Development of In Vitro Endothelialised Stents - Review.

Endovascular treatment is prevalent as a primary treatment for coronary and peripheral arterial diseases. Although the introduction of drug-eluting stents (DES) dramatically reduced the risk of in-stent restenosis, stent thrombosis persists as an issue. Notwithstanding improvements in newer generation DES, they are yet to address the urgent clinical need to abolish the late stent complications that result from in-stent restenosis and are associated with late thrombus formation. These often lead to acute coronary syndromes with high mortality in coronary artery disease and acute limb ischemia with a high risk of limb amputation in peripheral arterial disease. Recently, a significant amount of research has focused on alternative solutions to improve stent biocompatibility by using tissue engineering. There are two types of tissue engineering endothelialisation methods: in vitro and in vivo. To date, commercially available in vivo endothelialised stents have failed to demonstrate antithrombotic or anti-stenosis efficacy in clinical trials. In contrast, the in vitro endothelialisation methods exhibit the advantage of monitoring cell type and growth prior to implantation, enabling better quality control. The present review discusses tissue-engineered candidate stents constructed by distinct in vitro endothelialisation approaches, with a particular focus on fabrication processes, including cell source selection, stent material composition, stent surface modifications, efficacy and safety evidence from in vitro and in vivo studies, and future directions.

In Vitro Calcification of Bioprosthetic Heart Valves: Test Fluid Validation on Prosthetic Material Samples.

Calcification is a major failure mode of bioprosthetic heart valves. So far, cost and time saving in vitro analyses of calcification potentials are unreliable, mostly due to superficial or spontaneous precipitation of the applied fluids. In this study, we developed a near-physiological non-spontaneously precipitating fluid for an accelerated in vitro calcification assessment, and validated it by analyzing the calcification potential of two prosthetic materials within two reference-tests. The first test focused on the comparison of four calcification fluids under dynamic contact with n=12 commercial bovine pericardium patches. The second one focused on the validation of the most appropriate fluid by analyzing the calcification potential of pericardium vs. polyurethane. The patches were mounted in separate test compartments and treated simultaneously with the respective fluids at an accelerated test frequency. Calcification propensity and progression were detected macroscopically and microscopically. Structural analyses of all deposits indicated hydroxyapatite by X-ray powder diffraction, which is also most commonly observed in vivo. Histological examination by von Kossa staining showed matrix internal and superficial calcifications, depending on the fluid composition. The present study reveals promising results towards the development of a meaningful, cost and time saving in vitro analysis of the calcification potential of bioprosthetic heart valves.

Long-Term Venovenous Connection for Extracorporeal Carbon Dioxide Removal (ECCO2R)-Numerical Investigation of the Connection to the Common Iliac Veins.

PURPOSE: Currently used cannulae for extracorporeal carbon dioxide removal (ECCO2R) are associated with complications such as thrombosis and distal limb ischemia, especially for long-term use. We hypothesize that the risk of these complications is reducible by attaching hemodynamically optimized grafts to the patient's vessels. In this study, as a first step towards a long-term stable ECCO2R connection, we investigated the feasibility of a venovenous connection to the common iliac veins. To ensure its applicability, the drainage of reinfused blood (recirculation) and high wall shear stress (WSS) must be avoided. METHODS: A reference model was selected for computational fluid dynamics, on the basis of the analysis of imaging data. Initially, a sensitivity analysis regarding recirculation was conducted using as variables: blood flow, the distance of drainage and return to the iliocaval junction, as well as the diameter and position of the grafts. Subsequently, the connection was optimized regarding recirculation and the WSS was evaluated. We validated the simulations in a silicone model traversed by dyed fluid. RESULTS: The simulations were in good agreement with the validation measurements (mean deviation 1.64%). The recirculation ranged from 32.1 to 0%. The maximum WSS did not exceed 5.57 Pa. The position and diameter of the return graft show the highest influence on recirculation. A correlation was ascertained between recirculation and WSS. Overall, an inflow jet directed at a vessel wall entails not only high WSS, but also a flow separation and thereby an increased recirculation. Therefore, return grafts aligned to the vena cava are crucial. CONCLUSION: In conclusion, a connection without recirculation could be feasible and therefore provides a promising option for a long-term ECCO2R connection.

