Tailoring ZE21B Alloy with Nature-Inspired Extracellular Matrix Secreted by Micro-Patterned Smooth Muscle Cells and Endothelial Cells to Promote Surface Biocompatibility.

Delayed surface endothelialization is a bottleneck that restricts the further application of cardiovascular stents. It has been reported that the nature-inspired extracellular matrix (ECM) secreted by the hyaluronic acid (HA) micro-patterned smooth muscle cells (SMC) and endothelial cells (EC) can significantly promote surface endothelialization. However, this ECM coating obtained by decellularized method (dECM) is difficult to obtain directly on the surface of degradable magnesium (Mg) alloy. In this study, the method of obtaining bionic dECM by micro-patterning SMC/EC was further improved, and the nature-inspired ECM was prepared onto the Mg-Zn-Y-Nd (ZE21B) alloy surface by self-assembly. The results showed that the ECM coating not only improved surface endothelialization of ZE21B alloy, but also presented better blood compatibility, anti-hyperplasia, and anti-inflammation functions. The innovation and significance of the study is to overcome the disadvantage of traditional dECM coating and further expand the application of dECM coating to the surface of degradable materials and materials with different shapes.

Investigation of Mg-xLi-Zn alloys for potential application of biodegradable bone implant materials.

Implant therapy after osteosarcoma surgery is a major clinical challenge currently, especially the requirements for mechanical properties, degradability of the implants, and their inhibition of residual tumor cells. Biodegradable magnesium (Mg) alloy as medical bone implant material has full advantages and huge potential development space. Wherein, Mg-lithium (Li) based alloy, as an ultra-light alloy, has good properties for implants under certain conditions, and both Mg and Li have inhibitory effects on tumor cells. Therefore, Mg-Li alloy is expected to be applied in bone implant materials for mechanical supporting and inhibiting tumor cells simultaneously. In this contribution, the Mg-xLi-Zinc (Zn) series alloys (x = 3 wt%, 6 wt%, 9 wt%) were prepared to study the influence of different elements and contents on the structure and properties of the alloy, and the biosafety of the alloy was also evaluated. Our data showed that the yield strength, tensile strength, and elongation of as-cast Mg-xLi-Zn alloy were higher than those of as-cast Mg-Zn alloy; Mg-xLi-Zn alloy can kill osteosarcoma cells (MG-63) in a concentration-dependent manner, wherein Mg-3Li-Zn alloy (x = 3 wt%) and Mg-6Li-Zn alloy (x = 6 wt%) promoted the proliferation of osteoblasts (MC3T3) at a certain concentration of Li. In summary, our study demonstrated that the Mg-6Li-Zn alloy could be potentially applied as a material of orthopedic implant for its excellent multi-functions.

Micro-/Nano-Scales Direct Cell Behavior on Biomaterial Surfaces.

Cells are the smallest living units of a human body's structure and function, and their behaviors should not be ignored in human physiological and pathological metabolic activities. Each cell has a different scale, and presents distinct responses to specific scales: Vascular endothelial cells may obtain a normal function when regulated by the 25 µm strips, but de-function if the scale is removed; stem cells can rapidly proliferate on the 30 nm scales nanotubes surface, but stop proliferating when the scale is changed to 100 nm. Therefore, micro and nano scales play a crucial role in directing cell behaviors on biomaterials surface. In recent years, a series of biomaterials surface with micro and/or nano scales, such as micro-patterns, nanotubes and nanoparticles, have been developed to control the target cell behavior, and further enhance the surface biocompatibility. This contribution will introduce the related research, and review the advances in the micro/nano scales for biomaterials surface functionalization.

Platelet Adhesion and Activation on Chiral Surfaces: The Influence of Protein Adsorption.

