Different assembly of type IV collagen on hydrophilic and hydrophobic substrata alters endothelial cells interaction.
Considering the structural role of type IV collagen (Col IV) in the assembly of the basement membrane (BM) and the perspective of mimicking its organization for vascular tissue engineering purposes, we studied the adsorption pattern of this protein on model hydrophilic (clean glass) and hydrophobic trichloro(octadecyl)silane (ODS) surfaces known to strongly affect the behavior of other matrix proteins. The amount of fluorescently labeled Col IV was quantified showing saturation of the surface for concentration of the adsorbing solution of about 50microg/ml, but with approximately twice more adsorbed protein on ODS. AFM studies revealed a fine - nearly single molecular size - network arrangement of Col IV on hydrophilic glass, which turns into a prominent and growing polygonal network consisting of molecular aggregates on hydrophobic ODS. The protein layer forms within minutes in a concentration-dependent manner. We further found that human umbilical vein endothelial cells (HUVEC) attach less efficiently to the aggregated Col IV (on ODS), as judged by the significantly altered cell spreading, focal adhesions formation and the development of actin cytoskeleton. Conversely, the immunofluorescence studies for integrins revealed that the fine Col IV network formed on hydrophilic substrata is better recognized by the cells via both alpha1 and alpha2 heterodimers which support cellular interaction, apart from these on hydrophobic ODS where almost no clustering of integrins was observed.
Institut de Bioenginyeria de Catalunya, Barcelona, Spain.
N Miranda
NM
Coelho
C
C
González-García
J A
JA
Planell
M
M
Salmerón-Sánchez
G
G
Altankov
D001808:Blood Vessels / Q000166:cytology; D002448:Cell Adhesion; D002450:Cell Communication / Q000187:drug effects*; D024141:Collagen Type IV / Q000378:metabolism*; D042783:Endothelial Cells / Q000166:cytology / Q000502:physiology*; D004730:Endothelium, Vascular / Q000166:cytology; D005898:Glass; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D011485:Protein Binding; D012821:Silanes / Q000494:pharmacology; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds
D024141:Collagen Type IV; D012821:Silanes