PDF(Pubmed)
Abstract：
The topology of the basement membrane (BM) affects cell physiology and pathology, and BM thickening is associated with various chronic lung diseases. In addition, the topology of commercially available poly (ethylene terephthalate) (PET) membranes, which are used in preclinical in vitro models, differs from that of the human BM, which has a fibrous and elastic structure. In this study, we verified the effect of BM thickness on the differentiation of normal human bronchial epithelial (NHBE) cells. To evaluate whether the thickness of poly-ε-carprolactone (PCL) mesh affects the differentiation of NHBE cells, cells were grown on thin- (6-layer) and thick-layer (80-layer) meshes consisting of electrospun PCL nanofibers using an air-liquid interface (ALI) cell culture system. It was found that the NHBE cells formed a normal pseudostratified epithelium composed of ciliated, goblet, and basal cells on the thin-layer PCL mesh; however, goblet cell hyperplasia was observed on the thick-layer PCL mesh. Differentiated NHBE cells cultured on the thick-layer PCL mesh also demonstrated increased epithelial-mesenchymal transition (EMT) compared to those cultured on the thin-layer PCL mesh. In addition, expression of Sox9, nuclear factor (NF)-κB, and oxidative stress-related markers, which are also associated with goblet cell hyperplasia, was increased in the differentiated NHBE cells cultured on the thick-layer PCL mesh. Thus, the use of thick electrospun PCL mesh led to NHBE cells differentiating into hyperplastic goblet cells via EMT and the oxidative stress-related signaling pathway. Therefore, the topology of the BM, for example, thickness, may affect the differentiation direction of human bronchial epithelial cells.
摘要：
基底膜（BM）的拓扑结构影响细胞生理和病理，BM增厚与各种慢性肺部疾病有关。此外,可商购的聚(对苯二甲酸乙二醇酯)(PET)膜的拓扑结构，用于临床前体外模型，不同于人类的BM，具有纤维状和弹性结构。在这项研究中,我们验证了BM厚度对正常人支气管上皮（NHBE）细胞分化的影响。评价聚ε-己内酯(PCL)网片厚度是否影响NHBE细胞分化,使用气液界面（ALI）细胞培养系统，在由电纺PCL纳米纤维组成的薄（6层）和厚（80层）网格上生长细胞。发现NHBE细胞形成由纤毛，高脚杯,和薄层PCL网格上的基底细胞；然而，在厚层PCL网格上观察到杯状细胞增生。与在薄层PCL网上培养的细胞相比，在厚层PCL网上培养的分化NHBE细胞还显示出上皮-间充质转化（EMT）增加。此外,Sox9,核因子(NF)-κB,和氧化应激相关的标志物，它们也与杯状细胞增生有关，在厚层PCL网上培养的分化NHBE细胞中增加。因此，使用厚的电纺PCL网导致NHBE细胞通过EMT和氧化应激相关信号通路分化为增生性杯状细胞。因此,BM的拓扑结构，例如,厚度,可能影响人支气管上皮细胞的分化方向。
