The Effects of Fibroblast Growth Factor-2 (FGF-2) on Achilles Tendon Injury
Authors: Fujikawa Yuki , Nishimura Akinobu , Senga Yoshiyuki , Sudo Akihiro 
Published: December 2023

Category: Basic Sciences/Biologics; Ankle Introduction/Purpose: Achilles tendon defects are a challenging problem, and there is a need for new treatments. We investigated the use of Platelet-rich fibrin (PRF), a dense fibrin scaffold containing many growth factors and cytokines, for Achilles tendon injuries. The study found that PRF accelerates healing by promoting the proliferation and activation of tenocytes via FGF receptor/AKT signaling and TGF-β/SMAD3 signaling. While FGF-2, a growth factor in PRF, has been shown to promote tenocyte differentiation and proliferation, its effect on Achilles tendon injuries has not been reported. Therefore, this study aimed to investigate the effect of FGF-2 on Achilles tendon healing when administered alone and to determine its specific contribution to the healing process. Methods: We used a rat model of Achilles tendon defect. A 4-mm portion of the right Achilles tendon, 4 mm proximal to the calcaneal insertion, was resected to create a tendon defect. Gelatin hydrogel sheet (MedGel PI5) as a scaffold for PBS, FGF-2 (Rigroth®), and FGF receptor inhibitor (FGFRI: Pemigatinib) was placed into the gap before suturing it with nylon sutures. We observed HE-stained specimens at 200x and assessed histological healing by counting cells and vessels and grading cell morphology. The motor function was evaluated by the Basso, Beattie, and Bresnahan (BBB) score and the treadmill test. We compared the effects of FGF-2 on tenocytes isolated from rat Achilles tendons in the control, FGF2, and FGFRI groups. The proliferation and migration of tenocytes were measured by MTS assay and wound closure assay, respectively. Protein expression, such as Tenomodulin (Tnmd), Collagen Ⅰ, Ⅲ, and Screlaxis-A in cells, were evaluated by Western Blotting. Results: In the rat model, the number of cells, vessels and cell morphology scores at the defect sites increased in the FGF-2 group and decreased in the FGFRI group at 2 and 4 weeks postoperatively. In the motor functional evaluation, the FGF-2 group showed earlier improvement in the BBB score and the treadmill test compared to the control and FGFRI groups. The MTS assay showed that the number of viable cells was higher in the FGF-2 group and lower in the FGFRI group than in the control. Wound closure assay showed tenocyte migration was increased in the FGF-2 group and decreased in the FGFRI group. Regarding protein expression, Tnmd, Collagen I, III, and Screlaxis-A were increased in the FGF-2 group and decreased in the FGFRI group. Conclusion: In vivo, FGF-2 promoted healing of Achilles tendon injury, and FGFRI impeded it in terms of motor function and histology. Additionally, in vitro, FGF-2 promoted the proliferation and migration of tenocytes and enhanced extracellular matrix production, while FGFRI diminished these effects. Although native FGF-2 is produced at the injured site, further administration of FGF-2 accelerated Achilles tendon injury healing, while inhibiting the FGF receptor delayed recovery in this study. Therefore, FGF- 2 is very important in Achilles tendon injury healing and may promote recovery by enhancing angiogenesis, proliferation, and extracellular matrix production of tenocytes.