Adsorbed proteins and their conformational change on blood-contacting biomaterials will determine their final hemocompatibility. It has frequently been reported that surface chirality of biomaterials may highly influence their protein adsorption behavior. Here, lysine and tartaric acid with different chirality were immobilized onto TiO2 films respectively, and the influence of surface chirality on protein adsorption, platelet adhesion, and activation was also investigated. It showed that the l- and d-molecule grafted samples had almost the same grafting density, surface topography, chemical components, and hydrophilicity in this study. However, biological behaviors such as protein adsorption, platelet adhesion, and activation were quite different. The d-lysine grafted surface had a greater ability to inhibit both bovine serum albumin and fibrinogen adsorption, along with less degeneration of fibrinogen compared to the l-lysine anchored surface. However, the d-tartaric acid grafted surface adsorbed more protein but with less denatured fibrinogen compared to the l-tartaric acid grafted one. Further studies showed that the secondary structural change of the adsorbed albumin and fibrinogen on all surfaces with deduction of the α-helix content and increase of disordered structure, while the changing degree was apparently varied. As a result, the d-lysine immobilized surface absorbed less platelets and red blood cells and achieved slightly increased platelet activation. For tartaric acid anchored surfaces, a larger number of platelets adhered to the D-surface but were less activated compared to the L-surface. In conclusion, the surface chirality significantly influenced the adsorption and conformational change of blood plasma protein, which in turn influenced both platelet adhesion and activation.

Surface modification of esophageal stent materials by a polyethylenimine layer aiming at anti-cancer function.

Esophageal cancer is difficult to cure globally and possesses high mortality rate, and it is generally accepted that palliative care such as stent implantation is the main therapy method for esophageal cancer in later period. However, the restenosis caused by tumor cells and inflammatory cells seriously interferes the stent clinical application and limits its long-term services. To solve this problem, series of drug delivery stents were developed and proven rather effective in the early stage of implantation, but more serious restenosis occurred after the drug delivery was over, which endangered the patients' life. Therefore, endowing the esophageal stent continuous anti-cancer function become an ideal strategy for inhibiting the restenosis. In this contribution, the functional layer composed of polydopamine (PDA) and Poly-ethylenimine (PEI) with series of molecular weights (MW, 1.8 × 103, 1 × 104, 2.5 × 104 and 7 × 104 Da) were fabricated onto the esophageal stent material 317L stainless steel (317L SS) surface. The surface characterization including amine quantitative, atomic force microscopy (AFM) and water contact angle measurement indicated successful preparation of the PDA/PEI layer. The Eca109 cells culture results proved that the PDA/PEI layers significantly improve Eca109 cells apoptosis and necrosis, suggesting excellent anti-cancer function. In addition, we also found that the anti-cancer function of the PDA/PEI layers was positively correlated to the immobilized PEIs' MW. All the results demonstrated the potential application of the PDA/PEI layers on the surface modification of esophageal stent for continuous anti-cancer function. It is generally accepted that the restenosis caused by tumor cells seriously interferes the esophageal stent clinical application. Thus, endowing the esophageal stent continuous anti-cancer function is the ideal strategy for inhibiting the restenosis. In this work, we fabricated functional layers composed of polydopamine (PDA) and Poly-ethylenimine (PEI) with series of molecular weights (MW, 1.8 × 103, 1 × 104, 2.5 × 104 and 7 × 104 Da) onto the esophageal stent material 317L stainless steel (317L SS) surface to inhibit the tumor cells growth, and this function was related to the PEIs' molecular weights. The functional PDA/PEI layers were expected potentially applied for surface modification of esophageal stent materials.

Constructing bio-layer of heparin and type IV collagen on titanium surface for improving its endothelialization and blood compatibility.

The modification of cardiovascular stent surface for a better micro-environment has gradually changed to multi-molecule, multi-functional designation. In this study, heparin (Hep) and type IV collagen (IVCol) were used as the functional molecule to construct a bifunctional micro-environment of anticoagulation and promoting endothelialization on titanium (Ti). The surface characterization results (AFM, Alcian Blue 8GX Staining and fluorescence staining of IVCol) indicated that the bio-layer of Hep and IVCol were successfully fabricated on the Ti surface through electrostatic self-assembly. The APTT and platelet adhesion test demonstrated that the bionic layer possessed better blood compatibility compared with Ti surface. The adhesion, proliferation, migration and apoptosis tests of endothelial cells proved that the Hep/IVCol layer was able to enhance the endothelialization of the Ti surface. The in vivo animal implantation results manifested that the bionic surface could encourage new endothelialization. This work provides an important reference for the construction of multifunction micro-environment on the cardiovascular scaffold surface.

Co-culture of endothelial cells and patterned smooth muscle cells on titanium: construction with high density of endothelial cells and low density of smooth muscle cells.

