Shanghai Burn Institute, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Int Wound J. 2020 Oct;17(5):1428-1438. doi: 10.1111/iwj.13423. Epub 2020 Jun 9.
Negative pressure wound therapy (NPWT) has been widely used in various lesions. This study aimed to explore the biological effects of negative pressure on the polymorphonuclear neutrophils (PMNs), macrophages, and epidermal keratinocyte cells involved in wound healing. PMNs differentiated from HL-60, macrophages were derived from THP-1 monocytes, and keratinocytes were cultured in vitro, and they were treated with 0, -0.03 mp, and -0.05 mp, respectively. Cell ultrastructure; viability; apoptosis; and protein factors such as tumour necrosis factor-α (TNF-α), interferon-γ (IFN-γ), epidermal growth factor (EGF), epidermal growth factor receptor (EGFR), interleukin-17 (IL-17), and cell division cycle 42 (Cdc42) were determined by transmission electron microscopy (TEM), CCK8, flow cytometry (FCM), ELISA, and simple Western assays, respectively. After negative pressure stimulation, the cell ultrastructure of PMNs and macrophages cells was presented with a marked increase of lysosomes and a relative decrease of mitochondria. In addition, the cell viability was enhanced in PMNs and macrophages in a pressure-dependent manner and apoptosis ratios were significantly reduced in PMNs and macrophages. In addition, under -0.05 negative pressure, IFN-γ and IL-17 were significantly increased in PMNs or macrophages. Moreover, increased EGF and EGFR and Cdc42 levels in keratinocytes induced by the -0.05 mpa were detected, indicating that the migration chemotaxis of keratinocyte cells was enhanced. Negative pressure might promote cell proliferation, accelerate inflammatory responses, and promote epithelialisation during wound healing by increasing IFN-γ, IL-17, Cdc42, EGF, and EGFR in PMNs, macrophages, or keratinocytes under different negative pressures.
负压伤口疗法（NPWT）已广泛应用于各种病变。本研究旨在探讨负压对参与伤口愈合的多形核粒细胞（PMN）、巨噬细胞和表皮角质形成细胞的生物学效应。PMN 由 HL-60 分化而来，巨噬细胞由 THP-1 单核细胞衍生而来，角质形成细胞在体外培养，并分别用 0、-0.03mp 和-0.05mp 处理。采用透射电子显微镜（TEM）、CCK8、流式细胞术（FCM）、酶联免疫吸附试验（ELISA）和简单 Western 分析分别测定细胞超微结构、活力、凋亡以及肿瘤坏死因子-α（TNF-α）、干扰素-γ（IFN-γ）、表皮生长因子（EGF）、表皮生长因子受体（EGFR）、白细胞介素-17（IL-17）和细胞分裂周期蛋白 42（Cdc42）等蛋白因子。负压刺激后，PMN 和巨噬细胞的细胞超微结构呈现出溶酶体明显增加和线粒体相对减少的特征。此外，PMN 和巨噬细胞的细胞活力呈压力依赖性增强，PMN 和巨噬细胞的凋亡比例显著降低。此外，在-0.05 负压下，PMN 或巨噬细胞中的 IFN-γ和 IL-17 显著增加。此外，还检测到-0.05mpa 诱导的角质形成细胞中 EGF 和 EGFR 以及 Cdc42 水平增加，表明角质形成细胞的迁移趋化性增强。通过在不同负压下增加 PMN、巨噬细胞或角质形成细胞中的 IFN-γ、IL-17、Cdc42、EGF 和 EGFR，负压可能通过促进细胞增殖、加速炎症反应和促进上皮化来促进伤口愈合。
