Tissue-engineered Vascular Grafts Research Articles

Hyaluronan promotes the regeneration of vascular smooth muscle with potent contractile function in rapidly biodegradable vascular grafts

The regeneration of smooth muscle with physiological functions has been a key challenge in vascular tissue engineering. Hyaluronan (HA), as a major component of the extracellular matrix, plays a vital role in regulating tissue injury and repair. In this study, a biomimetic vascular graft was prepared by co-electrospinning of synthetic degradable polymers and native ECM components including collagen type-I as well as low and high molecular weight HA (LMW HA and HMW HA). Upon implantation in the rat abdominal aorta, the grafts exhibited sustained HA release that effectively enhanced the regeneration of vascular smooth muscle. Besides, LMW HA loaded vascular grafts demonstrated rapid endothelialization compared to the other groups. More importantly, HA-loaded poly(L-lactide-co-caprolactone) grafts demonstrated an optimal vascular media layer accompanied by well-organized elastin fibers after long-term implantation (6 months), and they maintained potent physiological function up to 1/3 that of the native artery. In contrast, inadequate smooth muscle regeneration was observed in poly(ε-caprolactone) grafts due to slow degradation restricting the regeneration. The mechanism was further investigated and explained by the HA-induced migration of smooth muscle cell (SMC) via CD44-mediated signaling. Besides, low molecular weight HA can promote the migration of vascular progenitor cells that further differentiate into SMCs. These results highlight the importance of HA in the regeneration of functional vascular smooth muscle, and provide a new insight into the fabrication of tissue engineering vascular grafts (TEVGs) via combining rapidly degradable polymers and bioactive ECM components that hold great translational potential.

In-Vivo Assessment of a Tissue Engineered Vascular Graft Computationally Optimized for Target Vessel Compliance

Compliance mismatch between a bypass graft and the host vasculature can lead to in-vivo graft failure. Specifically, hypocompliant (stiff) grafts are believed to provide a hemodynamic environment that promotes graft failure via thrombus formation and the development of intimal hyperplasia. Tissue engineered vascular grafts(TEVGs) offer an opportunity to control the compliance of a graft to match that of the native target vasculature and minimize these undesirable outcomes. Our group has recently developed a technique to computationally and experimentally optimize the compliance of a TEVG to a wide range of target values. In this work, acellular TEVGs were created using our validated computational optimization approach that were either compliance matched (CMgel- high gelatin content) or hypocompliant (HYPOpcl- high polycaprolactone content) relative to rat abdominal aorta.A third acellular hypocompliant graft (HYPOgen- increased genipin crosslinking) was also fabricated using the CMgel material composition but crosslinked with an increased genipin concentration such that it's compliance matched that of HYPOpcl grafts. All TEVGs were implanted interpositionally into the abdominal aorta of 21 Sprague Dawley rats(n=7 for each group, males=11, females=10) for 28 days, imaged in-vivo using ultrasound, explanted, and assessed for cellular/extracellular markers using immunofluorescence and two photon excitation fluorescence imaging. In-vivo ultrasound demonstrated that CMgel grafts remained compliance-matched while hypocompliant grafts remained stiff throughout the 4 week implantation period (p<0.05). Overall, the constructs with higher gelatin content (CMgel & HYPOgen) showed increased cellular infiltration and degradation compared to the HYPOpcl grafts. The compliance matched grafts showed an increase in the amount of contractile smooth muscle cell markers in the proximal portion of the graft compared to hypocompliant grafts (0.058±0.015 CMgel vs 0.033±0.009 HYPOpcl(p=0.007) & 0.022±0.018 HYPOgen, p=0.04). The middle section (along the axial length) of compliance matched grafts displayed an increase in the ratio of CD163 to CD68 macrophages (CMgel=1.83±1.82) compared to the HYPOpcl grafts (0.13±0.06, p=0.03)suggesting a reduced pro-inflammatory response in these optimized grafts. Our results suggest that computational optimization may improve the acute (28 day) in-vivo remodeling of a biopolymer TEVG. To the authors' knowledge,this is the first in-vivo rat study comparing electrospun acellular grafts that have been computationally optimized for target levels of compliance.

