RESUMEN
Iodinated contrast media (ICM) are widely used for diagnostic and interventional procedures in radiology and cardiology. Ideally, they should not interact with blood cells or vascular wall cells to avoid deteriorations of the blood circulation. However, it is well known that ICM can affect erythrocytes as well as endothelial cells which consequently might perturb especially the microcirculation. In former studies the influence of two ICM (iodixanol versus iopromide) on the vascular system, the development of blood stasis, on changes in renal resistive index (RRI) and vascular diameters, and on the post-mortem distribution of iodine as marker for ICM in the explanted kidneys was examined. The modus of ICM application into the supra-renal aorta followed the regime in interventional cardiology, so that 10 bolus injections were administered at steady intervals (iopromide 4,32âml / iodixanol 5âml) accompanied by infusion of 500âml isotonic NaCl-solution.In the present study, the post-mortem X-ray analysis revealed that there were no differences in iodine content in the regions of the mid-cortex and the medullo-pelvic transition zone of the kidneys after application of both ICM. Remarkable differences, however, were found in the region of the capsule-near cortex, where the application of iopromide led to a significantly lower iodine content in the microcirculation. This is in good agreement with former studies, in which a maldistribution in this area, presumably due to a decrease in arteriolar inflow as a result of stasis/occlusion was shown.
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There is growing evidence that COVID-19 not only affects the lungs but beyond that the endothelial system. Recent studies showed that this can lead to microcirculatory impairments and in consequence to functional disorders of all inner organs. The combination of endothelial dysfunction with a generalized inflammatory state and complement elements may together contribute to the overall pro-coagulative state described in COVID-19 patients leading to venular as well as to arteriolar occlusions.
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Betacoronavirus/patogenicidad , Infecciones por Coronavirus/patología , Endotelio Vascular/virología , Neumonía Viral/patología , COVID-19 , Infecciones por Coronavirus/virología , Endotelio Vascular/patología , Humanos , Pandemias , Neumonía Viral/virología , SARS-CoV-2RESUMEN
Repeated injections of iodinated contrast media (CM) can lead to a deterioration of the renal blood flow, can redistribute blood from the renal cortex to other parts of the kidney and can cause small decreases of the blood flow in cortical capillaries, a significant reduction in blood flow in peritubular capillaries and a significant reduction in blood flow in the vasa recta. Therefore, a study in pigs was designed, to show whether the repeated injection of CM boli, alone, can cause a reduction of oxygenation in the cortico-medullar renal tissue - the region with the highest oxygen demand in the kidney - of pigs.While the mean pO2-value had only decreased by 0.3 mmHg from 29.9±4.3 mmHg to 29.6±4.3 mmHg (pâ=â0.8799) after the tenth Iodixanol bolus, it decreased by 5.9 mmHg from 34.0±4.3 mmHg to 28.1±4.3 mmHg after the tenth Iopromide bolus (pâ=â0.044). This revealed a remarkable difference in the influence of these CM on the oxygen partial pressure in the kidney.Repeated applications of CM had a significant influence on the renal oxygen partial pressure. In line with earlier studies showing a redistribution of blood from the cortex to other renal areas, this study revealed that Iodixanol - in contrast to Iopromide - induced no changes in the pO2 in the cortico-medullar region which confirms that Iodixanol did not hinder the flow of blood through the renal micro-vessels. These results are in favor of a hypothesis from Brezis that a microcirculatory disorder might be the basis for the development of CI-AKI.
