The Distensibility of the Human Vena Cava and Its Importance to In Vitro Studies of Venous Compression Syndromes: A Search for a Suitable Polymer for 3-Dimensional Printing.

BACKGROUND: Venous compression syndromes are clinical conditions in which the large veins are compressed by other anatomical structures. Laboratory simulations may help us better understand the hemodynamics in venous compressions by creating situations similar to those seen in vivo. The aim of this study is to produce a model of the caval bifurcation using a polymer with distensibility similar to the human vena cava. METHODS: Fragments of the inferior vena cava were collected from 13 deceased kidney donors (aged 15-37 years) and were tested for deformation (strain) when subjected to distension at 50 N/cm2. Strips of 5 different polymers-thermic polyurethane and Agilus30 with Vero Magenta (AV) (in 3 different hardnesses) and silicone-were subjected to the same biomechanical tests and compared with the vena cava. A model of the caval bifurcation was produced with 3-D printing. RESULTS: The deformation (strain) of the vena cava wall was 0.16 ± 0.9 when submitted to stress close to 50 N/cm2. Silicone showed a strain higher than the standard deviation of venous fragments. The strain of AV resin 95 Shore was lower than the standard deviation of the venous fragments. AV Resins 70 and 85 Shore showed strains within the standard deviation of the venous specimen, with 70 Shore being closest to the mean venous strain. Therefore, this material was selected for modeling the caval bifurcation. The computed tomography scan image generated a computer model of the caval bifurcation and was printed in 3 dimensions. In addition, the segments of 2 adjacent vertebrae were also printed to reference the compression site. CONCLUSIONS: The 3-D printing of large veins can produce models with anatomy and biomechanics similar to those of human veins and opens a field of investigation into the hemodynamics of venous compression syndromes. Polymers with Shore A70 appear to have biomechanical properties similar to those of the vena cava wall. The model obtained in this study can be used in several in vitro studies of May-Thurner Syndrome.

Conditioning of hiPSC-derived cardiomyocytes using surface topography obtained with high throughput technology.

Surface functionalization of polymers aims to introduce novel properties that favor bioactive responses. We have investigated the possibility of surface functionalization of polyethylene terephthalate (PET) sheets by the combination of laser ablation with hot embossing and the application of such techniques in the field of stem cell research. We investigated the response of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) to topography in the low micrometer range. HiPSC-CMs are expected to offer new therapeutic tools for myocardial replacement or regeneration after an infarct or other causes of cardiac tissue loss. However, hiPSC-CMs are phenotypically immature compared to myocytes in the adult myocardium, hampering their clinical application. We aimed to develop and test a high-throughput technique for surface structuring that would improve hiPSC-CMs structural maturation. We used laser ablation with a ps-laser source in combination with nanoimprint lithography to fabricate large areas of homogeneous micron- to submicron line-like pattern with a spatial period of 3 µm on the PET surface. We evaluated cell morphology, alignment, sarcomeric myofibrils assembly, and calcium transients to evaluate phenotypic changes associated with culturing hiPSC-CMs on functionalized PET. Surface functionalization through hot embossing was able to generate, at low cost, low micrometer features on the PET surface that influenced the hiPSC-CMs phenotype, suggesting improved structural and functional maturation. This technique may be relevant for high-throughput technologies that require conditioning of hiPSC-CMs and may be useful for the production of these cells for drug screening and disease modeling applications with lower costs.

Early changes in myocyte contractility and cardiac function in streptozotocin-induced type 1 diabetes in rats.