Endothelialization has been considered a promising method to improve the biocompatibility of vascular implanted biomaterials. However, little is known about the anti-coagulation, anti-inflammatory, anti-atherosclerosis and anti-shedding property of the attached endothelial cells (ECs) and the relationship with their bio-environment and material-environment, which are both important evaluations to the cardiovascular biomaterials designed for tissue engineering applications and in vivo implantation. In this in vitro study, a novel co-culture model was built, where vascular smooth muscle cells (SMCs) were cultured on the hyaluronic acid (HA) micro-strip patterned titanium (Ti) surface on a low density to biomimetic the EC pericyte environment. Subsequently, the EC number and its functional factor, including nitric oxide (NO), prostacyclin (PGI2), tissue factor pathway inhibitor (TFPI), thrombomodulin (TM), and the inflammatory induced factor, endothelial leukocyte adhesion molecule-1 (E-selectin) were quantified, respectively. The anti-shedding property was also assessed by the blood flow shear stress (BFSS) acting. The results showed that the novel co-culture model possessed better EC coverage, functional factor release and anti-shedding functions than the control.

The enhanced antibacterial effect of BNNS_Van@CS/MAO coating on Mg alloy for orthopedic applications.

The development of multifunctional Mg-based active implants with controllable degradation and antibacterial capabilities has become a hotspot in the research field of biodegradable metallic materials. To this end, a BN nanosheets (BNNS) _vancomycin (Van) @chitosan (CS) nanocomposite coating containing two antibacterial components (BNNS and Van) was prepared on Mg alloys via a micro-arc oxidation (MAO) pre-treatment combined with following electrodeposition. The related characterizations of the coating show that the composite coating has a high roughness, hydrophobicity and fair corrosion resistance. In vitro antibacterial experiments show that the BNNS_Van@CS/MAO composite coating have obvious inhibitory effect on the growth of both E. coli and S. aureus. The antibacterial effect of the BNNS_Van@CS/MAO composite coating was attributed to the synergistic effect of CS, BNNS and Van. This study provides a valuable surface modification strategy for developing multifunctional Mg-based implants with good corrosion resistance and antibacterial properties.

Surface modification of biodegradable magnesium alloy with poly (L-lactic acid) and sulfonated hyaluronic acid nanoparticles for cardiovascular application.

Magnesium (Mg) and its alloys have attracted extensive attention of researchers in the field of cardiovascular implants due to their good mechanical properties and biosafety. Constructing a multifunctional hybrid coating seems to be an effective strategy to address the insufficient endothelialization and poor corrosion resistance of Mg alloy vascular stents. In this study, a dense layer of magnesium fluoride (MgF2) was prepared on the surface of Mg alloy aiming at better corrosion resistance; Thereafter, sulfonated hyaluronic acid (S-HA) was made into small sized nanoparticles (NP) which were deposited on the MgF2 surface by self-assembly method, followed with poly-L-lactic acid (PLLA) coating preparation by one-step pulling method. The blood and cell tests showed that the composite coating had good blood compatibility, pro-endothelial, anti-hyperplasia and anti-inflammatory functions. Compared to current clinical PLLA@ Rapamycin coating, our PLLA/NP@S-HA coating showed better functions of promoting endothelial cells growth. These results strongly furnished a promising and feasible strategy for the surface modification of Mg-based degradable cardiovascular stents.

Hydrogel-coated needles prevent puncture site bleeding in arteriovenous fistula and arteriovenous grafts in rats.

INTRODUCTION: Imperfect hemostasis after arteriovenous fistula (AVF) and arteriovenous graft (AVG) cannulation can cause a hematoma or pseudoaneurysm and leads to poor satisfaction. We hypothesized that a hydrogel-coated needle would effectively and rapidly stop bleeding after vascular cannulation in a rat AVF and AVG model. METHOD: A hydrogel comprised of sodium alginate (SA), hyaluronic acid (HA), and calcium carbonate was coated onto the surface of suture needles using a rotating system. The needles were observed using scanning electron microscopy (SEM) and immunofluorescence. Rat AVF with or without renal failure and AVG were punctured using bare and hydrogel-coated needles. The tissues were examined by histology. RESULT: The hydrogel was successfully coated onto the surface of 30 G needles and confirmed by SEM. Hydrogel-coated needles rapidly stopped bleeding after AVF and AVG cannulation in rat. CONCLUSION: In this preliminary animal research, hydrogel-coated needles can stop AVF and AVG puncture-site bleeding; but additional clinical studies are needed to justify whether it is still effective in clinical.