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Lesión Renal Aguda/inducido químicamente , Ácidos Triyodobenzoicos/química , Animales , Medios de Contraste , Hemodinámica , Masculino , Microcirculación , PorcinosRESUMEN
Osteoarthritis (OA) of the knee is a widespread disease, often resulting in pain, restricted mobility and a reduction of activities and participation. Initial studies gave hints that Acoustic Emission Analysis (AEA) is capable of detecting early changes in cartilage structure. However, up to date no in vivo validation studies have been conducted. A prospective pilot study was conducted to investigate this diagnostic capability and the accuracy of the AEA, using magnetic resonance imaging (MRI) as a reference standard. Additionally, potential factors influencing false positive or negative results were studied. Twenty-eight patients, receiving MRI due to discomfort of the knee, were examined with AEA. Sensitivity was 0.92 for the whole knee and 0.86 to 1 for different parts of the knee. The specificity was 0.7 and 0.59 to 0.78, respectively. Confidence intervals varied between 0 and 0.33 for sensitivity and 0.1 and 0.24 for specificity. The diagnostic accuracy of the AEA was shown to be good to very good. However, because of the relatively small number of patients involved, interpretation of the data should be handled with care. Future studies with greater sample sizes have to be conducted to confirm the results of this investigation.
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Acústica , Osteoartritis de la Rodilla/diagnóstico , Adulto , Estudios de Cohortes , Femenino , Humanos , Imagen por Resonancia Magnética , Masculino , Osteoartritis de la Rodilla/diagnóstico por imagenRESUMEN
BACKGROUND: The laser Doppler fluxmetry (LDF) is a non-invasive method to assess skin blood perfusion, measuring the flow of blood cells inside a tissue volume without harming the tissue. In the diagnosis of skin circulation disorders, the results of the LDF measurement are generally used in such a way that "normal" (or non-ill) or "pathological" values are achieved by comparison with a reference sample, for example of apparently healthy subjects. MATERIAL AND METHODS: In this study, the values of LDF for the diagnosis of microcirculatory disorders in patients with coronary artery disease (nâ=â20) or in patients with microcirculatory disorders, already diagnosed by capillary microscopy (nâ=â46), were examined. RESULTS: The mean values of LD amplitudes in the four frequency windows for patients with coronary artery disease were in the reference range. However, some of the patients showed reduced LD values: in eleven of the twenty patients, one or more mean LD amplitudes were below the reference range. Four of the eleven patients had pathologically decreased capillary erythrocyte velocities of veryâ=â0.09-0.21 [mm/s], while the other seven patients had normal blood circulation at rest.For all patients with a proven cutaneous microcirculatory disorder, the mean LD amplitude in at least one of the frequency windows FF2 to FF4 was pathologically reduced. CONCLUSION: The Laser-Doppler fluxmetry method used in the study allows the reliable diagnosis of cutaneous microcirculatory disorders.
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Flujometría por Láser-Doppler/métodos , Microcirculación/genética , Adulto , Enfermedad de la Arteria Coronaria , Femenino , Humanos , Masculino , Persona de Mediana EdadRESUMEN
BACKGROUND: Contrast-induced acute kidney injury (CI-AKI), a potentially life-threatening complication of iodinated contrast media in patients with impaired renal function, has attracted increasing attention in recent years. There is overwhelming evidence that the most important pre-disposing factor for a contrast-medium induced nephropathy is the pre-existence of a renal impairment. METHODS: The registry was performed as a part of a quality management project in the Dresden-Friedrichstadt heart catheter laboratory. In compliance with the Declaration of Helsinki/Somerset West, 9,026 patients were included between 2010 and 2015. 100 patients of these were participants in a chronic dialysis program. All patients were dialyzed on the day before angiography. In all patients a coronary angiography, in 28 patients a stent implantation and in 12 patients a surgical reconstruction had to be performed. Prior to the intervention and one, two and three days thereafter the serum creatinine was measured. RESULTS: Up to the third day after application of the iodinated contrast medium no significant changes of the serum creatinine (baseline value: 423.3±42.6µmol/l) occurred (ANOVA for repeated measures: pâ=â0.507). On average, a slight decrease of the serum creatinine was found.All patients remained in their routine dialysis-program. 18 out of 100 died during the next three months after the procedure. CONCLUSION: The study revealed that the coronary angiography using Iodixanol as iodinated contrast medium did not result in an increase of serum creatinine, which was drastically elevated in these patients before application of the iodinated contrast medium.
