Hard Block Degradable Polycarbonate Urethanes: Promising Biomaterials for Electrospun Vascular Prostheses.

We report biodegradable thermoplastic polyurethanes for soft tissue engineering applications, where frequently used carboxylic acid ester degradation motifs were substituted with carbonate moieties to achieve superior degradation properties. While the use of carbonates in soft blocks has been reported, their use in hard blocks of thermoplastic polyurethanes is unprecedented. Soft blocks consist of poly(hexamethylene carbonate), and hard blocks combine hexamethylene diisocyanate with the newly synthesized cleavable carbonate chain extender bis(3-hydroxypropylene)carbonate (BHPC), mimicking the motif of poly(trimethylene carbonate) with highly regarded degradation properties. Simultaneously, the mechanical benefits of segmented polyurethanes are exploited. A lower hard block concentration in BHPC-based polymers was more suitable for vascular grafts. Nonacidic degradation products and hard block dependent degradation rates were found. Implantation of BHPC-based electrospun degradable vascular prostheses in a small animal model revealed high patency rates and no signs of aneurysm formations. Specific vascular graft remodeling and only minimal signs of inflammatory reactions were observed.

Long Term Evaluation of Nanofibrous, Bioabsorbable Polycarbonate Urethane Grafts for Small Diameter Vessel Replacement in Rodents.

OBJECTIVE: Biodegradable materials for in situ vascular tissue engineering could meet the increasing clinical demand for sufficient synthetic small diameter vascular substitutes in aortocoronary bypass and peripheral vascular surgery. The aim of this study was to design a new degradable thermoplastic polycarbonate urethane (dPCU) with improved biocompatibility and optimal biomechanical properties. Electrospun conduits made from dPCU were evaluated in short and long term follow up and compared with expanded polytetrafluoroethylene (ePTFE) controls. METHODS: Both conduits were investigated prior to implantation to assess their biocompatibility and inflammatory potential via real time polymerase chain reaction using a macrophage culture. dPCU grafts (n = 28) and ePTFE controls (n = 28) were then implanted into the infrarenal abdominal aorta of Sprague-Dawley rats. After seven days, one, six, and 12 months, grafts were analysed by histology and immunohistochemistry (IHC) and assessed biomechanically. RESULTS: Anti-inflammatory signalling was upregulated in dPCU conduits and increased significantly over time in vitro. dPCU and ePTFE grafts offered excellent long and short term patency rates (92.9% in both groups at 12 months) in the rat model without dilatation or aneurysm formation. In comparison to ePTFE, dPCU grafts showed transmural ingrowth of vascular specific cells resulting in a structured neovessel formation around the graft. The graft material was slowly reduced, while the compliance of the neovessel increased over time. CONCLUSION: The newly designed dPCU grafts have the potential to be safely applied for in situ vascular tissue engineering applications. The degradable substitutes showed good in vivo performance and revealed desirable characteristics such as biomechanical stability, non-thrombogenicity, and minimal inflammatory response after long term implantation.

Dynamic measurement of centering forces on transvalvular cannulas.

In heart failure therapy, minimally invasive devices (transcatheter valves, catheter-based cannulas or pumps) are increasingly used. The interaction with the valve is of special importance as valve damage, backflow, and thrombus formation are known complications. Therefore, the aim of this in vitro study was to characterize the forces acting on different sized transvalvular cannulas at various transvalvular pressures for four different valves. In a pulsatile setup radial and tangential forces on transvalvular cannulas were measured for bioprosthetic, artificial pericardial tissue, fresh, and fixated porcine valves. The cannula position was varied from a central position to the wall in 10° rotational steps for the whole circular range and the use of different cannula diameters (4, 6, and 8 mm) and transvalvular pressures (40-100 mmHg). Centering forces of four different aortic valve types were identified and the three leaflets were visible in the force distribution. At the mid of the cusps and at the largest deflection the forces were highest (up to 0.8 N) and lowest in the commissures (up to 0.2 N). Whereas a minor influence of the cannula diameter was found, the transvalvular pressure linearly increased the forces but did not alter the force patterns. Centering forces that act on transvalvular cannulas were identified in an in vitro setup for several valves and valve types. Lowest centering forces were found in the commissures and highest forces were found directly at the cusps. At low pressures, low centering forces and an increased cannula movement can be expected.

