Vascular Graft Applications Research Articles

Expanded polytetrafluoroethylene/silk fibroin/salicin vascular graft fabrication for improved endothelialization and anticoagulation

Clinical application of small-diameter artificial vascular grafts has been hampered by intimal blockages owing to the lack of an endothelial layer and the formation of thrombosis. In this study, an expanded polytetrafluoroethylene (ePTFE) vascular graft was fabricated using a biofriendly ethanol/water mixture as a lubricant and drug carrier. Natural silk fibroin (SF) hydrogel and white willow extract salicin were embedded into ePTFE graft successfully via the ethanol/water lubricant and drug carrier as verified by Fourier-transform infrared spectroscopy (FTIR) and X-ray photoelectron spectroscopy (XPS). The ePTFE graft functionalized by SF hydrogel improved the proliferation of human umbilical vein endothelial cell (HUVEC) due to SF’s hydrophilia and cytocompatibility as measured by the live/dead and cell counting kit-8 (CCK-8) assays. The rapid endothelialization of ePTFE graft helped reduced intimal blockages in vitro. Furthermore, the salicin-modified ePTFE graft exhibited an anticoagulation property as demonstrated by platelet adhesion and activated partial thromboplastin time (APTT) tests. These results suggest that ePTFE graft fabricated with silk fibroin and salicin can be an attractive candidate for vascular grafts.

Polyurethane/polycaprolactone membrane grafted with conjugated linoleic acid for artificial vascular graft application

ABSTRACT Constructing satisfied small-diameter vascular graft (diameter less than 6 mm) remains an unsolvable challenge in vascular tissue engineering. This study described the fabrication of electrospun polyurethane/polycaprolactone (PU/PCL) membranes chemically grafted with various densities of conjugated linoleic acid (CLA) – an antithrombotic fatty acid – for making small-diameter blood vessel. Differences in mechanical, antithrombotic properties and biocompatibility of the membranes resulting from the CLA-grafting procedure were the focus of the study. Investigation of mechanical properties relevant to vascular graft application revealed that these properties of the membranes remained unaffected and satisfied clinical criteria following the CLA graft. Blood–membrane interaction assays showed that the CLA-grafted membranes mitigated the adhesion of blood cells, as well as preventing blood coagulation. These effects were also commensurate with increasing density of CLA, suggesting an effective approach to improve antithromboticity. Cellular tests suggested that CLA has an optimal density at which it promoted cell proliferation on the surface of the membranes; however, excessive presence of CLA might cause undesirable inhibition on cells. In conclusion, PU/PCL membrane grafted with CLA could be a prospective material for vascular tissue engineering with further development and investigation.

One-Pot Covalent Grafting of Gelatin on Poly(Vinyl Alcohol) Hydrogel to Enhance Endothelialization and Hemocompatibility for Synthetic Vascular Graft Applications.

Cardiovascular diseases remain the leading cause of death worldwide. Patency rates of clinically-utilized small diameter synthetic vascular grafts such as Dacron® and expanded polytetrafluoroethylene (ePTFE) to treat cardiovascular disease are inadequate due to lack of endothelialization. Sodium trimetaphosphate (STMP) crosslinked PVA could be potentially employed as blood-compatible small diameter vascular graft for the treatment of cardiovascular disease. However, PVA severely lacks cell adhesion properties, and the efforts to endothelialize STMP-PVA have been insufficient to produce a functioning endothelium. To this end, we developed a one-pot method to conjugate cell-adhesive protein via hydroxyl-to-amine coupling using carbonyldiimidazole by targeting residual hydroxyl groups on crosslinked STMP-PVA hydrogel. Primary human umbilical vascular endothelial cells (HUVECs) demonstrated significantly improved cells adhesion, viability and spreading on modified PVA. Cells formed a confluent endothelial monolayer, and expressed vinculin focal adhesions, cell-cell junction protein zonula occludens 1 (ZO1), and vascular endothelial cadherin (VE-Cadherin). Extensive characterization of the blood-compatibility was performed on modified PVA hydrogel by examining platelet activation, platelet microparticle formation, platelet CD61 and CD62P expression, and thrombin generation, which showed that the modified PVA was blood-compatible. Additionally, grafts were tested under whole, flowing blood without any anticoagulants in a non-human primate, arteriovenous shunt model. No differences were seen in platelet or fibrin accumulation between the modified-PVA, unmodified PVA or clinical, ePTFE controls. This study presents a significant step in the modification of PVA for the development of next generation in situ endothelialized synthetic vascular grafts.

