Jingan Li

Surface modification of implanted cardiovascular metal stents: From antithrombosis and antirestenosis to endothelialization

Driven by the complications occurring with bare metal stents and drug-eluting stents, concerns have been raised over strategies for long-term safety, with respect to preventing or inhibiting stent thrombosis, restenosis, and in-stent restenosis in particularly. Surface modification is very important in constructing a buffer layer at the interface of the organic and inorganic materials and in ultimately obtaining long-term biocompatibility. In this review, we summarize the developments in surface modification of implanted cardiovascular metal stents. This review focuses on the modification of metal stents via coating drugs or biomolecules to enhance antithrombosis, antirestenosis, and/or endothelialization. In addition, we indicate the probable future work involving the modification of the metallic blood-contacting surfaces of stents and other cardiovascular devices that are under development.

Multifunctional Coating Based on Hyaluronic Acid and Dopamine Conjugate for Potential Application on Surface Modification of Cardiovascular Implanted Devices.

Surface modification by conjugating 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 inducing desirable vascular cell-material interaction. In the present work, the multicoatings of hyaluronic acid (HA) and dopamine (PDA) were prepared onto 316L stainless steel (316L SS) via chemical conjugation (Michael addition, Schiff base reaction, and electrostatic adsorption). The results of platelet adhesion and activation and the whole blood tests indicated that the HA/PDA coatings obtained better hemocompatibility compared with the bare 316L SS and HA or PDA immobilized on 316L SS. The HA/PDA coatings also inhibited the proliferation of smooth muscle cells and adhesion/activation of macrophages effectively, whereas not all the HA/PDA coatings improved surface endothelialization rapidly and the effects of the multifunctional coatings on endothelial cell growth depend on the HA amounts (1.0, 2.0, and 5.0 mg/mL, labeled as PDA-HA-1, PDA-HA-2, and PDA-HA-5 respectively). Herein the PDA-HA-1 and PDA-HA-2 coatings were found to improve endothelial cell adhesion and proliferation significantly. The tissue compatibility of the HA/PDA coatings also depends on the HA amounts, and the PDA-HA-2 coating was proved to cause milder in vivo tissue response. Additionally, the mechanism of the HA molecular weight change and in vivo tissue response was also explored. These results effectively suggested that the HA/PDA coating might be promising when serving as a cardiovascular implanted device coating.

Research of smooth muscle cells response to fluid flow shear stress by hyaluronic acid micro-pattern on a titanium surface

The morphology of vascular smooth muscle cells (SMCs) in the normal physiological state depends on cytoskeletal distribution and topology beneath, and presents vertical to the direction of blood flow shear stress (FFSS) although SMCs physiologically are not directly exposed to the shear conditions of blood flow. However, this condition is relevant for arteriosclerotic plaques and the sites of a vascular stent, and little of this condition in vitro has been studied and reported till now. It is unclear what will happen to SMC morphology, phenotype and function when the direction of the blood flow changed. In this paper, the distribution of SMCs in a specific area on Ti surface was regulated by micro-strips of hyaluronic acid (HA). Cell morphology depended on the distribution of the cytoskeleton extending along the micrographic direction. Simulated vascular FFSS was perpendicular or parallel to the direction of the cytoskeleton distribution. Based on investigating the morphology, apoptotic number, phenotypes and functional factors of SMCs, it was obtained that SMCs of vertical groups showed more apoptosis, expressed more contractile types and secreted less TGF-β1 factor compared with SMCs of parallel groups, the number of ECs cultured by medium from SMCs of parallel groups was larger than vertical groups. This study could help to understand the effect of direction change of FFSS on patterned SMC morphology, phenotype and function.

A novel coating of type IV collagen and hyaluronic acid on stent material-titanium for promoting smooth muscle cell contractile phenotype

The method of stent implantation is currently considered an effective means of treating atherosclerosis. However, implanting of cardiovascular stent often leads to intimal breakage and hyperplasia. The phenomenon that vascular smooth muscle cells (SMCs) transform from contractile to synthetic phenotype becomes a serious obstacle to intimal recovery. To improve how SMCs transform from a synthetic to contractile phenotype, a technique of coimmobilization was used to form type IV collagen (CoIV) and hyaluronic acid (HA) coating on the widely used stent material, titanium (Ti). In this work, several bio-functional coatings made of CoIV/HA mixtures in different ratios were fabricated on the Ti surface. The quantitative characterization of CoIV showed that introducing HA could enhance the amount of the immobilized CoIV on the alkali activated Ti (TiOH) surface. The immunofluorescence staining results of myosin heavy chain (MHC) and DAPI showed that the coating of CoIV/HA in ratios of 200 μg/ml (M200) and 500 μg/ml (M500) also could promote SMCs expressing more contractile phenotype compared with TiOH/CoIV control samples, while the AO/PI staining results indicated that SMCs on the M200 and M500 samples showed less apoptosis ratio. Thus, we hope that this study can provide more helpful exploration and application for promoting the SMC contractile phenotype on the cardiovascular stents.