In Vitro Calcification of Bioprosthetic Heart Valves: Investigation of Test Fluids.

Calcification is a major reason for the failure of bioprosthetic heart valves. Therefore, several attempts towards an accelerated in vitro model were undertaken in order to provide a cost- and time-saving method for the analysis of calcification processes. Due to the problem of superficial or spontaneous precipitation, which occurred in the fluids applied, we focused our study on the development of a near-physiological calcification fluid. The desired fluid should not precipitate spontaneously and should neither promote nor inhibit calcification. Eleven different fluid compositions were tested without contact to potentially calcifying materials. Crucial factors regarding the fluid properties were the ionic product, the ionic strength, and the degree of supersaturation concerning dicalciumphosphate-dihydrate, octacalciumphosphate, and hydroxyapatite. The fluids were kept in polyethylene bottles and exposed to a slight vibration within a durability tester at 37 °C. The precipitation propensity was monitored optically and colorimetrically. A structural analysis of the deposits was carried out by x-ray powder diffraction and IR-spectroscopy, which showed the development of the crystal phases that are relevant in vivo. Only two of the fluids did not precipitate. Resulting from the computations of the effective fluid contents, the saturation degree concerning dicalciumphosphate-dihydrate seems to be the key factor for spontaneous precipitation.

[Current CO2 angiography]. / Aktuelle CO2-Angiographie.

BACKGROUND: Carbon dioxide (CO2) is a very good and validated alternative to iodine-containing contrast media in diagnostic and interventional angiography. Nevertheless, its routinely use is still limited to specialized centers. OBJECTIVE: The presentation of the current role and limitations of CO2 in diagnostic and interventional angiography (venous and arterial). MATERIAL AND METHODS: A comprehensive review of the literature regarding CO2 angiography (physical features, indications, contraindications, applications) was carried out. RESULTS: The results show that CO2 can be used as a reliable alternative for diagnostic angiography and to support interventions in many arterial and venous vascular territories. An exception is the intra-arterial use of CO2 above the diaphragm due to the risk of severe myocardial and cerebral complications, which represents the most important contraindication. Moreover, due to its very low viscosity, CO2 is more effective than iodine-containing contrast media in detecting bleeding arising from small vessels. CONCLUSION: The CO2 angiography is a safe and effective technique and can be used as valid alternative. In several clinical cases it even offers some advantages compared to iodine-containing contrast media.

Reliability of left atrial pressure estimation from left ventricular filling measurement in a total artificial heart.

A physiological control of a total artificial heart (TAH) requires reliable information on left arterial pressure (LAP). When LAP is derived indirectly from intrinsic TAH parameters like end diastolic volume (EDV) and diastole duration (Td), the transfer function and associated uncertainties need to be well understood.We derived a computational equivalent to a hydraulic model consisting of the venous compliance, the heart valve and the pump chamber, and studied the filling phase in cases of different venous compliance. We calculated a family of curves of pump chamber volume as a function of time for different venous compliances and LAP. To visualize the LAP transfer function and uncertainties associated to EDV, Td measurement error and unknown venous compliance a family of similar curves in the vicinity of assumed measurement was found and visualised in the parameter space.Results were in a realistic absolute range and showed expected trends despite some simplifications in the simulation model. The venous compliance has no significant influence on LAP values extracted from EDV and Td, except at very low values. The uncertainty in the extracted LAP is particularly high for high EDV and short Td.A physiological regulation therefore does not have to be individually adapted to the patient's venous compliances, but has to deal with uncertainties in the input values like blood pressures extracted from intrinsic device parameters.

Efficacy of magnetic thermoablation using SPIO in the treatment of osteoid osteoma in a bovine model compared to radiofrequency and microwave ablation.