Study of the Young's modulus and failure strength of binary polymer composite structures based on polyurethane based on stress-strain curve for tissue engineering vascular graft application

.The artery diseases such as the coronary arteries are of the important cardiovascular diseases. The autograft as a common surgical is the main treatment for this problem, but in many patients, the autografts are not. So, due to a large number of requirements, it needs to find suitable replacements for diseases of blood vessels. Tissue engineering at the nanoscale level is a promising approach to the design and fabrication of artificial blood vessels. Nanomaterial structures are highly contributive in tissue engineering vascular scaffolds (TEVS) due to their ability in mimicking the nanoscale dimension of the natural extracellular matrix (ECM) and the existing mechanical match between the native vessel and the scaffold. The aim of this research was developing and mechanically improving nano-fibrous composite scaffolds using blend electrospinning methods with different ratios of the polyethylene terephthalate (PET), polyurethane (PU) and polycaprolactone (PCL). The morphological and mechanical properties all structures were evaluated using SEM, FTIR and tensile properties. The neat and composite structures were completely intact with randomly oriented fibers and without any beads. Results showed that the average fiber diameter, porosity, stress and Young’s modulus changes’ range in composite structures (PCL/PU and PET/PU) obtained 343 ± 94 to 382 ± 83 nm, 58.6 ± 3.12 to 81 ± 1.7 %, 2.66 ± 0.39 to 19.05 ± 3.2 MPa and 3.18 ± 0.09 to 41.4± 3.31 MPa, respectively. The fabricated scaffolds and especially PET/PU structure exhibited suitable mechanical and biological properties and clinical requirements as a small-diameter vascular graft.

Development of double porous poly (ε - caprolactone)/chitosan polymer as tissue engineering scaffold.

Polymer blend made from poly(ε - caprolactone)/chitosan (PCL/CHT) offers interesting opportunities for biological applications. The paper presents a new way to fabricate PCL/CHT double-porosity (macrovoids with interconnected microporosity) membrane materials from a chemical optimization of the solvent and non-solvent phases and from a modified phase inversion technique. By varying the PCL/CHT proportion, it is shown that it is possible to improve the chemical and physical properties of the CHT carbohydrate polymer. The PCL/CHT membranes are fully characterized in term of physico-chemical properties (ATR-FTIR, XRD and DSC) to understand the miscibility of the two-polymer blend. Morphological characterization by SEM shows that by increasing CHT wt% in the blend, the size of the macrovoids was increasing. Rapid enzymatic degradation of PCL from all the blend was found by using lipase (from P. cepacia). The mechanisms at the origin of the morphological structuration of the material is also discussed. To test the ability to operate these materials as small diameter vascular scaffolds, cell culture with human umbilical vein endothelial cells (HUVECs) were carried out on the membrane and the results analyzed with laser scanning confocal microscopy (LSCM). Data suggest that the blend membrane with higher concentration of CHT polymer wt% have suitable properties that promote high number of cells on the surface by maintaining cellular cytoskeleton integrity within 3 days. The blend membrane with a double porous morphology could be potentially applicable in future for small diameter vascular graft application. The surface macrovoids (20-90 μm) could be useful for three-dimensional cellular adhesion and proliferation and interconnected microporous spongy network (7-20 μm) is expected to transfer essential nutrients, oxygen, growth factor between the macrovoids and the supernatant.

Influence of Electrical Field Collector Positioning and Motion Scheme on Electrospun Bifurcated Vascular Graft Membranes.

Currently, electrospinning membranes for vascular graft applications has been limited, due to random fiber alignment, to use in mandrel-spun, straight tubular shapes. However, straight, circular tubes with constant diameters are rare in the body. This study presents a method to fabricate curved, non-circular, and bifurcated vascular grafts based on electrospinning. In order to create a system capable of electrospinning membranes to meet specific patient needs, this study focused on characterizing the influence of fiber source, electrical field collector position (inside vs. outside the mandrel), and the motion scheme of the mandrel (rotation vs. rotation and tilting) on the vascular graft membrane morphology and mechanical properties. Given the extensive use of poly(ε-caprolactone) (PCL) in tubular vascular graft membranes, the same material was used here to facilitate a comparison. Our results showed that the best morphology was obtained using orthogonal sources and collector positioning, and a well-timed rotation and tilting motion scheme. In terms of mechanical properties, our bifurcated vascular graft membranes showed burst pressure comparable to that of tubular vascular graft membranes previously reported, with values up to 5126 mmHg. However, the suture retention strength shown by the bifurcated vascular graft membranes was less than desired, not clinically viable values. Process improvements are being contemplated to introduce these devices into the clinical range.

