Ghaffari Abdi, Jalili Reza, Ghaffari Mazyar, Miller Chris, Ghahary Aziz
Wound Healing Lab, Department of Surgery, Vancouver, British Columbia, Canada.
Wound Repair Regen. 2007 May-Jun;15(3):368-77. doi: 10.1111/j.1524-475X.2007.00239.x.
Bacterial burden significantly interferes with the healing process in chronic ulcers. Nitric oxide (NO) plays a key role in regulating skin's response to infection and wound healing. In previous studies, we demonstrated that exogenous NO gas (gNO) at 200 parts per million (ppm) exhibits potent antimicrobial effects against a representative range of pathogens. The aim of the present study is to explore the antimicrobial properties of gNO in vivo and to determine skin cells' sensitivity to the cytotoxic effects of gNO. To test gNO's antimicrobial effects, full-thickness wounds were infected with Staphylococcus aureus on the dorsal skin surface of New Zealand White rabbit and treated with 200 ppm gNO for 8 hours/day for 3 consecutive days. Significant reduction in wound bacterial content was observed in the presence of gNO. In a separate experiment, primary cultures of human fibroblasts, keratinocytes, and endothelial cells were established to test gNO's cytotoxicity in the skin. Methyl thiazolyl tetrazolium proliferation assays demonstrated that human skin cells, unlike bacterial cells, exhibited significant resistance toward gNO cytotoxicity. In vitro migration studies on keratinocytes and endothelial cells revealed that gNO treatment does not seem to interfere with reepithelialization and angiogenesis during the process of wound healing. Following 24 hours of gNO treatment, fibroblasts expressed significantly higher levels of procollagen and, to a lesser degree, a decrease in matrix metalloproteinase -1 mRNA. In conclusion, the present study provides evidence for the potential application of high doses of gNO as an antimicrobial agent for the treatment of infection in chronic nonhealing ulcers or burn patients, without compromising the viability, and function of skin cells.
细菌负荷严重干扰慢性溃疡的愈合过程。一氧化氮(NO)在调节皮肤对感染的反应和伤口愈合中起关键作用。在先前的研究中,我们证明200 ppm的外源性NO气体(gNO)对一系列代表性病原体具有强大的抗菌作用。本研究的目的是探索gNO在体内的抗菌特性,并确定皮肤细胞对gNO细胞毒性作用的敏感性。为了测试gNO的抗菌效果,在新西兰白兔背部皮肤表面用金黄色葡萄球菌感染全层伤口,并用200 ppm gNO连续3天每天处理8小时。在存在gNO的情况下,观察到伤口细菌含量显著降低。在另一项实验中,建立了人成纤维细胞、角质形成细胞和内皮细胞的原代培养物,以测试gNO在皮肤中的细胞毒性。甲基噻唑基四氮唑增殖试验表明,与细菌细胞不同,人皮肤细胞对gNO细胞毒性表现出显著抗性。对角质形成细胞和内皮细胞的体外迁移研究表明,gNO处理似乎不会干扰伤口愈合过程中的再上皮化和血管生成。在gNO处理24小时后,成纤维细胞表达的前胶原水平显著升高,基质金属蛋白酶-1 mRNA水平略有下降。总之,本研究为高剂量gNO作为抗菌剂用于治疗慢性不愈合溃疡或烧伤患者感染的潜在应用提供了证据,且不损害皮肤细胞的活力和功能。