Activated Schwann Cell-Like Cells on Aligned Fibrin-Poly(Lactic-Co-Glycolic Acid) Structures: A Novel Construct for Application in Peripheral Nerve Regeneration.

Tissue engineering approaches in nerve regeneration search for ways to support gold standard therapy (autologous nerve grafts) and to improve results by bridging nerve defects with different kinds of conduits. In this study, we describe electrospinning of aligned fibrin-poly(lactic-co-glycolic acid) (PLGA) fibers in an attempt to create a biomimicking tissue-like material seeded with Schwann cell-like cells (SCLs) in vitro for potential use as an in vivo scaffold. Rat adipose-derived stem cells (rASCs) were differentiated into SCLs and evaluated with flow cytometry concerning their differentiation and activation status [S100b, P75, myelin-associated glycoprotein (MAG), and protein 0 (P0)]. After receiving the proliferation stimulus forskolin, SCLs expressed S100b and P75; comparable to native, activated Schwann cells, while cultured without forskolin, cells switched to a promyelinating phenotype and expressed S100b, MAG, and P0. Human fibrinogen and thrombin, blended with PLGA, were electrospun and the alignment and homogeneity of the fibers were proven by scanning electron microscopy. Electrospun scaffolds were seeded with SCLs and the formation of Büngner-like structures in SCLs was evaluated with phalloidin/propidium iodide staining. Carrier fibrin gels containing rASCs acted as a self-shaping matrix to form a tubular structure. In this study, we could show that rASCs can be differentiated into activated, proliferating SCLs and that these cells react to minimal changes in stimulus, switching to a promyelinating phenotype. Aligned electrospun fibrin-PLGA fibers promoted the formation of Büngner-like structures in SCLs, which also rolled the fibrin-PLGA matrix into a tubular scaffold. These in vitro findings favor further in vivo testing.

Mass spectrometric imaging of in vivo protein and lipid adsorption on biodegradable vascular replacement systems.

Cardiovascular diseases present amongst the highest mortality risks in Western civilization and are frequently caused by arteriosclerotic vessel failure. Coronary artery and peripheral vessel reconstruction necessitates the use of small diameter systems that are mechanically stress-resistant and biocompatible. Expanded polytetrafluorethylene (ePTFE) is amongst the materials used most frequently for non-degradable and bio-degradable vessel reconstruction procedures, with thermoplastic polyurethanes (TPU) representing a promising substitute. The present study describes and compares the biological adsorption and diffusion occurring with both materials following implantation in rat models. Gel electrophoresis and thin-layer chromatography, combined with mass spectrometry and mass spectrometry imaging, were utilized to identify the adsorbed lipids and proteins. The results were compared with the analytes present in native aorta tissue. It was revealed that both polymers were severely affected by biological adsorption after 10 min in vivo. Proteins associated with cell growth and migration were identified, especially on the luminal graft surface, while lipids were found to be located on both the luminal and abluminal surfaces. Lipid adsorption and cholesterol diffusion were found to be correlated with the polymer modifications identified on degradable thermoplastic urethane graft samples, with the latter revealing extensive cholesterol adsorption. The present study demonstrates an interaction between biological matter and both graft materials, and provides insights into polymer changes, in particular, those observed with thermoplastic urethanes already after 10 min in vivo exposure. ePTFE demonstrated minor polymer modifications, whereas several different polymer signals were observed for TPU, all were co-localized with biological signals.

Two-photon lithography for customized microstructured surfaces and their influence on wettability and bacterial load.