Vascular endothelial growth factor immobilized on mussel-inspired three-dimensional bilayered scaffold for artificial vascular graft application: In vitro and in vivo evaluations

Currently, there is a great clinical demand for biocompatible and robust tissue-engineered tubular scaffolds for use as artificial vascular graft materials. Despite considerable research on vascular scaffolds, there has still been only limited development of scaffold materials possessing both sufficient mechanical strengths and biological effects for vascular application. In this work, we designed a mechanically robust, bilayered scaffold and manufactured it by combining electrospinning (ELSP) and three-dimensional (3D) printing techniques. This material was coated with polydopamine (PDA) and vascular endothelial growth factor (VEGF) was grafted directly on the scaffold surface to induce potent angiogenic activity. We confirmed that the coated-PDA layer was evenly deposited on the bare polycaprolactone (PCL) scaffold and could enable abundant VEGF immobilization with enhanced hydrophilicity. The VEGF immobilized porous tubular scaffold was well prepared without mechanical weakness induced by surface modification steps. During in vitro and in vivo testing, VEGF immobilized scaffolds elicited markedly enhanced vascular cell proliferation and angiogenic differentiation, as compared to non-treated groups. These results demonstrate that the developed scaffolds may represent an innovative paradigm in vascular tissue engineering by inducing angiogenesis as a means of remodeling and healing vascular defects for use in restorative procedures.

Active wrinkles to drive self-cleaning: A strategy for anti-thrombotic surfaces for vascular grafts

The inner surfaces of arteries and veins are naturally anti-thrombogenic, whereas synthetic materials placed in blood contact commonly experience thrombotic deposition that can lead to device failure or clinical complications. Presented here is a bioinspired strategy for self-cleaning anti-thrombotic surfaces using actuating surface topography. As a first test, wrinkled polydimethylsiloxane planar surfaces are constructed that can repeatedly transition between smooth and wrinkled states. When placed in contact with blood, these surfaces display markedly less platelet deposition than control samples. Second, for the specific application of prosthetic vascular grafts, the potential of using pulse pressure, i.e. the continual variation of blood pressure between systole and diastole, to drive topographic actuation was investigated. Soft cylindrical tubes with a luminal surface that transitioned between smooth and wrinkled states were constructed. Upon exposure to blood under continual pressure pulsation, these cylindrical tubes also showed reduced platelet deposition versus control samples under the same fluctuating pressure conditions. In both planar and cylindrical cases, significant reductions in thrombotic deposition were observed, even when the wrinkles had wavelengths of several tens of μm, far larger than individual platelets. We speculate that the observed thrombo-resistance behavior is attributable to a biofilm delamination process in which the bending energy within the biofilm overcomes interfacial adhesion. This novel strategy to reduce thrombotic deposition may be applicable to several types of medical devices placed into the circulatory system, particularly vascular grafts.

Preparation and mechanical performance of small-diameter PHBHHx vascular graft by electrospinning

The straight and corrugated PHBHHx (Poly(3-hydroxybutyrate-co-3-hydroxhexanoate)) vascular grafts with 6 mm inner diameters were prepared successfully by electrospinning. Moreover, their mechanical behaviors were investigated compared with the commercial vascular grafts (SW-ePTFE woven). Electrospun PHBHHx fibers had a submicron scale. Straight grafts and SW-ePTFE grafts exhibited higher Young’s modulus and tensile breaking strength in the circumferential direction than the corrugated grafts, but the break and elastic recovery rate were lower. Both the straight and corrugated samples had lower suture strength than the SW-ePTFE samples, but still satisfied the requirement of transplantation of small diameter vascular. The compliance of corrugated grafts was similar to SW-ePTFE grafts and lower than straight grafts. The results of hemolytic and cytotoxicity tests demonstrated that both the straight and corrugated grafts had good hemocompatibility and no cytotoxicity. Therefore, the results of this study showed the potential application of electrospun PHBHHX vascular grafts as small diameter vascular grafts.

Poly-L-lactic Acid (PLLA)-Chitosan-Collagen Electrospun Tube for Vascular Graft Application.