The endothelialization and hemocompatibility of the functional multilayer on titanium surface constructed with type IV collagen and heparin

The type IV collagen/heparin (IVCol/Hep) multilayer was developed on amino-silanized titanium (Ti) surface layer by layer self-assembly. Ti, TiOH, TiOHA and TiOHA(HC)3H were characterized by Fourier transform infrared spectroscopy (FTIR), water contact angle measurements and scanning electron microscopy (SEM), respectively. Alcian Blue 8GX staining and immunofluorescence staining were used to characterize the heparin (Hep) and type IV collagen (IVCol), respectively. The blood compatibilities of Ti and the treated Ti were evaluated by platelet adhesion test and clotting time using PRP. Blood compatibility tests reveal that the assembled functional multilayer displayed less platelets adhesion and prolonged APTTs time compared with the controlled Ti. Endothelial cells (ECs) culture results showed more attached and proliferated ECs on the TiOHA(HC)3H than that on Ti, especially compared with that on TiOH and TiOHA. Thus, the assembled Hep and IVCol multilayer can improve the cell compatibility and the blood compatibility. We anticipate that this IVCol/Hep functional multilayer will be beneficial to enhance the biocompatibility of the Ti-based biomaterial devices.

Tailoring of the titanium surface by preparing cardiovascular endothelial extracellular matrix layer on the hyaluronic acid micro-pattern for improving biocompatibility

It has been proved that high molecular weight hyaluronic acid (HMW-HA, 1×10(6) Da) micro-strips 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±0.5×10(5) cells/cm2>2.2±0.7×10(5) cells/cm2 on ECM/TiOH, 7.5±1.3×10(5) cells/cm2 on TiOH, 3.4±0.9×10(5) cells/cm2 on TiOH/HAP and 3.6±1.2×10(5) 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±0.1×10(3)<9.2±1.5×10(3) cells/cm2 on ECM/TiOH, 8.8±0.3×10(3) cells/cm2 on TiOH, 7.3±0.7×10(3) cells/cm2 on TiOH/HAP and 9.6±0.9×10(3) 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.

Guidance of Stem Cells to a Target Destination in Vivo by Magnetic Nanoparticles in a Magnetic Field

Stem cells contribute to physiological processes such as postischemic neovascularization and vascular re-endothelialization, which help regenerate myocardial defects or repair vascular injury. However, therapeutic efficacy of stem cell transplantation is often limited by inefficient homing of systemically administered cells, which results in a low number of cells accumulating at sites of pathology. In this study, anti-CD34 antibody-coated magnetic nanoparticles (Fe3O4@PEG-CD34) are shown to have high affinity to stem cells. The results of hemolysis rate and activated partial thromboplastin time (APTT) tests indicate that such nanoparticle may be used safely in the blood system. In vitro studies showed that a nanoparticle concentration of 100 μg/mL gives rise to a significant increase in cell retention using an applicable permanent magnet, exerting minimal negative effect on cell viability and migration. Subsequent in vivo studies indicate that nanopartical can specifically bind stem cells with good magnetic response. Anti-CD34 antibody coated magnetic nanoparticle may be used to help deliver stem cells to a lesion site in the body for better treatment.

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.

Effect of micropatterned TiO2 nanotubes thin film on the deposition of endothelial extracellular matrix: For the purpose of enhancing surface biocompatibility

The vascular endothelial cells (EC) extracellular matrix (ECM) on the biomaterial surface can significantly improve the blood compatibility and cell compatibility of the cardiovascular materials. In the present study, two types of micropatterned TiO2 nanotubes surfaces (gronano and toponano) were fabricated on the titanium surface by photolithography and two-step anodizing technology, for the purpose of enhancing the deposition and loading ability of the EC ECM. The effect of the micropatterned nanotubes on EC ECM deposition and loading was investigated by qualitative and quantitative characterizations of type IV collagen (CoIV). The blood compatibility of the deposited ECM layers was evaluated by platelet adhesion and activation tests, and the endothelialization function of the deposited ECM layers was investigated by EC culture for 3 days. As a result, there was more CoIV on the toponano surface compared with the control. Meanwhile, the ECM loaded toponano (ECM/toponano) possessed better blood compatibility and better endothelialization than the control. This ECM loaded micro-/nanocomposite thin film was anticipated for the potential application of the surface modification of cardiovascular devices based on its excellent biocompatibility.

Controlling Molecular Weight of Hyaluronic Acid Conjugated on Amine-rich Surface: Toward Better Multifunctional Biomaterials for Cardiovascular Implants

The molecular weights (MWs) of hyaluronic acid (HA) in extracellular matrix secreted from both vascular endothelial cells (VECs) and vascular smooth muscle cells (VSMCs) play crucial roles in the cardiovascular physiology, as HA with appropriate MW influences important pathways of cardiovascular homeostasis, inhibits VSMC synthetic phenotype change and proliferation, inhibits platelet activation and aggregation, promotes endothelial monolayer repair and functionalization, and prevents inflammation and atherosclerosis. In this study, HA samples with gradients of MW (4 × 103, 1 × 105, and 5 × 105 Da) were prepared by covalent conjugation to a copolymerized film of polydopamine and hexamethylendiamine (PDA/HD) as multifunctional coatings (PDA/HD-HA) with potential to improve the biocompatibility of cardiovascular biomaterials. The coatings immobilized with high-MW-HA (PDA/HD-HA-2: 1 × 105 Da; PDA/HD-HA-3: 5 × 105 Da) exhibited a remarkable suppression of platelet activation/aggregation and thrombosis under 15 dyn/cm2 blood flow and simultaneously suppressed the adhesion and proliferation of VSMC and the adhesion, activation, and inflammatory cytokine release of macrophages. In particular, PDA/HD-HA-2 significantly enhanced VEC adhesion, proliferation, migration, and functional factors release, as well as the captured number of endothelial progenitor cells under dynamic condition. The in vivo results indicated that the multifunctional surface (PDA/HD-HA-2) created a favorable microenvironment of endothelial monolayer formation and functionalization for promoting reendothelialization and reducing restenosis of cardiovascular biomaterials.