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

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 I 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 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,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.

人体皮肤等效材料弹性性能的测试方法研究

针对皮肤等效材料力学性能的特异性,我们对人体皮肤等效材料弹性性能的测试方法进行了研究,提出静载压入试验法进行等效材料的弹性测试,并设计实验装置进行实验,实验结果表明该测试方法正确可行。

组织工程皮肤的现状和展望

皮肤损伤及修复是临床工作者面临的重大难题。自体皮肤移植因供体不足而受限,而异体移植则面临免疫排斥反应。利用组织工程技术制作组织工程皮肤因而成为有效的解决途径。随着种子细胞和支架材料等的不断完善,组织工程皮肤的基础研究将更加丰富,临床应用的前景更加广阔。本文从组织工程皮肤的结构、分类、应用、产品等方面对该领域的国内外最新研究现状进行综述,分析了组织工程皮肤目前存在的问题,并提出未来发展方向即再生出含有毛囊、汗腺等附属器官的完整有功能的人类皮肤,对今后组织工程皮肤的研究和应用具有一定的参考价值。

敷料与人工皮肤技术研究进展

综述了目前常见的烧伤敷料的类型、结构、性能指标、应用及进展 ,介绍了普通敷料、生物敷料与合成敷料 ,特别是合成敷料中的薄膜、泡沫海绵膜、凝胶膜的特点 ,并分析了目前正在发展的复合型敷料 ,特别是利用人体组织工程制备的具有生物活性的人工皮肤的发展趋势。

细胞共培养人体皮肤类似物超微结构观察及其渗透性测试

目的观察人体皮肤类似物模型形态结构，对其屏障功能进行初步评价。方法通过胶原蛋白包被成纤维细胞，制备真皮层，真皮层表面接种角质细胞，浸没培养1周后，气一液界面培养以促进角质细胞的分化形成角质层，3周后收获皮肤模型；通过肉眼观察、电镜观察皮肤类似物的宏观结构和超微结构；以吲哚美辛作为模型药物，进行体外透皮实验考察细胞共培养皮肤类似物模型的屏障功能。结果皮肤类似物形成与人类皮肤相似的真皮层和表皮层，表皮层表面形成完整的角质层，具有与人类皮肤类似的超微结构；24h内渗透性实验结果显示皮肤类似物屏障功能略低于大鼠腹部皮肤，但22—24h，皮肤类似物对模型药的透过速率远小于大鼠腹部皮肤（约为其28％），且2种皮肤模型对吲哚美辛经皮渗透的稳态流速没有显著性差异（P〉0．05）。结论皮肤类似物有与人体皮肤相类似的形态结构特征，与大鼠腹部皮肤相比对吲哚美辛有相似的屏障作用，有望替代动物皮肤成为经皮给药系统评价的新型皮肤模型。