Scaffolds For Vascular Tissue Engineering Research Articles

Tailoring the Heterogeneous Structure of Macro-Fibers Assembled by Bacterial Cellulose Nanofibrils for Tissue Engineering Scaffolds.

Bacterial cellulose/oxidized bacterial cellulose nanofibrils (BC/oxBCNFs) macro-fibers are developed as a novel scaffold for vascular tissue engineering. Utilizing a low-speed rotary coagulation spinning technique and precise solvent control, macro-fibers with a unique heterogeneous structure with dense surface and porous core are created. Enhanced by a polydopamine (PDA) coating, these macro-fibers offer robust mechanical integrity, high biocompatibility, and excellent cell adhesion. When cultured with endothelial cells (ECs) and smooth muscle cells (SMCs), the macro-fibers support healthy cell proliferation and exhibit a unique spiral SMC alignment, demonstrating their vascular suitability. This innovative strategy opens new avenues for advances in tissue engineering.

Biological Materials for Tissue-Engineered Vascular Grafts: Overview of Recent Advancements.

The clinical demand for tissue-engineered vascular grafts is still rising, and there are many challenges that need to be overcome, in particular, to obtain functional small-diameter grafts. The many advances made in cell culture, biomaterials, manufacturing techniques, and tissue engineering methods have led to various promising solutions for vascular graft production, with available options able to recapitulate both biological and mechanical properties of native blood vessels. Due to the rising interest in materials with bioactive potentials, materials from natural sources have also recently gained more attention for vascular tissue engineering, and new strategies have been developed to solve the disadvantages related to their use. In this review, the progress made in tissue-engineered vascular graft production is discussed. We highlight, in particular, the use of natural materials as scaffolds for vascular tissue engineering.

Flexible, Suturable, and Leak-free Scaffolds for Vascular Tissue Engineering Using Melt Spinning.

Coronary artery disease affects millions worldwide. Bypass surgery remains the gold standard; however, autologous tissue is not always available. Hence, the need for an off-the-shelf graft to treat these patients remains extremely high. Using melt spinning, we describe here the fabrication of tubular scaffolds composed of microfibers aligned in the circumferential orientation mimicking the organized extracellular matrix in the tunica media of arteries. By variation of the translational extruder speed, the angle between fibers ranged from 0 to ∼30°. Scaffolds with the highest angle showed the best performance in a three-point bending test. These constructs could be bent up to 160% strain without kinking or breakage. Furthermore, when liquid was passed through the scaffolds, no leakage was observed. Suturing of native arteries was successful. Mesenchymal stromal cells were seeded on the scaffolds and differentiated into vascular smooth muscle-like cells (vSMCs) by reduction of serum and addition of transforming growth factor beta 1 and ascorbic acid. The scaffolds with a higher angle between fibers showed increased expression of vSMC markers alpha smooth muscle actin, calponin, and smooth muscle protein 22-alpha, whereas a decrease in collagen 1 expression was observed, indicating a positive contractile phenotype. Endothelial cells were seeded on the repopulated scaffolds and formed a tightly packed monolayer on the luminal side. Our study shows a one-step fabrication for ECM-mimicking scaffolds with good handleability, leak-free property, and suturability; the excellent biocompatibility allowed the growth of a bilayered construct. Future work will explore the possibility of using these scaffolds as vascular conduits in in vivo settings.

Bioinspired PLCL/Elastin Nanofibrous Vascular Tissue Engineering Scaffold Enhances Endothelial Cells and Inhibits Smooth Muscle Cells.

In vascular tissue engineering, a scaffold that can enhance the proliferation of endothelial cells (ECs) while inhibiting the synthetic differentiation of smooth muscle cells (SMCs) is crucial to prevent thrombus and restenosis after graft implantation. However, it is always challenging to incorporate both properties simultaneously in a vascular tissue engineering scaffold. In this study, a novel composite material was developed by combining a synthetic biopolymer of poly(l-lactide-co-caprolactone) (PLCL) and a natural biopolymer of elastin through electrospinning. Cross-linking of the PLCL/elastin composite fibers using EDC/NHS was performed to stabilize the elastin component. The incorporation of elastin into PLCL was found to enhance the hydrophilicity and biocompatibility of the resulting PLCL/elastin composite fibers, as well as the mechanical properties. Additionally, as a natural component of the extracellular matrix, elastin displayed antithrombotic properties reducing platelet adhesion and improving blood compatibility. Results of cell culture experiments with human umbilical vein ECs (HUVECs) and human umbilical artery SMCs (HUASMCs) showed that the composite fiber membrane had high cell viability, promoting the proliferation and adhesion of HUVECs and inducing a contractile phenotype in HUASMCs. These results indicate that the PLCL/elastin composite material has great potential for use in vascular graft applications due to its favorable properties and rapid endothelialization and contractile phenotypes of cells.

