Chemical Vapor Deposition Fabrication of Nano- and Micro-scale Biomimetic Surfaces

本研究的目的是开发一种肠道细胞培养基质，以提高体外药物转运研究的准确性，并为肠道组织再生提供支架。将开发一种完全模拟肠道基底膜自然形态的培养底物，为体外培养的细胞提供必要的发育线索。化学气相沉积(CVD)将被用来创建一种完全模拟天然纳米和微地形的肠道基底膜的二氧化硅模板。CVD将被用来用一种多孔的生物相容聚合物(如聚(甲基丙烯酸羟乙酯)(PHEMA))来处理硅胶模具。这种聚合物底物将从硅胶模具中分离出来，用于细胞培养。扫描电子显微镜将用来表征二氧化硅和PHEMA的拓扑结构，并将其与本地问题进行比较。我们将通过多种分析方法，将所开发设备的表面形态对肠道细胞表型和运输特性的影响与在非表面修饰表面培养的细胞进行比较。这项工作将导致开发一种基于细胞的工具，用于研究肠道上皮的运输，从而节省药物研究的成本和时间，并对正常和疾病(如结肠癌)状态下细胞-生物材料的相互作用和自然肠道功能有一个基本的了解。如果成功，这项研究将大大减少将药物推向市场的时间和开发成本，并为治疗包括短肠综合征、克罗恩病和结肠癌在内的多种肠道疾病做出重大贡献。它通过创造准确模拟体内发现的重要特征的培养底物，为当前的细胞培养技术提供了一种极大的改进方法，并为纳米和微观形貌对肠细胞表型的影响提供了基本的科学见解。

The objective of this proposed research is to develop an intestinal cell culture substrate to improve the accuracy of in vitro drug transport studies and provide a scaffold for intestinal tissue regeneration. A culture substrate that exactly mimics the natural topography of intestinal basement membrane will be developed, providing essential developmental cues to cells cultured in vitro. Chemical vapor deposition (CVD) will be used to create a silica template of intestinal basement membrane that exactly mimics the native nano- and micro-topography. CVD will be used to caoat the silica mold with a porous biocompatible polymer such as poly(2-hydroxyethyl methacrylate) (pHEMA). This polymer substrate will be separated from the silica mold and used for cell culture. SEM will be used to characterize the silica and pHEMA topology and compare it to native issue. The influence of the topography of the developed device on enterocyte cell phenotype and transport properties will be compared with cells cultured on non-topographically modified surfaces via a variety of assays.This work will result in the development of a cell-based tool for studying transport across the intestinal epithelium, leading to cost and time savings in drug studies, as well as a fundamental understanding of cell-biomaterial interactions and native intestinal function in normal and disease (e.g. colon cancer) states. If successful, this research will significantly reduce the time and development costs for bringing pharmaceuticals to market, as well as make serious contributions to the treatment of numerous intestinal illnesses, including short bowel syndrome, Crohn's disease, and colon cancer. It offers a greatly improved approach to current cell culture techniques through the creation of culture substrates that exactly mimic important features found in vivo, and provides basic scientific insight into the influence of nano- and micro-topography on enterocyte phenotype.