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Lesión Renal Aguda/inducido químicamente , Medios de Contraste/efectos adversos , Fallo Renal Crónico/complicaciones , Ácidos Triyodobenzoicos/efectos adversos , Lesión Renal Aguda/patología , Anciano , Anciano de 80 o más Años , Medios de Contraste/farmacología , Femenino , Humanos , Fallo Renal Crónico/tratamiento farmacológico , Fallo Renal Crónico/patología , Masculino , Persona de Mediana Edad , Ácidos Triyodobenzoicos/farmacologíaRESUMEN
Laser tissue soldering (LTS) based on indocyanine green (ICG)-mediated heat-denaturation of proteins might be a promising alternative technique for micro-suturing, but up to now the problem of too weak shear strength of the solder welds in comparison to sutures is not solved. Earlier reports gave promising results showing that solder supported by carrier materials can enhance the cohesive strength of the liquid solder. In these studies, the solder was applied to the carriers by dip coating. Higher reliability of the connection between the solder and the carrier material is expected when the solder is bound covalently to the carrier material. In the present study a poly(ether imide) (PEI) membrane served as carrier material and ICG-supplemented albumin as solder substrate. The latter was covalently coupled to the carrier membrane under physiological conditions to prevent structural protein changes. As laser source a diode continuous-wave laser emitting at 808ânm with intensities between 250âmW and 1500âmW was utilized. The albumin functionalized carrier membrane was placed onto the tunica media of explanted pig thoracic aortae forming an overlapping area of approximately 0.5×0.5âcm2. All tests were performed in a dry state to prevent laser light absorption by water. Infrared spectroscopy, spectro-photometrical determination of the secondary and primary amine groups after acid orange II staining, contact angle measurements, and atomic force microscopy proved the successful functionalization of the PEI membrane with albumin. A laser power of 450âmW LTS could generate a membrane-blood vessel connection which was characterized by a shear strength of 0.08±0.002âMPa, corresponding to 15% of the tensile strength of the native blood vessel. Theoretically, an overlapping zone of 4.1âmm around the entire circumference of the blood vessel could have provided shear strength of the PEI membrane-blood vessel compound identical to the tensile strength of the native blood vessel. These in-vitro results confirmed the beneficial effects of solder reinforcement by carrier membranes, and suggest LTS with covalently bound solders on PEI substrates for further studies in animal models.
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Albúminas/metabolismo , Vasos Sanguíneos/metabolismo , Terapia por Láser/métodosRESUMEN
The Laser Doppler technique (Laser-Doppler-Fluxmetry, LDF), a noninvasive method to estimate skin blood flow (LDF), is frequently used in research and clinical routine [1]. Here, the measurements were carried out with a new Laser Doppler system, the DOP-system, which allows to measure frequency spectra in four different frequency windows according to the velocities in venules (low velocity), capillaries (low to medium velocities), and in arteries (with high and very high velocities). However, the diagnostic reliability or the effectiveness of the LDF has not yet been evaluated sufficiently, which is indispensable, where medical diagnostics and therapy controls are concerned. For a valid interpretation of LDF values of individual patients, the knowledge of the reference range and the variability of the measured parameters is required.In four successive studies the reference range (62 apparently healthy subjects), the circadian variability (8 subjects), the variability from day-to-day (6 subjects) and over one year with monthly measurements (6 subjects) were evaluated.With the knowledge of the reference range, microcirculatory disorders can now be diagnosed using the DOP method. Following a standard measurement procedure there was no dependence of the measured data on the day or season of measurement.