Biocompatibility of silk methacrylate/gelatin-methacryloyl composite hydrogel and its feasibility as a vascular tissue engineering scaffold

Silk methacrylate (SilMA) has been studied extensively due to its ability to modify Silk fibroin (SF) by increasing the water solubility and enhancing the mechanical properties of SF hydrogels. However, SilMA hydrogels are generally soft with weak mechanical properties. In order to enhance the mechanical properties of hydrogel scaffolds, we used liquid nitrogen to modify SilMA to obtain a novel N2-SilMA/gelatin-methacryloyl (GelMA) composite hydrogel. N2-SilMA was successfully detected by Fourier transform infrared (FTIR) spectroscopy and 1H nuclear magnetic resonance. Scanning electron microscope showed that the composite hydrogel still had certain arrangement characteristics of SF and dense pores which met the necessary conditions for the cell scaffold. The mechanical tests showed that the mechanical properties of SilMA were greatly enhanced after modification at ultra-low temperature. We evaluated its cytocompatibility and biocompatibility, and the results showed that the composite scaffold promoted the growth of cells. Different types of composite hydrogels were injected into ICR mice and the results showed a stable scaffold structure in vivo, suggesting their ability to promote angiogenesis. In conclusion, the N2-SilMA/GelMA composite hydrogel had better mechanical properties, excellent cytocompatibility, and biological properties compared to the other groups.

Biomimetic and Bioactive Small Diameter Tubular Scaffolds for Vascular Tissue Engineering.

The present work aimed at the production and characterization of small caliber biomimetic and bioactive tubular scaffolds, which are able to favor the endothelialization process, and therefore potentially be suitable for vascular tissue engineering. The tubular scaffolds were produced using a specially designed mold, starting from a gelatin/gellan/elastin (GGE) blend, selected to mimic the composition of the extracellular matrix of native blood vessels. GGE scaffolds were obtained through freeze-drying and subsequent cross-linking. To obtain systems capable of promoting endothelization, the scaffolds were functionalized using two different bioactive peptides, Gly-Arg-Gly-Asp-Ser-Pro (GRGSDP) and Arg-Glu-Asp-Val (REDV). A complete physicochemical, mechanical, functional, and biological characterization of the developed scaffolds was performed. GGE scaffolds showed a good porosity, which could promote cell infiltration and proliferation and a dense external surface, which could avoid bleeding. Moreover, developed scaffolds showed good hydrophilicity, an elastic behavior similar to natural vessels, suitability for sterilization by an ISO accepted treatment, and an adequate suture retention strength. In vitro cell culture tests showed no cytotoxic activity against 3T3 fibroblasts. The functionalization with the REDV peptide favored the adhesion and growth of endothelial cells, while GRGDSP-modified scaffolds represented a better substrate for fibroblasts.

In Vitro Comparison Study of Plasma Treated Bilayer PGS/PCL and PGS/PLA Scaffolds for Vascular Tissue Engineering

In Vitro Comparison Study of Plasma Treated Bilayer PGS/PCL and PGS/PLA Scaffolds for Vascular Tissue Engineering

Correction to: Electrospun cellulose Nano fibril reinforced PLA/PBS composite scaffold for vascular tissue engineering

Correction to: Electrospun cellulose Nano fibril reinforced PLA/PBS composite scaffold for vascular tissue engineering

A review on developments of in-vitro and in-vivo evaluation of hybrid PCL-based natural polymers nanofibers scaffolds for vascular tissue engineering

