ltd 01.08.2021; Elsevier Science Ltd
• Subjects: Adipogenesis - drug effects; Adiponectin; Adipose-derived stem cells; Advanced-platelet-rich fibrin extract; Alizarin; Biological materials; Biomedical materials; Blood; Cbfa-1 protein; Cell culture; Cell differentiation; Cell Differentiation - drug effects; Cell proliferation; Complex Mixtures - chemistry; Complex Mixtures - pharmacology; Cytokines; Differentiation; Differentiation (biology); Dose-Response Relationship, Drug; Enzyme-linked immunosorbent assay; Fibrin; Gene expression; Hsp70 protein; Humans; Hypoxia-inducible factor 1a; Immunoregulation; Insulin-like growth factors; Interleukin 8; Mesenchymal Stem Cells - metabolism; Osteogenesis - drug effects; Paracrine; Paracrine signalling; Peripheral blood; Platelet-derived growth factor; Platelet-derived growth factor BB; Platelet-Rich Fibrin - chemistry; Platelets; Polymerase chain reaction; Regeneration (physiology); Regenerative medicine; Reverse transcription; Stem cells; Tissue engineering; Vascular endothelial growth factor; Paracrine; Adipose-derived stem cells; Advanced-platelet-rich fibrin extract; Differentiation
• ISSN: 0040-8166; 1532-3072; 1532-3072
• DOI: 10.1016/j.tice.2021.101506

•Adipose-derived stem cells (ASCs) possess the function of polydifferentiation.•A-PRF gradually secreted extract containing various cytokines such as VEGF, TGF-β1 and IL-1β, and the levels peaked at day 7.•A-PRFe significantly promote proliferation, paracrine function and adipogenic/osteogenic differentiation of ASCs in a dose-dependent manner. 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.