Mast Cells Tryptase Promotes Intestinal Fibrosis in Natural Decellularized Intestinal Scaffolds
Mast Cells Tryptase Promotes Intestinal Fibrosis in Natural Decellularized Intestinal Scaffolds
复制标题
DOI：
10.1007/s13770-022-00433-9
复制
发表时间：
2022-02
影响因子：
3.6
通讯作者：
Jian Wan;Tianqi Wu;Ying Liu;Mu-qing Yang;J. Fichna;Yibing Guo;Lu Yin;Chunqiu Chen
中科院分区：
文献类型：
--
作者：
Jian Wan;Tianqi Wu;Ying Liu;Mu-qing Yang;J. Fichna;Yibing Guo;Lu Yin;Chunqiu Chen
背景标准二维 (2D) 培养已证实肥大细胞 (MC) 在炎症性肠病 (IBD) 发病机制中的机制，但 MC 信号反应的调节在三维 (3D) 微环境中可能有很大不同。本研究的目的是开发基于脱细胞肠支架（DIS）的3D培养模型，并在3D模型中验证MC如何影响成纤维细胞表型。方法使用去污剂技术实现DIS，并通过组织学分析、定量和扫描电镜验证细胞外基质（ECM）成分。将人结肠成纤维细胞重新细胞化到支架中并通过 MCs 类胰蛋白酶和 TGFβ1 激活后，研究了 3D 成纤维细胞激活过程中基因和信号通路的变化，并与塑料板上 2D 细胞培养的变化进行比较。结果脱细胞过程有效去除了天然细胞碎片，同时保留了天然 ECM 成分和结构。移植的成纤维细胞可以渗入支架并保持其表型。无论成纤维细胞是2D还是3D培养，MCs类胰蛋白酶和转化生长因子β1（TGF-β1）都可以通过Akt和Smad2/3信号通路促进成纤维细胞分化为纤维化表型肌成纤维细胞。此外，3D培养中肌成纤维细胞的前胶原1α1和纤连蛋白合成高于2D培养。结论我们的结果表明DIS可以作为肠道纤维化研究的生物活性微环境，为未来肠道疾病建模以及基因和信号通路的筛选提供创新平台。
BackgroundStandard two-dimensional (2D) culture has confirmed the mechanism of mast cells (MCs) in the pathogenesis of inflammatory bowel disease (IBD), but the regulation of signaling responses of MCs may well differ in three-dimensional (3D) microenvironments. The aim of the study was to develop a 3D culture model based on decellularized intestinal scaffolds (DIS) and verify how MCs influenced fibroblasts phenotype in the 3D model.MethodsDIS were achieved using the detergent technique and extracellular matrix (ECM) components were verified by histologic analysis, quantification and scanning electron microscope. After human colon fibroblasts recellularized into the scaffolds and activated by MCs tryptase and TGFβ1, the changes in genes and signaling pathways during fibroblasts activation in 3D were studied and compared with the changes in 2D cell culture on plastic plates.ResultsDecellularization process effectively removed native cell debris while retaining natural ECM components and structure. The engrafted fibroblasts could penetrate into the scaffolds and maintain its phenotype. No matter whether fibroblasts were cultured in 2D or 3D, MCs tryptase and transforming growth factor β1 (TGF-β1) could promote the differentiation of fibroblasts into fibrotic-phenotype myofibroblasts through Akt and Smad2/3 signaling pathways. Furthermore, the pro-collagen1α1 and fibronectin synthesis of myofibroblasts in 3D was higher than in 2D culture.ConclusionOur results demonstrated that the DIS can be used as a bioactive microenvironment for the study of intestinal fibrosis, providing an innovative platform for future intestinal disease modeling and screening of genes and signaling pathways.