The main objective of the present review article is to investigate the in-vitro and in-vivo biocompatibility behavior of hybrid PCL-based scaffolds with collagen, gelatin, and chitosan to improve endothelialization for VTE applications. It is reported that the high-rate failure of small diameters vascular grafts (SDVGs, <6 mm) due to adhesion of platelets and plasma protein and aggregation, over-proliferation of smooth muscle cells (SMCs), and neointimal hyperplasia at the implantation site is main challenge in vascular tissue engineering (VTE). The fast re-establishment of a functional endothelial cell (EC) layer representing a crucial importance strategy has been proposed to reduce these adverse outcomes. Polycaprolactone (PCL), with optimum mechanical properties, it showed great potential in biomedical applications, but its biocompatibility is still highly concerned in VTE. Modifications of the PCL vascular grafts by developing the hybrid structures using natural polymers with optimum hydrophilicity and biocompatibility properties in order to speed up the re-endothelialization process have been proposed over the last years. Analyzing the mentioned results in the present study can offer a better understanding of the benefits and challenges of applying the natural polymer as a thrombotic response upon implantation in clinical trials. Also, it can suggest the PCL-Collagen as the best framework for the fabrication of rationally designed SDVGs for VTE.

Cyclopeptide-β-cyclodextrin/γ-glycerol methoxytrimethoxysilane film for potential vascular tissue engineering scaffolds

The mortality rate of cardiovascular diseases is the highest among all mortality rates worldwide. Allotransplantation and autotransplantation are limited by rejection reaction and availability. Tissue engineering provides new avenues for the treatment of cardiovascular diseases. However, the current small-diameter (<6 mm) vascular tissue-engineered scaffolds have many challenges, including thrombosis, stenosis, and infection. Small-diameter vascular scaffolds have structural and compositional requirements such as biocompatibility, porosity, and appropriate phase separation. We used liquid-crystal cyclopeptide（CYC）to modify β-cyclodextrin and mixed it with γ-glycerol methoxytrimethoxysilane (GPTMS) to prepare CYC-β-cyclodextrin (βCD)/GPTMS film by sol-gel. The chemical structure of CYC-βCD was confirmed by Fourier transform infrared spectroscopy and 1H-nuclear magnetic resonance. The chemical characterization of CYC-βCD/GPTMS film was performed by differential scanning calorimetry, X-ray diffraction, and small-angle X-ray scattering. The surface morphology and phase separation microstructure of the film were determined by scanning electron microscopy and atomic force microscopy, and the image of polarizing microscopy showed the liquid-crystal structure of the film. Cell culture experiments showed that CYC-βCD/GPTMS film had good cytocompatibility and induced growth and proliferation of cells. These results indicated the potential applications of CYC-βCD/GPTMS film in tissue engineering scaffolds.

3D printed GelMA/carboxymethyl chitosan composite scaffolds for vasculogenesis

The establishment of vascularization is the basic prerequisite for the survival and functionalization of three-dimensional (3 D) tissue-engineered constructs. The application of hydrogel to tissue regeneration has promoted the research and development of regenerative medicine. In this study, we fabricated the composite scaffold composed of gelatin methacryloyl (GelMA) and carboxymethyl chitosan (CMCS) and proved the potential of the scaffold to promote angiogenesis. To characterize the physical properties of the composite scaffold, the mechanical strength, swelling ratio and internal microstructure were tested. The biocompatibility of the GelMA/CMCS scaffold was evaluated by live/dead test and CCK-8. To evaluate the cell distribution, CD31 expression and endothelial differentiation of seeded rat bone marrow mesenchymal stem cells (BMSCs) on the scaffolds, the cytoskeletal staining, immunofluorescence staining and real-time PCR were performed. The composite GelMA/CMCS scaffolds seeded with BMSCs showed excellent mechanical properties, CD31 expression and vasculogenesis genes expression, compared with the pure GelMA scaffold. Furthermore, the activity of BMSCs in 3 D printed GelMA/CMCS scaffolds proved the feasibility of the bioprinting of cells-laden GelMA/CMCS composite scaffolds. In conclusion, our study demonstrated the potential of GelMA/CMCS composite scaffolds for vascular tissue engineering.

Biodegradable highly porous interconnected poly(ε‐caprolactone)/poly(L‐lactide‐co‐ε‐caprolactone) scaffolds by supercritical foaming for small‐diameter vascular tissue engineering

