Rifang Luo

The covalent immobilization of heparin to pulsed-plasma polymeric allylamine films on 316L stainless steel and the resulting effects on hemocompatibility

For an improved hemocompatibility of 316L stainless steel (SS), we develop a facile and effective approach to fabricating a pulsed-plasma polymeric allylamine (P-PPAm) film that possesses a high cross-linking degree and a high density of amine groups, which is used for subsequent bonding of heparin. The P-PPAm film as a stent coating shows good resistance to the deformation behavior of compression and expansion of a stent. Using deionized water as an aging medium, it is demonstrated that the heparin-immobilized P-PPAm (Hep-P-PPAm) surface has a good retention of heparin. The systematic in vitro hemocompatibility evaluation reveals lower platelet adhesion, platelet activation and fibrinogen activation on the Hep-P-PPAm surface, and the activated partial thromboplastin time prolongs for about 15 s compared with 316L SS. The P-PPAm surface significantly promotes adhesion and proliferation of endothelial cells (ECs). For the Hep-P-PPAm, although EC adhesion and proliferation is slightly suppressed initially, after cultivation for 3 days, the growth behavior of ECs is remarkably improved over 316L SS. In vivo results indicate that the Hep-P-PPAm surface successfully restrain thrombus formation by growing a homogeneous and intact shuttle-like endothelium on its surface. The Hep-P-PPAm modified 316L SS shows a promising application for vascular devices.

In Vitro Investigation of Enhanced Hemocompatibility and Endothelial Cell Proliferation Associated with Quinone-Rich Polydopamine Coating

Recent investigations have demonstrated that polydopamine (PDA)-modified surfaces were beneficial to the proliferation of endothelial cells (ECs). In this work, PDA coated 316L stainless steels (316L SS) were thermally treated at 50, 100, and 150 °C respectively (hereafter designated as Th50, Th100, and Th150) and consequently produced diverse surface chemical components. In vitro hemocompatibility and vascular cell-material interactions with ECs and smooth muscle cells (SMCs) affected by surface characteristics have been investigated. The Th150, rich in quinone, showed the best hemocompatibility and could effectively inhibit platelet adhesion, activation, and fibrinogen conformation transition. The polydopamine-modified surfaces were found to induce dramatic cell-material interaction with enhanced ECs proliferation, viability and migration, release of nitric oxide (NO), and reduced SMCs proliferation. The inhibitory effect of SMCs proliferation might be associated with the surface catechol content. The coating on Th150 showed a good resistance to the deformation of compression and expansion of vascular stents. These results effectively suggested that the Th150 coating might be promising when served as a stent coating platform.

Mussel‐Inspired Coating of Polydopamine Directs Endothelial and Smooth Muscle Cell Fate for Re‐endothelialization of Vascular Devices

Polydopamine (PDAM), a mussel adhesive protein inspired coating that can be easily deposited onto a wide range of metallic, inorganic, and organic materials, gains interest also in the field of biomaterials. In this work, PDAM is applied as coating on 316L stainless steel (SS) stents and the response of cells of the blood vessel wall, human umbilical vein endothelial cell (HUVEC), and human umbilical artery smooth muscle cell (HUASMC) as predictors for re-endothelialization is tested. It is found that the PDAM-modified surface significantly enhances HUVEC adhesion, proliferation, and migration, release of nitric oxide (NO), and secretion of prostaglandin I(2) (PGI(2) ). Additionally, the PDAM-modified surface shows a remarkable ability to decrease the adhesion and proliferation of HUASMCs. As a blood-contacting material, the PDAM tends to improve the hemocompatibility compared with the substrate 316L SS. It is noteworthy that the PDAM coating shows good resistance to the deformation behavior of compression and expansion of a stent. These data suggest the potential of PDAM as a blood-contacting material for the application in vascular stents or grafts.

