A novel coating method to reduce membrane infolding through pre-crimping of covered stents - Computational and experimental evaluation.

A covered stent has been used to treat carotid artery stenosis to reduce the chance of embolization, as it offers improved performance over bare-metal stents. However, membrane infolding of covered stents can affect efficiency and functionality for treating occlusive disease of first-order aortic branches. In order to mitigate the degree of infolding of the stent once it was re-expanded, we proposed a new coating method performed on the pre-crimped stent. A systematic study was carried out to evaluate this new coating technique: a) in vivo animal testing to determine the degree of membrane infolding; b) structural finite element modeling and simulation were used to evaluate the mechanical performance of the covered stent; and c) computational fluid dynamics (CFD) to evaluate hemodynamic behavior of the stents and risk of thrombosis after stent deployment. The degree of infolding was substantially reduced as demonstrated by the in vivo deployment of the pre-crimped stent compared to a conventional dip-coated stent. The structural analysis results demonstrated that the membrane of the covered stent manufactured by conventional dip-coating resulted in a large degree of infolding but this could be minimized by our new pre-crimped coating method. CFD studies showed that the new coating method reduced the risk of thrombosis compared to the conventional coating method. In conclusion, both simulation and in vivo testing demonstrate that our new pre-crimped coating method reduces membrane infolding compared with the conventional dip-coating method and may reduce risk of thrombosis.

Measurement of the Luminal Diameter of Peripheral Arterial Vasculature in Yorkshire×Landrace Swine by Using Ultrasonography and Angiography.

To select animals of appropriate size for preclinical studies of cardiovascular devices, reference knowledge of the cardiovascularanatomy relative to body weight is crucial. We measured the luminal diameters of the arteries (carotid, femoral, and iliac arteries) that are the common access vessels for endovascular and vascular procedures in Yorkshire×Landrace swine. Measurements were performed by using both ultrasound and angiographic methods and were correlated with body weight. Results showed no statistically significant difference between the left and right vessels in the diameters of the carotid,femoral, and iliac arteries. The diameters of the measured arteries showed high correlation with animal weight in pigs thatweighed less than 70 kg.

Optimization of a Novel Preferential Covered Stent through Bench Experiments and in Vitro Platelet Activation Studies.

Bare metal stenting (BMS) does not adequately address the atheroembolic characteristic of carotid artery stenosis. While simple covered stents (CS) may prevent dislodged fragments of the atherosclerotic plaque from entering the blood stream, they also block blood flow into the major branches of the artery alongside the lesion, which is not desirable. Preferential covered stents (PCS) behave as a covered stent in a tubular part of a vessel but maintain side-branch flow over the bifurcation region by means of slits in the membrane. Stent design, membrane material, and slits configuration are the three main components contributing to stent performance. Optimization of PCS designs was conducted and tested. METHODS: A newly designed BMS was developed and compared to a commercially available peripheral stent. Two materials (expanded poly(tetrafluoroethylene)) and silicone polyurethane co-polymers (Elast-eon E2A) were used as stent coverings with slits applied using various cutting methods to form the PCS. These PCS samples were tested for physical resilience, flexibility, ability to preserve side-branch flow, slit edge roughness, and platelet activation. RESULTS: Fabrication of E2A-coated stents required pretreatment of the stent with poly(ethylene glycol) to achieve firm attachment. The newly designed BMS with nine crowns design and larger cell size showed higher flexibility than commercially available stents. A combination of a larger stent cell size, E2A membrane coating, and three slits per stent cell unit configuration resulted in preserved side-branch flow similar to physiological conditions in the flow experiment. Slit edge roughness changed with different cutting methods and laser machine cutting parameters. In vitro studies showed platelet activation was minimal with lower slit edge roughness samples. CONCLUSION: An optimized PCS prototype was developed consisting of a newly designed stent, E2A membrane, and a three-slit pattern created by specific femtosecond laser cutting.

Epoxy cross-linked collagen and collagen-laminin Peptide hydrogels as corneal substitutes.