Small-Diameter Synthetic Vascular Graft Immobilized with the REDV Peptide Reduces Early-Stage Fibrin Clot Deposition and Results in Graft Patency in Rats.

Early-stage hemocompatibility is indispensable for manufacturing tissue-engineered vascular grafts used in regenerative medicine. In this study, we report the in vivo blood response and patency of small-diameter synthetic vascular grafts modified with the Arg-Glu-Asp-Val (REDV) peptide. Vascular grafts were prepared by casting REDV-conjugated poly(depsipeptide-co-caprolactone) on a stainless-steel mandril (diameter: 1.8 mm). After implanting the grafts into the abdominal aorta of rats for 24 h, all three control grafts without the peptide and three out of the four REDV (control sequence) peptide-modified grafts showed occlusion. The luminal surfaces of these grafts were covered with thick thrombi. In contrast, all the grafts containing the REDV peptide were patent, and their luminal surfaces were covered with a thin layer of fibrin. These results indicated that the REDV peptide on the luminal surface effectively reduced early-stage fibrin clot deposition and formed the pseudo-endothelium layer in a peptide sequence-specific manner, resulting in graft patency.

Automated Bioreactor System for the Cultivation of Autologous Tissue-Engineered Vascular Grafts.

In an aging society, diseases associated with irreversible damage of organs are frequent. An increasing percentage of patients requires bioartificial tissue or organ substitutes. Tissue engineering products depend on a well-defined process to ensure successful cultivation while meeting high regulatory demands. The goal of the presented work is the development of a bioreactor system for the cultivation of tissue-engineered vascular grafts (TEVGs) for autologous implantation and transition from a lab scale setup to standardized production. Key characteristics include (i) the automated reliable monitoring and control of a wide-range of parameters regarding implant conditioning, (ii) easy and sterile setup and operation, (iii) reasonable costs of disposables, and (iv) parallelization of automated cultivation processes. The presented prototype bioreactor system provides comprehensive physiologically conditioning, sensing, and imaging functionality to meet all requirements for the successful cultivation of vascular grafts on a productional scale.

Vascular Tissue Engineering: Advanced Techniques and Gene Editing in Stem Cells for Graft Generation.

The common occurrence of cardiovascular diseases and the lack of proper autologous tissues prompt and promote the pressing development of tissue-engineered vascular grafts (TEVGs). Current progress on scaffold production, genetically modified cells, and use of nanotechnology-based monitoring has considerably improved the long-term patency of engineered tissue grafts. However, challenges abound in the autologous materials and manipulation of genes and cells for tissue engineering. This review overviews current development in TEVGs and discusses recent improvements in scaffolding techniques and the efficiency of gene-editing tools and their ability to fill the existing gaps in stem cell and regenerative therapies. Current advances in three-dimensional printing approaches for fabrication of engineered tissues are also reviewed together with specific biomaterials for vascular tissues. In addition, the natural and synthetic polymers that hold increasing significance for vascular tissue engineering are highlighted. Both animal models and nanotechnology-based monitoring are proposed for preclinical evaluation of engineered grafts in view of their historical significance in tissue engineering. The ultimate success of tissue regeneration, which is yet to be fully realized, depends on the optimal performance of culture systems, biomaterial constructs, and stem cells in a suitable artificial physiological environment.

Antibody-Conjugated Electrospun Vascular Scaffolds to Enhance In Situ Endothelialization.

Tissue-engineered vascular grafts (TEVGs) are promising alternatives to small-diameter prosthetic grafts. Previous methods of seeding tubular scaffolds with autologous vascular cells have been successful; however, these methods require significant preparation time. Endothelial cell (EC) growth on the luminal surface of vascular scaffolds may be critical for the integration of a TEVG to the host environment. An alternative approach for TEVGs includes the in situ endothelialization of acellular scaffolds by capturing circulating endothelial progenitor cells (EPCs) and ECs from the bloodstream through the biofunctionalization of the vascular scaffolds. In this study, fibrous scaffolds were electrospun with a 1:1 poly(ε-caprolactone) (PCL)/collagen blend solution. The electrospun fibrous scaffolds were surface-modified by immobilizing EC-specific antibodies: CD31, vascular endothelial cadherin (VE-CAD), vascular endothelial growth factor receptor 2 (VEGFR2), and von Willebrand factor (vWF). Antibodies most efficacious at capturing ECs were then paired to examine their potential synergistic cell-capturing capabilities. The study demonstrated that vascular scaffolds bioconjugated with dual antibodies demonstrated synergistic capture efficacy compared to bioconjugation with a single antibody. The capture of circulating EPCs and ECs can be optimized with bioconjugation of one or more antibodies on the luminal surface of TEVGs.