BACKGROUND: Device-related bacterial infections account for a large proportion of hospital-acquired infections. The ability of bacteria to form a biofilm as a protective shield usually makes treatment impossible without removal of the implant. Topographic surfaces have attracted considerable attention in studies seeking antibacterial properties without the need for additional antimicrobial substances. As there are still no valid rules for the design of antibacterial microstructured surfaces, a fast, reproducible production technique with good resolution is required to produce test surfaces and to examine their effectiveness with regard to their antibacterial properties. METHODS: In this work various surfaces, flat and with microcylinders in different dimensions (flat, 1, 3 and 9 µm) with a surface area of 7 × 7 mm were fabricated with a nanoprinter using two-photon lithography and evaluated for their antibiofilm effect. The microstructured surfaces were cultured for 24 h with different strains of Pseudomonas aeruginosa and Staphylococcus aureus to study bacterial attachment to the patterned surfaces. In addition, surface wettability was measured by a static contact angle measurement. RESULTS: Contact angles increased with cylinder size and thus hydrophobicity. Despite the difference in wettability, Staphylococcus aureus was not affected by the microstructures, while for Pseudomonas aeruginosa the bacterial load increased with the size of the cylinders, and compared to a flat surface, a reduction in bacteria was observed for one strain on the smallest cylinders. CONCLUSIONS: Two-photon lithography allowed rapid and flexible production of microcylinders of different sizes, which affected surface wettability and bacterial load, however, depending on bacterial type and strain.

Biodegradable, Self-Reinforcing Vascular Grafts for In Situ Tissue Engineering Approaches.

Clinically available small-diameter synthetic vascular grafts (SDVGs) have unsatisfactory patency rates due to impaired graft healing. Therefore, autologous implants are still the gold standard for small vessel replacement. Bioresorbable SDVGs may be an alternative, but many polymers have inadequate biomechanical properties that lead to graft failure. To overcome these limitations, a new biodegradable SDVG is developed to ensure safe use until adequate new tissue is formed. SDVGs are electrospun using a polymer blend composed of thermoplastic polyurethane (TPU) and a new self-reinforcing TP(U-urea) (TPUU). Biocompatibility is tested in vitro by cell seeding and hemocompatibility tests. In vivo performance is evaluated in rats over a period for up to six months. Autologous rat aortic implants serve as a control group. Scanning electron microscopy, micro-computed tomography (µCT), histology, and gene expression analyses are applied. TPU/TPUU grafts show significant improvement of biomechanical properties after water incubation and exhibit excellent cyto- and hemocompatibility. All grafts remain patent, and biomechanical properties are sufficient despite wall thinning. No inflammation, aneurysms, intimal hyperplasia, or thrombus formation are observed. Evaluation of graft healing shows similar gene expression profiles of TPU/TPUU and autologous conduits. These new biodegradable, self-reinforcing SDVGs may be promising candidates for clinical use in the future.

Electrospun small-diameter polyurethane vascular grafts: ingrowth and differentiation of vascular-specific host cells.

No small-diameter synthetic graft has yet shown comparable performance to autologous vessels. Synthetic conduits fail due to their inherent surface thrombogenicity and the development of intimal hyperplasia. In addressing these shortcomings, electrospinning offers an interesting alternative to other nanostructured, cardiovascular substitutes because of the close match of electrospun materials to the biomechanical and structural properties of native vessels. In this study, we investigated the in vivo behavior of electrospun, small-diameter conduits in a rat model. Vascular grafts composed of polyurethane were fabricated by electrospinning. Prostheses were implanted into the abdominal aorta in 40 rats for either 7 days, 4 weeks, 3 months, or 6 months. Retrieved specimens were evaluated by histology, immunohistochemical staining, confocal laser scanning microscopy, and scanning electron microscopy. At all time points, we found no evidence of foreign body reaction or graft degradation. The overall patency rate of the intravascular implants was 95%. Within 7 days, grafts revealed ingrowth of host cells. CD34+ cells increased significantly from 7 days up to 6 months of implantation (P < 0.05). Myofibroblasts and myocytes showed increasing cell numbers up to 3 months (P < 0.05). Ki67 staining indicated unaltered cell proliferation during the whole follow-up period. Besides biomechanical benefits, electrospun polyurethane grafts exhibit excellent biocompatibility in vivo. Cell immigration and differentiation seems to be promoted by the nanostructured artificial matrix.

