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采用微生物活检和共聚焦显微镜技术,基于氧化应激水平,建立微创临床模型测试防晒霜毒性——概念验证研究。

A minimally invasive clinical model to test sunscreen toxicity based on oxidative stress levels using microbiopsy and confocal microscopy - a proof of concept study.

机构信息

Future Industries Institute, University of South Australia, Adelaide, Australia.

The University of Queensland Diamantina Institute, School of Medicine, The University of Queensland, Dermatology Research Centre, Brisbane, Australia.

出版信息

Int J Cosmet Sci. 2020 Oct;42(5):462-470. doi: 10.1111/ics.12646. Epub 2020 Aug 30.

DOI:10.1111/ics.12646
PMID:32619281
Abstract

OBJECTIVE

This proof-of-concept study demonstrated that using minimally invasive skin microsampling could enable significantly higher throughput of cosmetic testing in volunteers than conventional biopsy. Nanoparticle sunscreen was used as a model to test toxicity based on oxidative stress using microbiopsy and confocal imaging.

METHODS

Six volunteers were recruited for this study (3 males and 3 females). Zinc oxide nanoparticle containing topical formulation was prepared at 10% w/v. Each volunteer had 3 areas of 4 cm each mapped on each inner forearm for a total of 6 treatment areas (intact/ tape-stripped and with/without treatment). The topical zinc-nanoparticle formulation was applied directly to volunteer skin (2mg/cm ) for 2 hrs. Microbiopsied tissue from each treatment group was stained for reactive oxygen and nitrogen species in addition to mitochondrial superoxide. The stained samples were then imaged using confocal microscopy prior to image analysis.

RESULTS

Skin exposed to zinc oxide nanoparticles did not show any significant increases in oxidative stress. Zinc oxide nanoparticle tape-stripped skin resulted in signal significantly lower (P < 0.001) oxidative stress levels than t-butylated hydroxytoluene treated tape-stripped skin for oxidative stress markers. Topically applied zinc oxide nanoparticles had no detectable effect on the oxidative status in volunteer skin. No adverse reactions or effects were observed after all treatments including microbiopsy.

CONCLUSION

The data support the hypothesis that microbiopsy is a viable approach to study cosmeceutical- skin interactions in volunteers with capacity for molecular assays and high throughput with very low risk to the volunteer.

摘要

目的

本概念验证研究表明,使用微创皮肤微采样可以比传统活检显著提高志愿者的化妆品测试通量。纳米粒子防晒霜被用作模型,通过微生物活检和共焦成像来测试基于氧化应激的毒性。

方法

本研究招募了 6 名志愿者(3 名男性和 3 名女性)。制备了 10%w/v 的含氧化锌纳米颗粒的局部制剂。每位志愿者的每个内前臂有 3 个 4cm 区域,总共 6 个治疗区域(完整/胶带剥离和有/无治疗)。将局部锌纳米粒子制剂直接涂于志愿者皮肤(2mg/cm )2 小时。对每个治疗组的微生物活检组织进行活性氧和氮物种以及线粒体超氧化物的染色。染色后的样本使用共焦显微镜进行成像,然后进行图像分析。

结果

暴露于氧化锌纳米颗粒的皮肤未显示任何氧化应激的显著增加。与 t-丁基对苯二酚处理的胶带剥离皮肤相比,氧化锌纳米粒子胶带剥离皮肤的氧化应激标志物的氧化应激水平显著降低(P<0.001)。局部应用的氧化锌纳米粒子对志愿者皮肤的氧化状态没有可检测的影响。所有治疗包括微生物活检后,均未观察到不良反应或影响。

结论

数据支持这样一种假设,即微生物活检是一种可行的方法,可以在志愿者中研究化妆品-皮肤相互作用,具有分子检测的能力和高通量,对志愿者的风险非常低。

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