Department of Biomedical Materials Engineering , Kangwon National University , Chuncheon 24341 , Republic of Korea.
Institute of Bioscience and Biotechnology , Kangwon National University , Chuncheon 24341 , Republic of Korea.
ACS Appl Mater Interfaces. 2018 Dec 19;10(50):43503-43511. doi: 10.1021/acsami.8b17740. Epub 2018 Dec 5.
Bilayered cell sheets were fabricated with coaxial hydro-nanofibrils for three-dimensional (3D) cultivation in a biomimetic environment. Polycaprolactone (PCL) was electrospun and hydrolyzed to release fragmented nanofibrils (NF) in an alkaline condition. Methacrylated gelatin (GelMA) was adsorbed and phototethered on the surface of the fibrils to prepare coaxial NF composed of hydrophilic shells and hydrophobic cores. GelMA layers on the NF were characterized by X-ray photoemission spectroscopy, Fourier-transform infrared spectroscopy, and thermogravimetric analysis. The GelMA showed higher decoration level on NF compared to that on native gelatin. GelMA-decorated NF significantly enhanced cell proliferation rate and phenotypic expression of human dermal fibroblasts when spontaneous formation of cell sheets was observed for 7 days. HaCaT cells were layered on top of the fibroblast sheets and further cultivated in air-water interfaces to prepare bilayered skin sheets. After 21 days of incubation, the top layers of the bilayered sheets showed higher expression of pan-keratin, and the dermal cells showed higher proliferation in the GelMA-decorated NF.
双层细胞片通过同轴水纳米纤维在仿生环境中进行 3D 培养。聚己内酯（PCL）通过静电纺丝和在碱性条件下水解，释放出碎片状纳米纤维（NF）。甲基丙烯酰化明胶（GelMA）被吸附并光键合在纤维表面上，制备由亲水壳和疏水芯组成的同轴 NF。通过 X 射线光电子能谱、傅里叶变换红外光谱和热重分析对 NF 上的 GelMA 进行了表征。与天然明胶相比，GelMA 在 NF 上的修饰水平更高。当观察到 7 天自发形成细胞片时，GelMA 修饰的 NF 显著提高了人皮肤成纤维细胞的增殖率和表型表达。HaCaT 细胞分层在成纤维细胞片的顶部，并进一步在气-液界面中培养以制备双层皮肤片。孵育 21 天后，双层片的顶层显示出更高的全角蛋白表达，并且在 GelMA 修饰的 NF 中真皮细胞表现出更高的增殖。