A rat arteriovenous graft model using decellularized vein.

The high rate of clinical failure of prosthetic arteriovenous grafts continues to suggest the need for novel tissue-engineered vascular grafts. We tested the hypothesis that the decellularized rat jugular vein could be successfully used as a conduit and that it would support reendothelialization as well as adaptation to the arterial environment. Autologous (control) or heterologous decellularized jugular vein (1 cm length, 1 mm diameter) was sewn between the inferior vena cava and aorta as an arteriovenous graft in Wistar rats. Rats were sacrificed on postoperative day 21 for examination. All rats survived, and grafts had 100% patency in both the control and decellularized groups. Both control and decellularized jugular vein grafts showed similar rates of reendothelialization, smooth muscle cell deposition, macrophage infiltration, and cell turnover. The outflow veins distal to the grafts showed similar adaptation to the arteriovenous flow. Both CD34, CD90 and nestin positive cells, as well as M1-type and M2-type macrophages accumulated around the graft. This model shows that decellularized vein can be successfully used as an arteriovenous graft between the rat aorta and the inferior vena cava. Several types of cells, including progenitor cells and macrophages, are present in the host response to these grafts in this model. This model can be used to test the application of arteriovenous grafts before conducting large animal experiments.

Histology and Mechanics of In Vivo Tissue-Engineered Vascular Graft for Children

This study sought to evaluate the histologic and mechanical properties of autologous invivo tissue-engineered vascular grafts (invivo TEVGs) used for pediatric heart surgery. Molds of invivo TEVGs made of silicone drain tubes were embedded into subcutaneous spaces in 2 boys during their first operation and were used as patch materials to treat pulmonary artery stenosis during the second operation. The remaining pieces of the patches were evaluated histologically and mechanically. Invivo TEVGs had very smooth luminal surfaces, and their walls mainly comprised collagen fibers and small numbers of fibroblasts. Mean wall thickness was 200 μm, mean suture retention strength was 2.26 N, and burst pressure was 3057 mm Hg. Human invivo TEVGs mainly comprise collagen fibers, and their mechanical properties prove them safe for pulmonary arterioplasty. Therefore, human invivo TEVGs may be promising alternatives to autologous pericardium for pediatric cardiovascular surgical procedures that often require multistage operations.

Fibrin – a promising material for vascular tissue engineering

This review looks at the use of fibrin in vascular tissue engineering (VTE). Autologous fibrin is one of the most affordable biopolymers because it can be obtained from peripheral blood by simple techniques. A description and comparative analysis of the methods and approaches for producing fibrin gel is provided. The ability of fibrin to promote cell attachment and migration, survival and angiogenesis, to accumulate growth factors and release them in a controlled manner, are unique and extremely useful in VTE. Fibrin gels can serve as a three-dimensional matrix molded in different sizes and shapes to be applied in a variety of ways, including as a scaffold, coating, or impregnation material. Fibrin’s high porosity and biodegradability allows controllable release of growth factors, yet fibrinolysis must be tightly regulated to avoid side effects. We discuss the main methods of regulating the rate of fibrinolysis, as well as possible side effects of such exposure. Low mechanical strength is the main limitation in using fibrin as a scaffold for vascular tissue engineering. Possible options for increasing the strength properties of fibrin matrix and evaluating their effectiveness are presented. We propose that unique biocompatibility and ideal biodegradation profile of fibrin justify its use as a scaffold material for developing an ideal fully autologous small-diameter tissue-engineered vascular graft.