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Flujometría por Láser-Doppler/métodos , Microcirculación/fisiología , Piel/irrigación sanguínea , Adulto , Femenino , Voluntarios Sanos , Humanos , Masculino , Persona de Mediana Edad , Valores de Referencia , Reproducibilidad de los Resultados , Adulto JovenRESUMEN
Ultrasound contrast agents (USCA) allows the dynamic detection of blood flow of both the macro and microvasculature. An obvious prerequisite for USCAs is the unhindered passage of clinically relevant dose levels through the microcirculation especially of the lungue, where they have to pass capillaries with diameters of around 4 µm. While smaller microbubbles rapidly passed through the microcirculation along with the red blood cells, larger microbubbles, however, were observed to coalesce and interrupt the blood flow. Whether this might influence the tissue oxygen tension is unclear up to now.To examine this question a bolus of 2.4âml SonoVue™ was injected into the suprarenal aorta at a flow rate of 10âml/s (a dosage usually applied in the clinic). The pO2 in the outer medulla of the kidney was continuously measured using a flexible pO2 microcatheter. In addition, the SonoVue™ injection and its passage through the renal vasculature were documented by the CEUS technology to assess whether the microbubbles passed the kidney.The study revealed that SonoVue™ induced no changes of the mean oxygen partial pressure in the outer medulla which confirms that these microbubbles on their way through the medullar capillaries did not hinder the co-flow of blood through the renal microvessels in a big animal model with a renal morphology and function comparable to human kidneys. These results demonstrate that the CEUS diagnostic itself did not influence the system to be examined which is a most important prerequisite for any diagnostic method.
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Medios de Contraste/uso terapéutico , Riñón/irrigación sanguínea , Microburbujas/uso terapéutico , Ultrasonografía/métodos , Humanos , Riñón/patologíaRESUMEN
In drug eluting stents the cytostatic drugs Sirolimus or Tacrolimus are used to inhibit blood vessel restenosis by limiting the proliferation of smooth muscle cells. However, the cytostatic activity of both drugs was shown to be not cell specific and could also affect the stent endothelialisation, respectively. Currently, only limited in vitro data are available about the impact of Sirolimus and Tacrolimus on endothelial cell proliferation over a broad concentration range. To answer this question the following study was performed.Commercially obtained HUVEC were expanded with DMEM cell culture medium (GIBCO, Germany) supplemented with 5 vol% fetal calf serum on non-coated regular polystyrene-based 24-multiwell plates. For drug testings 2×104 cells/cm2 were seeded and grown for 24âh until 30-40% of the multiwell surfaces were covered and then exposed to Sirolimus (1.0×10-11 - 1.0×10-5âmol/l) or Tacrolimus (2.0×10-8 - 6.2×10-5âmol/l), both dissolved in DMSO. 12, 24 and 48âh after adding the drugs cell numbers per area were quantified by counting the cells in six wells with four fields of view per well, representing 0.6âmm2, using a confocal laser microscope.After 48âh of cell growth in the drug-free cell culture medium, the HUVEC number increased from 2.0×104 to 3.55×104 cells/cm2 (mean cell doubling time: 53.6âh, nâ=â6). At lower concentrations (≤2.0×10-6âmol/l) Tacrolimus reduced the number of adherent HUVEC significantly less than Sirolimus (pâ<â0.05). However, at higher concentrations (≥2.07×10-5âmol/l) the effect of Tacrolimus on the number of adherent endothelial cells was significantly greater than that of Sirolimus (pâ<â0.05). At the highest concentration applied (6.22×10-5âmol/l), Tacrolimus induced detachment of all HUVECs within 12âh after drug application. The number of adherent HUVEC decreased only slightly (about 9%) after Sirolimus application at the highest concentration (1.09×10-5âmol/l).These data show that in a non-flow model the cytostatic drug Tacrolimus reduced the number of adherent endothelial cells less than Sirolimus, as long as the drug concentration did not surpass 10-6âmol/l. At the limits of solubility, Sirolimus (1×10-5âmol/l) reduced the number of adherent endothelial cells less than Tacrolimus (6×10-5âmol/l), which induced detachment of endothelial cells.
