Studies of a novel human thrombomodulin immobilized substrate: surface characterization and anticoagulation activity evaluation.
Immobilization of the anticoagulative or antithrombogenic biomolecule has been considered as one of the important methods to improve the blood compatibility of artificial biomaterials. In this study, a novel immobilization reaction scheme was utilized to incorporate the human thrombomodulin, an endothelial cell associated glycoprotein, onto the cover glass surface with an aim to develop an anticoagulative substrate. Trichlorotriazine and amino-terminated silane were employed as the coupling agents, while the polyethylene glycol with a molecular weight of 1500 was used as the spacer in this reaction scheme. Protein C activation assay indicated the immobilized human thrombomodulin still has this coenzymatic activity but is lower, possibly due to the conformation variation by the coupling agents. In vitro platelet adhesion assay has demonstrated the surface with immobilized human thrombomodulin is much less platelet-activating than others. Therefore, the novel reaction scheme proposed here is very promising for future development of an anticoagulative silicon or cover glass substrate (e.g. implantable sensor or biochip) by the immobilization of antithrombogenic protein, such as the human thrombomodulin in this study.
Han
H S
HS
Department of Medical Technology, Chung Hwa Institute of Technology, Tainan, Taiwan, ROC.
Yang
S L
SL
Yeh
H Y
HY
Lin
J C
JC
Wu
H L
HL
Shi
G Y
GY
D000925:Anticoagulants / Q000378:metabolism; D001672:Biocompatible Materials; D006801:Humans; D066298:In Vitro Techniques; D008422:Materials Testing; D010973:Platelet Adhesiveness; D011486:Protein C / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D013057:Spectrum Analysis; D013499:Surface Properties; D018180:Thrombomodulin / Q000378:metabolism*; D014965:X-Rays
D000925:Anticoagulants; D001672:Biocompatible Materials; D011486:Protein C; D011994:Recombinant Proteins; D018180:Thrombomodulin