Hyaluronic acid/polyethyleneimine nanoparticles loaded with copper ion and disulfiram for esophageal cancer.

In the clinical treatment of cancer, improving the effectiveness and targeting of drugs has always been a bottleneck problem that needs to be solved. In this contribution, inspired by the targeted inhibition on cancer from combination application of disulfiram and divalent copper ion (Cu2+), we optimized the concentration of disulfiram and Cu2+ ion for inhibiting esophageal cancer cells, and loaded them in hyaluronic acid (HA)/polyethyleneimine (PEI) nanoparticles with specific scales, in order to improve the effectiveness and targeting of drugs. The in vitro cell experiments demonstrated that more drug loaded HA/PEI nanoparticles accumulated to the esophageal squamous cell carcinoma (Eca109) and promoted higher apoptosis ratio of Eca109. Both in vitro and in vivo biological assessment verified that the disulfiram/Cu2+ loaded HA/PEI nanoparticles promoted the apoptosis of cancer cells and inhibited the tumor proliferation, but had no toxicity on other normal organs.

Inhibition of programmed death-1 decreases neointimal hyperplasia after patch angioplasty.

Neointimal hyperplasia remains an obstacle after vascular interventions. Programmed death-1 (PD-1) antibody treatment decreases tumor cell proliferation and secretion of inflammatory factors, and several antineoplastic drugs show efficacy against neointimal hyperplasia. We hypothesized that inhibition of PD-1 inhibits neointimal hyperplasia in a rat patch angioplasty model. In a rat aorta patch angioplasty model, four groups were compared: the control group without treatment, a single dose of humanized PD-1 antibody (4 mg/kg) injected immediately after patch angioplasty, PD-1 antibody-coated patches, and BMS-1 (PD-1 inhibitor)-coated patches. Patches were harvested (Day 14) and analyzed. After patch angioplasty, PD-1-positive cells were present. Inhibition of PD-1 using both intraperitoneal injection of humanized PD1 antibody as well as using patches coated with humanized PD1 antibody significantly decreased neointimal thickness (p = 0.0199). There were significantly fewer PD-1 (p = 0.0148), CD3 (p = 0.0072), CD68 (p = 0.0001), CD45 (p = 0.001), and PCNA (p < 0.0001)-positive cells, and PCNA/α-actin dual positive cells (p = 0.0005), in the treated groups. Patches coated with BMS-1 showed similarly decreased neointimal thickness and accumulation of inflammatory cells. Inhibition of PD-1 using PD-1 antibody or its inhibitor BMS-1 can significantly decrease neointimal thickness in vascular patches. Inhibition of the PD-1 pathway may be a promising therapeutic strategy to inhibit neointimal hyperplasia.

Conjugating heparin, Arg-Glu-Asp-Val peptide, and anti-CD34 to the silanic Mg-Zn-Y-Nd alloy for better endothelialization.

Magnesium alloy is generally accepted as a potential cardiovascular stent material due to its good mechanical properties, biocompatibility, and biodegradability, and has become one of the research hotspots in this field. However, too fast degradation rate and delayed surface endothelialization have been the bottleneck of further application of magnesium alloy stent. In this study, we selected Mg-Zn-Y-Nd, a kind of biodegradable magnesium alloy for cardiovascular stent, and passivated its surface by alkali heat treatment and silane treatment to improve the corrosion resistance, subsequently conjugated Arg-Glu-Asp-Val (REDV) peptide and anti-CD34 to promote endothelial cells adhesion and capture endothelial progenitor cells respectively, further improving surface endothelialization. In addition, the heparin was also immobilized to the Mg-Zn-Y-Nd surface for the consideration of anti-coagulation and anti-inflammation. Systematic material characterization and biological evaluation show that we have successfully developed this composite surface on Mg-Zn-Y-Nd alloy, and achieved multiple functions such as corrosion resistance, promoting endothelialization, and inhibiting platelet/macrophage adhesion.

The mechanism of PDA/PEI/5-Fu coated esophageal stent material on inhibiting cancer associated pathological cells.