Improved immobilization of biomolecules to quinone-rich polydopamine for efficient surface functionalization

Polydopamine (PDA), a bio-inspired polymer, has been very attractive for diverse functional applications by immobilizing biomolecules. In this work, a novel approach of using PDA for improved biomolecule immobilization was developed. A thin PDA layer was strategically coated onto 316L stainless steel and thermally treated at 150°C (PDA-Th150). Subsequently, amino-terminated polyethylene glycol (mPEG-NH2) or vascular endothelial growth factor (VEGF) was immobilized onto PDA and PDA-Th150 surface. The results of attenuated total reflection-Fourier transform-infrared spectroscopy (ATR-FTIR) and X-ray photoelectron spectroscopy (XPS) showed higher coverage of quinine on PDA-Th150 surface. The functionalized PDA-Th150 significantly improved its ability of immobilizing mPEG-NH2 or VEGF, as shown by platelet adhesion test and endothelial cell proliferation experiments. This novel approach may also be used for efficient immobilization of biomolecules on different metal devices.

A simple one-step modification of various materials for introducing effective multi-functional groups

Covalent immobilization of various biomolecules is a desired strategy for bio-multifunctional surface modification. Multi-functionalization of a material surface is considered to be the premise of immobilizing a variety of biomolecules. However, currently adopted methods, used to introduce proper reactive functional groups on material surfaces, mostly are hard to be carried out and frequently can only introduce insufficient functional groups. In this work, we successfully develop the films (GAHD films) prepared via the simple copolymerization of gallic acid (GA) and hexamethylenediamine (HD), which can be deposited on different kinds of material surfaces including metals, ceramics and polymers by a one-step dip-coating method. Moreover, these copolymerized GAHD films possess high concentration of multi-functional groups like carboxyl (COOH), primary amine (-NH2) and quinone groups on the surfaces. Fourier transform infrared spectroscopy (FTIR) and X-ray photoelectron spectroscopy (XPS) results prove either the occurrence of Michael addition reaction, Schiff base reaction in the film-forming process, or the existence of COOH, NH2 and quinone groups on the surfaces. The maximum contents of carboxyl and amine on the GAHD film are 24.9 nmol/cm(2) and 31.7 nmol/cm(2) respectively. After dynamical immersion for 30 days, slight swellings can be observed, which reveals that the GAHD films possess good stability. Moreover, Heparin (Hep), fibronectin (Fn) and laminin (Ln) are successfully immobilized on the GAHD film surfaces. The results of cell counting kit-8 (CCK-8) and rhodamine fluorescence photograph indicate that the 1:1.62 GAHD film has good cytocompatibility.

Effects of polydopamine functionalized titanium dioxide nanotubes on endothelial cell and smooth muscle cell.

Previous investigations have demonstrated that TiO2 nanotubes (NTs) with particular structure cues could control the behavior of different types of cells, including endothelial cells (ECs) and smooth muscle cells (SMCs). Besides, polydopamine (PDA) modified surfaces were reported to be beneficial to increase the proliferation and viability of ECs and meanwhile could inhibit the proliferation of SMCs. The TiO2 nanotubes (NTs) were functionalized with polydopamine (PDA) (PDA/NTs) to study the synergetic effect of both nanotopography (NTs) and chemical cues (PDA) of TiO2 nanotubes on the regulation of cellular behavior of ECs and SMCs. The PDA-modified TiO2 nanotubes were subjected to field-emission scanning electron microscopy (FE-SEM), X-ray photoelectron spectroscopy (XPS), and water contact angle (WCA) analysis. In vitro cell culture tests confirmed that, comparing with flat titanium (Ti) and TiO2 nanotubes, PDA/NTs surface synergistically promoted ECs attachment, proliferation, migration and release of nitric oxide (NO). Meanwhile, the PDA/NTs performed well in reducing SMCs adhesion and proliferation. This novel approach might provide a new platform to investigate the synergistic effect of local chemistry and topography, as well as the applications for the development of titanium-based implants for enhanced endothelialization.

