Effect of extracellular matrix extract of decellularized extracellular matrix material on immune characteristics of RAW264.7 macrophages

Objective This study aims to investigate how extracellular matrix material extract affects macrophage activity, migration, phagocytosis, the expression of proinflammatory factors, and ROS production.Methods We assessed the effect of dECM extract on macrophage activity using the CCK-8 method under a light microscope. We used the Transwell migration assay to evaluate the effects of dECM extracts on the recruitment and chemotaxis of macrophages. pHrodo staining was employed to examine the impact of dECM extracts on macrophage phagocytosis. We utilized RT-qPCR and immunofluorescence staining to assess how dECM extract affects the expression of proinflammatory genes in macrophages. Additionally, we examined the expression of specific proinflammatory functional molecules. The DCFH-DA fluorescent probe method was used to detect the effect of dECM material extract on ROS production in macrophages.Results Compared with DMEM complete medium, CCK-8 results showed that dECM material extract could increase the activity of macrophages; RT-qPCR results indicated that dECM extracts could down-regulate the gene expression of proinflammatory cytokines and specific functional molecules; Flow cytometry results demonstrated that the extract of dECM material could reduce the production of ROS by macrophages. The results indicated that the dECM material extract exhibited favorable compatibility and beneficial biological properties for macrophages.

### 研究目的
本研究旨在探讨细胞外基质材料提取物对巨噬细胞活性、迁移能力、吞噬作用、促炎因子表达以及活性氧（reactive oxygen species，ROS）生成的影响。
### 研究方法
本研究采用CCK-8法结合光学显微镜，评估脱细胞细胞外基质（decellularized extracellular matrix，dECM）提取物对巨噬细胞活性的影响；通过Transwell迁移实验，评价dECM提取物对巨噬细胞募集与趋化作用的效果；采用pHrodo染色法，检测dECM提取物对巨噬细胞吞噬功能的影响；利用RT-qPCR与免疫荧光染色，评估dECM提取物对巨噬细胞促炎基因的表达调控作用；此外，本研究还检测了特定促炎功能分子的表达水平；采用DCFH-DA荧光探针法，检测dECM材料提取物对巨噬细胞ROS生成的影响。
### 研究结果
与DMEM完全培养基对照组相比，CCK-8检测结果显示，dECM材料提取物可提升巨噬细胞活性；RT-qPCR结果表明，dECM提取物能够下调促炎细胞因子及特定功能分子的基因表达水平；流式细胞术结果证实，dECM材料提取物可降低巨噬细胞的ROS生成量。综合上述结果，dECM材料提取物对巨噬细胞表现出良好的生物相容性与有益的生物学特性。