Metal stent implantation is usually applied to alleviate nonoperative palliative esophageal obstruction for esophageal cancer in the later period. However, in-stent restenosis after stent implantation limits the esophageal stents' performance due to lack of effective suppression of pathological cells from cancer microenvironment. In previous work, we modified the esophageal stent material 317L stainless steel (317LSS) surface with a poly-dopamine/poly-ethylenimine/5-fluorouracil layer (PDA/PEI/5-Fu), which had strong anti-tumor and anti-restenosis functions. Nevertheless, the mechanism of PDA/PEI/5-Fu layer against tumor and inflammation remains unclear. In this work, we revealed the mechanism of PDA/PEI/5-Fu suppressing the esophageal cancer related pathological cells (esophageal tumor cells, epithelial cells, and fibroblast) and inflammatory cells (macrophages) via series of experiments. Our data suggested that the PEI inhibited viability and E-cadherin expression of the pathological cells, and blocked the NF-κB signal pathway (reducing levels of p-NF-κB proteins). The loaded 5-Fu inhibited the inflammatory factors (TNF-α and IL-1ß) release and promoted the anti-inflammation/anti-tumor factors (IL-10 and IL-4) release from macrophages, and also suppressed pathological cells migration; both the PEI and 5-Fu contributed to the upregulation of Bax and Caspase-3 (pro-tumor-apoptosis factor), as well as the downregulation of Bcl-2 (anti-tumor-apoptosis factor) in esophageal tumor cells. All the results showed that PDA/PEI/5-Fu coating had potential multipath anti-cancer and anti-inflammatory effects in the surface modification of esophageal stents.

Preparing a novel magnesium-doped hyaluronan/polyethyleneimine nanoparticle to improve endothelial functionalisation.

Nowadays, tissue engineering vascularisation has become an important means of organ repair and treatment of major traumatic diseases. Vascular endothelial layer regeneration and endothelial functionalisation are prerequisites and important components of tissue engineering vascularisation. The present researches of endothelial functionalisation mainly focus on tissue engineering scaffold preparation and implant surface modification. Few studies have reported the interaction of endothelial functionalisation and scaled materials, especially the nanomaterials. Magnesium (Mg), as an essential cytotropic active element in the human body, should promote the growth of endothelial cells. However, the authors' previous work found that the Mg in the alloys had a defect of delayed endothelialisation, which may be attributed to the non-uniform scales of the degradation products from Mg alloys. To validate this hypothesis and fabricate a novel nanomaterial for tissue engineering vascularisation, the authors prepared Mg-doped hyaluronan (HA)/polyethyleneimine (PEI) nanoparticles for endothelial cells testing. Their data showed that the Mg-doped HA/PEI nanoparticle with small scales (diameter <150 nm) presented better ability on improving endothelial cells growth, functionalisation and nitric oxide release.

Enhancing biocompatibility and corrosion resistance of biodegradable Mg-Zn-Y-Nd alloy by preparing PDA/HA coating for potential application of cardiovascular biomaterials.

In this paper the poly-dopamine (PDA)/hyaluronic acid (HA) coatings with different HA molecular weight (MW, 4 × 103, 1 × 105, 5 × 105 and 1 × 106 Da) were prepared onto the NaOH passivated Mg-Zn-Y-Nd alloy aiming at potential application of cardiovascular implants. The characterization of weight loss, polarization curves and surface morphology indicated that the coatings with HA MW of 1 × 105 (PDA/HA-2) and 1 × 106 Da (PDA/HA-4) significantly enhanced the corrosion resistance of Mg-Zn-Y-Nd. In vitro biological test also suggested better hemocompatibility, pro-endothelialization, anti-hyperplasia and anti-inflammation functions of the PDA/HA-2- and PDA/HA-4-coated Mg-Zn-Y-Nd alloy. Nevertheless, the in vivo implantation of SD rats' celiac artery demonstrated that the PDA/HA-2 had preferable corrosion resistance and biocompatibility.

Tailoring of cardiovascular stent material surface by immobilizing exosomes for better pro-endothelialization function.

