Koppes Abigail N, Kamath Megha, Pfluger Courtney A, Burkey Daniel D, Dokmeci Mehmet, Wang Lin, Carrier Rebecca L
Northeastern University, Chemical Engineering, Boston, MA, USA.
Biofabrication. 2016 Aug 22;8(3):035011. doi: 10.1088/1758-5090/8/3/035011.
Native small intestine possesses distinct multi-scale structures (e.g., crypts, villi) not included in traditional 2D intestinal culture models for drug delivery and regenerative medicine. The known impact of structure on cell function motivates exploration of the influence of intestinal topography on the phenotype of cultured epithelial cells, but the irregular, macro- to submicron-scale features of native intestine are challenging to precisely replicate in cellular growth substrates. Herein, we utilized chemical vapor deposition of Parylene C on decellularized porcine small intestine to create polymeric intestinal replicas containing biomimetic irregular, multi-scale structures. These replicas were used as molds for polydimethylsiloxane (PDMS) growth substrates with macro to submicron intestinal topographical features. Resultant PDMS replicas exhibit multiscale resolution including macro- to micro-scale folds, crypt and villus structures, and submicron-scale features of the underlying basement membrane. After 10 d of human epithelial colorectal cell culture on PDMS substrates, the inclusion of biomimetic topographical features enhanced alkaline phosphatase expression 2.3-fold compared to flat controls, suggesting biomimetic topography is important in induced epithelial differentiation. This work presents a facile, inexpensive method for precisely replicating complex hierarchal features of native tissue, towards a new model for regenerative medicine and drug delivery for intestinal disorders and diseases.
天然小肠具有独特的多尺度结构（如隐窝、绒毛），而传统的二维肠道药物递送和再生医学培养模型中并未包含这些结构。结构对细胞功能的已知影响促使人们探索肠道拓扑结构对培养上皮细胞表型的影响，但天然肠道不规则的宏观到亚微米尺度特征在细胞生长基质中精确复制具有挑战性。在此，我们利用聚对二甲苯C在脱细胞猪小肠上的化学气相沉积，制造出含有仿生不规则多尺度结构的聚合物肠道复制品。这些复制品被用作具有宏观到亚微米肠道拓扑特征的聚二甲基硅氧烷（PDMS）生长基质的模具。所得的PDMS复制品呈现出多尺度分辨率，包括宏观到微观尺度的褶皱、隐窝和绒毛结构，以及底层基底膜的亚微米尺度特征。在PDMS基质上进行10天的人上皮结肠直肠细胞培养后，与平坦对照相比，包含仿生拓扑特征使碱性磷酸酶表达提高了2.3倍，这表明仿生拓扑结构在诱导上皮分化中很重要。这项工作提出了一种简便、廉价的方法，用于精确复制天然组织的复杂层次特征，朝着为肠道疾病开发再生医学和药物递送新模型的方向迈进。