Electrospinning of aligned fibers with adjustable orientation using auxiliary electrodes.

A conventional electrospinning setup was upgraded by two turnable plate-like auxiliary high-voltage electrodes that allowed aligned fiber deposition in adjustable directions. Fiber morphology was analyzed by scanning electron microscopy and attenuated total reflection Fourier transform infrared spectroscopy (FTIR-ATR). The auxiliary electric field constrained the jet bending instability and the fiber deposition became controllable. At target speeds of 0.9 m s-1 90% of the fibers had aligned within 2°, whereas the angular spread was 70° without the use of auxiliary electrodes. It was even possible to orient fibers perpendicular to the rotational direction of the target. The fiber diameter became smaller and its distribution narrower, while according to the FTIR-ATR measurement the molecular orientation of the polymer was unaltered. This study comprehensively documents the feasibility of directed fiber deposition and offers an easy upgrade to existing electrospinning setups.

Electrospinning of small diameter vascular grafts with preferential fiber directions and comparison of their mechanical behavior with native rat aortas.

Conventional electrospun small diameter vascular grafts have a random fiber orientation. In order to achieve mechanical characteristics similar to a native blood vessel, a controllable fiber orientation is of interest. In this study the electrospinning jet was directly controlled by means of an auxiliary, changeable electrostatic field, so that the fibers could be deposited in adjustable orientations. Prostheses with circumferentially, axially, fenestrated and randomly aligned fibers were electrospun on Ø2mm mandrels out of a thermoplastic polyurethane (PUR) and a polylactid acid (PLLA). The impact of the materials and the various preferential fiber orientations on the resulting biomechanics was investigated and compared with that of the native rat aorta in quasistatic and dynamic hoop tensile tests. The test protocol included 3000 dynamic loading cycles in the physiological blood pressure range and ended with a quasistatic tensile test. Any orientation of the fibers in a particular direction resulted in a significant reduction in scaffold porosity for both materials. The standard randomly oriented PUR grafts showed the highest compliance of 29.7 ± 5.5 [%/100 mmHg] and were thus closest to the compliance of the rat aortas, which was 37.2 ± 6.5 [%/100 mmHg]. The maximum tensile force was increased at least 6 times compared to randomly spun grafts by orienting the fibers in the circumferential direction. During the 3000 loading cycles, creeping of the native rat aorta was below 1% whereas the electrospun grafts showed creeping up to 2.4 ± 1.2%. Although the preferred fiber orientations were only partially visible in the scanning electron micrographs, the mechanical effects were evident. The investigations suggest a multi-layer wall structure of the vascular prosthesis, since none of the preferred fiber directions and the materials used could imitate the typical j-shaped mechanical characteristics of the rat aorta.

Graphene Oxide Coating Improves the Mechanical and Biological Properties of Decellularized Umbilical Cord Arteries.

The lack of small-diameter vascular grafts (inner diameter <5 mm) to substitute autologous grafts in arterial bypass surgeries has a massive impact on the prognosis and progression of cardiovascular diseases, the leading cause of death globally. Decellularized arteries from different sources have been proposed as an alternative, but their poor mechanical performance and high collagen exposure, which promotes platelet and bacteria adhesion, limit their successful application. In this study, these limitations were surpassed for decellularized umbilical cord arteries through the coating of their lumen with graphene oxide (GO). Placental and umbilical cord arteries were decellularized and perfused with a suspension of GO (C/O ratio 2:1) with ∼1.5 µm lateral size. A homogeneous GO coating that completely covered the collagen fibers was obtained for both arteries, with improvement of mechanical properties being achieved for umbilical cord decellularized arteries. GO coating increased the maximum force in 27%, the burst pressure in 29%, the strain in 25%, and the compliance in 10%, compared to umbilical cord decellularized arteries. The achieved theoretical burst pressure (1960 mmHg) and compliance (13.9%/100 mmHg) are similar to the human saphenous vein and mammary artery, respectively, which are used nowadays as the gold standard in coronary and peripheral artery bypass surgeries. Furthermore, and very importantly, coatings with GO did not compromise the endothelial cell adhesion but decreased platelet and bacteria adhesion to decellularized arteries, which will impact on the prevention of thrombosis and infection, until full re-endothetialization is achieved. Overall, our results reveal that GO coating has an effective role in the improvement of decellularized umbilical cord artery performance, which is a huge step toward their application as a small-diameter vascular graft.

