Li Hui, Yang Hefeng, Ma Bo, Qiao Jia, Chen Fanfan, Wang Pinwen, Yu Riyue, Sun Jingjing, Chen Yuanwei
Department of Stomatology, Beijing Shijitan Hospital, Capital Medical University, Beijing, China.
Department of Prosthodontics, Kunming Medicine University School and Hospital of Stomatology, Kunming, China.
J Mater Chem B. 2025 Jul 30;13(30):9182-9202. doi: 10.1039/d5tb00474h.
Functional tissue repair is often constrained by inflammation and fibrosis. Alternatively activated M2 macrophages have emerged as promising therapeutic targets for optimizing graft-to-host interactions; however, efficient induction methods are required. Presumably, the outcome of regenerative wound healing or scar formation/fibrosis might be dependent on the balance between M2a and M2c sub-phenotypes. This study utilized dual-functionalized graphene oxide (GO) as a DNA delivery agent to induce M2a and M2c macrophage polarization. Mechanistically, molecular characteristics were analyzed using RNA sequencing. We designed GO with polyethyleneimine (PEI) modification and subsequently conjugated it with polyethylene glycol (PEG)-folate (FA) to target human THP-1-derived macrophage activation. Specifically, the resulting GO-PEI-PEG-FA (GPPF) compound effectively activated CD206CD209M2a and CD163MerTKM2c phenotype polarization. The efficient delivery of IL4 or IL10 plasmid DNA using GPPF (GPPF/pIL4 or GPPF/pIL10) significantly enhanced macrophage cellular elongation and reduced MHC-II-associated antigen presentation. M2a(GPPF/pIL4) and M2c(GPPF/pIL10) were validated as negative regulators of the immune response and positive regulators of Th2 effectors. Up-regulated genes in M2a(GPPF/pIL4) even inhibited type I interferon production and restricted the innate immune response. Supplemental to the established data, M2a(GPPF/pIL4) behaved similar to IFN-responsive macrophages, restricting viral life cycles and promoting myogenesis and osteogenesis. Meanwhile, M2c(GPPF/pIL10) was characterized using IL10 signaling, anti-fibrosis, and neutrophil-mediated suppression of the LPS-bacterial response. Regarding the tissue remodeling process, the two subsets attenuated negative-regulated BMP signaling to facilitate osteoinduction and up-regulated NAMPT to establish a transient stem cell-activating niche for tissue regeneration. This study underscored the potential of functionalized GO-induced M2 sub-phenotypes as modulators in regenerative medicine.
功能性组织修复常常受到炎症和纤维化的限制。交替活化的M2巨噬细胞已成为优化移植物与宿主相互作用的有前景的治疗靶点；然而，需要有效的诱导方法。据推测，再生性伤口愈合或瘢痕形成/纤维化的结果可能取决于M2a和M2c亚表型之间的平衡。本研究利用双功能化氧化石墨烯（GO）作为DNA递送剂来诱导M2a和M2c巨噬细胞极化。从机制上讲，使用RNA测序分析分子特征。我们设计了用聚乙烯亚胺（PEI）修饰的GO，随后将其与聚乙二醇（PEG）-叶酸（FA）偶联，以靶向人THP-1来源的巨噬细胞活化。具体而言，所得的GO-PEI-PEG-FA（GPPF）化合物有效地激活了CD206CD209 M2a和CD163MerTK M2c表型极化。使用GPPF（GPPF/pIL4或GPPF/pIL10）有效递送IL4或IL10质粒DNA显著增强了巨噬细胞的细胞伸长并减少了MHC-II相关的抗原呈递。M2a（GPPF/pIL4）和M2c（GPPF/pIL10）被证实为免疫反应的负调节因子和Th2效应器的正调节因子。M2a（GPPF/pIL4）中上调的基因甚至抑制I型干扰素的产生并限制先天性免疫反应。作为已建立数据的补充，M2a（GPPF/pIL4）的行为类似于IFN反应性巨噬细胞，限制病毒生命周期并促进肌生成和成骨。同时，M2c（GPPF/pIL10）通过IL10信号传导、抗纤维化和中性粒细胞介导的对LPS-细菌反应的抑制来表征。关于组织重塑过程，这两个亚群减弱了负调节的BMP信号传导以促进骨诱导，并上调NAMPT以建立用于组织再生的瞬时干细胞激活微环境。本研究强调了功能化GO诱导的M2亚表型作为再生医学调节剂的潜力。