AbstractBiodegradable ϕ4 mm tubular porous poly(ε‐caprolactone)/poly(L‐lactide‐co‐ε‐caprolactone) (PCL/PLCL) scaffolds are fabricated successfully via one‐step microcellular supercritical carbon dioxide foaming process. The effect of blending ratio on the rheology, pore structures, mechanical property, wettability, and biocompatibility of PCL/PLCL blends tubular scaffold are reported. Rheological results show that PCL matrix and PLCL dispersed phase has good compatibility. The melt strength of PCL can be enhanced obviously by adding PLCL. With an increase of PLCL content from 10 to 30 wt%, the pore size increases from 7.6 to 24.9 μm due to the homogeneous nucleation effect. The maximum open‐cell content can reach 77% for PCL/PLCL foamed sample. Cyclical tensile and compliance tests show that few content of dispersed PLCL (10–20 wt%) improves the flexibility and recoverability. Cell viability results demonstrate that human umbilical vein endothelial cells (HUVECs) cultured on all PCL/PLCL porous scaffolds exhibit a typical spindle‐like cell morphology. Moreover, HUVECs have a higher density and spreading areas on surface of 10% PLCL scaffold. The results gathered in this paper may open a new perspective for the fabrication of small‐diameter vascular tissue engineering scaffold.

Degradation and Characterisation of Electrospun Polycaprolactone (PCL) and Poly(lactic-co-glycolic acid) (PLGA) Scaffolds for Vascular Tissue Engineering.

The current study aimed to evaluate the characteristics and the effects of degradation on the structural properties of Poly(lactic-co-glycolic acid) (PLGA)- and polycaprolactone (PCL)-based nanofibrous scaffolds. Six scaffolds were prepared by electrospinning, three with PCL 15% (w/v) and three with PLGA 10% (w/v), with electrospinning processing times of 30, 60 and 90 min. Both types of scaffolds displayed more robust mechanical properties with increased spinning times. The tensile strength of both scaffolds with 90-min electrospun membranes did not show a significant difference in their strengths, as the PCL and PLGA scaffolds measured at 1.492 MPa ± 0.378 SD and 1.764 MPa ± 0.7982 SD, respectively. All membranes were shown to be hydrophobic under a wettability test. A degradation behaviour study was performed by immersing all scaffolds in phosphate-buffered saline (PBS) solution at room temperature for 12 weeks and for 4 weeks at 37 °C. The effects of degradation were monitored by taking each sample out of the PBS solution every week, and the structural changes were investigated under a scanning electron microscope (SEM). The PCL and PLGA scaffolds showed excellent fibre structure with adequate degradation, and the fibre diameter, measured over time, showed slight increase in size. Therefore, as an example of fibre water intake and progressive degradation, the scaffold’s percentage weight loss increased each week, further supporting the porous membrane’s degradability. The pore size and the porosity percentage of all scaffolds decreased substantially over the degradation period. The conclusion drawn from this experiment is that PCL and PLGA hold great promise for tissue engineering and regenerative medicine applications.

Assessment of Electrospun Pellethane-Based Scaffolds for Vascular Tissue Engineering.

We examined the physicochemical properties and the biocompatibility and hemocompatibility of electrospun 3D matrices produced using polyurethane Pellethane 2363-80A (Pel-80A) blends Pel-80A with gelatin or/and bivalirudin. Two layers of vascular grafts of 1.8 mm in diameter were manufactured and studied for hemocompatibility ex vivo and functioning in the infrarenal position of Wistar rat abdominal aorta in vivo (n = 18). Expanded polytetrafluoroethylene (ePTFE) vascular grafts of similar diameter were implanted as a control (n = 18). Scaffolds produced from Pel-80A with Gel showed high stiffness with a long proportional limit and limited influence of wetting on mechanical characteristics. The electrospun matrices with gelatin have moderate capacity to support cell adhesion and proliferation (~30–47%), whereas vascular grafts with bivalirudin in the inner layer have good hemocompatibility ex vivo. The introduction of bivalirudin into grafts inhibited platelet adhesion and does not lead to a change hemolysis and D-dimers concentration. Study in vivo indicates the advantages of Pel-80A grafts over ePTFE in terms of graft occlusion, calcification level, and blood velocity after 6 months of implantation. The thickness of neointima in Pel-80A–based grafts stabilizes after three months (41.84 ± 20.21 µm) and does not increase until six months, demonstrating potential for long-term functioning without stenosis and as a suitable candidate for subsequent preclinical studies in large animals.

Using Wet Electrospun PCL/Gelatin/CNT Yarns to Fabricate Textile-Based Scaffolds for Vascular Tissue Engineering.