PURPOSE: To evaluate heating efficacy of superparamagnetic iron oxide nanoparticles (SPIO) for electromagnetic ablation (EMA) of osteoid osteoma (OO) using an ex vivo model compared to radiofrequency ablation (RFA) and microwave ablation (MWA). METHODS: A model for OO using sliced bovine tibia and sliced muscle tissue was developed. A bone cavity filled with either a mixture of SPIO and agarose or pure agarose (control group) was established. EMA was performed using an experimental system, RFA and MWA using clinically approved systems, and the ablation protocols recommended by the vendor. For temperature measurements, fiberoptic temperature probes were inserted inside the cavity, on the outside of the periosteum, and at a 5 mm distance to the periosteum. RESULTS: Maximum temperatures with or without SPIO in the nidus were as follows: EMA: 79.9 ± 2.5/22.3 ± 0.7 °C; RFA: 95.1 ± 1.8/98.6 ± 0.9 °C; MWA: 85.1 ± 10.8/83.4 ± 9.62 °C. In RFA with or without SPIO significantly higher temperatures were achieved in the nidus compared to all other groups (p < 0.05). In MWA significantly higher temperatures were observed in the 5 mm distance to the periosteum compared to EMA and RFA with or without SPIO (p < 0.05). In MWA temperature decrease between nidus and the 5 mm distance to the periosteum was significantly lower than in RFA with or without SPIO (p < 0.0001). In MWA without SPIO temperature decrease was significantly lower than in the EMA group (p < 0.05). CONCLUSION: In the experimental setting, ablation of OO is safe and effective using EMA. It is less invasive than RFA and MWA, and it theoretically allows repeated treatments without repeated punctures. In comparison, the highest temperatures in the nidus are reached using RFA.

Effects of post-processing treatments on sensory quality and Shiga toxigenic Escherichia coli reductions in dry-fermented sausages.

The effects of post-processing treatments on sensory quality and reduction of Shiga toxigenic Escherichia coli (STEC) in three formulations of two types of dry-fermented sausage (DFS; salami and morr) were evaluated. Tested interventions provided only marginal changes in sensory preference and characteristics. Total STEC reductions in heat treated DFS (32°C, 6days or 43°C, 24h) were from 3.5 to >5.5 log from production start. Storing of sausages (20°C, 1month) gave >1 log additional STEC reduction. Freezing and thawing of sausages in combination with storage (4°C, 1month) gave an additional 0.7 to 3.0 log reduction in STEC. Overall >5.5 log STEC reductions were obtained after storage and freezing/thawing of DFS with increased levels of glucose and salt. This study suggests that combined formulation optimisation and post-process strategies should be applicable for implementation in DFS production to obtain DFS with enhanced microbial safety and high sensory acceptance and quality.

Non-destructive analysis of extracellular matrix development in cardiovascular tissue-engineered constructs.

In the field of tissue engineering, there is an increasing demand for non-destructive methods to quantify the synthesis of extracellular matrix (ECM) components such as collagens, elastin or sulphated glycosaminoglycans (sGAGs) in vitro as a quality control before clinical use. In this study, procollagen I carboxyterminal peptide (PICP), procollagen III aminoterminal peptide (PIIINP), tropoelastin and sGAGs are investigated for their potential use as non-destructive markers in culture medium of statically cultivated cell-seeded fibrin gels. Measurement of PICP as marker for type I collagen synthesis, and PIIINP as marker of type III collagen turnover, correlated well with the hydroxyproline content of the fibrin gels, with a Pearson correlation coefficient of 0.98 and 0.97, respectively. The measurement of tropoelastin as marker of elastin synthesis correlated with the amount of elastin retained in fibrin gels with a Pearson correlation coefficient of 0.99. sGAGs were retained in fibrin gels, but were not detectable in culture medium at any time of measurement. In conclusion, this study demonstrates the potential of PICP and tropoelastin as non-destructive culture medium markers for collagen and elastin synthesis. To our knowledge, this is the first study in cardiovascular tissue engineering investigating the whole of here proposed biomarkers of ECM synthesis to monitor the maturation process of developing tissue non-invasively, but for comprehensive assessment of ECM development, these biomarkers need to be investigated in further studies, employing dynamic cultivation conditions and more complex tissue constructs.

Carbon dioxide-contrasted computed tomography angiography: high pitch protocols and adapted injection parameters improve imaging quality.

