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pH 中性超氧化溶液对人皮肤成纤维细胞的体外作用

Effects of pH-neutral, super-oxidised solution on human dermal fibroblasts in vitro.

作者信息

González-Espinosa Diana, Pérez-Romano Leafar, Guzmán-Soriano Beatriz, Arias Edmundo, Bongiovanni Cheryl M, Gutiérrez Andrés A

机构信息

Cell Therapy Unit, Instituto Nacional de Rehabilitación, Mexico City, México.

出版信息

Int Wound J. 2007 Sep;4(3):241-50. doi: 10.1111/j.1742-481X.2007.00331.x.

Abstract

For the past 20 years, super-oxidised solutions (SOSs) have been shown to be potent antimicrobials and disinfectants via oxidative damage. However, the potential toxicity of SOSs on eukaryotic cells has not been documented in vitro. This is relevant because oxygen and chlorine reactive species may possibly induce ageing and irreversible cellular dysfunctions that eventually produce cell death. The present study investigates the cytotoxicity and oxidative stress induced by a novel, pH-neutral SOS (i.e. Microcyn, MCN) on young, primary diploid - human dermal fibroblast (HDF) cultures. For this purpose, hydrogen peroxide (HP) was used as a positive control of oxidative damage. When these solutions were used at concentrations indicated for wound care (i.e. undiluted MCN or 880 mM HP), HP was significantly more toxic than MCN. After 5 and 30 minutes of exposure, cell viability was 38% and 5%, respectively, in 880 mM HP-treated cells versus 75% and 70% in MCN-treated populations, respectively. HP induced both apoptosis and necrosis, whereas MCN induced only necrosis. Genotoxic and ageing studies were then conducted at sublethal HP concentrations as previously reported in the literature. Cellular DNA and RNA were partially degraded only in HDFs exposed to 500 microM HP for 30 minutes but not in those exposed to undiluted MCN. At this same concentration, HP induced the formation of 8-hydroxy-2'deoxyguanosine adducts in HDFs but this effect was neither observed in control- nor observed in MCN-treated cells. HDFs were further exposed to 5 microM HP or 10% MCN for 1 month. The expression of senescence-associated-beta-galactosidase was only significantly elevated in cells chronically exposed to 5 microM HP. Altogether, these results show that MCN is significantly less cytotoxic than antiseptic HP concentrations (i.e. 880 mM) and that, in vitro, it does not induce genotoxicity or accelerated ageing.

摘要

在过去20年中,超氧化溶液(SOSs)已被证明是通过氧化损伤发挥作用的强效抗菌剂和消毒剂。然而,SOSs对真核细胞的潜在毒性在体外尚未见报道。这一点很重要,因为氧和氯活性物质可能会诱导衰老和不可逆的细胞功能障碍,最终导致细胞死亡。本研究调查了一种新型的pH中性SOS(即Microcyn,MCN)对年轻的原代二倍体人皮肤成纤维细胞(HDF)培养物的细胞毒性和氧化应激。为此,使用过氧化氢(HP)作为氧化损伤的阳性对照。当这些溶液以伤口护理指示的浓度使用时(即未稀释的MCN或880 mM HP),HP的毒性明显高于MCN。暴露5分钟和30分钟后,880 mM HP处理的细胞的细胞活力分别为38%和5%,而MCN处理的细胞群体的细胞活力分别为75%和70%。HP诱导了凋亡和坏死,而MCN仅诱导了坏死。然后按照文献中先前报道的方法,在亚致死性HP浓度下进行了遗传毒性和衰老研究。仅在暴露于500 microM HP 30分钟的HDF中,细胞DNA和RNA出现部分降解,而暴露于未稀释MCN的细胞中未出现这种情况。在相同浓度下,HP诱导HDF中8-羟基-2'-脱氧鸟苷加合物的形成,但在对照细胞和MCN处理的细胞中均未观察到这种效应。HDF进一步暴露于5 microM HP或10% MCN中1个月。衰老相关β-半乳糖苷酶的表达仅在长期暴露于5 microM HP的细胞中显著升高。总之,这些结果表明,MCN的细胞毒性明显低于防腐用HP浓度(即880 mM),并且在体外,它不会诱导遗传毒性或加速衰老。

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