Incorporating conductive materials in scaffolds has shown advantages in regulating adhesion, mitigation, and proliferation of electroactive cells for tissue engineering applications. Among various conductive materials, carbon nanotubes (CNTs) have shown great promises in tissue engineering because of their good mechanical properties. However, the broad application of CNTs in tissue engineering is limited by current methods to incorporate CNTs in polymers that require miscible solvents to dissolve CNTs and polymers or CNT surface modification. These methods either limit polymer selections or adversely affect the properties of polymer/CNT composites. Here, we report a novel method to fabricate polymer/CNT composite yarns by electrospinning polycaprolactone/gelatin into a bath of CNT dispersion and extracting electrospun fibers out of the bath. The concentration of CNTs in the bath affects the thermal and mechanical properties and the yarns' degradation behavior. In vitro biological test results show that within a limited range of CNT concentrations in the bath, the yarns exhibit good biocompatibility and the ability to guide cell elongation and alignment. We also report the design and fabrication of a vascular scaffold by knitting the yarns into a textile fabric and combining the textile fabric with gelatin. The scaffold has similar mechanical properties to native vessels and supports cell proliferation. This work demonstrates that the wet electrospun polymer/CNT yarns are good candidates for constructing vascular scaffolds and provides a novel method to incorporate CNTs or other functional materials into biopolymers for tissue engineering applications.

Three-dimensional silk fibroin microsphere-nanofiber scaffolds for vascular tissue engineering

A significant limitation in the engineering of artificial small-diameter vascular scaffolds is that the number of endothelial cells (ECs) is not sufficient to generate a confluent coverage of the vascular scaffolds, so that the surfaces of vascular scaffolds form thrombus via platelet adhesion and aggregation. Thrombus decrease relies on three-dimensional (3D) scaffolds to mimic the natural extracellular matrix (ECM) as templates to regulate cell behavior and facilitate tissue maturation. Here, we developed 3D scaffolds consisting of silk fibroin (SF) nanofibers and homogeneous microspheres by electrospinning and microfluidics. The nanofibers with diameters ranging from 250 to 350 ​nm doped with microspheres (2–10 ​μm) formed bridge-shaped structures. ECs were seeded and maintained on the 3D microsphere-nanofiber scaffolds with a mean fiber diameter of 300 ​nm. A 10% higher ratio of cell proliferation on 3D microsphere-nanofiber SF scaffolds was noted as compared to that on microporous and sponge-like SF scaffolds with small surface network fabricated by freeze-drying. Moreover, the gene transcript levels including CD146, VE-C and PECAM-1 were better preserved on 3D microsphere-nanofiber SF scaffolds than those on freeze-dried scaffolds. Thus, the developed 3D microsphere-nanofiber structure may have a myriad of applications in vascular tissue engineering scaffolds and cardiovascular devices.

On the biocompatibility of graphene oxide towards vascular smooth muscle cells

Graphene and its derivatives have shown fascinating potential in biomedical applications. However, the biocompatibility of graphene with vascular smooth muscle cells (VSMCs) and applications to vascular engineering have not been explored extensively. Using a rat aortic smooth muscle cell line, A7r5, as a VSMC model, we have explored the effects of graphene oxide (GO) on the growth and behaviours of VSMCs. Results demonstrated that GO had no obvious toxicity to VSMCs. Cells cultured on GO retained the expression of smooth muscle cell-specific markers CNN1, ACTA2 and SMTN, on both mRNA and protein levels. A wound healing assay demonstrated no effect of GO on cell migration. We also found that small-flaked GO favoured the proliferation of VSMCs, suggesting a potential of using surface chemistry or physical properties of GO to influence cell growth behaviour. These results provide insight into the suitability of GO as a scaffold for vascular tissue engineering.

Strategies to Improve Nanofibrous Scaffolds for Vascular Tissue Engineering.

The biofabrication of biomimetic scaffolds for tissue engineering applications is a field in continuous expansion. Of particular interest, nanofibrous scaffolds can mimic the mechanical and structural properties (e.g., collagen fibers) of the natural extracellular matrix (ECM) and have shown high potential in tissue engineering and regenerative medicine. This review presents a general overview on nanofiber fabrication, with a specific focus on the design and application of electrospun nanofibrous scaffolds for vascular regeneration. The main nanofiber fabrication approaches, including self-assembly, thermally induced phase separation, and electrospinning are described. We also address nanofibrous scaffold design, including nanofiber structuring and surface functionalization, to improve scaffolds’ properties. Scaffolds for vascular regeneration with enhanced functional properties, given by providing cells with structural or bioactive cues, are discussed. Finally, current in vivo evaluation strategies of these nanofibrous scaffolds are introduced as the final step, before their potential application in clinical vascular tissue engineering can be further assessed.

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.

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.