Poly-L-Lactic acid (PLLA) blended with chitosan and collagen was used to fabricate a conduit for blood vessel engineering through an electrospinning process. Various concentrations of chitosan were used in the blend in order to study its effect on the morphology, chemical bond, tensile strength, burst pressure, hemocompatibility, and cell viability (cytotoxicity) of the tube.In vitro assessments indicated that addition of chitosan-collagen could improve cell viability and hemocompatibility. Best results were demonstrated by the conduit with 10% PLLA, 0.5% chitosan, and 1% collagen. Tensile strength reached 2.13 MPa and burst pressure reached 2593 mmHg, both values that are within the range value of native blood vessel. A hemolysis percentage of 1.04% and a cell viability of 86.2% were obtained, meeting the standards of high hemocompatibility and low cytotoxicity for vascular graft material. The results are promising for further development toward vascular graft application.

A novel polyurethane modified with biomacromolecules for small-diameter vascular graft applications

There is a growing demand for small-caliber tissue-engineered vascular grafts to replace damaged vessels. Fabricated scaffolds are unable to precisely mimic the mechanical properties of native vessels, provide long-term patency and support cell adhesion and growth, in particular support endothelialization. In this study, a new biodegradable poly(ether ester urethane) urea (PEEUU) was synthesized. The synthesized polyurethane was then functionalized by introducing free amino groups through aminolysis for further surface modification by immobilization of biomacromolecules on the surface of vascular grafts. The modified surfaces were then characterized using attenuated total reflectance-Fourier transform infrared spectroscopy, water contact angle measurement and atomic force microscopy. The mechanical properties of the fabricated scaffolds were analyzed, revealing mechanical properties close to that of the natural vessels. Surface modifications led to improved cell–scaffold interactions, showing appropriate cell attachment and function on the scaffolds. A confluent layer of endothelial cells was formed on biomacromolecule-immobilized PEEUU vascular grafts. The preliminary results of this study demonstrated that the new polyurethane modified with biomacromolecules can be considered as a candidate material for vascular tissue engineering application with capability to support endothelialization of fabricated vascular grafts.

Regulation of macrophage polarization and promotion of endothelialization by NO generating and PEG-YIGSR modified vascular graft

As an effective clinic treatment for cardiovascular disease, vascular transplantation gains much acceptance recently. However, due to the acute thrombosis and intimal hyperplasia, long-term failure of synthetic grafts after implanted in small diameter blood vessel decelerates its commercial use. The continued acute inflammation and delayed endothelialization have been considered as fundamental reasons. To enhance the adhesion and organization of endothelial cells (ECs) and improve the vascular remodeling process, we have constructed a vascular graft based on electrospun polycaprolactone (PCL) matrix, on which organoselenium-immobilized polyethyleneimine (SePEI) for in situ nitric oxide (NO) generation and hyaluronic acid (HA) grafted with poly (ethylene glycol) (PEG) modified Tyr-Ile-Gly-Ser-Arg (YIGSR) for antifouling and EC adhesion were deposited through electrostatic layer-by-layer assembly. The in vitro results showed that SePEI deposited on the grafts could catalyze stable generation of NO. After in situ implantation in rats for 4 and 8weeks, the graft promoted the transformation of macrophages into an anti-inflammatory phenotype (M2), which helped endothelium remodeling. YIGSR on the outmost layer facilitated more rapid and organized EC adhesion compared to PCL and non-modified grafts. PEG polymer chain on the outmost layer mitigated nonspecific adsorption of undesirable blood components. In our study, we first demonstrated the regulation of macrophage polarization by an NO-generating vascular graft. The results indicated that the approach of anti-inflammatory macrophage polarization and enhanced endothelialization through NO generation and PEG-modified YIGSR in our study may provide a new perspective for the clinic application of cell-free small-diameter vascular grafts.

Production and hemocompatibility assessment of novel electrospun polyurethane nanofibers loaded with dietary virgin coconut oil for vascular graft applications

To develop biodegradable polymer scaffolds suitable for vascular tissue engineering applications, the bioengineering community has invested an extensive effort. The most common cause for the failure of vascular graft scaffolds is thrombosis. In this work, the scaffold based on polyurethane and virgin coconut oil was produced by electrospinning process for vascular tissue engineering applications with improved antithrombogenicity. The diameter of this electrospun polyurethane/virgin coconut oil composite was found to be reduced in the range of 886 ± 207 nm compared to pristine polyurethane which was in the range of 969 ± 217 nm. The Fourier transform infrared spectroscopy analysis revealed the interaction between polyurethane and virgin coconut oil as indicated by phase shifting of CH bond along with the formation of hydrogen bond. The contact angle measurement of fabricated composites was found to be increased owing to hydrophobic nature and also exhibited enhanced thermal stability as noted in thermogravimetric analysis. The atomic force microscopy analysis insinuated the increased surface roughness of the composite in comparison with the pure polyurethane. Developed scaffold resulted in delayed blood clotting as revealed by activated partial thromboplastin time and partial thromboplastin time assay. The hemolytic index of fabricated composites was found to be low indicating the enhanced safety of red blood cells. Hence, the newly developed nanofibrous composite scaffold could open the door for a suitable alternative for vascular graft applications.

