TY - JOUR
T1 - Substrate-induced assembly of fibronectin into networks
T2 - Influence of surface chemistry and effect on osteoblast adhesion
AU - Rico, Patricia
AU - Hernández, José Carlos Rodríguez
AU - Moratal, David
AU - Altankov, George
AU - Pradas, Manuel Monleón
AU - Salmerón-Sánchez, Manuel
N1 - Copyright:
Copyright 2014 Elsevier B.V., All rights reserved.
PY - 2009/11/1
Y1 - 2009/11/1
N2 - The influence of surface chemistry-substrates with controlled surface density of-OH groups-on fibronectin (FN) conformation and distribution is directly observed by atomic force microscopy (AFM). FN fibrillogenesis, which is known to be a process triggered by interaction with integrins, is shown in our case to be induced by the substrate (in absence of cells), which is able to enhance FN-FN interactions leading to the formation of a protein network on the material surface. This phenomenon depends both on surface chemistry and protein concentration. The level of the FN fibrillogenesis was quantified by calculating the fractal dimension of the adsorbed protein from image analysis of the AFM results. The total amount of adsorbed FN is obtained by making use of a methodology that employs Western blotting combined with image analysis of the corresponding protein bands, with the lowest sensitivity threshold equal to 15ng of adsorbed protein. Further, FN adsorption is correlated to human osteoblast adhesion through morphology and actin cytoskeleton formation. Actin polymerization is in need of the formation of the protein network on the substrate's surface. Cell morphology is more rounded (as quantified by calculating the circularity of the cells by image analysis) when the degree of FN fibrillogenesis on the substrate is lower.
AB - The influence of surface chemistry-substrates with controlled surface density of-OH groups-on fibronectin (FN) conformation and distribution is directly observed by atomic force microscopy (AFM). FN fibrillogenesis, which is known to be a process triggered by interaction with integrins, is shown in our case to be induced by the substrate (in absence of cells), which is able to enhance FN-FN interactions leading to the formation of a protein network on the material surface. This phenomenon depends both on surface chemistry and protein concentration. The level of the FN fibrillogenesis was quantified by calculating the fractal dimension of the adsorbed protein from image analysis of the AFM results. The total amount of adsorbed FN is obtained by making use of a methodology that employs Western blotting combined with image analysis of the corresponding protein bands, with the lowest sensitivity threshold equal to 15ng of adsorbed protein. Further, FN adsorption is correlated to human osteoblast adhesion through morphology and actin cytoskeleton formation. Actin polymerization is in need of the formation of the protein network on the substrate's surface. Cell morphology is more rounded (as quantified by calculating the circularity of the cells by image analysis) when the degree of FN fibrillogenesis on the substrate is lower.
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U2 - 10.1089/ten.tea.2009.0141
DO - 10.1089/ten.tea.2009.0141
M3 - Article
C2 - 19382854
AN - SCOPUS:72649084943
VL - 15
SP - 3271
EP - 3281
JO - Tissue Engineering - Part A.
JF - Tissue Engineering - Part A.
SN - 1937-3341
IS - 11
ER -