Abstract

It has been proved that high molecular weight hyaluronic acid (HMW-HA, 1×106 Da) microstrips on titanium (Ti) surface can elongate the human vascular endothelial cell (EC) morphology, subsequently enhance endothelial extracellular matrix (ECM) deposition in our previous work. The HMW-HA micro-strips were anticipated to possess good hemocompatibility and EC compatibility simultaneously. However, the single HMW-HA micro-strips on Ti substrate showed bad hemocompatibility. To solve this problem, a method combining HA micro-pattern and EC decellularization was developed, and the endothelial extracellular matrix layer on the HA micro-pattern (ECM/HAP) showed excellent hemocompatibility and endothelial progenitor cells (EPCs) compatibility (cell number: 14.3±0.5×105 cells/cm2 > 2.2±0.7×105 cells/cm2 on ECM/TiOH,7.5±1.3×105 cells/cm2 on TiOH, 3.4±0.9×105 cells/cm2 on TiOH/HAP and 3.6±1.2×105 cells/cm2 on Ti). We also found that the ECM/HAP coating could significantly inhibit the excessive proliferation of smooth muscle cells (SMCs) (cck-8 absorption: 0.25±0.06 < 1.18±0.09 A.U. on ECM/TiOH,0.87±0.15 A.U. on TiOH and 1.55±0.11 A.U. on Ti) and the attachment of macrophages (cell number: 1.3±0.1×103 < 9.2±1.5×103 cells/cm2 on ECM/TiOH, 8.8±0.3×103 cells/cm2 on TiOH,7.3±0.7×103 cells/cm2 on TiOH/HAP and 9.6 ±0.9×103 cells/cm2 on Ti in 12 h). These data suggest that the multifunctional ECM/HAP coating can be used to build the bionic human endothelial ECM on the biomaterials surface, which might provide a potential and effective method for surface modification of cardiovascular devices.

2. Materials and methods

2.1. Materials and reagents 

mouse monoclonal antihuman fibrinogen (FGN)

 chain antibody(ab119948) and HRP conjugated goat antihuman fibrinogen anti-body (ab64664) were purchased from abcam (UK). Prostacyclin(PGI2) ELISA kit was provided by Wuhan ColorfulGene BiologicalTechnology Co. Ltd., China. All the other reagents were analyticalof grade.