2010 Volume No 19 – pages 262-272
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Title: Different assembly of type IV collagen on hydrophilic and hydrophobic substrata alters endothelial cells interaction |
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Author: NM Coelho, C González-García, JA Planell, M Salmerón-Sánchez, G Altankov |
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Address: ICREA – Molecular Dynamics, Feixa Llarga Pavelo Govern Planta 1 No 1112, Bellvitge Barcelona Hospitalet de Llobregat, 08907 Barcelona, Spain |
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E-mail: george.altankov at icrea.es |
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Key Words: Collagen type IV, adsorption, assembly, hydrophilic, hydrophobic, surfaces. |
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Publication date: June 9th 2010 |
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Abstract: 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 50μg/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 α1 and α2 heterodimers which support cellular interaction, apart from these on hydrophobic ODS where almost no clustering of integrins was observed. |
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Article download: Pages
262-272 (PDF file) |
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