Polyethylene oxide surfaces of variable chain density by chemisorption of PEO-thiol on gold: adsorption of proteins from plasma studied by radiolabelling and immunoblotting.
The mechanisms involved in the inhibition of protein adsorption by polyethylene oxide (PEO) are not completely understood, but it is believed that PEO chain length, chain density and chain conformation all play a role. In this work, surfaces formed by chemisorption of PEO-thiol to gold were investigated: the effects of PEO chain density, chain length (600, 750, 2000 and 5000 MW) and end-group (-OH, -OCH3) on protein adsorption from plasma are reported. Similar to previous single protein adsorption studies (L.D. Unsworth et al., Langmuir 2005;21:1036-41) it was found that, of the different surfaces investigated, PEO layers formed from solutions near the cloud point adsorbed the lowest amount of fibrinogen from plasma. Layers of hydroxyl-terminated PEO of MW 600 formed under these low solubility conditions showed almost complete suppression (versus controls) of the Vroman effect, with 20+/-1 ng/cm2 adsorbed fibrinogen at the Vroman peak and 6.7+/-0.6 ng/cm2 at higher plasma concentration. By comparison, Vroman peak adsorption was 70+/-20 and 50+/-3 ng/cm2, respectively, for 750-OCH3 and 2000-OCH3 layers formed under low solubility conditions; adsorption on these surfaces at higher plasma concentration was 16+/-9 and 12+/-3 ng/cm2. Thus in addition to the effect of solution conditions noted previously, the results of this study also suggest a chain end group effect which inhibits fibrinogen adsorption to, and/or facilitates displacement from, hydroxyl terminated PEO layers. Fibrinogen adsorption from plasma was not significantly different for surfaces prepared with PEO of molecular weight 750 and 2000 when the chain density was the same ( approximately 0.5 chains/nm2) supporting the conclusion that chain density may be the key property for suppression of protein adsorption. The proteins eluted from the surfaces after contact with plasma were investigated by SDS-PAGE and immunoblotting. A number of proteins were detected on the various surfaces including fibrinogen, albumin, C3 and apolipoprotein A-I. The blot responses were zero or weak for all four proteins of the contact system; some complement activation was observed on all of the surfaces studied.
Unsworth
Larry D
LD
Department of Chemical Engineering, McMaster University, 1280 Main Street West, Hamilton, Ont., Canada L8S 4L7.
Sheardown
Heather
H
Brash
John L
JL
D000327:Adsorption; D000418:Albumins / Q000737:chemistry; D016632:Apolipoprotein A-I / Q000737:chemistry; D001672:Biocompatible Materials / Q000737:chemistry*; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D005340:Fibrinogen / Q000737:chemistry; D006046:Gold / Q000737:chemistry*; D006801:Humans; D015151:Immunoblotting; D007457:Iodine Radioisotopes / Q000737:chemistry; D007477:Ions; D011092:Polyethylene Glycols / Q000737:chemistry*; D013438:Sulfhydryl Compounds / Q000737:chemistry*; D013499:Surface Properties; D013696:Temperature; D013997:Time Factors
D000418:Albumins; D016632:Apolipoprotein A-I; D001672:Biocompatible Materials; D007457:Iodine Radioisotopes; D007477:Ions; D013438:Sulfhydryl Compounds; D011092:Polyethylene Glycols; D006046:Gold; D005340:Fibrinogen