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Merck

Gastrointestinal synthetic epithelial linings.

Science translational medicine (2020-08-28)
Junwei Li, Thomas Wang, Ameya R Kirtane, Yunhua Shi, Alexis Jones, Zaina Moussa, Aaron Lopes, Joy Collins, Siddartha M Tamang, Kaitlyn Hess, Rameen Shakur, Paramesh Karandikar, Jung Seung Lee, Hen-Wei Huang, Alison Hayward, Giovanni Traverso
摘要

Epithelial tissues line the organs of the body, providing an initial protective barrier as well as a surface for nutrient and drug absorption. Here, we identified enzymatic components present in the gastrointestinal epithelium that can serve as selective means for tissue-directed polymerization. We focused on the small intestine, given its role in drug and nutrient absorption and identified catalase as an essential enzyme with the potential to catalyze polymerization and growth of synthetic biomaterial layers. We demonstrated that the polymerization of dopamine by catalase yields strong tissue adhesion. We characterized the mechanism and specificity of the polymerization in segments of the gastrointestinal tracts of pigs and humans ex vivo. Moreover, we demonstrated proof of concept for application of these gastrointestinal synthetic epithelial linings for drug delivery, enzymatic immobilization for digestive supplementation, and nutritional modulation through transient barrier formation in pigs. This catalase-based approach to in situ biomaterial generation may have broad indications for gastrointestinal applications.

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Sigma-Aldrich
Protease Inhibitor Cocktail, for use with mammalian cell and tissue extracts, DMSO solution
Sigma-Aldrich
D -(+)-葡萄糖, ≥99.5% (GC)
Sigma-Aldrich
总蛋白提取细胞裂解缓冲液
Sigma-Aldrich
Triton X-100, for molecular biology
Sigma-Aldrich
磷酸酶抑制剂混合物3, DMSO solution
Sigma-Aldrich
过氧化氢酶 来源于牛肝脏, powder, suitable for cell culture, 2,000-5,000 units/mg protein
USP
盐酸多巴胺 盐酸盐, United States Pharmacopeia (USP) Reference Standard
USP
Lactase, United States Pharmacopeia (USP) Reference Standard
Sigma-Aldrich
过氧化氢酶 来源于人类红细胞, ≥90% (SDS-PAGE), buffered aqueous solution, ≥30,000 units/mg protein