A bi-functional epoxy-based cross-linker, 1,4-Butanediol diglycidyl ether (BDDGE), was investigated in the fabrication of collagen based corneal substitutes. Two synthetic strategies were explored in the preparation of the cross-linked collagen scaffolds. The lysine residues of Type 1 porcine collagen were directly cross-linked using l,4-Butanediol diglycidyl ether (BDDGE) under basic conditions at pH 11. Alternatively, under conventional methodology, using both BDDGE and 1-Ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC)/N-hydroxysuccinimide (NHS) as cross-linkers, hydrogels were fabricated under acidic conditions. In this latter strategy, Cu(BF4)2·XH2O was used to catalyze the formation of secondary amine bonds. To date, we have demonstrated that both methods of chemical cross-linking improved the elasticity and tensile strength of the collagen implants. Differential scanning calorimetry and biocompatibility studies indicate comparable, and in some cases, enhanced properties compared to that of the EDC/NHS controls. In vitro studies showed that human corneal epithelial cells and neuronal progenitor cell lines proliferated on these hydrogels. In addition, improvement of cell proliferation on the surfaces of the materials was observed when neurite promoting laminin epitope, IKVAV, and adhesion peptide, YIGSR, were incorporated. However, the elasticity decreased with peptide incorporation and will require further optimization. Nevertheless, we have shown that epoxy cross-linkers should be further explored in the fabrication of collagen-based hydrogels, as alternatives to or in conjunction with carbodiimide cross-linkers.

Adhesion, proliferation, and gene expression profile of human umbilical vein endothelial cells cultured on bilayered polyelectrolyte coatings composed of glycosaminoglycans.

This study characterized human umbilical vein endothelial cell (HUVEC) adhesion, proliferation, and gene expression on bilayered polyelectrolyte coatings composed of an outermost layer of glycosaminoglycans (hyaluronan, heparin, or chondroitin sulfate), with an underlying layer of poly-L-lysine or chitosan. The proportion of cells that adhered to the various polyelectrolyte coatings after 1 and 2 h incubations was quantified by the WST-8 assay. Interchanging poly-L-lysine with chitosan resulted in significant differences in cellular adhesion to the outermost glycosaminoglycan layer after 1 h, but these differences became insignificant after 2 h. The proliferation of HUVEC on the various bilayered polyelectrolyte coatings over 10 days was characterized using the WST-8 assay. Regardless of whether the underlying layer was poly-L-lysine or chitosan, HUVEC proliferation on the hyaluronan outermost layer was significantly less than on heparin or chondroitin sulfate. Additionally, it was observed that there was more proliferation with poly-L-lysine as the underlying layer, compared to chitosan. Subsequently, real-time polymerase chain reaction was used to analyze the expression of seven genes related to adhesion, migration, and endothelial function (VWF, VEGFR, VEGFA, endoglin, integrin-α5, ICAM1, and ICAM2) by HUVEC cultured on the various bilayered polyelectrolyte coatings for 3 days. With poly-L-lysine as the underlying layer, biologically significant differences (greater than twofold) in the expression of VWF, VEGFR, VEGFA, endoglin, and ICAM1 were observed among the three glycosaminoglycans. With chitosan as the underlying layer, all three glycosaminoglycans displayed biologically significant differences in the expression of VWF and VEGFR compared to the chitosan control. CT-HA displayed the highest level of expression of VWF, whereas expression levels of VEGFR were almost similar among the three glycosaminoglycans.

Conformational behavior of fibrinogen on topographically modified polymer surfaces.

The influence of topographical surface features at the submicron scale on the structural changes in the surface-adsorbed fibrinogen was investigated on poly(lactic-co-glycolic-acid) (PLGA) films. Attenuated total reflection-Fourier transform infrared spectroscopy (ATR-FTIR) was employed in this study for the induced conformational change of fibrinogen over various adsorption times, while the adsorption kinetics of fibrinogen was quantified by the enzyme linked immunosorbent assay (ELISA). When a PLGA surface is modified topographically, the adsorbed fibrinogen undergoes less conformational change when compared to adsorption on the pristine PLGA surface. The extent of conformational change is related to platelet adhesion. Reduced thrombogenicity was demonstrated by the higher ratios of alpha-helix to beta-turn and beta-sheet to beta-turn structures on the topographic PLGA film, which suggests that topographical manipulation of surfaces is a viable approach to influence the thrombogenicity of surfaces.

