Arai Kenichi, Okabe Motonori, Kobashi Daisuke, Ichimura Kenji, Fathy Moustafa, Oba Jiro, Furuichi Etsuko, Yoshida Satoshi, Yoshida Toshiko
Department of Clinical Biomaterial Applied Science, Faculty of Medicine, University of Toyama, Toyama, Japan.
Department of Systems Function and Morphology, Faculty of Medicine, University of Toyama, Toyama, Japan.
J Burn Care Res. 2023 Jan 5;44(1):146-157. doi: 10.1093/jbcr/irac161.
Wound healing evaluation methods in a third-degree burn injury model are categorized as histological (re-epithelialization and granulation tissue formation) and molecular (quantitative polymerase chain reaction). In general, mRNA expression is normalized to those of the housekeeping gene. Although the housekeeping gene expression is generally stable, it has been reported that the stability of these genes depends on the wound healing process and treatment method. In this study, we identified the most stable housekeeping gene (TATA-binding protein) for studying gene expression in a third-degree burn injury model, in which wound healing was promoted by grafting human amnion-derived mesenchymal cells. We investigated the wound healing effect of human amnion-derived mesenchymal cells in the injury model. The formation of granulation tissue, the differentiation from fibroblasts to myofibroblasts, and functional vascular structure were promoted in the full-thickness skin excision site by treatment with these cells. The expression of angiogenic, pro-inflammatory and anti-inflammatory related mRNA was measured and normalized to that of the housekeeping gene, showing that treatment with the cells promoted the infiltration of endothelial cells and differentiation of M1 and M2 macrophages. In conclusion, wound healing in a third-degree burn injury model can be accurately analyzed using the optimized housekeeping gene.
三度烧伤损伤模型中的伤口愈合评估方法分为组织学方法（再上皮化和肉芽组织形成）和分子学方法（定量聚合酶链反应）。一般来说，mRNA表达量会以内参基因的表达量进行标准化。尽管内参基因的表达通常是稳定的，但据报道，这些基因的稳定性取决于伤口愈合过程和治疗方法。在本研究中，我们确定了在三度烧伤损伤模型中研究基因表达时最稳定的内参基因（TATA结合蛋白），该模型通过移植人羊膜间充质细胞促进伤口愈合。我们在该损伤模型中研究了人羊膜间充质细胞的伤口愈合效果。通过用这些细胞进行处理，全层皮肤切除部位的肉芽组织形成、成纤维细胞向肌成纤维细胞的分化以及功能性血管结构均得到促进。测量了血管生成、促炎和抗炎相关mRNA的表达，并以内参基因的表达进行标准化，结果表明用这些细胞处理可促进内皮细胞的浸润以及M1和M（此处原文有误，推测是M2）巨噬细胞的分化。总之，使用优化的内参基因可以准确分析三度烧伤损伤模型中的伤口愈合情况。