Glycocalyx‐Like Hydrogel Coatings for Small Diameter Vascular Grafts

AbstractNovel biological vascular conduits, such as decellularized tissue engineered vascular grafts (TEVGs) are hindered by high thrombogenicity. To mimic the antithrombogenic surface of native vessels with a continuous glycosaminoglycan layer that is present on endothelial cells (ECs), a hyaluronic acid (HA) modified surface is established, to effectively shield blood platelets from collagen‐triggered activation. Using the amine groups present on 4 mm diameter decellularized TEVGs, a continuous HA hydrogel coating is built via a bifunctional thiol‐reactive cross‐linker, thereby avoiding nonspecific collagen matrix cross‐linking. The HA hydrogel layer recreates a luminal wall, “hiding” exposed collagen from the bloodstream. In vitro blood tests show that adhered platelets, fibrinogen absorption, and fibrin formation on HA‐coated decellularized TEVGs are significantly lower than on uncoated decellularized TEVGs. The HA surface also inhibits macrophage adhesion in vitro. HA‐coated decellularized syngeneic rat aortae (≈1.5 mm diameter), and TEVGs in rat and canine models, respectively, are protected from aggressive thrombus formation, and preserve normal blood flow. Re‐endothelialization is also observed. HA‐coated TEVGs may be an off‐the‐shelf small‐diameter vascular graft with dual benefits: antithrombogenic protection and promotion of endothelium.

A Dense Fibrillar Collagen Scaffold Differentially Modulates Secretory Function of iPSC-Derived Vascular Smooth Muscle Cells to Promote Wound Healing.

The application of human-induced pluripotent stem cells (hiPSCs) to generate vascular smooth muscle cells (hiPSC-VSMCs) in abundance is a promising strategy for vascular regeneration. While hiPSC-VSMCs have already been utilized for tissue-engineered vascular grafts and disease modeling, there is a lack of investigations exploring their therapeutic secretory factors. The objective of this manuscript was to understand how the biophysical property of a collagen-based scaffold dictates changes in the secretory function of hiPSC-VSMCs while developing hiPSC-VSMC-based therapy for durable regenerative wound healing. We investigated the effect of collagen fibrillar density (CFD) on hiPSC-VSMC’s paracrine secretion and cytokines via the construction of varying density of collagen scaffolds. Our study demonstrated that CFD is a key scaffold property that modulates the secretory function of hiPSC-VSMCs. This study lays the foundation for developing collagen-based scaffold materials for the delivery of hiPSC-VSMCs to promote regenerative healing through guiding paracrine signaling pathways.

Approaches to antithrombotic modification of vascular implants

Vascular implants in contact with blood must have high thrombotic resistance. However, in some cases, their implantation is associated with thrombosis and subsequent impaired patency of the blood vessel. Most often, this problem affects implants intended for reconstruction of small diameter vessels, which is associated with hemodynamic features in this part of the bloodstream. These include blood vessel prostheses, tissue-engineered vascular grafts, and endovascular stents. The features of the implant material are of great importance when choosing a method for its modification in order to improve biocompatibility and thromboresistance. The review analyzes current experience in using various methods of immobilizing drugs to the surface of vascular prostheses and endovascular stents made from stable and biodegradable polymers. The prospects of creating thromboresistant vascular grafts and stents by joint immobilization on the surface of the polymer material of drugs with antithrombogenic activity and biologically active molecules that regulate the reaction to a foreign body and implant remodeling were evaluated. Numerous studies in the review demonstrating a wide range of ways to modify blood vessel prostheses, tissue-engineered vascular grafts, and endovascular stents with antithrombotic drugs to increase their thrombosis resistance. The main approaches of antithrombotic modification include conjugation of drugs and biologically active molecules on the implant surface. At the same time, new technologies are aimed not only at inhibiting the process of thrombus formation, but also at reducing the intensity of the inflammation process and stimulating the reparation of vascular tissue.

Catalytic Generation of Nitric Oxide from Poly(ε-caprolactone)/Phosphobetainized Keratin Mats for a Vascular Tissue Engineering Scaffold.

