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

doi:10.1016/j.jot.2020.01.003

Fibroblast growth factor 2-induced human amniotic mesenchymal stem cells combined with autologous platelet rich plasma augmented tendon-to-bone healing

J Orthop Translat. 2020 Feb 12:24:155-165. doi: 10.1016/j.jot.2020.01.003. eCollection 2020 Sep.

Authors

Jun Zhang¹, Ziming Liu², Jingfeng Tang¹, Yuwan Li³, Qi You¹, Jibin Yang¹, Ying Jin¹, Gang Zou¹, Zhen Ge¹, Xizhong Zhu¹, Qifan Yang¹, Yi Liu¹

Affiliations

¹ Department of Orthopaedic Surgery, Affiliated Hospital of Zunyi Medical University, China.
² Institute of Sports Medicine, Beijing Key Laboratory of Sports Injuries, Peking University Third Hospital, China.
³ Department of Orthopaedic Surgery, The First Affiliated Hospital of Chongqing Medical University, Chongqing.

Abstract

Objective: 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.

Methods: 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.

Results: 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.

Conclusion: hAMSCs transfected with the FGF-2 gene combined with autologous PRP could augment tendon-to-bone healing in a rabbit extra-articular model.

The translational potential of this article: 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.

Keywords: Human amniotic mesenchymal stem cells; Platelet-rich plasma; Tendon-to-bone healing; Tendon-to-bone junction.