S-nitroso human serum albumin as a nitric oxide donor in drug-eluting vascular grafts: Biofunctionality and preclinical evaluation.

Currently available synthetic small diameter vascular grafts reveal low patency rates due to thrombosis and intimal hyperplasia. Biofunctionalized grafts releasing nitric oxide (NO) in situ may overcome these limitations. In this study, a drug-eluting vascular graft was designed by blending polycaprolactone (PCL) with S-nitroso-human-serum-albumin (S-NO-HSA), a nitric oxide donor with prolonged half-life. PCL-S-NO-HSA grafts and patches were fabricated via electrospinning. The fabrication process was optimized. Patches were characterized in vitro for their morphology, drug release, biomechanics, inflammatory effects, cell proliferation, and expression of adhesion molecules. The selected optimized formulation (8%PCL-S-NO-HSA) had superior mechanical/morphological properties with high protein content revealing extended NO release (for 28 days). 8%PCL-S-NO-HSA patches significantly promoted endothelial cell proliferation while limiting smooth muscle cell proliferation. Expression of adhesion molecules (ICAM-1, VCAM-1) and pro-inflammatory macrophage/cytokine markers (CD80, IL-1α, TNF-α) was significantly reduced. 8%PCL-S-NO-HSA patches had superior immunomodulatory properties by up-regulating anti-inflammatory cytokines (IL-10) and M2 macrophage marker (CD163) at final time points. Grafts were further evaluated in a small rodent model as aortic implants up to 12 weeks. Grafts were assessed by magnetic resonance imaging angiography (MRI) in vivo and after retrieval by histology. All grafts remained 100 % patent with no signs of thrombosis or calcification. 8%PCL-S-NO-HSA vascular grafts supported rapid endothelialization, whereas smooth muscle cell proliferation was hampered in earlier phases. This study indicates that 8%PCL-S-NO-HSA grafts effectively support long-term in situ release of bioactive NO. The beneficial effects observed can be promising features for long-term success of small diameter vascular grafts. STATEMENT OF SIGNIFICANCE: Despite extensive research in the field of small diameter vascular graft replacement, there is still no appropriate substitute to autografts yet. Various limitations are associated with currently available synthetic vascular grafts such as thrombogenicity and intimal hyperplasia. Therefore, developing new generations of such conduits has become a major focus of research. One of the most significant signaling molecules that are involved in homeostasis of the vascular system is nitric oxide. The new designed nitric-oxide eluting vascular grafts described in this study induce rapid surface endothelialization and late migration of SMCs into the graft wall. These beneficial effects have potential to improve current limitations of small diameter vascular grafts.

Impact of the testing protocol on the mechanical characterization of small diameter electrospun vascular grafts.

