Arrangement of type IV collagen on NH 2 and COOH functionalized surfaces

Nuno Miranda Coelho, Cristina González-García, Manuel Salmerón-Sánchez, George Altankov

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)3009-3018
Number of pages10
JournalBiotechnology and Bioengineering
Volume108
Issue number12
DOIs
Publication statusPublished - Dec 1 2011
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Fingerprint Dive into the research topics of 'Arrangement of type IV collagen on NH <sub>2</sub> and COOH functionalized surfaces'. Together they form a unique fingerprint.

  • Cite this

    Coelho, N. M., González-García, C., Salmerón-Sánchez, M., & Altankov, G. (2011). Arrangement of type IV collagen on NH 2 and COOH functionalized surfaces. Biotechnology and Bioengineering, 108(12), 3009-3018. https://doi.org/10.1002/bit.23265