Tokuyama Eijiro, Nagai Yusuke, Takahashi Ken, Kimata Yoshihiro, Naruse Keiji
The Department of Plastic and Reconstructive Surgery, Okayama University Graduate School of Medicine, Okayama, Japan.
Menicon Co., Ltd., Aichi, Japan.
PLoS One. 2015 Nov 3;10(11):e0141989. doi: 10.1371/journal.pone.0141989. eCollection 2015.
All previous reports concerning the effect of stretch on cultured skin cells dealt with experiments on epidermal keratinocytes or dermal fibroblasts alone. The aim of the present study was to develop a system that allows application of stretch stimuli to human skin equivalents (HSEs), prepared by coculturing of these two types of cells. In addition, this study aimed to analyze the effect of a stretch on keratinization of the epidermis and on the basement membrane. HSEs were prepared in a gutter-like structure created with a porous silicone sheet in a silicone chamber. After 5-day stimulation with stretching, HSEs were analyzed histologically and immunohistologically. Stretch-stimulated HSEs had a thicker epidermal layer and expressed significantly greater levels of laminin 5 and collagen IV/VII in the basal layer compared with HSEs not subjected to stretch stimulation. Transmission electron microscopy revealed that the structure of the basement membrane was more developed in HSEs subjected to stretching. Our model may be relevant for extrapolating the effect of a stretch on the skin in a state similar to an in vivo system. This experimental system may be useful for analysis of the effects of stretch stimuli on skin properties and wound healing and is also expected to be applicable to an in vitro model of a hypertrophic scar in the future.
以往所有关于拉伸对培养的皮肤细胞影响的报告，都仅涉及对表皮角质形成细胞或真皮成纤维细胞的实验。本研究的目的是开发一种系统，该系统能够对通过这两种细胞共培养制备的人皮肤等效物（HSE）施加拉伸刺激。此外，本研究旨在分析拉伸对表皮角质化和基底膜的影响。HSE在硅胶腔室中由多孔硅胶片形成的槽状结构中制备。经过5天的拉伸刺激后，对HSE进行组织学和免疫组织学分析。与未接受拉伸刺激的HSE相比，拉伸刺激的HSE具有更厚的表皮层，并且在基底层中层粘连蛋白5和胶原蛋白IV/VII的表达水平显著更高。透射电子显微镜显示，接受拉伸的HSE中基底膜的结构更发达。我们的模型可能与推断在类似于体内系统的状态下拉伸对皮肤的影响有关。该实验系统可能有助于分析拉伸刺激对皮肤特性和伤口愈合的影响，并且有望在未来应用于肥厚性瘢痕的体外模型。