AIM: For the proper function of small diameter vascular grafts their mechanical properties are essential. A variety of testing methods and protocols exists to measure tensile strength, compliance and viscoelastic material behavior. In this study the impact of the measurement protocol in hoop tensile tests on the measured compliance and tensile strength was investigated. METHODS: Vascular grafts made out of two different materials, a thermoplastic polyurethane (PUR) and polylactid acid (PLLA), with three different wall thicknesses were produced by electrospinning. Samples were tested with a measurement protocol that allowed the comparison of dynamic sample loading to a common quasistatic tensile test. Influence of measurement temperature, preconditioning cycles and the influence of a high number of loading cycles was also investigated. Compliance and tensile strength were evaluated and compared between the different samples and the different load cases. RESULTS: In all samples a significant difference in the measured compliance was seen between an unloaded sample and a sample that was already in a preloaded state. For example in the PUR group with 100 µm wall thickness at 37 °C, the first compliance was 32.6 ± 9.6%/100 mmHg, which reduced to 15.4 ± 2.9%/100 mmHg at preloaded state. The PLLA group showed 7.5 ± 4.3%/100 mmHg vs. 0.94 ± 0.11%/100 mmHg respectively. The measurements showed the importance of dynamic testing, as the samples viscoelastic behavior had a considerable influence on the measured compliance. The quasistatic ultimate tensile test alone was not able to predict the sample's in vivo compliance. The measurement temperature had a significant influence on tensile strength and compliance. Both, the number of preconditioning cycles and the high number of loading cycles had a minor influence on the sample's compliance. CONCLUSION: With a quasistatic tensile tests alone, overestimated compliance values are measured in viscoelastic electrospun vascular samples, therefore dynamic loading cycles are required. Measurements at 37 °C are mandatory, as temperature has a significant influence on the mechanical properties.

Riboflavin-mediated photooxidation to improve the characteristics of decellularized human arterial small diameter vascular grafts.

Vascular grafts with a diameter of less than 6 mm are made from a variety of materials and techniques to provide alternatives to autologous vascular grafts. Decellularized materials have been proposed as a possible approach to create extracellular matrix (ECM) vascular prostheses as they are naturally derived and inherently support various cell functions. However, these desirable graft characteristics may be limited by alterations of the ECM during the decellularization process leading to decreased biomechanical properties and hemocompatibility. In this study, arteries from the human placenta chorion were decellularized using two distinct detergents (Triton X-100 or SDS), which differently affect ECM ultrastructure. To overcome biomechanical strength loss and collagen fiber exposure after decellularization, riboflavin-mediated UV (RUV) crosslinking was used to uniformly crosslink the collagenous ECM of the grafts. Graft characteristics and biocompatibility with and without RUV crosslinking were studied in vitro and in vivo. RUV-crosslinked ECM grafts showed significantly improved mechanical strength and smoothening of the luminal graft surfaces. Cell seeding using human endothelial cells revealed no cytotoxic effects of the RUV treatment. Short-term aortic implants in rats showed cell migration and differentiation of host cells. Functional graft remodeling was evident in all grafts. Thus, RUV crosslinking is a preferable tool to improve graft characteristics of decellularized matrix conduits.

Assessment of a long-term in vitro model to characterize the mechanical behavior and macrophage-mediated degradation of a novel, degradable, electrospun poly-urethane vascular graft.

An assessment tool to evaluate the degradation of biodegradable materials in a more physiological environment is still needed. Macrophages are critical players in host response, remodeling and degradation. In this study, a cell culture model using monocyte-derived primary macrophages was established to study the degradation, macro-/micro-mechanical behavior and inflammatory behavior of a new designed, biodegradable thermoplastic polyurethane (TPU) scaffold, over an extended period of time in vitro. For in vivo study, the scaffolds were implanted subcutaneously in a rat model for up to 36 weeks. TPU scaffolds were fabricated via the electrospinning method. This technique provided a fibrous scaffold with an average fiber diameter of 1.39 ± 0.76 µm and an average pore size of 7.5 ± 1.1 µm. The results showed that TPU scaffolds supported the attachment and migration of macrophages throughout the three-dimensional matrix. Scaffold degradation could be detected in localized areas, emphasizing the role of adherent macrophages in scaffold degradation. Weight loss, molecular weight and biomechanical strength reduction were evident in the presence of the primary macrophage cells. TPU favored the switch from initial pro-inflammatory response of macrophages to an anti-inflammatory response over time both in vitro and in vivo. Expression of MMP-2 and MMP-9 (the key enzymes in tissue remodeling based on ECM modifications) was also evident in vitro and in vivo. This study showed that the primary monocyte-derived cell culture model represents a promising tool to characterize the degradation, mechanical behavior as well as biocompatibility of the scaffolds during an extended period of observation.

A passive magnetically and hydrodynamically suspended rotary blood pump.

