Mo Qingyun, Zheng Haotian, Liu Chuanquan, Sun Yuzhi, Cao Zhicheng, Sheng Renwang, Zhang Yanan, Wang Mingyue, Kuang Baian, Backman Ludvig J, Li Jiaxiang, Zhang Wei, Chen Jialin
School of Medicine, Southeast University, 210009, Nanjing, China; Center for Stem Cell and Regenerative Medicine, Southeast University, 210009, Nanjing, China.
Center for Stem Cell and Regenerative Medicine, Southeast University, 210009, Nanjing, China; Department of Orthopaedic Surgery, Institute of Digital Medicine, Nanjing First Hospital, Nanjing Medical University, 210006, Nanjing, China.
Acta Biomater. 2025 Jun 15;200:432-451. doi: 10.1016/j.actbio.2025.05.040. Epub 2025 May 26.
The dysregulation of the inflammatory microenvironment following tendon injury significantly hinders regeneration. In this study, we developed an all-silk-derived functional scaffold (rKL@MPs-ASF) by integrating silk fibroin (SF) microspheres (MPs) loaded with the anti-inflammatory protein recombinant α-Klotho (rKL) into a biomimetic aligned SF (ASF) scaffold. This scaffold is designed to regulate the inflammatory microenvironment and facilitate tendon regeneration. Proteomic analysis revealed that rKL preserves the tenogenic differentiation potential of tendon stem/progenitor cells (TSPCs) by mitigating the oxidative stress response in a tumor necrosis factor-alpha-induced inflammatory microenvironment in vitro. The rKL@MPs-ASF scaffold demonstrated good drug loading/release capabilities and biocompatibility in vitro. In a rat full-thickness Achilles tendon defect model, the rKL@MPs-ASF scaffold reduced inflammatory cells infiltration and promoted fibroblast infiltration compared to the PBS@MPs-ASF group at 4 weeks post-operation. At 8 weeks post-operation, rKL@MPs-ASF-treated tendons showed increased collagen fiber deposition and reduced heterogeneous ossification, facilitating tendon regeneration and functional recovery. In conclusion, this all-silk-derived functional system creates a conducive microenvironment for tendon regeneration. STATEMENT OF SIGNIFICANCE: Regulation of the inflammatory microenvironment plays a crucial role in modulating the differentiation of TSPCs, thereby promoting tendon tissue regeneration. In this study, we demonstrate that rKL effectively preserves the tenogenic differentiation potential of TSPCs by mitigating oxidative stress within an inflammatory microenvironment. We developed an innovative, all-silk-based functional system (rKL@MPs-ASF), which integrates rKL-loaded SF MPs into an aligned silk fibroin scaffold. This system enables the controlled release of rKL, thereby modulating inflammation, promoting collagen fiber deposition, inhibiting heterotopic ossification, and ultimately improving tendon regeneration and functional recovery. Our findings highlight the potential of the rKL@MPs-ASF system, which combines structural and biological properties with a versatile drug-delivery platform, as a promising strategy for enhancing tendon repair and regenerative outcomes.
肌腱损伤后炎症微环境的失调显著阻碍了再生。在本研究中，我们通过将负载抗炎蛋白重组α-klotho（rKL）的丝素蛋白（SF）微球（MPs）整合到仿生排列的SF（ASF）支架中，开发了一种全丝源功能支架（rKL@MPs-ASF）。该支架旨在调节炎症微环境并促进肌腱再生。蛋白质组学分析表明，rKL通过减轻体外肿瘤坏死因子-α诱导的炎症微环境中的氧化应激反应，保留了肌腱干/祖细胞（TSPCs）的成腱分化潜能。rKL@MPs-ASF支架在体外表现出良好的药物负载/释放能力和生物相容性。在大鼠全层跟腱缺损模型中，术后4周时，与PBS@MPs-ASF组相比，rKL@MPs-ASF支架减少了炎症细胞浸润并促进了成纤维细胞浸润。术后8周时，经rKL@MPs-ASF处理的肌腱胶原纤维沉积增加，异位骨化减少，促进了肌腱再生和功能恢复。总之，这种全丝源功能系统为肌腱再生创造了有利的微环境。重要性声明：炎症微环境的调节在调节TSPCs的分化中起着关键作用，从而促进肌腱组织再生。在本研究中，我们证明rKL通过减轻炎症微环境中的氧化应激有效地保留了TSPCs的成腱分化潜能。我们开发了一种创新的、全丝基功能系统（rKL@MPs-ASF），它将负载rKL的SF MPs整合到排列的丝素蛋白支架中。该系统能够控制rKL的释放，从而调节炎症、促进胶原纤维沉积、抑制异位骨化，并最终改善肌腱再生和功能恢复。我们的研究结果突出了rKL@MPs-ASF系统的潜力，该系统将结构和生物学特性与多功能药物递送平台相结合，作为增强肌腱修复和再生效果的一种有前途的策略。