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Stents Liberadores de Fármacos/estadística & datos numéricos , Células Endoteliales/metabolismo , Inmunosupresores/uso terapéutico , Miocitos del Músculo Liso/metabolismo , Sirolimus/uso terapéutico , Tacrolimus/uso terapéutico , Proliferación Celular , Humanos , Inmunosupresores/farmacología , Sirolimus/farmacología , Tacrolimus/farmacologíaRESUMEN
The endothelialization of cardiovascular prostheses is known to improve their haemocompatibility. As such body-foreign materials often do not endothelialize spontaneously. A lot of in vitro studies are ongoing how endothelialization of biomaterials can be improved. In this study the influence of different components of a tissue-typical extracellular matrix (ECM) like laminin, fibronectin or gelatin on the formation of an endothelial cell monolayer and on the shear resistance of adherent cells on these substrates was studied.The study revealed that the density of human venous endothelial cells (HUVEC) monolayers differed markedly between cells grown on a natural ECM and cells grown on singularized components of an ECM (pâ<â0.001). Only HUVEC grown on laminin showed similar densities and a stress fiber pattern comparable to HUVEC grown on the ECM. HUVEC grown on gelatin- or fibronectin-coated coverslips were less firmly attached to the substrate; frequently individual HUVEC and even groups of cells detached.Concluding it seems that coating of implants with laminin supports the formation of shear resistant endothelial cell (EC) monolayer - superior to other ECM components.
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Células Endoteliales/efectos de los fármacos , Endotelio Vascular/efectos de los fármacos , Matriz Extracelular/efectos de los fármacos , Células Cultivadas , HumanosRESUMEN
INTRODUCTION: Clinical complications like thrombosis or anaphylaxis have been described to go along with the intra-venous or intra-arterial injection of iodinated contrast media (CM). It has been suggested that the administration of CM affects rheological parameters and thereby causes reduced blood velocity in microvessels. In vitro studies revealed significant buckling of endothelial cells after exposure to CM reducing the lumen of vessels. The aim of this study was to test the influence of CM on three-dimensional microvascular tubules with open lumina within an organotypic soft-tissue co-culture assay in vitro. This model, which is based on the co-culture of endothelial cells and fibroblasts, allows the analysis and quantitation of different parameters of microvascular endothelial capillary structures. MATERIAL AND METHODS: Human dermal fibroblasts and human dermal microvascular endothelial cells were co-cultured for 10 days. Fibroblasts were adapted to the endothelial cell medium before co-culture and allowed to proliferate as well as produce extracellular matrix. The co-cultures were exposed to three different CM, i.e., Iomeprol (Imeron 400MCT), Iodixanol (Visipaque 320) or Iohexol (Accupaque 350) for 1.5 minutes or 5.0 minutes, respectively. For this, a mixture of CM and cell culture medium in a ratio of 30% CM by volume was prepared. After fixation in methanol/acetone, the endothelial cells were immunolabeled with the endothelial marker anti-CD31 and the tubular structures were assessed morphometrically. RESULTS: In the organotypic soft-tissue co-cultures with fibroblasts, the endothelial cells developed three-dimensional capillary-like structures which expanded via sprouting branches. After incubation with the different CM, the numbers of endothelial tubes (pâ=â0.001) and their lengths (pâ=â0.003) were significantly lower after the 5 minutes incubation time, when compared to the 1.5 minutes incubation time. The tubular diameters were significantly reduced after 5 minutes (pâ<â0.001), when compared to the 1.5 minutes incubation duration. Interestingly, Iomeprol and Iodixanol induced an elongation of the tubular branches during incubation duration of 1.5 minutes (pâ=â0.015). However, after 5 minutes incubation, the tubular branches were drastically shorter in the presence of Iomeprol and Iodixanol than the tubular branches of the control (pâ=â0.007). SUMMARY AND CONCLUSION: All CM exerted a negative effect on the parameters of in vitro blood vessel development.
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Técnicas de Cocultivo/métodos , Células Endoteliales/metabolismo , Técnicas de Cultivo de Célula , Medios de Contraste , Células Endoteliales/citología , Fibroblastos/efectos de los fármacos , HumanosRESUMEN
Radiographic contrast media (RCM) can initiate microcirculatory disorders. This study was performed to investigate effects of Ioxaglate on the cutaneous microcirculation. The investigation was carried out as prospective randomized double-blind comparison in parallel-group design on two groups of nâ=â10 patients each who had to undergo a diagnostic coronary angiography.The confirmatory parameter of the study was mean erythrocyte capillary velocity [vRBC in mm/sec]. VRBC in the ipsilateral nail-fold capillaries was recorded continuously for 3âmin before and 6âmin after injection of RCM or isotonic saline solution in the A. axillaris respectively, and was evaluated off-line.VRBC in nailfold capillaries was found to be decreased by Ioxaglate by 34% 150 seconds after injection, while isotonic NaCl solution immediately induced a slight increase of 14%.