Immobilization of serum albumin and peptide aptamer for EPC on polydopamine coated titanium surface for enhanced in-situ self-endothelialization

Restenosis and thrombosis are two major complications associated with vascular stents and grafts. The homing of circulating endothelial progenitor cells (EPCs) onto implant surfaces brings a new strategy to solve these problems by accelerating self -endothelialization in situ. Peptide aptamers with high affinity and specific recognition of EPCs can be immobilized to capture EPCs from the circulating blood. In this study, a biotinylated peptide aptamer (TPSLEQRTVYAK-GGGC-K-Biotin) for EPC, and bovine serum albumin (BSA) were co-immobilized onto titanium surface through avidin-biotin recognition to endow the surface with specific affinity for EPC and anti-platelet adhesion properties. Quartz crystal microbalance with dissipation (QCM-D), X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM) and water contact angle measuring were adopted for coating characterization. EPC affinity and hemocompatibility of the coating were also investigated in vitro. The results demonstrated that aptamer and BSA co-immobilized surface significantly reduced platelet adhesion and fibrinogen adsorption/activation. Besides, such functional surface could remarkably enhance EPC adhesion, without affecting the behavior of endothelial cells (ECs) and smooth muscle cells (SMCs) obviously. The result shows the possibility of utilizing such a multifunctional surface in cardiovascular implants.

Cooperative control of blood compatibility and re-endothelialization by immobilized heparin and substrate topography.

A wide variety of environmental cues provided by the extracellular matrix, including biophysical and biochemical cues, are responsible for vascular cell behavior and function. In particular, substrate topography and surface chemistry have been shown to regulate blood and vascular compatibility individually. The combined impact of chemical and topographic cues on blood and vascular compatibility, and the interplay between these two types of cues, are subjects that are currently being explored. In the present study, a facile polydopamine-mediated approach is introduced for immobilization of heparin on topographically patterned substrates, and the combined effects of these cues on blood compatibility and re-endothelialization are systematically investigated. The results show that immobilized heparin and substrate topography cooperatively modulate anti-coagulation activity, endothelial cell (EC) attachment, proliferation, focal adhesion formation and endothelial marker expression. Meanwhile, the substrate topography is the primary determinant of cell alignment and elongation, driving in vivo-like endothelial organization. Importantly, combining immobilized heparin with substrate topography empowers substantially greater competitive ability of ECs over smooth muscle cells than each cue individually. Moreover, a model is proposed to elucidate the cooperative interplay between immobilized heparin and substrate topography in regulating cell behavior.

Proliferation and functionality of human umbilical vein endothelial cells on angiopoietin-1 immobilized 316L stainless steel

Angiopoietin-1 (Ang-1), a vascular-specific growth factor secreted from periendothelial cells, has drawn increasing attention in clinical applications because it can promote the reconstruction of blood vessels and has an anti-inflammatory effect compared with vascular endothelial growth factor (VEGF). In this study, Ang-1 was firstly covalently conjugated onto polydopamine (PDA) coated 316L stainless steel (SS), aiming at developing an Ang-l modified surface for endothelialization. The results of Fourier transform infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy (XPS) and water contact angle (WCA) measurements confirmed the successful immobilization of Ang-1. Quartz crystal microbalance with dissipation (QCM-D) studies demonstrated that ∼154 ng cm-2 of Ang-1 were bonded onto the PDA surface. To confirm its functionality, the effects of the Ang-1-modified coating on the growth behavior of human umbilical vein endothelial cells (HUVECs) were studied. As a result, the Ang-1 functionalized surface significantly enhanced endothelial cell adhesion, proliferation and migration. It was also found the Ang-1 functionalized coating could promote the release of nitric oxide (NO), secretion of prostacyclin-2 (PGI2) and inhibit the apoptosis of HUVECs. These data effectively suggested angiopoietin-1 could potentially be applied not only in neovascularization such as ischemic reperfusion and vascularization of tissue engineering scaffolds, but also in surface modification of cardiovascular implant materials for re-endothelialization.