Diabetes can elicit direct deleterious effects on the myocardium, independent of coronary artery disease or hypertension. These cardiac disturbances are termed diabetic cardiomyopathy showing increased risk of heart failure with or without reduced ejection fraction. Presently, there is no specific treatment for this type of cardiomyopathy and in the case of type I diabetes, it may start in early childhood independent of glycemic control. We hypothesized that alterations in isolated myocyte contractility and cardiac function are present in the early stages of experimental diabetes in rats before overt changes in myocardium structure occur. Diabetes was induced by single-dose injection of streptozotocin (STZ) in rats with data collected from control and diabetic animals 3 weeks after injection. Left ventricle myocyte contractility was measured by single-cell length variation under electrical stimulation. Cardiac function and morphology were studied by high-resolution echocardiography with pulsed-wave tissue Doppler imaging (TDI) measurements and three-lead surface electrocardiogram. Triglycerides, cholesterol and liver enzyme levels were measured from plasma samples obtained from both groups. Myocardial collagen content and perivascular fibrosis of atria and ventricle were studied by histological analysis after picrosirius red staining. Diabetes resulted in altered contractility of isolated cardiac myocytes with increased contraction and relaxation time intervals. Echocardiography showed left atrium dilation, increased end-diastolic LV and posterior wall thickness, with reduced longitudinal systolic peak velocity (S') of the septum mitral annulus at the apical four-chamber view obtained by TDI. Triglycerides, aspartate aminotransferase and alkaline phosphatase were elevated in diabetic animals. Intertitial collagen content was higher in atria of both groups and did not differ among control and diabetic animals. Perivascular intramyocardial arterioles collagen did not differ between groups. These results suggest that alterations in cardiac function are present in the early phase in this model of diabetes type 1 and occur before overt changes in myocardium structure appear as evaluated by intersticial collagen deposition and perivascular fibrosis of intramyocardial arterioles.

Computational tool for morphological analysis of cultured neonatal rat cardiomyocytes.

This study describes the development and evaluation of a semiautomatic myocyte edge-detector using digital image processing. The algorithm was developed in Matlab 6.0 using the SDC Morphology Toolbox. Its conceptual basis is the mathematical morphology theory together with the watershed and Euclidean distance transformations. The algorithm enables the user to select cells within an image for automatic detection of their borders and calculation of their surface areas; these areas are determined by adding the pixels within each myocyte's boundaries. The algorithm was applied to images of cultured ventricular myocytes from neonatal rats. The edge-detector allowed the identification and quantification of morphometric alterations in cultured isolated myocytes induced by 72 hours of exposure to a hypertrophic agent (50 µM phenylephrine). There was a significant increase in the mean surface area of the phenylephrine-treated cells compared with the control cells (p<;0.05), corresponding to cellular hypertrophy of approximately 50%. In conclusion, this edge-detector provides a rapid, repeatable and accurate measurement of cell surface areas in a standardized manner. Other possible applications include morphologic measurement of other types of cultured cells and analysis of time-related morphometric changes in adult cardiac myocytes.

Changes in cardiac heparan sulfate proteoglycan expression and streptozotocin-induced diastolic dysfunction in rats.

BACKGROUND: Changes in the proteoglycans glypican and syndecan-4 have been reported in several pathological conditions, but little is known about their expression in the heart during diabetes. The aim of this study was to investigate in vivo heart function changes and alterations in mRNA expression and protein levels of glypican-1 and syndecan-4 in cardiac and skeletal muscles during streptozotocin (STZ)-induced diabetes. METHODS: Diabetes was induced in male Wistar rats by STZ administration. The rats were assigned to one of the following groups: control (sham injection), after 24 hours, 10 days, or 30 days of STZ administration. Echocardiography was performed in the control and STZ 10-day groups. Western and Northern blots were used to quantify protein and mRNA levels in all groups. Immunohistochemistry was performed in the control and 30-day groups to correlate the observed mRNA changes to the protein expression. RESULTS: In vivo cardiac functional analysis performed using echocardiography in the 10-day group showed diastolic dysfunction with alterations in the peak velocity of early (E) diastolic filling and isovolumic relaxation time (IVRT) indices. These functional alterations observed in the STZ 10-day group correlated with the concomitant increase in syndecan-4 and glypican-1 protein expression. Cardiac glypican-1 mRNA and skeletal syndecan-4 mRNA and protein levels increased in the STZ 30-day group. On the other hand, the amount of glypican in skeletal muscle was lower than that in the control group. The same results were obtained from immunohistochemistry analysis. CONCLUSION: Our data suggest that membrane proteoglycans participate in the sequence of events triggered by diabetes and inflicted on cardiac and skeletal muscles.

