Effect of the conformation and orientation of adsorbed fibronectin on endothelial cell spreading and the strength of adhesion

D. J. Iuliano, Steven S Saavedra, G. A. Truskey

Research output: Contribution to journalArticle

214 Citations (Scopus)

Abstract

The effect of surface hydrophobicity upon the conformation of the cell binding domain of fibronectin (Fn) and the influence of Fn conformation on bovine aortic endothelial cell (BAEC) adhesion were examined. The free sulfhydryl group of Fn located near the cell binding domain was selectively labeled with acrylodan, a polarity sensitive fluor. Fluorescence emission was monitored in solution and upon adsorption to hydrophilic glass and hydrophobic silanized glass. The acrylodan-labeled Fn emission maximum shifted to longer wavelengths upon adsorption and the shift was greater for acrylodan-labeled Fn adsorbed to hydrophilic glass than hydrophobic silane, suggesting that the acrylodan was in a more solvent accessible environment on glass than silane. BAEC, suspended in serum-free medium, attached for 15 or 120 min onto glass or silane surface containing pre-adsorbed Fn, after which cell spreading and the strength of adhesion in a parallel plate flow chamber were measured. Cell spreading was similar on both surfaces after 15 min attachment, but BAECs were more spread on glass than silane after 120 min. At low surface concentrations of Fn, BAECs were more adherent on glass than silane. At higher surfaces concentrations, adhesion was similar. After a 2-h incubation in serum-free medium, cells on glass showed more extensive development of focal contacts as determined by immunofluorescent staining for vinculin. Cell adhesion under flow was reduced on silane by inhibition of protein synthesis with cycloheximide, suggesting that cell attachment to silane was promoted by cellular synthesis of Fn. The results indicate that changes in the conformation of the Fn cell binding domain affect Fn affinity for its cell surface receptor.

Original languageEnglish (US)
Pages (from-to)1103-1113
Number of pages11
JournalJournal of Biomedical Materials Research
Volume27
Issue number8
StatePublished - Aug 1993
Externally publishedYes

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Endothelial cells
Fibronectins
Crystal orientation
Silanes
Conformations
Adhesion
Glass
Cell adhesion
Serum-Free Culture Media
Adsorption
Vinculin
Hydrophobicity
Cell Surface Receptors
Cycloheximide
Fluorescence
Proteins
Wavelength
acrylodan

ASJC Scopus subject areas

  • Biomedical Engineering
  • Biomaterials

Cite this

Effect of the conformation and orientation of adsorbed fibronectin on endothelial cell spreading and the strength of adhesion. / Iuliano, D. J.; Saavedra, Steven S; Truskey, G. A.

In: Journal of Biomedical Materials Research, Vol. 27, No. 8, 08.1993, p. 1103-1113.

Research output: Contribution to journalArticle

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