Kocsis Dorottya, Sztankovics Dániel, Józsa Liza, Németh Afrodité, Garay Tamás, Naszlady Márton Bese, Lengyel Miléna, Vecsernyés Miklós, Antal István, Sebestyén Anna, Erdő Franciska
Faculty of Information Technology and Bionics, Pázmány Péter Catholic University, Práter u. 50a., 1083 Budapest, Hungary.
Department of Pathology and Experimental Cancer Research, Semmelweis University, Üllői út 26., 1085 Budapest, Hungary.
Bioengineering (Basel). 2025 Feb 24;12(3):230. doi: 10.3390/bioengineering12030230.
In recent times, with the need for a reduction, refinement, and replacement of in vivo animal testing, there has been an increasing demand for the use of relevant in vitro human cell systems in drug development. There is also a great demand for the replacement of skin tissue in various wounds and burns. Furthermore, human skin cell-based in vitro systems can be used to investigate the side effects (toxicity and irritation) and tissue penetration of topical preparations. In this study, exploratory experiments were performed to produce artificial epidermis using two hydrogel scaffolds, alginate and GelMA C. The amount of keratinocytes added to the matrix (10-50-100 × 10/mL) and the duration of tissue maturation (fresh, 1-3-4 weeks) were optimized in an extensive study. The behavior and structure of the two hydrogels were functionally and morphologically assessed. The permeability order for caffeine in the tested barriers was the following: alginate > GelMA C > cellulose acetate membrane > rat skin. It was concluded that GelMA C matrix provides a more favorable environment for cell survival and tissue differentiation (as demonstrated by histology and immunohistochemistry) than alginate. The 3-week incubation and 50 × 10/mL cell number proved to be the most beneficial in the given system. This study provides data for the first time on the multifactorial optimization of two potential skin substitutes for tissue manufacturing. In order to use these results in tissue engineering, the fabricated artificial epidermis preparations must also be optimized for biocompatibility and from physical and mechanical point of views.
近年来，随着减少、优化和替代体内动物试验的需求，药物研发中对使用相关体外人类细胞系统的需求日益增加。在各种伤口和烧伤中替代皮肤组织的需求也很大。此外，基于人类皮肤细胞的体外系统可用于研究局部制剂的副作用（毒性和刺激性）以及组织渗透性。在本研究中，进行了探索性实验，使用两种水凝胶支架（海藻酸盐和甲基丙烯酰化明胶C）制备人工表皮。在一项广泛的研究中，优化了添加到基质中的角质形成细胞数量（10 - 50 - 100×10/mL）和组织成熟的持续时间（新鲜、1 - 3 - 4周）。对两种水凝胶的行为和结构进行了功能和形态学评估。咖啡因在测试屏障中的渗透顺序如下：海藻酸盐＞甲基丙烯酰化明胶C＞醋酸纤维素膜＞大鼠皮肤。得出的结论是，与海藻酸盐相比，甲基丙烯酰化明胶C基质为细胞存活和组织分化提供了更有利的环境（组织学和免疫组织化学证明）。在给定系统中，3周的孵育和50×10/mL的细胞数量被证明是最有益的。本研究首次提供了关于两种潜在皮肤替代物用于组织制造的多因素优化数据。为了将这些结果应用于组织工程，制备的人工表皮制剂还必须在生物相容性以及物理和力学方面进行优化。
