Fibrin Substrate Supports Chondrogenesis of Adipose-Derived Stem Cells Supplemented by Human Platelet-Rich Plasma and L-Ascorbic Acid 2-Phosphate

Abstract

Background & Aim Cartilage engineering study is globally rising nowadays to overcome the need of cartilage tissue. Induced chondrogenesis of adipose-derived stem cells (ADSCs) supplemented by platelet-rich plasma (PRP) or L-ascorbic acid 2-phosphate (LAA2P) has been reported. The PRP contains TGF-β to induce chondrogenesis while LAA2P induced chondrogenesis through sodium vitamin C transporter 2 (SCVT-2). Type 2 collagen (T2Col) is chondrogenesis marker and type 1 collagen (T1Col) is osteogenesis marker. However, the use of fibrin clot PRP as a substrate for chondrogenesis is still lack of data. This research aimed to study chondrogenesis of ADSCs supplemented by 10% PRP or LAA2P (50µg/mL) with fibrin as substrate. Methods, Results & Conclusion The ADSCs was isolated from human lipoaspirates while PRP was isolated from human blood. The fibrin clot substrate was fabricated trough platelet activation as a waste form. The characteristic of ADSCs, PRP composition and wet weight of fibrin substrate were conducted. Then, the ADSCs cultured in fibrin substrate with different groups of supplementation which were 10% fetal bovine serum (FBS), 10% PRP, and combination of 10%FBS-LAA2P (50µg/mL). Chondrogenesis of ADSCs on day 12 were evaluated by glycosaminoglycans (GAG) and gene expression of T2Col and T1Col. The results showed that the cell was fibroblast-like shape with the viability more than 85% and the doubling time about 2.1 days. The PRP contains highest level of platelet but lack of red blood cell (RBC), white blood cell (WBC), lymphocyte and neutrophils. The GAG value of PRP and LAA2P groups were higher than FBS group. Furthermore, the T2Col and T1Col expression of PRP was significantly higher (p <0.05) than LAA2P and FBS groups, respectively. Those results showed that PRP induced chondrogenesis of ASCs faster than LAA2P. The fastest process of chondrogenesis which regulated by PRP was suggested caused by TGF-β ligand-receptor interaction. It concluded that the fibrin substrate supports chondrogenesis of ASCs which occurred faster in 10% PRP of supplementation than LAA2P (50µg/mL). Cartilage engineering study is globally rising nowadays to overcome the need of cartilage tissue. Induced chondrogenesis of adipose-derived stem cells (ADSCs) supplemented by platelet-rich plasma (PRP) or L-ascorbic acid 2-phosphate (LAA2P) has been reported. The PRP contains TGF-β to induce chondrogenesis while LAA2P induced chondrogenesis through sodium vitamin C transporter 2 (SCVT-2). Type 2 collagen (T2Col) is chondrogenesis marker and type 1 collagen (T1Col) is osteogenesis marker. However, the use of fibrin clot PRP as a substrate for chondrogenesis is still lack of data. This research aimed to study chondrogenesis of ADSCs supplemented by 10% PRP or LAA2P (50µg/mL) with fibrin as substrate. The ADSCs was isolated from human lipoaspirates while PRP was isolated from human blood. The fibrin clot substrate was fabricated trough platelet activation as a waste form. The characteristic of ADSCs, PRP composition and wet weight of fibrin substrate were conducted. Then, the ADSCs cultured in fibrin substrate with different groups of supplementation which were 10% fetal bovine serum (FBS), 10% PRP, and combination of 10%FBS-LAA2P (50µg/mL). Chondrogenesis of ADSCs on day 12 were evaluated by glycosaminoglycans (GAG) and gene expression of T2Col and T1Col. The results showed that the cell was fibroblast-like shape with the viability more than 85% and the doubling time about 2.1 days. The PRP contains highest level of platelet but lack of red blood cell (RBC), white blood cell (WBC), lymphocyte and neutrophils. The GAG value of PRP and LAA2P groups were higher than FBS group. Furthermore, the T2Col and T1Col expression of PRP was significantly higher (p <0.05) than LAA2P and FBS groups, respectively. Those results showed that PRP induced chondrogenesis of ASCs faster than LAA2P. The fastest process of chondrogenesis which regulated by PRP was suggested caused by TGF-β ligand-receptor interaction. It concluded that the fibrin substrate supports chondrogenesis of ASCs which occurred faster in 10% PRP of supplementation than LAA2P (50µg/mL).