Chen Rui, Li Yujie, Zhuang Yangyang, Zhang Yiming, Wu Hailong, Lin Tao, Chen Shixuan
Jiangsu Key Laboratory of Marine Bioresources and Environment, Jiangsu Ocean University, Lianyungang, China.
Zhejiang Engineering Research Center for Tissue Repair Materials, Wenzhou Institute, University of Chinese Academy of Sciences, Wenzhou, China.
Front Bioeng Biotechnol. 2023 Mar 24;11:1159068. doi: 10.3389/fbioe.2023.1159068. eCollection 2023.
Immune evaluation of biomaterials for tissue regeneration is a critical preclinical evaluation. The current evaluation criterion (ISO 10993-1 or GB/T 16886) uses rodents to perform the immune evaluation. However, the immune system of rodents is different from humans, the obtained results may not be reliable, which could lead directly to the failure of clinical trials. Granulocyte-macrophage colony-stimulating factor (GM-CSF) shows a great potential application in tissue regeneration by regulating local immune responses. The presented work combines the advantages of GM-CSF (immunoregulation) and hierarchically 3D nanofiber scaffolds (tissue regeneration). Firstly, we fabricated GM-CSF loaded 3D radially aligned nanofiber scaffolds, and then subcutaneous implantation was performed in humanized mice. The whole scaffold and surrounding tissue were harvested at each indicated time point. Finally, the cell infiltration and local immune responses were detected by histological observations, including H&E and Masson staining and immunochemistry. We found significant cell migration and extracellular matrix deposition within the 3D radially aligned nanofiber scaffold after subcutaneous implantation. The locally released GM-CSF could accelerate the expression of human dendritic cells (CD11c) only 3 days after subcutaneous implantation. Moreover, higher expression of human cytotoxic T cells (CD3/CD8), M1 macrophages (CD68/CCR7) was detected within GM-CSF loaded radially aligned nanofiber scaffolds and their surrounding tissues. The 3D radially aligned scaffold can accelerate cell migration from surrounding tissues to regenerate the wound area. And the locally released GM-CSF enhances dendritic cell recruitment and activation of cytotoxic T cells and M1 macrophages. Taken together, the GM-CSF loaded 3D radially aligned nanofiber scaffolds have a promising potential for achieving tissue regeneration.
用于组织再生的生物材料的免疫评估是一项关键的临床前评估。当前的评估标准（ISO 10993-1或GB/T 16886）使用啮齿动物进行免疫评估。然而，啮齿动物的免疫系统与人类不同，所获得的结果可能不可靠，这可能直接导致临床试验失败。粒细胞-巨噬细胞集落刺激因子（GM-CSF）通过调节局部免疫反应在组织再生中显示出巨大的潜在应用价值。本研究结合了GM-CSF（免疫调节）和分级3D纳米纤维支架（组织再生）的优点。首先，我们制备了负载GM-CSF的3D径向排列纳米纤维支架，然后在人源化小鼠中进行皮下植入。在每个指定时间点收集整个支架及其周围组织。最后，通过组织学观察检测细胞浸润和局部免疫反应，包括苏木精-伊红（H&E）和Masson染色以及免疫化学。我们发现在皮下植入后，3D径向排列的纳米纤维支架内有显著的细胞迁移和细胞外基质沉积。局部释放的GM-CSF在皮下植入后仅3天就能加速人类树突状细胞（CD11c）的表达。此外，在负载GM-CSF的径向排列纳米纤维支架及其周围组织中检测到人类细胞毒性T细胞（CD3/CD8）、M1巨噬细胞（CD68/CCR7）的表达更高。3D径向排列的支架可以加速细胞从周围组织迁移到伤口区域进行再生。局部释放的GM-CSF增强了树突状细胞的募集以及细胞毒性T细胞和M1巨噬细胞的激活。综上所述，负载GM-CSF的3D径向排列纳米纤维支架在实现组织再生方面具有广阔的应用前景。
