成纤维细胞生长因子2诱导的人羊膜间充质干细胞联合自体富血小板血浆促进腱骨愈合。

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

作者信息

Zhang Jun, Liu Ziming, Tang Jingfeng, Li Yuwan, You Qi, Yang Jibin, Jin Ying, Zou Gang, Ge Zhen, Zhu Xizhong, Yang Qifan, Liu Yi

机构信息

Department of Orthopaedic Surgery, Affiliated Hospital of Zunyi Medical University, China.

Institute of Sports Medicine, Beijing Key Laboratory of Sports Injuries, Peking University Third Hospital, China.

出版信息

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

DOI:10.1016/j.jot.2020.01.003

PMID:33101966

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7548348/

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 ( < 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.

摘要

目的

本研究旨在探讨成纤维细胞生长因子2（FGF-2）对人羊膜间充质干细胞（hAMSCs）体外胶原纤维形成和成骨分化的影响，以及FGF-2诱导的hAMSCs联合自体富血小板血浆（PRP）对体内腱骨愈合的影响。

方法

体外实验中，用不同浓度的FGF-2（0、10、20和40 ng/ml）诱导hAMSCs 14天，通过定量实时逆转录PCR、蛋白质免疫印迹法、免疫荧光和天狼星红染色检测韧带分化和成骨分化的结果。此外，使用携带FGF-2基因的慢病毒转染hAMSCs，并通过定量实时逆转录PCR（qRT-PCR）和蛋白质免疫印迹法检测转染效率。体内实验中，通过组织学检查、生物力学分析和影像学分析，检测转染FGF-2基因的hAMSCs联合自体PRP对腱骨愈合的影响。

结果

体外实验中，不同浓度的FGF-2（10、20和40 ng/ml）均促进了hAMSCs的韧带分化和成骨分化，低浓度的FGF-2（10 ng/ml）对分化有良好效果。此外，携带FGF-2基因的慢病毒以最佳感染复数（MOI）（50）成功转染至hAMSCs，且成功制备了自体PRP。体内实验中，转染FGF-2基因的hAMSCs联合自体PRP对腱骨愈合的效果优于其他组（P<0.05），组织学检查、生物力学分析和影像学分析均证实了这一点。