Tissue-engineered vascular graft (TEVG) is a promising alternative to meet the clinical demand of organ shortages. Herein, human hair keratin was extracted by the reduction method, followed by modification with zwitterionic 2-methacryloyloxyethyl phosphorylcholine (MPC) through thiol-Michael addition to improve blood clotting nature. Then, phosphobetainized keratin (PK) was coelectrospun with poly(ε-caprolactone) (PCL) to afford PCL/PK mats with a ratio of 7:3. The surface morphology, chemical structure, and wettability of these mats were characterized. The biocomposite mats selectively enhanced adhesion, migration, and growth of endothelial cells (ECs) while suppressed proliferation of smooth muscle cells (SMCs) in the presence of glutathione (GSH) and GSNO due to the catalytic generation of NO. In addition, these mats exhibited good blood anticoagulant activity by reducing platelet adhesion, prolonging blood clotting time, and inhibiting hemolysis. Taken together, these NO-generating PCL/PK mats have potential applications as a scaffold for vascular tissue engineering with rapid endothelialization and reduced SMC proliferation.

Tissue‐engineered Vascular Grafts for Congenital Heart Surgery

ObjectiveWe developed a tissue engineered vascular graft (TEVG) formed by seeding autologous bone marrow‐derived mononuclear cells onto a biodegradable tubular scaffold fabricated from a knitted polyglycolic acid core and coated with a 50:50 copolymer of poly lactic acid and polycaprolactone. Once implanted, the scaffold degrades and neotissue forms giving rise to neovessel which is structurally and functionally similar to the native vessel into which it was implanted. Herein we will describe the natural history of the evolution of the mechanical properties of the TEVG after implantation and evaluate the impact of the mechanical properties of the TEVG on graft performance.MethodsWe utilized an ovine intrathoracic inferior vena cava model in order to evaluate the evolving mechanical properties of the TEVG and determine their impact on graft performance. We implanted TEVG into 24 juvenile lambs and serially assessed graft morphometry and function using serial angiography and intravascular ultrasound (IVUS) over a 6 ‐month to 1‐year period. We serially measured changes in graft size using both angiography and IVUS. We also assessed the graft hemodynamics at angiography. We measured graft displacement over both the cardiac and respiratory cycle. Finally, a subset of TEVGs were harvested and underwent biaxial mechanical testing.ResultsThe TEVG undergoes significant remodeling when after implantation. As the scaffold degrades, the TEVG transforms from a relatively stiff conduit to a highly compliant neovessel with growth capacity.ConclusionsThe transformation of the TEVG from a relatively stiff vascular conduit to a more compliant neovessel enables the graft to function as a capacitance vessel improving its performance when used in the venous circulation.Support or Funding InformationNIH R01HL139796

Spontaneous reversal of stenosis in tissue-engineered vascular grafts.

We developed a tissue-engineered vascular graft (TEVG) for use in children and present results of a U.S. Food and Drug Administration (FDA)-approved clinical trial evaluating this graft in patients with single-ventricle cardiac anomalies. The TEVG was used as a Fontan conduit to connect the inferior vena cava and pulmonary artery, but a high incidence of graft narrowing manifested within the first 6 months, which was treated successfully with angioplasty. To elucidate mechanisms underlying this early stenosis, we used a data-informed, computational model to perform in silico parametric studies of TEVG development. The simulations predicted early stenosis as observed in our clinical trial but suggested further that such narrowing could reverse spontaneously through an inflammation-driven, mechano-mediated mechanism. We tested this unexpected, model-generated hypothesis by implanting TEVGs in an ovine inferior vena cava interposition graft model, which confirmed the prediction that TEVG stenosis resolved spontaneously and was typically well tolerated. These findings have important implications for our translational research because they suggest that angioplasty may be safely avoided in patients with asymptomatic early stenosis, although there will remain a need for appropriate medical monitoring. The simulations further predicted that the degree of reversible narrowing can be mitigated by altering the scaffold design to attenuate early inflammation and increase mechano-sensing by the synthetic cells, thus suggesting a new paradigm for optimizing next-generation TEVGs. We submit that there is considerable translational advantage to combined computational-experimental studies when designing cutting-edge technologies and their clinical management.