Application of omentum as an in vivo bioreactor for regeneration of decellularized human internal mammary artery.

This study investigates the extent of cell seeding as well as lumen patency in acellular Internal mammary artery (IMA) scaffolds that have been re-cellularized by omentum as a natural bioreactor. Sixteen Virgin female Wistar rats were selected for implantation of acellular scaffold in omentum. Following laparotomy omentum was retracted to the outside of the abdomen and the more vascularized portion of it was selected for the experiment. The scaffold was wrapped by omentum and placed between two layers of rectus muscles that have been previously dissected. Samples were taken from scaffolds at 2 and 3 months for histopathological evaluation. All the grafts were explanted at 2 and 3 months and the lumens were completely patent compared to the native scaffold. The histology of implanted IMA after 2 and 3 months showed progressive recellularization especially by smooth muscle cells over the media layer. CD 34 staining was positive adjacent to the grafts which showed well angiogenesis. Trichrome and Movat's Pentachrome staining showed normal collagen formation in the inner media layer. Promising results obtained from the introduced protocol for in vivo recellularization, including patent lumen and proper cell seeding, encourages application of the mentioned technique for experimental vascular graft application. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2685-2693, 2017.

Anti-acute thrombogenic surface using coaxial electrospraying coating for vascular graft application

Improvement of thrombo-resistant properties is vital for vascular substitutes. Heparin, an effective natural anticoagulant, attracted concerns about its incorporate way, releasing profile, and the biological function when applied onto vascular substitutes. Coaxial electrospraying technology was used to develop and form PCL-PEG encapsulated heparin microspheres in this study. Heparin can maintain its activity and sustained release in 15days with the package and protection of shell layer. When the microspheres were sprayed onto the PCL porous membrane, the materials showed sufficient property in avoiding the platelet adhesion and blood coagulation.

Biomimetic coating of cross-linked gelatin to improve mechanical and biological properties of electrospun PET: A promising approach for small caliber vascular graft applications.

Electrospun PET (ePET) is a promising material for small caliber vascular graft applications owing to its tunable mechanical properties, biocompatibility, and nanofibrous structure that mimic the morphology of natural extracellular matrix. However, the inherent inertness of PET impairs the adhesion and proliferation of endothelial cells on the inner surface of ePET tubular grafts, hindering the formation of a functional endothelium. Gelatin coatings, owing to their ability to promote endothelialization, are a valuable approach to overcome the limitations of ePET. Herein, a novel process for the deposition of stable biomimetic coatings of gelatin on ePET tubular grafts is proposed. Electrospun PET was first aminated by plasma treatment and then coated with a gelatin hydrogel cross-linked in situ by a Michael-type addition reaction. Amination provided a superhydrophilic behavior to the ePET surface, allowing easy gelatin interpenetration along the wall thickness of the tubular structure, and the obtainment of thin coatings that maintained the morphology of ePET fibers. Gelatin coating was stable at long term in a physiological-like environment, noncytotoxic and promoted in vitro cell adhesion and proliferation. Noteworthy, the mechanical properties of gelatin-coated ePET tubular grafts were improved in terms of elastic modulus, compliance, and elastic recoil, finally better matching the characteristics of native blood vessels. Altogether, the proposed coating technique successfully combines the advantages of ePET nanofibrous structure with cross-linked gelatin biological cues and mechanical reinforcement, and emerges as a promising strategy for the development of biocompatible small caliber vascular grafts with superior biomimetic and mechanical properties. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2405-2415, 2017.

Compositions Including Synthetic and Natural Blends for Integration and Structural Integrity: Engineered for Different Vascular Graft Applications.

