Layer-by-layer construction of the heparin/fibronectin coatings on titanium surface:stability and functionality

Abstract

Layer-by-layer assembly as a versatile bottom-up nanofabrication technique has been widely used in the development of biomimetic materials with superior mechanical and biological properties. In this study, layer-by-layer assembled heparin/fibronectin biofunctional films were fabricated on titanium (Ti) surface to enhance the blood anticoagulation and accelerate the endothelialization simultaneously. The wettability and chemical changes of the assembled films were investigated by static water contact angle measurement and fourier transform infrared spectroscopy (FTIR). The morphology of modified Ti surfaces were observed using scanning electron microscopy (SEM). The real time assembly process was in-situ monitored by quartz crystal microbalance with dissipation (QCM-D). The stability of the films was evaluated by measuring the changes in wettability and the quantity of heparin and fibronectin on the surfaces. The anticoagulation properties of the films were quantitatively rated using Activated partial thromboplastin time (APTT) analysis. New peaks of hydroxyl and amino group were observed on the assembled Ti srufaces by FTIR. The contact angles varied among the films with different bilayer numbers, indicating the successful graft of the heparin and fibronectin layer-by-layer. QCM-D results showed that the frequency shift increased with the bilayer numbers, and the heparin and fibronectin could form multilayers. The assembly films were stable after incubation in PBS for 24h based on the results of the contact angle measurement and the quantity of heparin and fibronectin analysis. APTT results suggested that the assembled films kept excellent antithrombotic properties. All these results revealed that the assembled heparin/fibronectin films with stabiltiy and anticoagulation property could be firmly formed on titanium surfaces.Our study further demonstrates that layer-by-layer assembly of heparin and fibronectin will provide a potential and effective tool for biomaterials surface modification.