Cell‐assembled Extracellular Matrix (CAM) as a Biomaterial for Building Vascular Grafts

Sheets of human CAM, produced by cultured cells in vitro, have been rolled to produce completely biological tissue‐engineered vascular grafts (TEVGs). Despite a rapid and successful progression to clinical trials, this approach remains complex. Our novel strategy is to produce TEVG with yarns produced from CAM sheets using a textile‐based approach. This strategy is faster, more reliable, more versatile and more easily automated. CAM sheets can be cut in ribbons of various widths, which can also be twisted to make threads. Yarn (ribbons or threads) can be processed in different ways: devitalized (freezing/dehydration/freezing/rehydration cycle), decellularized (8 mM CHAPS, 1 M NaCl, 25 mM EDTA, 0.12 M NaOH) and/or gamma‐sterilized (25 kGy). We have evaluated the effect of these processing steps on the histological and mechanical properties of the yarn. We have also assessed the impact of these steps on the remodeling (histological and mechanical) of the yarn after subcutaneous implantation in a nude rat for up to 6 months. Results showed that the human CAM‐based yarn creates very little inflammatory reaction and is essentially only mildly remodeled by the host response. However, some processing steps do lead to increased reactivity and decreased performance in vivo. Woven vessels display remarkable mechanical properties without the need for synthetic scaffolding or chemical crosslinking. An allogeneic large animal model is in development to address remodeling in an allogeneic setting in an immunologically competent recipient. Because it can be produced according to specific mechanical and physical specifications, and because it can be used with standard textile assembly methods, the CAM can truly be called a “bio‐material”. More importantly, because of its native‐like structure, this material has the potential to be accepted by the host without significant immune responses and to support a long‐term “physiological” remodeling process.Support or Funding InformationNational Institute of Health Small Business Innovation Research (SBIR) program (HL105010), the Ministère de la Recherche et de l’Enseignement Supérieur, the Chaire Senior de l’Initiative d’Excellence de l’Université de Bordeaux (IdEx Bordeaux), the Agence Nationale de la Recherche (ANR‐16‐CE18‐0024‐01), the Région Nouvelle Aquitaine (2016‐1R30402), and the European Research Council (Advanced Grant # 785908).A thread and a woven vessel made from human CAM.Figure 1

A computational bio-chemo-mechanical model of in vivo tissue-engineered vascular graft development.

Stenosis is the primary complication of current tissue-engineered vascular grafts used in pediatric congenital cardiac surgery. Murine models provide considerable insight into the possible mechanisms underlying this situation, but they are not efficient for identifying optimal changes in scaffold design or therapeutic strategies to prevent narrowing. In contrast, computational modeling promises to enable time- and cost-efficient examinations of factors leading to narrowing. Whereas past models have been limited by their phenomenological basis, we present a new mechanistic model that integrates molecular- and cellular-driven immuno- and mechano-mediated contributions to in vivo neotissue development within implanted polymeric scaffolds. Model parameters are inferred directly from in vivo measurements for an inferior vena cava interposition graft model in the mouse that are augmented by data from the literature. By complementing Bayesian estimation with identifiability analysis and simplex optimization, we found optimal parameter values that match model outputs with experimental targets and quantify variability due to measurement uncertainty. Utility is illustrated by parametrically exploring possible graft narrowing as a function of scaffold pore size, macrophage activity, and the immunomodulatory cytokine transforming growth factor beta 1 (TGF-β1). The model captures salient temporal profiles of infiltrating immune and synthetic cells and associated secretion of cytokines, proteases, and matrix constituents throughout neovessel evolution, and parametric studies suggest that modulating scaffold immunogenicity with early immunomodulatory therapies may reduce graft narrowing without compromising compliance.

Surface Modification by Nanobiomaterials for Vascular Tissue Engineering Applications.

