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Proteolytically Degradable Hydrogels with a Fluorogenic Substrate for Studies of Cellular Proteolytic Activity and Migration
Author(s) -
Lee SooHong,
Miller Jordan S.,
Moon James J.,
West Jennifer L.
Publication year - 2005
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1021/bp0502429
Subject(s) - self healing hydrogels , peg ratio , chemistry , biophysics , bovine serum albumin , covalent bond , ethylene glycol , substrate (aquarium) , proteolytic enzymes , confocal microscopy , cell migration , fluorescence microscope , biochemistry , polymer chemistry , fluorescence , cell , enzyme , microbiology and biotechnology , biology , organic chemistry , ecology , finance , economics , physics , quantum mechanics
We have developed proteolytically degradable hydrogels with covalently immobilized fluorogenic protease substrates to visualize extracellular proteolytic activity and cell migration in three dimensions. Dye quenched‐bovine serum albumin (DQ‐BSA), a quenched, proteolytically activated fluorogenic substrate, was conjugated to poly(ethylene glycol) (PEG)‐monoacrylate, and the product (DQ‐BSA‐PEG) was then covalently incorporated into proteolytically degradable and cell adhesive PEG hydrogels via photopolymerization. The DQ‐BSA‐PEG substrate in solution and incorporated into hydrogels exhibited significantly enhanced fluorescence after exposure to enzymes. Fibroblasts seeded within this hydrogel spread in three dimensions and extended lamellipodia. Cell migration and proteolytic activity were visualized using confocal microscopy. Proteolytic activity was concentrated near cell surfaces and remained present in the tracks where cell migration had occurred.