Regulation of extracellular matrix assembly and structure by hybrid M1/M2 macrophages.
Regulation of extracellular matrix assembly and structure by hybrid M1/M2 macrophages.
复制标题
混合M1/M2巨噬细胞对细胞外基质组装和结构的调节
DOI：
10.1016/j.biomaterials.2021.120667
复制
发表时间：
2021-03
期刊：
影响因子：
14
通讯作者：
Spiller KL
中科院分区：
文献类型：
--
作者：
Witherel CE;Sao K;Brisson BK;Han B;Volk SW;Petrie RJ;Han L;Spiller KL
植入生物材料周围的异常细胞外基质（ECM）组装是异物反应的标志，其中植入物被包封在厚纤维组织中，阻止其正常功能。虽然巨噬细胞是已知的成纤维细胞行为的调节剂，但其表型如何影响ECM组装和异物反应的进展知之甚少。在这项研究中，我们使用了生理相关的巨噬细胞表型的体外模型，以及从体内皮下植入的明胶水凝胶中控制释放巨噬细胞调节细胞因子，以研究巨噬细胞在ECM组装中的作用。原代人巨噬细胞被极化为四种不同的表型，每种表型都与纤维化相关，包括促炎性M1、促愈合M2和混合型M1/M2，混合型M1/M2是通过将巨噬细胞同时暴露于M1和M2促进刺激而产生的。此外，巨噬细胞首先极化为M1，然后极化为M2（M1→M2），以产生通常在正常伤口愈合期间观察到的表型。在巨噬细胞条件培养基中培养的人皮肤成纤维细胞上调了许多参与ECM组装调控的基因，特别是在M2条件培养基中。混合M1/M2巨噬细胞条件培养基导致成纤维细胞产生具有较厚和较少对齐的纤维的基质，而M2巨噬细胞条件培养基导致形成具有较薄纤维的更对齐的基质。明胶甲基丙烯酸酯水凝胶含有白细胞介素-4（IL-4）和IL-13加载的聚（乳酸-共-乙醇酸）（PLGA）微粒的设计，以促进小鼠皮下在体内模型中的M2表型。水凝胶外植体的NanoString多重基因表达分析显示，不含细胞因子的水凝胶导致大多数M1表型标志物在早期时间点（3天）高度表达，但IL 4 + IL 13的释放促进了M2标志物和与ECM组装调控相关的基因（如Col 5a 1和Col 6a 1）的上调。生化分析和二次谐波发生显微镜显示，IL 4 + IL 13的释放增加了总硫酸化糖胺聚糖含量并降低了原纤维排列，这通常与较少的纤维化组织相关。总之，这些结果表明，混合M1/M2巨噬细胞调节ECM组装，并且将平衡向M2转移可能促进ECM的结构和组成变化，并增强下游重塑的潜力。
Aberrant extracellular matrix (ECM) assembly surrounding implanted biomaterials is the hallmark of the foreign body response, in which implants become encapsulated in thick fibrous tissue that prevents their proper function. While macrophages are known regulators of fibroblast behavior, how their phenotype influences ECM assembly and the progression of the foreign body response is poorly understood. In this study, we used in vitro models with physiologically relevant macrophage phenotypes, as well as controlled release of macrophage-modulating cytokines from gelatin hydrogels implanted subcutaneously in vivo to investigate the role of macrophages in ECM assembly. Primary human macrophages were polarized to four distinct phenotypes, which have each been associated with fibrosis, including pro-inflammatory M1, prohealing M2, and a hybrid M1/M2, generated by exposing macrophages to M1- and M2-promoting stimuli simultaneously. Additionally, macrophages were first polarized to M1 and then to M2 (M1→M2) to generate a phenotype typically observed during normal wound healing. Human dermal fibroblasts that were cultured in macrophage-conditioned media upregulated numerous genes involved in regulation of ECM assembly, especially in M2-conditioned media. Hybrid M1/M2 macrophage-conditioned media caused fibroblasts to produce a matrix with thicker and less aligned fibers, while M2 macrophage-conditioned media caused the formation of a more aligned matrix with thinner fibers. Gelatin methacrylate hydrogels containing interleukin-4 (IL4) and IL13-loaded poly(lactic-co-glycolic acid) (PLGA) microparticles were designed to promote the M2 phenotype in a murine subcutaneous in vivo model. NanoString multiplex gene expression analysis of hydrogel explants showed that hydrogels without cytokines caused mostly M1 phenotype markers to be highly expressed at an early time point (3 days), but the release of IL4+IL13 promoted upregulation of M2 markers and genes associated with regulation of ECM assembly, such as Col5a1 and Col6a1. Biochemical analysis and second harmonic generation microscopy showed that the release of IL4+IL13 increased total sulfated glycosaminoglycan content and decreased fibril alignment, which is typically associated with less fibrotic tissue. Together, these results show that hybrid M1/M2 macrophages regulate ECM assembly, and that shifting the balance towards M2 may promote architectural and compositional changes in ECM with enhanced potential for downstream remodeling.
登录
查看更多内容
影响因子：
--
作者：
Bredfeldt JS;Liu Y;Conklin MW;Keely PJ;Mackie TR;Eliceiri KW
通讯作者：
Eliceiri KW
影响因子：
2.7
作者：
Benjamini, Yoav;Krieger, Abba M.;Yekutieli, Daniel
通讯作者：
Yekutieli, Daniel
影响因子：
6
作者：
Brisson, Becky K.;Mauldin, Elizabeth A.;Volk, Susan W.
通讯作者：
Volk, Susan W.
影响因子：
5
作者：
de Vries, HJC;Enomoto, DNH;Bos, JD
通讯作者：
Bos, JD
影响因子：
3.2
作者：
Digiacomo, Graziana;Tusa, Ignazia;Rovida, Elisabetta
通讯作者：
Rovida, Elisabetta