In vitro haemocompatibility and stability of two types of heparin-immobilized silicon surfaces

Abstract

Heparin was covalently immobilized onto a silicon surface by two different methods, carbodiimide-based immobilization and photo-immobilization. In the former method, a (3-aminopropyl) trimethoxysilane (APTMS) self-assembled monolayer (SAM) or multilayer was first coated onto the silicon surface as the bridging layer, and heparin was then attached to the surface in the presence of water-soluble carbodiimide. In the latter method, an octadecyltrichlorosilane (OTS) SAM was coated on the silicon surface as the bridging layer, and heparin was modified by attaching photosensitive aryl azide groups. Upon UV illumination, the modified heparin was then covalently immobilized onto the surface. The hydrophilicity of the silicon surface changed after each coating step, and heparin aggregates on APTMS SAM and OTS SAM were observed by atomic force microscopy (AFM). In vitro haemocompatibility assays demonstrated that the deposition of APTMS SAM, APTMS multilayer and OTS SAM enhanced the silicon's haemocompatibility, which was further enhanced by the heparin immobilization. There is no evident distinction regarding the haemocompatibility between the heparin-immobilized surfaces by both methods. However, heparin on silicon with APTMS SAM and multilayer as the bridging layers is very unstable when tested in vitro with a saline solution at 37 °C, due to the instability of APTMS SAM and multilayer on silicon. Meanwhile, photo-immobilized heparin on silicon with OTS SAM as the bridging layer showed superb stability.