Tissue engineering approaches for small-diameter arteries require a scaffold that simultaneously maintains patency by preventing thrombosis and intimal hyperplasia, maintains its structural integrity after grafting, and allows integration. While synthetic and extracellular matrix-derived materials can provide some of these properties individually, developing a scaffold that provides the balanced properties needed for vascular graft survival in the clinic has been particularly challenging. After 30 years of research, there are now several scaffolds currently in clinical trials. However, these products are either being investigated for large-diameter applications or they require pre-seeding of endothelial cells. This progress report identifies important challenges unique to engineering vascular grafts for high pressure arteries less than 4 mm in diameter (e.g., coronary artery), and discusses limitations with the current usage of the term "small-diameter." Next, the composition and processing techniques used for generating tissue engineered vascular grafts (TEVGs) are discussed, with a focus on the benefits of blended materials. Other scaffolds for non-tissue engineering approaches and stents are also briefly mentioned for comparison. Overall, this progress report discusses the importance of defining the most critical challenges for small diameter TEVGs, developing new scaffolds to provide these properties, and determining acceptable benchmarks for scaffold responses in the body.

Promoting Endothelialization of Polymeric Cardiovascular Biomaterials

The lack of a blood compatible synthetic interface is one of the largest unaddressed challenges in the field of biomaterials science. This technological shortcoming hinders the successful clinical application of small diameter vascular grafts and other cardiovascular devices such as stents and artificial heart valves. Therefore, intensive research activities are ongoing to develop polymer materials with improved blood compatibility. One attractive strategy to improve the blood compatibility of an interface is to design surfaces that promote the development of an endothelium, the monolayer of endothelial cells that line our native vasculature and is responsible for blood compatibility. This article describes the recent strategies that have been used to generate polymeric materials that promote the development of an endothelium, discusses shortcomings in the field, and proposes future directions of research that can be undertaken to design next generation polymeric biomaterial that promote endothelialization.image

Nano-scale clustering of integrin-binding ligands regulates endothelial cell adhesion, migration, and endothelialization rate: novel materials for small diameter vascular graft applications.

Blood contacting devices are commonly used in today's medical landscape. However, such devices (including small diameter vascular grafts) are limited by poor blood compatibility and may fail due to thrombosis. An attractive strategy for improving the blood compatibility of such devices is to generate biomaterials that foster a confluent and functioning endothelial cell layer. Synthesizing materials that display integrin-binding peptide ligands is a common way to promote endothelialization. However, in addition to integrin-ligand binding, integrin clustering is necessary to achieve intracellular signaling events that influence cellular phenotype. In this study, we explored the impact of nano-scale clustering of integrin-binding ligands on endothelial cell functions by designing novel materials that promote the clustering of integrin receptors. RGD-functionalized copolymers were prepared via reversible addition-fragmentation chain transfer (RAFT) polymerization and used for the preparation of random and nano-clustered surfaces spanning a range of global and local RGD densities (global densities 0.4-1.9 μg of peptide per mg of polymer and local densities of 1-2.4 ligands per nano-cluster). The adhesion and migration of endothelial cells was improved on nano-clustered surfaces compared to random surfaces. The highest adhesion and migration speed of endothelial cells and the rapid development of the endothelial monolayer were observed on surfaces with the highest local and global peptide density. These results indicate that the nano-clustering of peptide ligands is a promising strategy for next generation cardiovascular biomaterials, especially for small diameter vascular graft applications where the development of a confluent and functioning endothelium holds the potential to prevent device failure due to thrombosis.

Pericyte Seeded Dual Peptide Scaffold with Improved Endothelialization for Vascular Graft Tissue Engineering.

The development of synthetic vascular grafts for coronary artery bypass is challenged by insufficient endothelialization, which increases the risk of thrombosis, and the lack of native cellular constituents, which favors pathological remodeling. Here, a bifunctional electrospun poly(ε-caprolactone) (PCL) scaffold with potential for synthetic vascular graft applications is presented. This scaffold incorporates two tethered peptides: the osteopontin-derived peptide (Adh) on the "luminal" side and a heparin-binding peptide (Hep) on the "abluminal" side. Additionally, the "abluminal" side of the scaffold is seeded with saphenous vein-derived pericytes (SVPs) as a source of proangiogenic growth factors. The Adh peptide significantly increases endothelial cell adhesion, while the Hep peptide promotes accumulation of vascular endothelial growth factor secreted by SVPs. SVPs increase endothelial migration both in a transwell assay and a modified scratch assay performed on the PCL scaffold. Seeding of SVPs on the "abluminal"/Hep side of the scaffold further increases endothelial cell density, indicating a combinatory effect of the peptides and pericytes. Finally, SVP-seeded scaffolds are preserved by freezing in a xeno-free medium, maintaining good cell viability and function. In conclusion, this engineered scaffold combines patient-derived pericytes and spatially organized functionalities, which synergistically increase endothelial cell density and growth factor retention.