Banque de Tissus et Cellules, Hospices Civils de Lyon and LBTI, UMR 5305, Lyon, France,
Pierre Fabre, R&D PFDC, Département Pharmacologie, Toulouse, France,
Skin Pharmacol Physiol. 2020;33(1):17-29. doi: 10.1159/000503154. Epub 2019 Dec 18.
Among in vitro skin models, the scaffold-free skin equivalent (SFSE), without exogenous material, is interesting for pharmacotoxicological studies. Our aim was to adapt in vivo biophysical methods to study the structure, thickness, and extracellular matrix of our in vitro model without any chemical fixation needed as for histology.
We evaluated 3 batches of SFSE and characterized them by histology, transmission electron microscopy (TEM), and immunofluorescence. In parallel, we investigated 3 biophysical methods classically used for in vivo evaluation, optical coherence tomography (OCT), and laser scanning microscopy (LSM) imaging devices as well as the cutometer suction to study the biomechanical properties.
OCT allowed the evaluation of SFSE total thickness and its different compartments. LSM has a greater resolution enabling an evaluation at the cell scale and the orientation of collagen fibers. The viscoelasticity measurement by cutometry was possible on our thin skin model and might be linked with mature collagen bundles visible in TEM and LSM and with elastic fibers seen in immunofluorescence.
Our data demonstrated the simplicity and sensitivity of these different in vivo biophysical devices on our thin skin model. These noninvasive tools allow to study the morphology and the biomechanics of in vitro models.
在体外皮肤模型中,无外源性材料的无支架皮肤等效物(SFSE)对于药理毒理学研究很有趣。我们的目的是调整体内生物物理方法,以研究我们的体外模型的结构、厚度和细胞外基质,而无需像组织学那样进行任何化学固定。
我们评估了 3 批 SFSE,并通过组织学、透射电子显微镜(TEM)和免疫荧光进行了表征。同时,我们研究了 3 种经典的用于体内评估的生物物理方法,即光学相干断层扫描(OCT)和激光扫描显微镜(LSM)成像设备以及皮肤弹性测量仪的吸力,以研究生物力学特性。
OCT 允许评估 SFSE 的总厚度及其不同隔室。LSM 具有更高的分辨率,能够在细胞尺度上进行评估,并能够观察胶原纤维的取向。皮肤弹性测量仪的粘弹性测量在我们的薄皮模型上是可行的,并且可能与 TEM 和 LSM 中可见的成熟胶原束以及免疫荧光中可见的弹性纤维有关。
我们的数据证明了这些不同的体内生物物理设备在我们的薄皮模型上的简单性和敏感性。这些非侵入性工具允许研究体外模型的形态和生物力学。