Mineralized collagen (MC) is a biomaterial that is commonly used in the treatment of bone defects. However, the inflammatory response after biomaterial implantation is a recurrent problem that requires urgent attention. Our previous studies on MC-macrophage interactions were descriptive but we did not perform an in-depth analysis on a genetic level to investigate the underlying mechanisms. In this study, we cultured RAW264.7 cells on MC or collagen and examined the proliferation of the macrophages by Cell Counting Kit-8 assay. We sequenced the RNA of the cultured cells to discover differential gene expression patterns and found that a total of 1183 genes were differentially expressed between the MC- and collagen-cultured groups, of which 396 genes were upregulated and 787 were downregulated. Gene ontology analysis revealed that biological processes in MC-cultured cells, such as inflammation and innate immunity, were downregulated; whereas nucleosome assembly, megakaryocyte differentiation, and chromatin assembly were upregulated. We identified several pathways associated with immunity that were significantly enriched using the Kyoto Encyclopedia of Genes and Genomes. Furthermore, we validated the differentially expressed genes from RNA sequencing by quantitative real-time polymerase chain reaction. This study provides insight into the macrophage phenotype based on the microenvironment, which is the foundation for the clinical application of MC-based interventions.

中文翻译：

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矿化胶原蛋白巨噬细胞炎症和免疫基因表达谱

矿化胶原蛋白 (MC) 是一种常用于治疗骨缺损的生物材料。然而，生物材料植入后的炎症反应是一个需要紧急关注的反复出现的问题。我们之前对 MC-巨噬细胞相互作用的研究是描述性的，但我们没有在遗传水平上进行深入分析以研究潜在机制。在这项研究中，我们在 MC 或胶原蛋白上培养 RAW264.7 细胞，并通过 Cell Counting Kit-8 测定法检查巨噬细胞的增殖。我们对培养细胞的 RNA 进行测序以发现差异基因表达模式，发现在 MC 和胶原培养组之间共有 1183 个基因差异表达，其中 396 个基因上调，787 个下调。基因本体分析显示，MC 培养细胞中的生物过程，如炎症和先天免疫，被下调；而核小体组装、巨核细胞分化和染色质组装被上调。我们确定了几种与免疫相关的途径，这些途径使用京都基因和基因组百科全书得到了显着丰富。此外，我们通过定量实时聚合酶链反应验证了来自 RNA 测序的差异表达基因。本研究提供了基于微环境对巨噬细胞表型的洞察，这是基于 MC 的干预措施临床应用的基础。和染色质组装被上调。我们确定了几种与免疫相关的途径，这些途径使用京都基因和基因组百科全书得到了显着丰富。此外，我们通过定量实时聚合酶链反应验证了来自 RNA 测序的差异表达基因。本研究提供了基于微环境对巨噬细胞表型的洞察，这是基于 MC 的干预措施临床应用的基础。和染色质组装被上调。我们确定了几种与免疫相关的途径，这些途径使用京都基因和基因组百科全书得到了显着丰富。此外，我们通过定量实时聚合酶链反应验证了来自 RNA 测序的差异表达基因。本研究提供了基于微环境对巨噬细胞表型的洞察，这是基于 MC 的干预措施临床应用的基础。