Salerno Simona, De Santo Maria Penelope, Drioli Enrico, De Bartolo Loredana
Institute on Membrane Technology, National Research Council of Italy (CNR-ITM), Via P. Bucci, cubo 17/C, 87036 Rende, Italy.
Department of Physics and CNR-Nanotec, University of Calabria, Via P. Bucci, cubo 31/C, 87036 Rende, Italy.
Membranes (Basel). 2021 May 27;11(6):394. doi: 10.3390/membranes11060394.
The creation of partial or complete human epidermis represents a critical aspect and the major challenge of skin tissue engineering. This work was aimed at investigating the effect of nano- and micro-structured CHT membranes on human keratinocyte stratification and differentiation. To this end, nanoporous and microporous membranes of chitosan (CHT) were prepared by phase inversion technique tailoring the operational parameters in order to obtain nano- and micro-structured flat membranes with specific surface properties. Microporous structures with different mean pore diameters were created by adding and dissolving, in the polymeric solution, polyethylene glycol (PEG Mw 10,000 Da) as porogen, with a different CHT/PEG ratio. The developed membranes were characterized and assessed for epidermal construction by culturing human keratinocytes on them for up to 21 days. The overall results demonstrated that the membrane surface properties strongly affect the stratification and terminal differentiation of human keratinocytes. In particular, human keratinocytes adhered on nanoporous CHT membranes, developing the structure of the corneum epidermal top layer, characterized by low thickness and low cell proliferation. On the microporous CHT membrane, keratinocytes formed an epidermal basal lamina, with high proliferating cells that stratified and differentiated over time, migrating along the axis and forming a multilayered epidermis. This strategy represents an attractive tissue engineering approach for the creation of specific human epidermal strata for testing the effects and toxicity of drugs, cosmetics and pollutants.
构建部分或完整的人体表皮是皮肤组织工程的一个关键方面，也是主要挑战。这项工作旨在研究纳米和微米结构的壳聚糖（CHT）膜对人角质形成细胞分层和分化的影响。为此，采用相转化技术制备壳聚糖（CHT）的纳米多孔和微多孔膜，通过调整操作参数，以获得具有特定表面性质的纳米和微米结构的平板膜。通过在聚合物溶液中添加和溶解聚乙二醇（PEG Mw 10,000 Da）作为致孔剂，并改变CHT/PEG比例，制备出具有不同平均孔径的微孔结构。对所制备的膜进行表征，并通过在其上培养人角质形成细胞长达21天来评估其用于表皮构建的性能。总体结果表明，膜的表面性质强烈影响人角质形成细胞的分层和终末分化。具体而言，人角质形成细胞粘附在纳米多孔CHT膜上，形成角质层表皮顶层结构，其特点是厚度低且细胞增殖少。在微孔CHT膜上，角质形成细胞形成表皮基底层，其中高增殖细胞随着时间的推移进行分层和分化，沿轴迁移并形成多层表皮。这种策略代表了一种有吸引力的组织工程方法，用于创建特定的人体表皮层，以测试药物、化妆品和污染物的效果及毒性。
