Use of reconstructed skin model to assess the photoprotection afforded by three sunscreen products having different SPF values against DNA lesions and cellular alterations

D. Lelièvre , F. Canivet , F. Thillou , C. Tricaud , C. Le Floc'h , F. Bernerd
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Abstract

Introduction

Both UVB and UVA rays induce biological damages in the epidermis and the dermis that contribute to photo-carcinogenesis and photoaging. In the present study, the photoprotective effect of 2 ISO standard sunscreens, P3 (Sun Protection Factor [SPF]15) and P6 (SPF40) and of an SPF50+ labeled commercial sunscreen product was tested in reconstructed skin tissues exposed to increasing doses of UV Solar Simulated Radiation (UV-SSR). UV-induced damages were evaluated using several biological markers, including DNA lesions in the presence or absence of sunscreen protection.

Method

T-Skin™ model samples (EPISKIN), composed of a fibroblast-populated dermal equivalent and a fully differentiated epidermis, were protected with the test sunscreens (1.3 mg/cm² topically applied on molded polymethyl methacrylate plate) before being exposed to increasing UV doses (0 – 2.5 – 5 - 25 – 40 J/cm²). Twenty-four hours after exposure, tissues with and without sunscreen protection, were analyzed for skin viability and morphology, DNA lesions (cyclobutane pyrimidine dimer) and inflammatory mediator quantification. Results were compared to untreated exposed tissues using a Wilcoxon non-parametric test.

Results

For untreated tissues, UV-SSR exposure induced a dose-dependent decrease in epidermal and dermal viabilities, an increase in release of proinflammatory cytokines and matrix metalloproteinases and were associated with morphological damages at doses as low as 5 J/cm2. DNA lesions were even detected at the lowest dose of 2.5 J/cm2, and their number increased with the UV-SSR dose. In the samples protected with sunscreens, these abnormalities were partially or totally prevented with P6 providing a better protection compared to P3, and the SPF50+ sunscreen showing a trend for better protection than P6, for example against DNA damage.

Conclusions

This study demonstrates that photoprotective effects of different sunscreens can be discriminated and ranked on reconstructed skin tissues (T-Skin™ model) exposed to UV-SSR. Showing significant differences between the reference products P3 and P6 in line with their respective SPF values, such study allows the evaluation of epidermal and dermal damages at the tissue, cellular and molecular levels. It thus opens the way to a new model of integrated assessment of sunscreens. In line with its labeled 50+ SPF, the commercial test product confirmed its improved protection especially on DNA damage prevention.

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利用重建皮肤模型评估三种不同SPF值的防晒产品对DNA损伤和细胞改变的光保护作用
UVB和UVA射线都会引起表皮和真皮层的生物损伤,从而导致光致癌和光老化。在本研究中,测试了2种ISO标准防晒霜,P3(防晒系数[SPF]15)和P6 (SPF40)以及SPF50+标记的商业防晒霜产品在暴露于增加剂量的紫外线太阳模拟辐射(UV- ssr)下的重建皮肤组织中的光防护效果。使用几种生物标记来评估紫外线引起的损伤,包括在有或没有防晒霜保护的情况下的DNA损伤。方法t- skin™模型样品(EPISKIN)由成纤维细胞填充的真皮等量物和完全分化的表皮组成,在暴露于增加的紫外线剂量(0 - 2.5 - 5 - 25 - 40 J/cm²)之前,使用测试防晒霜(1.3 mg/cm²局部涂在模制的聚甲基丙烯酸甲酯板上)进行保护。暴露24小时后,对有和没有防晒霜保护的组织进行皮肤活力和形态学、DNA损伤(环丁烷嘧啶二聚体)和炎症介质定量分析。使用Wilcoxon非参数检验将结果与未处理的暴露组织进行比较。结果在未处理的组织中,低至5 J/cm2的UV-SSR暴露诱导表皮和真皮活力呈剂量依赖性下降,促炎细胞因子和基质金属蛋白酶的释放增加,并与形态学损伤相关。在最低剂量为2.5 J/cm2时也能检测到DNA损伤,且损伤数量随UV-SSR剂量的增加而增加。在受防晒霜保护的样本中,P6提供了比P3更好的保护,而SPF50+防晒霜显示出比P6更好的保护趋势,例如防止DNA损伤。结论不同防晒剂对暴露于UV-SSR的重建皮肤组织(T-Skin™模型)的光保护作用可以区分和分级。参考产品P3和P6的SPF值存在显著差异,该研究可以在组织、细胞和分子水平上评估表皮和真皮损伤。因此,它开辟了一种综合评估防晒霜的新模式。商业测试产品的SPF值为50+,证实其保护效果更好,尤其是在防止DNA损伤方面。
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