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Purpose: This study was designed to determine whether a novel anti-melanogenic agent, PF3758309, has the potential to cause acute cutaneous irritation using a human skin equivalent model(HSEM). Methods: Human skin equivalent was constructed through incubation of normal human derived epidermal keratinocytes(HEKs) on collagen matrix insert with proliferation media. Using constructed human skin equivalent, the irritation potential of PF3758309 were investigated whether the viability of this agent-treated HESM is under 50%(irritant) or not(non-irritant) using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay after applying the agent to the epidermal surface of the HSEM. Also, the PF3758309-mediated anti-pigmentation effects were analyzed using Fontana-Masson staining in the HSEM. Results: The integrity of the constructed HSEM was confirmed using immunohistochemical staining with differentiation markers of epidermis, and observed that Keratin 5, Keratin 1 and Involucrin were stained in basal, supra-basal and granular layers, respectively. In vitro irritation assay showed that the mean viabilities of the PF3758309 was 83.6%, 78.8% and 77.8% at the treatment doses of 0.2, 0.5 and 1 mg, respectively;however, the mean viability of the positive control(5% sodium dodecyl sulfate)-treated HESM was 2.8%. Also, in vitro corrosion assay showed that the mean viabilities of the PF3758309 was 95.3%, 95.0% and 94.2% at the treatment doses of 0.2, 0.5 and 1 mg, respectively. Furthermore, using a Fontana-Masson staining assay, the melanin levels in the PF3758309-treated HSEM was significantly decreased compared with the levels in control HSEM. Conclusion: The anti-melanogenic agent, PF3758309, has no skin irritation potential under the conditions used in this study.

关键词： Human skin equivalent PF3758309 Irritation Skin Anti-melanogenesis

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In Vitro Assessment of Irritation Potential of an Anti-Melanogenic Agent,PF3758309,in a Reconstructed Human Epidermis Model

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北京日化 2018年第4期 33-40页

作者： ayeong lee Yu Rim lee Ji Yea Kim Jin Hyuk Jung Young Kum Park Yu-Jung Kim Sung-Nae lee Sungkwan An Seunghee Bae Research Institute for Molecular-Targeted Drugs Department of Cosmetics EngineeringKonkuk UniversitySeoulKorea Korea Institute of Dermatological Sciences SeoulKorea Gragem Corporation Cheongju-siChuncheongbuk-doKorea Department of Beauty Yakson Care Yeoju Institute of TechnologyYeoju-siGyeongi-doKorea Department of Cosmetology Kyung-In Women's UniversityIncheonKorea

Purpose:This study was designed to determine whether a novel anti-melanogenic agent,PF3758309,has the potential to cause acute cutaneous irritation using a human skin equivalent model (HSEM).Methods:Human skin equivalent was constructed through incubation of normal human derived epidermal keratinocytes (HEKs)on collagen matrix insert with proliferation *** constructed human skin equivalent,the irritation potential of PF3758309 were investigated whether the viability of this agent-treated HESM is under 50%(irritant)or not (non-irritant)using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)assay after applying the agent to the epidermal surface of the ***,the PF3758309-mediated anti-pigmentation effects were analyzed using Fontana-Masson staining in the ***:The integrity of the constructed HSEM was confirmed using immunohistochemical staining with differentiation markers of epidermis,and observed that Keratin 5,Keratin I and Involucrin were stained in basal,supra-basal and granular layers,*** vitro irritation assay showed that the mean viabilities of the PF3758309 was 83.6%,78.8%and 77.8%at the treatment doses of 0.2,0.5 and I mg,respectively;however,the mean viability of the positive control (5%sodium dodecyl sulfate)-treated HESM was 2.8%.Also,in vitro corrosion assay showed that the mean viabilities of the PF3758309 was 95.3%,95.0%and 94.2% at the treatment doses of 0.2,0.5 and 1 mg,***,using a Fontana-Masson staining assay,the melanin levels in the PF3758309-treated HSEM was significantly decreased compared with the levels in control ***: The anti-melanogenic agent,PF3758309,has no skin irritation potential under the conditions used in this study.

关键词： Human skin equivalent PF3758309 Irritation,Skin Anti-melanogenesis

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