Surface modification with an antithrombin-heparin complex for anticoagulation: studies on a model surface with gold as substrate.
Gold was used as a substrate for immobilization of an antithrombin-heparin (ATH) covalent complex to investigate ATH as a surface modifier to prevent blood coagulation. Three different surface modification methods were used to attach ATH to gold: (i) direct chemisorption; (ii) using dithiobis(succinimidyl propionate) (DSP) as a linker molecule and (iii) using polyethylene oxide (PEO) as a linker/spacer. The ATH-modified surfaces were compared to analogous heparinized surfaces. Water contact angles and X-ray photoelectron spectroscopy confirmed the modifications and provided data on surface properties and possible orientation. Ellipsometry measurements showed that surface coverage of DSP and PEO was high. ATH and heparin densities were quantified using radioiodination and quartz crystal microbalance, respectively. The surface density of ATH was greatest on the DSP surface (0.17 microg cm(-2)) and lowest on the PEO (0.05 microg cm(-2)). The low uptake on the PEO surface was likely due to the protein resistance of the PEO component. Using radioiodinated antithrombin (AT), it was shown that ATH-immobilized surfaces bound significantly greater amounts from both buffer and plasma than the analogous heparinized surfaces. Immunoblot analysis of proteins adsorbed from plasma demonstrated that surfaces chemisorbed with PEO, whether or not subsequently modified with ATH, inhibited non-specific adsorption. The immunoblot response for AT was stronger on the DSP-ATH than on the heparin surfaces, thus confirming the results from radiolabelling. The ATH surfaces again showed higher selectivity for AT binding than analogous heparin-modified surfaces, indicating the enhanced anticoagulant potential of ATH for biomaterial surface modification.
Sask
Kyla N
KN
School of Biomedical Engineering, McMaster University, 1280 Main Street West, Hamilton, ON, Canada L8S 4K1.
Zhitomirsky
Igor
I
Berry
Leslie R
LR
Chan
Anthony K C
AK
Brash
John L
JL
D000327:Adsorption / Q000187:drug effects; D000925:Anticoagulants / Q000494:pharmacology*; D000991:Antithrombins / Q000494:pharmacology*; D001777:Blood Coagulation / Q000187:drug effects*; D001798:Blood Proteins / Q000378:metabolism; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D005340:Fibrinogen / Q000494:pharmacology; D006046:Gold / Q000494:pharmacology*; D006493:Heparin / Q000494:pharmacology*; D006801:Humans; D007457:Iodine Radioisotopes; D008422:Materials Testing / Q000379:methods*; D056951:Photoelectron Spectroscopy; D013499:Surface Properties / Q000187:drug effects; D014867:Water / Q000737:chemistry
D000925:Anticoagulants; D000991:Antithrombins; D001798:Blood Proteins; D007457:Iodine Radioisotopes; D014867:Water; D006046:Gold; D005340:Fibrinogen; D006493:Heparin