Fibrinogen adsorption, platelet adhesion and activation on mixed hydroxyl-/methyl-terminated self-assembled monolayers

doi:10.1016/j.biomaterials.2006.06.010

Biomaterials

Volume 27, Issue 31, November 2006, Pages 5357-5367

https://doi.org/10.1016/j.biomaterials.2006.06.010 Get rights and content

Abstract

The effect of surface wettability on fibrinogen adsorption, platelet adhesion and platelet activation was investigated using self-assembled monolayers (SAMs) containing different ratios of longer chain methyl- and shorter chain hydroxyl-terminated alkanethiols (C15CH3 vs. C11OH) on gold.

Protein adsorption studies were performed using radiolabeled human fibrinogen (HFG). Platelet adhesion and activation studies with and without pre-adsorbed fibrinogen, albumin and plasma were assessed using scanning electron microscopy (SEM) and a glutaraldehyde-induced fluorescence technique (GIFT). Results demonstrated a linear decrease of HFG adsorption with the increase of OH groups on the monolayer (increase of the hydrophilicity). Platelet adhesion and activation also decrease with increase of hydrophilicity of surface. Concerning SAMs pre-immersed in proteins, fibrinogen adsorption was related with high platelet adhesion and activation. The passivant effect of albumin on platelet adhesion and activation was only demonstrated on SAMs contained C11OH. When all the blood proteins are present (plasma) platelet adhesion was almost absent on SAMs with 65% and 100% C11OH. This could be explained by the higher albumin affinity of the SAMs with 65% C11OH and the lower total protein adsorption associated with SAMs with 100% C11OH.

Section snippets

Introdution

Thrombus formation, resulting from protein adsorption, platelet adhesion and platelet activation limits the success of blood-contacting medical devices. The adsorption of plasma proteins to the surface is one of the first events to occur and influences all subsequent responses to the biomaterial. Although the adsorption of fibrinogen is related to the adhesion and activation of platelets [1], [2], a monolayer of albumin appears to minimize these events avoiding subsequent thrombus formation [3]

Gold substrates

Gold substrates were prepared as described elsewhere [8]. Briefly, chromium (5 nm) and gold (25 nm) films were deposited by ion beam sputtering onto silicon wafers. The thin layer of chromium was used to improve adhesion of gold to silicon. Gold substrates (1×1 cm²) were used for the preparation of all SAMs except for the ones to be used in radiolabelling (0.5×0.5 cm²).

Monolayer formation

11-mercapto-1-undecanol (SH–(CH₂)₁₁OH; 97%, Aldrich; C11OH) and 1-hexadecanethiol (SH–(CH₂)₁₅CH₃; 92%, Aldrich; C15CH3) were used

Surface characterization

The surfaces of pure and mixed SAMs were characterized using ellipsometry. In Fig. 1, the monolayer thickness, measured by ellipsometry, and the contact angle are plotted as a function of the percentage of C11OH present on the surface, calculated by the method described by us before [8]. Briefly, the percentage of C11OH in the mixed SAMs was calculated using the O (1 s) peak intensity normalized to the O (1 s) peak intensity of the SAMs prepared from pure C11OH solutions. As observed in this

Discussion

The aim of the present work was to study the effect of surface wettability on protein adsorption, platelet adhesion and activation. Surfaces with a gradual range of wettabilities were prepared using SAMs with different percentages of a hydrophobic and a hydrophilic functional group (–CH₃ and –OH, respectively). Ellipsometry was applied as an accurate technique for determining the average monolayer thickness of the pure and mixed SAMs prepared from solutions of C11OH and C15CH3. Since these SAMs

Conclusions

Surfaces with a graded range of wettabilities were prepared using SAMs with different percentages of a hydrophobic and a hydrophilic functional group (–CH₃ and –OH, respectively). A linear decrease of fibrinogen adsorption with increase of surface hydrophilicity was observed. The increase of hydrophilicity was also correlated with a decrease of platelet adhesion and activation.

Fibrinogen adsorption was associated with high platelet adhesion and activation. However, results with gold and SAMs

Acknowledgements

The authors would like to Portuguese Blood Institute (IPS) for donating the platelet concentrate (PC) and the human plasma, by assistance and by enabling them to carry out the platelet adhesion tests. Inês Gonçalves is grateful to the Portuguese Foundation for Science and Technology (FCT) for awarding her the Grant SFRH/BD/188337/2004. Ma Cristina L. Martins is grateful to the Portuguese Foundation for Science and Technology (FCT) for awarding her a post-doctoral scholarship. Professor Ratner

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View full text

Fibrinogen adsorption, platelet adhesion and activation on mixed hydroxyl-/methyl-terminated self-assembled monolayers

Abstract

Section snippets

Introdution

Gold substrates

Monolayer formation

Surface characterization

Discussion

Conclusions

Acknowledgements

J Biol Chem

Biomaterials

Biomaterials

J Colloid Interf Sci

Ultramicroscopy

Colloid Surf

Biomaterials

J Biol Chem

Biomaterials

Platelet adhesion to polystyrene-based surfaces preadsorbed with plasmas selectively depleted in fibrinogen, fibronectin, vitronectin, or von Willebrand's factor

J Biomed Mater Res

Peptide, protein, and cellular interactions with self-assembled monolayer model surfaces

J Biomed Mater Res

Endothelial cell growth and protein adsorption on terminally functionalized, self-assembled monolayers of alkanethiolates on gold

Langmuir

All about albumin: biochemistry, genetics and medical applications

Protein adsorption on mixtures of hydroxyl- and methylterminated alkanethiols self-assembled monolavers

J Biomed Mater Res A

Glutaraldehyde induced fluorescence technique (GIFT): a new method for the imaging of platelet adhesion on biomaterials

J Biomed Mater Res

Protein adsorption on low-temperature isotropic carbon. 3. Isotherms, competitivity, desorption and exchange of human albumin and fibrinogen

Biomaterials