A combined hydrodynamic-magnetic bearing allows the design of rotary blood pumps that are not encumbered with mechanical bearings and magnets requiring sensors or electrical power. However, such pumps have so far needed very small and accurately manufactured gaps between rotor and housing to assure effective hydromagnetic bearing behavior. In order to use this concept in disposable pump heads, a design that allows larger rotor-housing gaps, and thus larger manufacturing tolerances, is needed. A pump with passive magnetic bearings and a gap between rotor and housing in the range of 0.5 mm was designed. Numerical simulations were performed to optimize the rotor geometry at low levels of shear stress. An experimental test stand was used to find a range of speeds and gap settings that resulted in low levels of vibration and useful pressure-flow relationships. Three different rotor geometries were tested using a viscosity-adjusted test fluid. Blood damage tests were conducted within the desirable range of speeds and gap settings. In this study stable pump performance was demonstrated at total gap widths between 0.3 and 0.7 mm at flows of 0-10 L/min, with afterloads up to 230 mm Hg. Best performance was achieved with rotors sliding on a fluid pillow between the rotor and the outer housing at a gap distance of 50 to 250 microm. The inner gap distance, between the rotor and the inner housing, could be as great as 500 microm. Hemolysis tests on the prototype within the chosen operating range showed lower values (NIH = 0.0029 +/- 0.0012 g/100 L) than the Biomedicus BP-80 pump (NIH = 0.0033 +/- 0.0011 g/100 L). In conclusion, it is possible to build rotary blood pumps with passive hydromagnetic bearings that have large gaps between their rotors and housings. Rotor behavior is sensitive to the position of the permanent magnetic drive unit. To minimize vibration and blood damage, the fluid gaps and the rotational speed have to be adjusted according to the desired operating point of the pump. Further study is needed to optimize the magnetic drive unit and to ascertain its ability to withstand inertial loads imposed by sudden movements and external shock.

Biomechanical properties of decellularized porcine pulmonary valve conduits.

Tissue-engineered heart valves constructed from a xenogeneic or allogeneic decellularized matrix might overcome the disadvantages of current heart valve substitutes. One major necessity besides effective decellularization is to preserve the biomechanical properties of the valve. Native and decellularized porcine pulmonary heart valve conduits (PPVCs) (with [n = 10] or without [n = 10] cryopreservation) were compared to cryopreserved human pulmonary valve conduits (n = 7). Samples of the conduit were measured for wall thickness and underwent tensile tests. Elongation measurement was performed with a video extensometer. Decellularized PPVC showed a higher failure force both in longitudinal (+73%; P < 0.01) and transverse (+66%; P < 0.001) direction compared to human homografts. Failure force of the tissue after cryopreservation was still higher in the porcine group (longitudinal: +106%, P < 0.01; transverse: +58%, P < 0.001). In comparison to human homografts, both decellularized and decellularized cryopreserved porcine conduits showed a higher extensibility in longitudinal (decellularized: +61%, P < 0.001; decellularized + cryopreserved: +51%, P < 0.01) and transverse (decellularized: +126%, P < 0.001; decellularized + cryopreserved: +118%, P < 0.001) direction. Again, cryopreservation did not influence the biomechanical properties of the decellularized porcine matrix.

Acellular vascular matrix grafts from human placenta chorion: Impact of ECM preservation on graft characteristics, protein composition and in vivo performance.

