A major barrier in the development of a clinically-useful small-diameter tissue engineered vascular graft (TEVG) is the scaffold component. Scaffold requirements include matching the mechanical and structural properties with those of native vessels and optimizing the microenvironment to foster cell integration, adhesion, and growth. We have developed a small-diameter, bi-layered, biodegradable, elastomeric scaffold based on a synthetic, biodegradable elastomer. The scaffold incorporates a highly porous inner layer, allowing cell integration and growth, and an external, fibrous reinforcing layer deposited by electrospinning. Scaffold morphology and mechanical properties were assessed, quantified, and compared to those of native vessels. Scaffolds were then seeded with adult stem cells via a rotational vacuum seeding device to obtain a TEVG, cultured in dynamic conditions for 7 days, and evaluated for cellularity. The scaffold showed a firm integration of the two polymeric layers with no delaminations. Mechanical properties were physiologically-consistent showing anisotropy, elastic modulus (1.4±0.4 MPa), and ultimate tensile stress (8.3±1.7 MPa) comparable with native vessels. Compliance and suture retention force were 4.6±0.5×10−4 mmHg−1 and 3.4±0.3 N, respectively. Seeding resulted in a rapid, uniform, bulk integration of cells, with a seeding efficiency of 92±1%. The scaffolds maintained a high level of cellular density throughout dynamic culture. This approach, combining artery-like mechanical properties and a rapid and efficient cellularization, might contribute to the future clinical translation of TEVGs.

Collagen is commonly used as a tissue-engineering scaffold, yet its in vivo applications are limited by a deficiency in mechanical strength. The purpose of this work was to explore the utilization of a unique enzymatic crosslinking procedure aimed at improving the mechanical properties of collagen-based scaffold materials. Type I bovine collagen gel was crosslinked by transglutaminase, which selectively mediates the chemical reaction between glutamine and lysine residues on adjacent protein fibers, thus providing covalent amide bonds that serve to reinforce the three-dimensional matrix. The degree of crosslinking was verified by thermal analysis and amine group content. The denaturation temperature of crosslinked collagen reached a maximum of 66 +/- 1 degrees C. The chemical reaction was confirmed to be noncytotoxic with respect to bone marrow stromal cells acquired from New Zealand White rabbits. Tube-shaped cellular constructs fashioned from crosslinked collagen and bone marrow stromal cells were found to have burst pressures significantly higher than their noncrosslinked analogs (71 +/- 4 mmHg vs. 46 +/- 3 mmHg; p < 0.01). Thus, the transglutaminase mediated reaction served to successfully strengthen collagen gels while remaining benign toward cells.

Arterial vein grafts (AVGs) often fail due to intimal hyperplasia, thrombosis, or accelerated atherosclerosis. Various approaches have been proposed to address AVG failure, including delivery of temporary mechanical support, many of which could be facilitated by peri-vascular placement of a biodegradable polymer wrap. The purpose of this work was to demonstrate that a polymer wrap can be applied to vein segments without compromising viability/function, and to demonstrate one potential application; i.e., gradually imposing the mid-wall circumferential wall stress (CWS) in wrapped veins exposed to arterial levels of pressure.Poly(ester urethane)urea, collagen, and elastin were combined in solution, and then electrospun onto freshly-excised porcine internal jugular vein segments. Tissue viability was assessed via Live/ Dead™ staining for necrosis, and vasomotor-challenge with epinephrine and sodium nitroprusside for functionality. Wrapped vein segments were also perfused for 24-hrs within an ex vivo vascular perfusion system under arterial conditions (pressure=120/80 mmHg; flow=100 mL/min), and CWS was calculated every hour.Our results showed that the electrospinning process had no deleterious effects on tissue viability, and that the mid-wall CWS vs. time profile could be dictated through the composition and degradation of the electrospun wrap. This may have important clinical applications by enabling the engineering of an improved AVG.

The host immune response to an implanted biomaterial, particularly the phenotype of infiltrating macrophages, is a key determinant of biocompatibility and downstream remodeling outcome. The present study used a subcutaneous rat model to compare the tissue response, including macrophage phenotype, remodeling potential, and calcification propensity of a biologic scaffold composed of glutaraldehyde‐fixed bovine pericardium (GF‐BP), the standard of care for heart valve replacement, with those of an electrospun polycarbonate‐based supramolecular polymer scaffold (ePC‐UPy), urinary bladder extracellular matrix (UBM‐ECM), and a polypropylene mesh (PP). The ePC‐UPy and UBM‐ECM materials induced infiltration of mononuclear cells throughout the thickness of the scaffold within 2 days and neovascularization at 14 days. GF‐BP and PP elicited a balance of pro‐inflammatory (M1‐like) and anti‐inflammatory (M2‐like) macrophages, while UBM‐ECM and ePC‐UPy supported a dominant M2‐like macrophage phenotype at all timepoints. Relative to GF‐BP, ePC‐UPy was markedly less susceptible to calcification for the 180 day duration of the study. UBM‐ECM induced an archetypical constructive remodeling response dominated by M2‐like macrophages and the PP caused a typical foreign body reaction dominated by M1‐like macrophages. The results of this study highlight the divergent macrophage and host remodeling response to biomaterials with distinct physical and chemical properties and suggest that the rat subcutaneous implantation model can be used to predict in vivo biocompatibility and regenerative potential for clinical application of cardiovascular biomaterials.

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Numerical algorithms for subspace system identification (N4SID) are a powerful tool for generating the state space (SS) representation of any system. The purpose of this work was to use N4SID to generate SS models of the flowrate and pressure generation within an ex vivo vascular perfusion system (EVPS). Accurate SS models were generated and converted to transfer functions (TFs) to be used for proportional integral and derivative (PID) controller design. By prescribing the pressure and flowrate inputs to the pumping components within the EVPS and measuring the resulting pressure and flowrate in the system,_four TFs were estimated;_two for a flowrate controller (H(RP,f) and H(RPP,f)) and two for a pressure controller (H(RP,p) and H(RPP,p)). In each controller,_one TF represents a roller pump (H(RP,f) and H(RP,p)),_and the other represents a roller pump and piston in series (H(RPP,f) and H(RPP,p)). Experiments to generate the four TFs were repeated five times (N=5) from which average TFs were calculated. The average model fits, computed as the percentage of the output variation (to_the_prescribed_inputs) reproduced by the model, were 94.93+/-1.05% for H(RP,p), 81.29+/-0.20% for H(RPP,p), 94.45+/-0.73% for H(RP,f), and 77.12+/-0.36% for H(RPP,f). The simulated step, impulse, and frequency responses indicate that the EVPS is a stable system and can respond to signals containing power of up to 70_Hz.

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