背景:皮肤伤口愈合代表了一种常见的基本现象,需要不同类型的细胞参与,并且是公众的主要关注点。有证据证实,使用近红外(NIR)的光生物调节(PBM)可以促进伤口愈合,但是所涉及的细胞和精确的分子机制仍然难以捉摸。
方法:在大鼠背部制作直径1.0cm的全层皮肤缺损,随机分为对照组,10J,15J,和30个J组。测量术后第4、8和12天的伤口愈合率。进行HE和Masson染色以揭示组织学特征。进行免疫荧光染色以标记表皮干细胞(ESC)和毛囊干细胞(HFSC)。Westernblot检测ESCs和HFSCs相关蛋白的表达。收集皮肤伤口组织用于RNA测序。进行了基因本体论和京都基因和基因组百科全书分析,使用CytoHubba鉴定了hub基因,并通过qRT-PCR进行了验证。
结果:PBM可以促进上皮再生,细胞外基质沉积,伤口愈合,增加KRT14+/PCNA+ESC和KRT15+/PCNA+HFSC的数量,并上调P63、Krt14和PCNA的蛋白表达。三百六十六个差异表达基因(DEGs)和7个hub基因,包括Sox9,Krt5,Epcam,Cdh1,Cdh3,Dsp,并鉴定了Pkp3。这些DEGs富含皮肤发育,细胞连接,和参与细胞-细胞粘附等的钙粘蛋白结合。,而这些hub基因与皮肤来源的干细胞和细胞粘附有关。
结论:PBM通过促进ESCs和HFSCs增殖,提高干细胞和细胞粘附相关基因的表达,增强上皮再生,从而加速伤口愈合。这可以通过调节皮肤来源的干细胞的增殖和调节与细胞粘附相关的基因来提供促进伤口愈合和再上皮化的有价值的替代策略。
Cutaneous wound healing represents a common fundamental phenomenon requiring the participation of cells of distinct types and a major concern for the public. Evidence has confirmed that photobiomodulation (PBM) using near-infrared (NIR) can promote wound healing, but the cells involved and the precise molecular mechanisms remain elusive.
Full-thickness skin defects with a diameter of 1.0 cm were made on the back of rats and randomly divided into the control group, 10 J, 15 J, and 30 J groups. The wound healing rate at days 4, 8, and 12 postoperatively was measured. HE and Masson staining was conducted to reveal the histological characteristics. Immunofluorescence staining was performed to label the epidermal stem cells (ESCs) and hair follicle stem cells (HFSCs). Western blot was performed to detect the expressions of proteins associated with ESCs and HFSCs. Cutaneous wound tissues were collected for RNA sequencing. Gene ontology and the Kyoto Encyclopedia of Genes and Genomes analysis was performed, and the hub genes were identified using CytoHubba and validated by qRT-PCR.
PBM can promote reepithelialization, extracellular matrix deposition, and wound healing, increase the number of KRT14+/PCNA+ ESCs and KRT15+/PCNA+ HFSCs, and upregulate the protein expression of P63, Krt14, and PCNA. Three hundred and sixty-six differentially expressed genes (DEGs) and 7 hub genes including Sox9, Krt5, Epcam, Cdh1, Cdh3, Dsp, and Pkp3 were identified. These DEGs are enriched in skin development, cell junction, and cadherin binding involved in cell-cell adhesion etc., while these hub genes are related to skin derived stem cells and cell adhesion.
PBM accelerates wound healing by enhancing reepithelialization through promoting ESCs and HFSCs proliferation and elevating the expression of genes associated with stem cells and cell adhesion. This may provide a valuable alternative strategy to promote wound healing and reepithelialization by modulating the proliferation of skin derived stem cells and regulating genes related to cell adhesion.