Stent intervention as available method in clinic has been widely applied for cardiovascular disease treatment for decades. However, the restenosis caused by late thrombosis and hyperplasia still limits the stents long-term application, and the essential cause is usually recognized as endothelial functionalization insufficiency of the stent material surface. Here, we address this limitation by developing a pro-endothelial-functionalization surface that immobilized a natural factors-loaded nanoparticle, exosome, onto the poly-dopamine (PDA) coated materials via electrostatic binding. This PDA/Exosome surface not only increased the endothelial cells number on the materials, but also improved their endothelial function, including platelet endothelial cell adhesion molecule-1 (CD31) expression, cell migration and nitric oxide release. The pro-inflammation macrophage (M1 phenotype) attachment and synthetic smooth muscle cell proliferation as the interference factors for the endothelialization were not only inhibited by the PDA/Exosome coating, while the cells were also regulated to anti-inflammation macrophage (M2 phenotype) and contractile smooth muscle cell, which may contribute to endothelialization. Thus, it can be summarized this method has potential application on surface modification of cardiovascular biomaterials.

Effects of degradation products of biomedical magnesium alloys on nitric oxide release from vascular endothelial cells.

Nitric oxide (NO) released by vascular endothelial cells (VECs), as a functional factor and signal pathway molecule, plays an important role in regulating vasodilation, inhibiting thrombosis, proliferation and inflammation. Therefore, numerous researches have reported the relationship between the NO level in VECs and the cardiovascular biomaterials' structure/functions. In recent years, biomedical magnesium (Mg) alloys have been widely studied and rapidly developed in the cardiovascular stent field for their biodegradable absorption property. However, influence of the Mg alloys' degradation products on VEC NO release is still unclear. In this work, Mg-Zn-Y-Nd, an Mg alloy widely applied on the biodegradable stent research, was investigated on the influence of the degradation time, the concentration and reaction time of degradation products on VEC NO release. The data showed that the degradation product concentration and the reaction time of degradation products had positive correlation with NO release, and the degradation time had negative correlation with NO release. All these influencing factors were controlled by the Mg alloy degradation behaviors. It was anticipated that it might make sense for the cardiovascular Mg alloy design aiming at VEC NO release and therapy.

Engineering Cardiovascular Implant Surfaces to Create a Vascular Endothelial Growth Microenvironment.

Cardiovascular disease (CVD) is generally accepted as the leading cause of morbidity and mortality worldwide, and an increasing number of patients suffer from atherosclerosis and thrombosis annually. To treat these disorders and prolong the sufferers' life, several cardiovascular implants have been developed and applied clinically. Nevertheless, thrombosis and hyperplasia at the site of cardiovascular implants are recognized as long-term problems in the practice of interventional cardiology. Here, we start this review from the clinical requirement of the implants, such as anti-hyperplasia, anti-thrombosis, and pro-endothelialization, wherein particularly focus on the natural factors which influence functional endothelialization in situ, including the healthy smooth muscle cells (SMCs) environment, blood flow shear stress (BFSS), and the extracellular matrix (ECM) microenvironment. Then, the currently available strategies on surface modification of cardiovascular biomaterials to create vascular endothelial growth microenvironment are introduced as the main topic, e.g., BFSS effect simulation by surface micro-patterning, ECM rational construction and SMCs phenotype maintain. Finally, the prospects for extending use of the in situ construction of endothelial cells growth microenvironment are discussed and summarized in designing the next generation of vascular implants.

The effect of anti-CD133/fucoidan bio-coatings on hemocompatibility and EPC capture.

Surface modification by immobilizing biomolecules has been widely proved to enhance biocompatibility of cardiovascular implanted devices. Here, we aimed at developing a multifunctional surface that not only provides good hemocompatibility but also functions well in capturing circulating endothelial progenitor cells (EPCs) in the blood flow to improve the surface endothelialization. In the present work, we preferred to chemically co-immobilize (Michael addition and Schiff base reaction) the anti-CD133 (EPC-specific antibody) and fucoidan (EPC-mobilization factor, which also contribute to better hemocompatibility) onto a polydopamine (PDA) film which is famous for its stability and endothelial cell (EC) compatibility. The quantality of anti-CD133 and other surface characterization (X-ray photoemission spectroscopy, atomic force microscopy and water contact angle measurement) demonstrated successful preparation of the CD133/fucoidan coating. The platelets adhesion/activation test suggested improved hemocompatibility of this bio-coating. The ex vivo experiment on New Zealand white rabbits showed that the anti-CD133/fucoidan coating had good ability on capture the circulating EPC. In addition, the quartz crystal microbalance-D indicated that the EPC behaviors, including adhesion, spreading and extracellular matrix re-molding, were related to the density of anti-CD133 in the coating. We hope this anti-CD133/fucoidan multi-functional coating may provide potential application on surface modification of cardiovascular biomaterials.