Late gadolinium enhancement magnetic resonance imaging in the diagnosis and prognosis of endomyocardial fibrosis patients.

BACKGROUND: Endocardial fibrous tissue (FT) deposition is a hallmark of endomyocardial fibrosis (EMF). Echocardiography is a first-line and the standard technique for the diagnosis of this disease. Although late gadolinium enhancement (LGE) cardiovascular magnetic resonance (CMR) allows FT characterization, its role in the diagnosis and prognosis of EMF has not been investigated. METHODS AND RESULTS: Thirty-six patients (29 women; age, 54±12 years) with EMF diagnosis after clinical evaluation and comprehensive 2-dimensional Doppler echocardiography underwent cine-CMR for assessing ventricular volumes, ejection fraction and mass, and LGE-CMR for FT characterization and quantification. Indexed FT volume (FT/body surface area) was calculated after planimetry of the 8 to 12 slices obtained in the short-axis view at end-diastole (mL/m(2)). Surgical resection of FT was performed in 16 patients. In all patients, areas of LGE were confined to the endocardium, frequently as a continuous streak from the inflow tract extending to the apex, where it was usually most prominent. There was a relation between increased FT/body surface area and worse New York Heart Association functional class and with increased probability of surgery (P<0.05). The histopathologic examination of resected FT showed typical features of EMF with extensive endocardial fibrous thickening, proliferation of small vessels, and scarce inflammatory infiltrate. In multivariate analysis, the patients with FT/body surface area >19 mL/m(2) had an increased mortality rate, with a relative risk of 10.8. CONCLUSIONS: Our study provides evidence that LGE-CMR is useful in the diagnosis and prognosis of EMF through quantification of the typical pattern of FT deposition.

A new approach to heart valve tissue engineering: mimicking the heart ventricle with a ventricular assist device in a novel bioreactor.

The 'biomimetic' approach to tissue engineering usually involves the use of a bioreactor mimicking physiological parameters whilst supplying nutrients to the developing tissue. Here we present a new heart valve bioreactor, having as its centrepiece a ventricular assist device (VAD), which exposes the cell-scaffold constructs to a wider array of mechanical forces. The pump of the VAD has two chambers: a blood and a pneumatic chamber, separated by an elastic membrane. Pulsatile air-pressure is generated by a piston-type actuator and delivered to the pneumatic chamber, ejecting the fluid in the blood chamber. Subsequently, applied vacuum to the pneumatic chamber causes the blood chamber to fill. A mechanical heart valve was placed in the VAD's inflow position. The tissue engineered (TE) valve was placed in the outflow position. The VAD was coupled in series with a Windkessel compliance chamber, variable throttle and reservoir, connected by silicone tubings. The reservoir sat on an elevated platform, allowing adjustment of ventricular preload between 0 and 11 mmHg. To allow for sterile gaseous exchange between the circuit interior and exterior, a 0.2 µm filter was placed at the reservoir. Pressure and flow were registered downstream of the TE valve. The circuit was filled with culture medium and fitted in a standard 5% CO(2) incubator set at 37 °C. Pressure and flow waveforms were similar to those obtained under physiological conditions for the pulmonary circulation. The 'cardiomimetic' approach presented here represents a new perspective to conventional biomimetic approaches in TE, with potential advantages.

Adenoviral expression of calmodulin antisense reduces hypertrophy in cultured cardiomyocytes.