PURPOSE: To systematically investigate the impact of image acquisition and contrast injection parameters for CO2-enhanced CT angiography (CTA) of the aorto-iliac and peripheral arteries in a pig model using commercially available equipment. The aim was to establish an imaging protocol that is ready for use in human subjects. MATERIALS AND METHODS: Three domestic swine underwent CO2-CTA with varying injection parameters: pitch (1.0, 3.0), injection pressure (0.7 bar, 1.0 bar, 1.3 bar) and scan delay (2 s, 4 s, 6 s). Objective (vessel diameter) and subjective (image quality) parameters and applied radiation doses were systematically evaluated. To ensure clinical applicability of the setting, only approved catheters/injectors and standard injection parameters were evaluated. RESULTS: The image quality scores were superior and the vessel diameter was larger with high pitch in comparison to standard pitch (diameters: 4.7 ± 2.0 mm vs. 3.6 ± 2.1 mm, p = 0.0040, scores: 2.6 ± 1.1 vs. 2.0 ± 1.1, p = 0.0038). High injection pressure (1.3 bar) improved the image quality as assessed by subjective and objective ratings (diameters: 3.6 ± 2.0 mm, 4.0 ± 2.1 mm and 4.6 ± 2.1 mm, for 0.7, 1.0 and 1.3 bar, p-values ≤ 0.0052, scores: 1.9 ± 1.1, 2.3 ± 1.1 and 2.7 ± 1.2, p-values ≤ 0.0017), the same was observed for a shorter injection delay (diameters: 3.5 ± 2.0 mm, 4.2 ± 2.1 mm and 4.8 ± 2.1 mm, for 6 s, 4 s, and 2 s, p ≤ 0.0022, scores: 1.9 ± 1.1, 2.3 ± 1.1 and 2.7 ± 1.1, p-values ≤ 0.0013). The dose length products were 239 ± 47 mGycm (high pitch) and 565 ± 63 mGycm (standard pitch, p-values < 0.0001). CONCLUSION: A higher pitch, shorter delay and higher injection pressure improve image quality in CO2-enhanced CTA. Since commercially available, clinically approved equipment was used. The protocol is now ready for use in human subjects.

Silicon carbide-enhanced microwave ablation in an ex-vivo bovine liver model - effects on heat distribution and ablation volume.

PURPOSE: Evaluation of the maximum temperatures and ablation volumes in microwave ablation (MWA) after injection of different concentrations of silicon carbide (SiC) particles in an ex-vivo bovine liver model. MATERIALS AND METHODS: 15 ml of different concentrations of SiC particles (20 vol% SiC; 50 vol% SiC) mixed with 2 % gelatin were injected into an ex-vivo bovine liver. As a reference group, 2 % gelatin without SiC was injected. MWA was performed using a clinical MWA system with different generator settings (10 - 45 W/10 minutes). The temperature was measured at a distance of 5 mm and 10 mm from the applicator. Afterwards the liver tissue was sliced along the short and long axis, the ablation zones were measured on the x, y and z-axis and the ablation volume was calculated. All experiments were performed 5 times (total: 40 experiments). RESULTS: The average maximum temperatures measured at a generator setting of 45 W at a distance of 5 mm from the applicator were 103.4 ± 4.6 °C (20 vol% SiC), 103.3 ± 6.5 °C (50 vol% SiC) and 96.0 ± 4.2 °C in the control group (0 vol% SiC). At 45 W, injection of 20 vol% SIC caused a significantly higher maximum temperature than that achieved in the control group (p = 0.016). No significant temperature increase compared to the control group could be measured using 50 vol% SiC. The mean ablation volumes at 45 W and 20 vol% SiC and 50 vol% SiC were significantly larger (172.7 ± 31.5 ml and 171.0 ± 34.7 ml, respectively) than those achieved in the control group (111.2 ± 23.8 ml) (p = 0.027 and p = 0.045). CONCLUSION: In an ex-vivo bovine liver model, the SiC particles demonstrated an enhancing effect of MWA with respect to maximum temperatures and ablation volume. Therefore, SiC is a promising candidate for enhancing MWA in vivo.

Investigation of superparamagnetic iron oxide nanoparticles for MR-visualization of surgical implants.

