Fibroblast growth factor 2–induced human amniotic mesenchymal stem cells combined with autologous platelet rich plasma augmented tendon-to-bone healing
Jun Zhang, Ziming Liu, Jingfeng Tang, Yuwan Li, Qi You, Jibin Yang, Ying Jin, Gang Zou, Zhen Ge, Xizhong Zhu, Qifan Yang, Yi Liu
Abstract
The purpose of this study was to explore the effect of fibroblast growth factor 2 (FGF-2) on collagenous fibre formation and the osteogenic differentiation of human amniotic mesenchymal stem cells (hAMSCs) in vitro, as well as the effect of FGF-2–induced hAMSCs combined with autologous platelet-rich plasma (PRP) on tendon-to-bone healing in vivo. In vitro, hAMSCs were induced by various concentrations of FGF-2 (0, 10, 20, and 40 ng/ml) for 14 days, and the outcomes of ligamentous differentiation and osteogenic differentiation were detected by quantitative real-time reverse transcription PCR, Western blot, immunofluorescence, and picrosirius red staining. In addition, a lentivirus carrying the FGF-2 gene was used to transfect hAMSCs, and transfection efficiency was detected by quantitative real time reverse transcription PCR (qRT-PCR) and Western blot. In vivo, the effect of hAMSCs transfected with the FGF-2 gene combined with autologous PRP on tendon-to-bone healing was detected via histological examination, as well as biomechanical analysis and radiographic analysis. In vitro, different concentrations of FGF-2 (10, 20, and 40 ng/ml) all promoted the ligamentous differentiation and osteogenic differentiation of hAMSCs, and the low concentration of FGF-2 (10 ng/ml) had a good effect on differentiation. In addition, the lentivirus carrying the FGF-2 gene was successfully transfected into hAMSCs with an optimal multiplicity of infection (MOI) (50), and autologous PRP was prepared successfully. In vivo, the hAMSCs transfected with the FGF-2 gene combined with autologous PRP had a better effect on tendon-to-bone healing than the other groups (p < 0.05), as evidenced by histological examination, biomechanical analysis, and radiographic analysis. hAMSCs transfected with the FGF-2 gene combined with autologous PRP could augment tendon-to-bone healing in a rabbit extra-articular model. hAMSCs transfected with the FGF-2 gene combined with autologous PRP may be a good clinical treatment for tendon-to-bone healing, especially for acute sports-related tendon–ligament injuries.