Small diameter vascular grafts from human placenta, decellularized with either Triton X-100 (Triton) or SDS and crosslinked with heparin were constructed and characterized. Graft biochemical properties, residual DNA, and protein composition were evaluated to compare the effect of the two detergents on graft matrix composition and structural alterations. Biocompatibility was tested in vitro by culturing the grafts with primary human macrophages and in vivo by subcutaneous implantation of graft conduits (n = 7 per group) into the flanks of nude rats. Subsequently, graft performance was evaluated using an aortic implantation model in Sprague Dawley rats (one month, n = 14). In situ graft imaging was performed using MRI angiography. Retrieved specimens were analyzed by electromyography, scanning electron microscopy, histology and immunohistochemistry to evaluate cell migration and the degree of functional tissue remodeling. Both decellularization methods resulted in grafts of excellent biocompatibility in vitro and in vivo, with low immunogenic potential. Proteomic data revealed removal of cytoplasmic proteins with relative enrichment of ECM proteins in decelluarized specimens of both groups. Noteworthy, LC-Mass Spectrometry analysis revealed that 16 proteins were exclusively preserved in Triton decellularized specimens in comparison to SDS-treated specimens. Aortic grafts showed high patency rates, no signs of thrombus formation, aneurysms or rupture. Conduits of both groups revealed tissue-specific cell migration indicative of functional remodeling. This study strongly suggests that decellularized allogenic grafts from the human placenta have the potential to be used as vascular replacement materials. Both detergents produced grafts with low residual immunogenicity and appropriate mechanical properties. Observed differences in graft characteristics due to preservation method had no impact on successful in vivo performance in the rodent model.

Biocompatibility Assessment of a New Biodegradable Vascular Graft via In Vitro Co-culture Approaches and In Vivo Model.

Following the implantation of biodegradable vascular grafts, macrophages and fibroblasts are the major two cell types recruited to the host-biomaterial interface. In-vitro biocompatibility assessment usually involves one cell type, predominantly macrophages. In this study, macrophage and fibroblast mono- and co-cultures, in paracrine and juxtacrine settings, were used to evaluate a new biodegradable thermoplastic polyurethane (TPU) vascular graft. Expanded-polytetrafluoroethylene (ePTFE) grafts served as controls. Pro/anti-inflammatory gene expression of macrophages and cytokines was assessed in vitro and compared to those of an in vivo rat model. Host cell infiltration and the type of proliferated cells was further studied in vivo. TPU grafts revealed superior support in cell attachment, infiltration and proliferation compared with ePTFE grafts. Expression of pro-inflammatory TNF-α/IL-1α cytokines was significantly higher in ePTFE, whereas the level of IL-10 was higher in TPU. Initial high expression of pro-inflammatory CCR7 macrophages was noted in TPU, however there was a clear transition from CCR7 to anti-inflammatory CD163 expression in vitro and in vivo only in TPU, confirming superior cell-biomaterial response. The co-culture models, especially the paracrine model, revealed higher fidelity to the immunomodulatory/biocompatibility behavior of degradable TPU grafts in vivo. This study established an exciting approach developing a co-culture model as a tool for biocompatibility evaluation of degradable biomaterials.

Biodegradable, thermoplastic polyurethane grafts for small diameter vascular replacements.

Biodegradable vascular grafts with sufficient in vivo performance would be more advantageous than permanent non-degradable prostheses. These constructs would be continuously replaced by host tissue, leading to an endogenous functional implant which would adapt to the need of the patient and exhibit only limited risk of microbiological graft contamination. Adequate biomechanical strength and a wall structure which promotes rapid host remodeling are prerequisites for biodegradable approaches. Current approaches often reveal limited tensile strength and therefore require thicker or reinforced graft walls. In this study we investigated the in vitro and in vivo biocompatibility of thin host-vessel-matched grafts (n=34) formed from hard-block biodegradable thermoplastic polyurethane (TPU). Expanded polytetrafluoroethylene (ePTFE) conduits (n=34) served as control grafts. Grafts were analyzed by various techniques after retrieval at different time points (1 week; 1, 6, 12 months). TPU grafts showed significantly increased endothelial cell proliferation in vitro (P<0.001). Population by host cells increased significantly in the TPU conduits within 1 month of implantation (P=0.01). After long-term implantation, TPU implants showed 100% patency (ePTFE: 93%) with no signs of aneurysmal dilatation. Substantial remodeling of the degradable grafts was observed but varied between subjects. Intimal hyperplasia was limited to ePTFE conduits (29%). Thin-walled TPU grafts offer a new and desirable form of biodegradable vascular implant. Degradable grafts showed equivalent long-term performance characteristics compared to the clinically used, non-degradable material with improvements in intimal hyperplasia and ingrowth of host cells.