For the development of a surgical mesh implant that is visible in magnetic resonance imaging (MRI), superparamagnetic iron oxides (SPIOs) are integrated into the material of the mesh. In order to get a high quality mesh regarding both mechanical and imaging properties, a narrow size distribution and homogenous spatial distribution, as well as a strong magnetization of SPIOs within the filament of the mesh are required. In this work, six different samples of SPIOs composed of a magnetite core are synthesized with and without stabilizing dodecanoic acid and analyzed using a superconducting quantum interference device (SQUID), transmission electron microscope (TEM) and a magnetic force microscope (MFM) to determine the properties that are beneficial for the assembly and imaging of the implant. These analyses show the feasibility of visualization of surgical implants with incorporated SPIOs and the influence of the agglomeration of SPIOs on their magnetization and on a homogenous spatial distribution within the polymer of the mesh.

[Bone biopsy needles: mechanical properties, needle design and specimen quality]. / Knochen-Biopsiesysteme: mechanische Eigenschaften, Nadeldesign und Probenqualität.

PURPOSE: To quantitatively analyze differences in mechanical properties, needle design including signs of wear, subjective handling and specimen quality of bone biopsy needles. MATERIALS AND METHODS: In this study 19 different bone biopsy systems (total 38; 2 /type) were examined. With each biopsy needle five consecutive samples were obtained from vertebral bodies of swine. During puncture a force-torques sensor measured the mechanical properties and subjective handling was assessed. Before and after each biopsy the needles were investigated using a profile projector and signs of wear were recorded. Afterwards, a pathologist semi-quantitatively examined the specimen regarding sample quality. The overall evaluation considered mechanical properties, needle wear, subjective handling and sample quality. Differences were assessed for statistical significance using ANOVA and t-test. RESULTS: Needle diameter (p = 0.003) as well as needle design (p = 0.008) affect the mechanical properties significantly. Franseen design is significantly superior to other needle designs. Besides, length reduction recorded by the profile projector, as a quality criterion showed notable distinctions in between the needle designs. CONCLUSION: Bone biopsy needles vary significantly in performance. Needle design has an important influence on mechanical properties, handling and specimen quality. Detailed knowledge of those parameters would improve selecting the appropriate bone biopsy needle.

Silicon carbide as a heat-enhancing agent in microwave ablation: in vitro experiments.

PURPOSE: Silicon carbide (SiC) is an inert compound material with excellent microwave absorption and heat-conducting properties. The aim of our study was to investigate the heat-enhancing effects of SiC in microwave ablation in an in vitro setting. MATERIALS AND METHODS: Different concentrations of SiC powder were mixed with 2% gelatin, producing a 20-ml mixture that was then heated with a clinical microwave ablation system (5 min/45 W). Temperature was measured continuously fiberoptically. Additional heating properties were assessed for the most heatable concentrations at different energy settings (10, 20, and 30 W). As reference, 2% gelatin without SiC was heated. Statistical evaluation by analysis of variance with post hoc Student-Newman-Keuls testing was performed. RESULTS: For the different SiC concentrations, maximum temperatures measured were 45.7 ± 1.2°C (0% SiC, control), 50.4 ± 3.6°C (2% SiC), 60.8 ± 1.8°C (10% SiC), 74.9 ± 2.6°C (20% SiC), and 83.4 ± 2.5°C (50% SiC). Differences between all data points were significant (P < 0.05). Maximum temperatures that used 20% SiC were 36.3 ± 2.76°C (10 W), 48.7 ± 4.18°C (20 W), and 50.6 ± 0.68°C (30 W). The use of 50% SiC maximum temperatures resulted in values of 46.2 ± 2.52°C (10 W), 70.1 ± 0.64°C (20 W), and 83.0 ± 4.69°C (30 W). With 20% SiC and 50% SiC mixtures, the 10 W maximum temperatures were significantly lower than at all other power levels, and maximum temperatures with 20 and 30 W were significantly lower when compared with 45 W (P < 0.05). CONCLUSIONS: SiC is a nontoxic, highly effective substance for enhancing microwave-induced heating with a microwave ablation system in vitro. These data suggest its usefulness for enhancement of ablative effects in percutaneous tumor therapy. Further investigations need to be performed to evaluate the ex vivo and in vivo ablation effects and the possible methods for administration of SiC particles.