The effect of topography of polymer surfaces on platelet adhesion.

In this study, the effect of surface topography on fibrinogen and platelet adsorption was investigated. High aspect ratio surface features, in the submicron to nanometer range, were constructed on the poly- (lactic-co-glycolic-acid) (PLGA) films. The topographic surfaces were fabricated by solvent-mediated polymer casting on a master template. Fibrinogen adsorption and platelets adhesion on these topographic surfaces were quantified by enzyme linked immunosorbent assay (ELISA) and lactate dehydrogenase (LDH) assay respectively, while the activation of platelets was quantified by flow cytometric analysis using fluorescein isothiocyanate (FITC) tagging. The lowest fibrinogen adsorption amount and platelet activity was observed on surfaces with specific topographical features in the submicron range with a significant reduction in adhesion when compared to the pristine PLGA films. The topographical parameters found to induce low levels of fibrinogen adsorption and platelet response were high aspect ratio structures (>3:1) with reduced interspacing (<200 nm) or high density. The results signify that topographical manipulation of thrombogenic surfaces of biodegradable polymers is a feasible approach for reducing their thrombogenicity.

A novel nanostructured poly(lactic-co-glycolic-acid)-multi-walled carbon nanotube composite for blood-contacting applications: thrombogenicity studies.

Composite films of poly(lactic-co-glycolic-acid) with multi-walled carbon nanotubes (PLGA-MWCNT) having two different nanotube orientations, namely random and vertically aligned, have been fabricated and characterized. The effect of these nanostructured surfaces on platelet adhesion is evaluated. In particular, the thrombogenicity of the nanostructured composite films is compared with that of pristine graphite (a low thrombogenic material) and PLGA film, in order to determine the influence of surface chemistry and topography on platelet adhesion. The results in this study show that the PLGA-MWCNT composite with vertically aligned nanotubes exhibits very low levels of fibrinogen adsorption and platelet adhesion, which can be attributed to both chemical and topographical effects. Platelet adhesion shows a good correlation with the presence of COOH groups and appears to be sensitive to the topographic features of the composite films. The results in this study suggest that in addition to chemistry, nanotopographical surface modifications could be an effective strategy in the development of low thrombogenic and hemocompatible materials.

Platelet adhesion studies on nanostructured poly(lactic-co-glycolic-acid)-carbon nanotube composite.

Design of blood-compatible surfaces is required to minimize platelet-surface interactions and increase the thromboresistance of foreign surfaces. Poly(lactic-co-glycolic-acid)-carbon nanotube (PLGA-CNT) composite is studied as a building material to fabricate artificial blood prostheses. This nanocomposite-based biomaterial is prepared by an electrostatic Layer-by-Layer (LbL) deposition technique, in which layers of CNTs are adsorbed onto a PLGA film. Before incubation in nonstimulated platelet-rich plasma (PRP) for platelet studies, fibrinogen is immobilized on PLGA-CNT composite. Interactions between the plasma proteins, e.g. fibrinogen and PRP, are investigated on the prepared PLGA-CNT composite. Contact angle measurements on the PLGA-CNT composite displayed a good resistance of platelets adhesion on a hydrophilic surface with an angle of 64.94 degrees as compared to pristine PLGA control with an angle of 93.43 degrees . A significant reduction of adhesion is observed on the PLGA-CNT composite, as well as the absence of platelet activation. On the contrary, both platelet adhesion and activation are observed on control samples. We inferred this suppression in secretion of granule contents in the platelet by the presence of the CNTs that resulted in the absence of platelet activation and its subsequent inhibition in the release of adhesive membrane receptors on the PLGA-CNT composite.