Zöller Nadja Nicole, Hofmann Matthias, Butting Manuel, Hrgovic Igor, Bereiter-Hahn Jürgen, Bernd August, Kaufmann Roland, Kippenberger Stefan, Valesky Eva
Department of Dermatology, Venereology, and Allergology, Goethe University Hospital, Frankfurt/Main, Germany.
Kinematic Cell Research Group, Goethe University, Frankfurt/Main, Germany.
Indian J Dermatol. 2019 Mar-Apr;64(2):85-89. doi: 10.4103/ijd.IJD_410_17.
Organotypic tissue-cultured skin equivalents are used for a broad range of applications either as possible substitute for animal tests or for transplantation in patient-centered care.
In this study, we implemented melanocytes in a tissue-cultured full-thickness skin equivalent, consisting of epidermis and dermis. The versatility of this skin-like model with respect to pigmentation and morphological criteria was tested.
Pigmented skin equivalents were morphologically characterized, and melanogenesis was evaluated after treatment with kojic acid - a tyrosinase inhibitor and forskolin - a well-known activator of the cyclic adenosine 3,5-monophosphate pathway. Pigmentation was measured either by determination of the extinction at 400 nm after melanin extraction with KOH correlated to a melanin standard curve or by reflectance colorimetric analysis, monitoring reflectance of 660 nm and 880 nm emitting diodes.
The morphological analysis revealed characteristic epidermal stratification with melanocytes located at the basal layer. Stimulation with forskolin increased the pigmentation, whereas treatment with kojic acid caused bleaching.
The present study demonstrates that the herein-introduced organotypic tissue-cultured skin equivalent is comparable to the normal human skin and its versatility in tests regarding skin pigmentation. Therefore, this model might help understand diseases with dysfunctional pigmentation such as melasma, vitiligo, and postinflammatory hyperpigmentation.
器官型组织培养皮肤替代物被广泛应用于多种用途，既可以作为动物试验的可能替代物，也可用于以患者为中心的护理中的移植。
在本研究中，我们将黑素细胞植入由表皮和真皮组成的组织培养全层皮肤替代物中。测试了这种类似皮肤的模型在色素沉着和形态学标准方面的多功能性。
对色素沉着的皮肤替代物进行形态学表征，并在用曲酸（一种酪氨酸酶抑制剂）和福斯可林（一种著名的环磷酸腺苷途径激活剂）处理后评估黑素生成。色素沉着的测量方法有两种，一种是用氢氧化钾提取黑色素后测定400nm处的吸光度，并与黑色素标准曲线相关联；另一种是通过反射比色分析，监测660nm和880nm发光二极管的反射率。
形态学分析显示出具有位于基底层的黑素细胞的特征性表皮分层。用福斯可林刺激可增加色素沉着，而用曲酸处理则导致色素减退。
本研究表明，本文引入的器官型组织培养皮肤替代物与正常人类皮肤相当，并且在皮肤色素沉着测试中具有多功能性。因此，该模型可能有助于理解色素沉着功能障碍的疾病，如黄褐斑、白癜风和炎症后色素沉着。
