中性粒细胞胞外诱捕网的治疗靶向作用可改善小鼠的一期和二期愈合。
Therapeutic Targeting of Neutrophil Extracellular Traps Improves Primary and Secondary Intention Wound Healing in Mice.
机构信息
Department of Pediatric Surgery, University Medical Center Hamburg-Eppendorf, Hamburg, Germany.
Children's Medical Research Institute, Sydney University, Westmead, NSW, Australia.
出版信息
Front Immunol. 2021 Feb 25;12:614347. doi: 10.3389/fimmu.2021.614347. eCollection 2021.
BACKGROUND
Neutrophils are the first responders in wound healing after injury that mediate pro- and anti-inflammatory activities i.a. through the formation of extracellular traps (NETs). However, excessive NETs presence in wound tissue can cause local hyperinflammation and -coagulation resulting in delayed wound healing. To improve wound healing, we aimed to examine the role of NETs and DNase1 on primary and secondary wound healing.
METHODS
The study included 93 C57BL/6 mice, with 3 different genotypes: wildtype, Pad4-, and DNase1-Knockout (KO). Pad4-KO mice show limited NETs formation, while DNase1-KO mice cannot disintegrate them. All 3 genotypes were included in (1) a laparotomy group and (2) a thermal injury group. Animals in both groups either received DNase1 or a vehicle i.p. post wound induction and wound assessment and euthanasia were conducted. Laparotomy and burn scars were assessed using the stony brook scar evaluation scale and modified Yeong scale respectively. Tissue was analyzed histologically using H&E staining. Ly6g, Collagen I and III, SMA, and Fibrinogen were visualized and neutrophils activation (NE, MPO) and NETs (H3cit) formation assessed.
RESULTS
All animals survived with no complications. DNase1 treatment led to a significantly improved scar appearance in both groups, which was also seen in Pad4-KO mice. In the laparotomy group DNase1 improved collagen deposition and fibrin concentration was significantly reduced by DNase1 treatment. Markers of neutrophil activation were significantly reduced in the treatment and Pad4-KO group. In the thermal injury group wound closure time was significantly reduced after DNase1 treatment and in the Pad4-KO group. Even though inflammation remained high in the thermal injury model over time, neutrophil activation and NETs formation were significantly reduced by DNase1 treatment compared to controls.
DISCUSSION
Primary and secondary intention wound healing is improved by targeting NETs through DNase1 treatment or genetic KO, as assessed by wound closure time and scar appearances. Additionally, wound stability was not affected by DNASE treatment. The results suggest that overall wound healing is accelerated and DNase1 appears to be a promising option to reduce scar formation; which should be evaluated in humans.
背景
中性粒细胞是损伤后伤口愈合的最初反应者，通过形成细胞外陷阱（NETs）来介导促炎和抗炎活性。然而，伤口组织中过多的 NETs 会导致局部过度炎症和凝血，从而导致伤口愈合延迟。为了改善伤口愈合，我们旨在研究 NETs 和 DNase1 在原发性和继发性伤口愈合中的作用。
方法
该研究包括 93 只 C57BL/6 小鼠，具有 3 种不同的基因型：野生型、Pad4-和 DNase1 敲除（KO）。Pad4-KO 小鼠显示 NETs 形成受限，而 DNase1-KO 小鼠无法使其解体。所有 3 种基因型均包含在（1）剖腹术组和（2）热损伤组中。两组动物均在伤口诱导后接受 DNase1 或载体腹腔内注射，并进行伤口评估和安乐死。使用石溪疤痕评估量表和改良 Yeong 量表分别评估剖腹术和烧伤疤痕。使用 H&E 染色对组织进行组织学分析。可视化 Ly6g、胶原蛋白 I 和 III、SMA 和纤维蛋白原，并评估中性粒细胞活化（NE、MPO）和 NETs（H3cit）的形成。
结果
所有动物均存活且无并发症。DNase1 治疗在两组中均显著改善了疤痕外观，在 Pad4-KO 小鼠中也观察到了这一点。在剖腹术组中，DNase1 改善了胶原蛋白沉积，DNase1 处理显著降低了纤维蛋白浓度。治疗组和 Pad4-KO 组中中性粒细胞活化标志物显著减少。在热损伤组中，DNase1 治疗和 Pad4-KO 组的伤口闭合时间显著缩短。尽管热损伤模型中的炎症随时间推移仍然很高，但与对照组相比，DNase1 治疗显著降低了中性粒细胞活化和 NETs 的形成。
讨论
通过 DNase1 治疗或遗传 KO 靶向 NETs，可改善原发性和继发性伤口愈合，通过伤口闭合时间和疤痕外观进行评估。此外，DNASE 治疗并不影响伤口稳定性。结果表明，整体伤口愈合加快，DNase1 似乎是减少疤痕形成的有前途的选择；这应该在人类中进行评估。
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