To investigate the therapeutic effect of human dental pulp stem cells (DPSCs) transfected with adenovirus expressing hepatocyte growth factor (HGF) in a mouse model of collagen-induced arthritis (CIA). DPSCs were modified with Ad-HGF to produce HGF-overexpressing DPSCs, DPSCs-HGF. In experimental mouse CIA model, DPSCs-HGF and DPSCs-Null (modified with Ad-Null) were engrafted via intravenously after disease onset, which was determined by the presence of joint swelling. The therapeutic effects on joints were evaluated at 49 days after collagen injection by histopathological analysis and microcomputed tomography imaging. The inflammatory cytokines were analyzed both in sera and joints via MILLIPLEX kit and immunohistochemical staining, respectively, and the regulatory T cells (Tregs) were analyzed in peripheral blood by using flow cytometry. Furthermore, primary fibroblast-like synoviocytes were isolated, colony formation analysis and FACS were performed to evaluate the effect of HGF on the proliferation and cell cycle of FLSs. Western blot assay was carried out to clarify the signal pathway of HGF-cMet. We found that without HGF modification, DPSC transfusion was helpful in controlling autoimmune status, local synovitis, and bone erosion after intravenous administration. However, HGF-modified DPSCs have dual role in rheumatoid arthritis (RA). In the early phase, HGF overexpression inhibited RA progression by its immunosuppressive effects, while in the late phase, HGF promoted synovitis by activating fibroblast-like synoviocytes to produce pathogenic IL-6, accelerating cell proliferation and inducing apoptosis resistance via phosphorylating the c-Met/Akt pathway. The overall effect of HGF modification attenuated the therapeutic effect of DPSCs. Our study provides a comprehensive evaluation of the therapeutic effect of DPSCs in the mouse model and a primary answer to the divergence of whether HGF is harmful or helpful in RA.

中文翻译：

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肝细胞生长因子过度表达的纸浆干细胞在类风湿性关节炎中表现出双重作用。

研究转染表达肝细胞生长因子（HGF）的腺病毒的人牙髓干细胞（DPSCs）在胶原性关节炎（CIA）小鼠模型中的治疗作用。用 Ad-HGF 修饰 DPSC 以产生过表达 HGF 的 DPSC、DPSC-HGF。在实验性小鼠 CIA 模型中，DPSCs-HGF 和 DPSCs-Null（用 Ad-Null 修饰）在疾病发作后通过静脉内移植，由关节肿胀的存在确定。在注射胶原蛋白后 49 天，通过组织病理学分析和微计算机断层扫描成像评估了对关节的治疗效果。分别通过MILLIPLEX试剂盒和免疫组织化学染色分析血清和关节中的炎性细胞因子，并使用流式细胞术分析外周血中的调节性T细胞（Tregs）。此外，分离原代成纤维细胞样滑膜细胞，进行集落形成分析和FACS，以评估HGF对FLSs增殖和细胞周期的影响。进行蛋白质印迹分析以阐明HGF-cMet的信号通路。我们发现，在没有 HGF 修饰的情况下，输注 DPSC 有助于控制静脉给药后的自身免疫状态、局部滑膜炎和骨侵蚀。然而，HGF 修饰的 DPSC 在类风湿关节炎 (RA) 中具有双重作用。在早期，HGF 过表达通过其免疫抑制作用抑制 RA 进展，而在晚期，HGF 通过激活成纤维细胞样滑膜细胞产生致病性 IL-6，加速细胞增殖并通过磷酸化 c-Met 诱导细胞凋亡抗性促进滑膜炎/Akt 通路。HGF修饰的整体效果减弱了DPSCs的治疗效果。我们的研究对 DPSCs 在小鼠模型中的治疗效果进行了全面评估，并对 HGF 对 RA 有害还是有益的分歧提供了主要答案。