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Arrangement of type IV collagen on NH 2 and COOH functionalized surfaces
Author(s) -
Coelho Nuno Miranda,
GonzálezGarcía Cristina,
SalmerónSánchez Manuel,
Altankov George
Publication year - 2011
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.23265
Subject(s) - focal adhesion , integrin , chemistry , adhesion , biophysics , basement membrane , cytoskeleton , type iv collagen , rap1 , protein adsorption , cell adhesion , umbilical vein , extracellular matrix , adsorption , laminin , microbiology and biotechnology , biochemistry , cell , signal transduction , in vitro , biology , organic chemistry
Apart from the paradigm that cell–biomaterials interaction depends on the adsorption of soluble adhesive proteins we anticipate that upon distinct conditions also other, less soluble ECM proteins such as collagens, associate with the biomaterials interface with consequences for cellular response that might be of significant bioengineering interest. Using atomic force microscopy (AFM) we seek to follow the nanoscale behavior of adsorbed type IV collagen (Col IV)—a unique multifunctional matrix protein involved in the organization of basement membranes (BMs) including vascular ones. We have previously shown that substratum wettability significantly affects Col IV adsorption pattern, and in turn alters endothelial cells interaction. Here we introduce two new model surfaces based on self‐assembled monolayers (SAMs), a positively charged –NH 2 , and negatively charged –COOH surface, to learn more about their particular effect on Col IV behavior. AFM studies revealed distinct pattern of Col IV assembly onto the two SAMs resembling different aspects of network‐like structure or aggregates (suggesting altered protein conformation). Moreover, the amount of adsorbed FITC‐labeled Col IV was quantified and showed about twice more protein on NH 2 substrata. Human umbilical vein endothelial cells attached less efficiently to Col IV adsorbed on negatively charged COOH surface judged by altered cell spreading, focal adhesions formation, and actin cytoskeleton development. Immunofluorescence studies also revealed better Col IV recognition by both α 1 and α 2 integrins on positively charged NH 2 substrata resulting in higher phosphorylated focal adhesion kinase recruitment in the focal adhesion complexes. On COOH surface, no integrin clustering was observed. Taken altogether these results, point to the possibility that combined NH 2 and Col IV functionalization may support endothelization of cardiovascular implants. Biotechnol. Bioeng. 2011;108: 3009–3018. © 2011 Wiley Periodicals, Inc.