Sustained myocardial hypertrophy is associated with an increased risk of sudden death and progression to heart failure. Multiple signal pathways are involved in cardiac hypertrophy and understanding their interaction may point to new therapeutic targets. In this work, we tested the hypothesis that adenovirus-mediated calmodulin (CaM) antisense expression will reduce the intracellular availability of CaM and inhibit the hypertrophic response. Three recombinant adenoviruses were constructed: AdASCaM, containing the AntiSense sequence of CaM and the enhanced green fluorescent protein (GFP) coding sequence; AdCaM, containing the coding sequence of CaM and the GFP sequence; and the AdGFP, containing the GFP coding sequence. Neonatal rat ventricular cardiomyocytes were infected with AdASCaM, AdCaM, or AdGFP and stimulated with phenylephrine (PE, 50 microM) or angiotensin II (AngII, 10 microM) for 48 h and cell surface area measured with planimetry. After PE treatment, the surface areas of cardiomyocytes infected with AdASCaM or AdGFP were 411 +/- 174.3 micro(2) and 832.6 +/- 372.3 micro(2), respectively (P < 0.01). After AngII treatment, the surface areas of cardiomyocytes infected with AdASCaM or AdGFP were 441.5 +/- 149.2 micro(2) and 726 +/- 328.3 micro(2), respectively (P < 0.01). Adenoviral expression of the CaM antisense (AdASCaM) significantly inhibited PE or AngII-induced cardiomyocyte hypertrophy. Cardiomyocytes infected with the AdCaM showed increased area when compared with those infected with the AdGFP. These results suggest that adenovirus-mediated changes in CaM expression may alter hypertrophy in cardiac myocytes.

Echocardiographic assessment of global ventricular function using the myocardial performance index in rats with hypertrophy.

Myocardial hypertrophy is the hallmark of chronic pressure overload and the myocardial performance index (MPI) is an easily recordable measurement of Doppler time intervals. In this study, the utility of using MPI to assess the progression of hypertrophy in the aortic-banded rat model was evaluated. Male Wistar rats (70-90 g) underwent ascending aorta constriction (n = 4) or a sham operation (n = 5). High-resolution echocardiography was performed 4, 7, 10, and 12 weeks after the surgery. Over this follow-up interval, animals in the aortic-banded group demonstrated an increase in their mean left ventricular (LV) mass and MPI compared with controls. MPI reflects ventricular performance in small animals with LV hypertrophy, showing alterations early after aorta constriction.

Noninvasive assessment of hemodynamic parameters in experimental stenosis of the ascending aorta.

We sought to noninvasively evaluate left ventricular (LV) function after cardiac hypertrophy induced by experimental stenosis of the ascending aorta. Male Wistar rats (70-90 g) underwent ascending aorta constriction by the surgical placement of a titanium clip (n=5) or sham operation (n=6). High-resolution bidimensional, pulsed-wave Doppler (PWD) and pulsed-wave tissue Doppler imaging (TDI) were performed 22 weeks after surgery. PWD was used to obtain mitral flow velocities, and TDI was used to obtain velocities along the septal mitral annulus and LV posterior wall. Clip placement produced myocardial hypertrophy with decreased systolic myocardial peak velocity in both the long and short axes. Increased myocardial mass, that is, posterior wall and septal thickness, was indicative of ventricular remodeling. Diastolic dysfunction was observed, with an increased early to late ratio of mitral velocities and increased left atrium dimension, consistent with a left ventricular restrictive filling pattern.

Um circuito simples com bomba única para circulação extracorpórea com oxigenação autógena / A simple circuit with only centrifugal pump for extracorporeal circulation with autogenous oxygenation

Material e Métodos: Foi testado em 30 cães um circuito capaz de promover circulação extracorpórea (CEC) com oxigenação autógena (OA) do sangue, usando apenas uma bomba centrífuga. Esta montagem dispensou bombeamento para o lado direito: o gradiente de pressão bastante para vencer a resistência arterial pulmonar foi vencido aumentando-se a pressão nas artérias pulmonares pela expansão da volemia e diminuindo-se a pressão do átrio esquerdo pela drenagem dessa câmara mediante um sifão. O coração foi mantido em ritmo de fibrilação ventricular durante o período de perfusão e ao seu término, o ritmo próprio foi recuperado mediante cardioversão elétrica.Resultados: Este circuito permitiu a manutenção de parâmetros hemodinâmicos e gases sangüíneos adequados durante a perfusão. O campo operatório e a mobilidade do coração foram similares aos proporcionados pela CEC convencional. Conclusão: Concluímos que o uso de bomba centrífuga única simplifica a OA, podendo tornar-se uma escolha prática nos procedimentos de revascularização do miocárdio(AU)#S#as#BR