Depertment of Oral and Maxillofacial Surgery, School and Hospital of Stomatology, Shandong University and Shandong Provincial Key Laboratory of Oral Tissue Regeneration and Shandong Engineering Laboratory for Dental Materials and Oral Tissue Regeneration, Jinan, Shandong, China.
Liaocheng People's Hospital, Medical College of Liaocheng University, Liaocheng, China.
J Biomed Mater Res A. 2021 Aug;109(8):1328-1336. doi: 10.1002/jbm.a.37124. Epub 2020 Nov 4.
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.
矿化胶原（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亟待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，但我们没有在基因水平上进行深入分析，以研究潜在的机制。在这项研究中，我们在 MC 或胶原上培养 RAW264.7 细胞，并通过细胞计数试剂盒-8 测定法检测巨噬细胞的增殖。我们对培养细胞的 RNA 进行测序，以发现差异表达基因的模式，发现 MC 培养组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 个基因下调。基因本体论分析显示，MC 培养细胞中的生物学过程，如炎症和先天免疫，下调；而核小体组装、巨核细胞分化和染色质组装上调。我们使用京都基因与基因组百科全书鉴定了几个与免疫相关的显著富集途径。此外，我们通过定量实时聚合酶链反应验证了 RNA 测序中差异表达的基因。这项研究基于微环境提供了对巨噬细胞表型的深入了解，为 MC 为基础的干预措施的临床应用奠定了基础。