Treatment of cardiovascular disease has achieved great success using artificial implants, particularly synthetic-polymer made grafts. However, thrombus formation and restenosis are the current clinical problems need to be conquered. New biomaterials, modifying the surface of synthetic vascular grafts, have been created to improve long-term patency for the better hemocompatibility. The vascular biomaterials can be fabricated from synthetic or natural polymers for vascular tissue engineering. Stem cells can be seeded by different techniques into tissue-engineered vascular grafts in vitro and implanted in vivo to repair the vascular tissues. To overcome the thrombogenesis and promote the endothelialization effect, vascular biomaterials employing nanotopography are more bio-mimic to the native tissue made and have been engineered by various approaches such as prepared as a simple surface coating on the vascular biomaterials. It has now become an important and interesting field to find novel approaches to better endothelization of vascular biomaterials. In this article, we focus to review the techniques with better potential improving endothelization and summarize for vascular biomaterial application. This review article will enable the development of biomaterials with a high degree of originality, innovative research on novel techniques for surface fabrication for vascular biomaterials application.

Stiffness of the aligned fibers affects structural and functional integrity of the oriented endothelial cells

Promoting healthy endothelialization of the tissue-engineered vascular grafts is of great importance in preventing the occurrence of undesired post-implantation complications including neointimal hyperplasia, late thrombosis, and neoatherosclerosis. Previous researches have demonstrated the crucial role of scaffold topography or stiffness in modulating the behavior of the monolayer endothelial cells (ECs). However, effects of the stiffness of scaffolds with anisotropic topography on ECs within vivo like oriented morphology has received little attention. In this study, aligned fibrous substrates (AFSs) with tunable stiffness (14.68–2141.72 MPa), similar to the range of stiffness of the healthy and diseased subendothelial matrix, were used to investigate the effects of fiber stiffness on ECs’ attachment, orientation, proliferation, function, remodeling and dysfunction. The results demonstrate that stiffness of the AFSs, capable of providing topographical cues, is a crucial endothelium-protective microenvironmental factor by maintaining stable and quiescent endothelium with in vivo like orientation and strong cell–cell junctions. Stiffer AFSs exacerbated the disruption of endothelium integrity, the occurrence of endothelial-to-mesenchymal transition (EndMT), and the inflammation-induced activation in the endothelial monolayer. This study provides new insights into the understanding on how the stiffness of biomimicking anisotropic substrate regulates the structural and functional integrity of the in vivo like endothelial monolayer, and offers essential designing parameters in engineering biomimicking small-diameter vascular grafts for the regeneration of viable blood vessels. Statement of significanceIn vascular tissue engineering, promoting endothelialization on scaffold surface has been considered as a paramount strategy to reduce post-implantation complications. Electrospun aligned fibers have been known to provide contact guidance effect in directing endothelial cells' oriented growth, however, whether the formed EC monolayer in ‘correct’ orientation shape is of ‘correct’ function hasn't been explored yet. Given the recognized important role of substrate stiffness in endothelial function, AFSs across physiologically relevant range of moduli (14.68–2141.72 MPa) while maintaining consistent surface chemistry and topographical features were employed to investigate the fiber stiffness effects on ECs function in anisotropic morphology. This study will provide more insightful perspectives in the physiologically remodeling progression of vascular endothelium and design of vascular scaffolds.

Shapeable large-pore electrospun polycaprolactam cotton facilitates the rapid formation of a functional tissue engineered vascular graft

Tissue engineered vascular grafts (TEVGs) with diameters <6 mm have broad applications. However, maintaining graft post-implantation patency and function is still difficult to achieve and of urgent necessity. In this study, we use an effective strategy to make the large-pore electrospun polycaprolactone (PCL) cotton material (Cotton) and utilized immunoreactive pre-cultivation to prepare a small diameter autologous TEVG. The material assessment via scanning electron microscopy revealed that Cotton has favorable characteristics that make it suitable for immunoreactive pre-cultivation. It also possesses great mechanical properties. Due to strong reshape ability of Cotton, designed vascular patch and vascular scaffold and others could be obtained. For vascular scaffold, Cotton was wrapped around a polytetrafluoroethylene (PTFE) tube and implanted subcutaneously in rats for up to 2 weeks to prepare autologous TEVGs. Autologous TEVGs were then transplanted into the abdominal aorta of the same rat. Cotton TEVG formed functional graft with favorable patency after transplantation for 4 weeks. Histological results showed that the TEVGs had characteristics similar to that of the native aorta. Our results indicate PCL cotton material could be a kind of shapeable material and used to create a promising biocompatible and functional vascular graft over a relatively short period of time.