Houcine A, Delalleau A, Heraud S, Guiraud B, Payre B, Duplan H, Delisle M-B, Damour O, Bessou-Touya S
Centre de Microscopie Electronique Appliquée à la Biologie, Faculté de Médecine Rangueil, Toulouse III, Université P. Sabatier, Toulouse, France.
Pixience, Toulouse, France.
Skin Res Technol. 2016 Aug;22(3):284-94. doi: 10.1111/srt.12259. Epub 2015 Oct 27.
The reliability of the biophysical properties of skin equivalents (SEs) remains a challenge for medical applications and for product efficacy tests following the European Directive 2003/15/EC2 on the prohibition of animal experiments for cosmetic products.
We propose to adapt the biophysical in vivo testing techniques to compare full thickness model growth vs. time. The interest in using such techniques lies in possible comparisons between in vivo and in vitro skin as well as monitoring samples over the culture time.
High frequency ultrasound technique, optical coherence tomography (OCT), and laser scanning microscopy were used to analyze SEs morphology at days D42 and D60 whereas their microstructure was assessed through transmission electron microscopy and classical histology. A correlation between these observations and mechanical measurements has been proposed so as to underline the consequence of both the development of the dermis elastic fibers and the epidermis differentiation.
Ultrasounds measurements show a highly homogeneous dermis whereas the OCT technique clearly distinguishes the stratum corneum and the living epidermis. The increase in the thicknesses of these layers as well as the growth in elastin and collagen fibers results in strong modifications of the samples mechanical properties.
对于医疗应用以及按照欧洲关于禁止化妆品动物实验的指令2003/15/EC2进行的产品功效测试而言，皮肤替代物（SEs）生物物理特性的可靠性仍然是一项挑战。
我们建议采用生物物理体内测试技术来比较全层模型的生长与时间的关系。使用此类技术的意义在于可以对体内和体外皮肤进行比较，以及在培养期间对样本进行监测。
在第42天和第60天使用高频超声技术、光学相干断层扫描（OCT）和激光扫描显微镜分析SEs的形态，而通过透射电子显微镜和经典组织学评估其微观结构。已提出这些观察结果与力学测量之间的相关性，以强调真皮弹性纤维发育和表皮分化的后果。
超声测量显示真皮高度均匀，而OCT技术能清晰区分角质层和活表皮。这些层厚度的增加以及弹性蛋白和胶原纤维的生长导致样本力学性能发生显著变化。