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Enfermedad de la Arteria Coronaria/tratamiento farmacológico , Ácido Yoxáglico/uso terapéutico , Microcirculación/efectos de los fármacos , Anciano , Enfermedad de la Arteria Coronaria/sangre , Método Doble Ciego , Femenino , Humanos , Ácido Yoxáglico/administración & dosificación , Masculino , Persona de Mediana Edad , Estudios ProspectivosRESUMEN
BACKGROUND: Intra-arterial administration of radiographic contrast media (CM) is discussed to impair renal perfusion. The pathogenesis of contrast-induced Nephropathy (CIN) is still not clarified. OBJECTIVE: This trial was performed to prove the effects of two CM with different molecular structure on renal perfusion. METHODS: A prospective, randomized study on 16 pigs was designed to compare the outcome after application of a low-osmolar iodinated CM (770âmOsm/kg H2O - Group1) and an iso-osmolar iodinated CM (290âmOsm/kg H2o - Group2).Color Coded Doppler Sonography (LOGIQ E9, GE, Milwaukee, USA) was applied for measuring the Renal Resistive Index (RRI) before and after the first, fifth, and tenth bolus of CM. Statistics was performed using analysis of variance for repeated measurements with the Factor "CM". RESULTS: All flow spectra were documented free of artifacts and Peak Systolic Velocity (PSV), Enddiastolic Velocity (EDV) and RRI respectively could be calculated. Mean PSV in Group 1 led to a decrease while in Group 2 PSV showed a significant increase after CM (pâ=â0,042). The course of the mean EDV in both groups deferred accordingly (pâ=â0,033). Mean RRI over time significantly deferred in both groups (pâ=â0,001). It showed a biphasic course in Group 2 and a decrease over time in Group 2. CONCLUSION: While iso-osmolar CM induced an increase of PSV and EDV together with a decrease of RRI, low-osmolar CM could not show this effect or rather led to the opposite.
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Medios de Contraste/uso terapéutico , Glomerulonefritis Membranosa/inducido químicamente , Riñón/efectos de la radiación , Animales , Velocidad del Flujo Sanguíneo , Humanos , Estudios Prospectivos , PorcinosRESUMEN
BACKGROUND: The evaluation of the interaction of human, venous endothelial cells (HUVEC) with body foreign materials on the cellular level cannot be performed in vivo, but is investigated in vitro under standard culture conditions. To maintain the vitality, proliferation and morphology of HUVEC seeded on body foreign substrates over days, the cell culture medium is usually exchanged every second day. It is well known, that alterations in the microenvironment of cells bear the risk of influencing cell morphology and function. In the current study the influence of cell culture medium exchange on HUVEC cytoskeletal microfilament structure and function was investigated. MATERIAL AND METHODS: HUVEC in the third passage were seeded on extracellular matrix (ECM) - which was secreted from bovine corneal endothelial cells on glass- until functional confluence was reached. The experiment started 11 days after HUVEC seeding with an exchange of the cell culture medium followed by a staining of the actin microfilaments with phalloidin-rhodamin 1.5 and 5 minutes after medium exchange. The microfilaments were documented by use of an Olympus microscope (IMT-2) equipped with a UV lamp and online connected to a TV chain (Sony XC 50 ST/monochrome) implying an OPTIMAS - Image analysis system. Prostacyclin was analysed in the cell culture supernatant. RESULTS: 1.5âmin after culture medium exchange in the functionally confluent cultures a slight disturbance of the actin microfilament structure with a broadening of the marginal filament band, a partial disconnection of cell-cell contacts and the appearance of intercellular fenestrations were observed. 5 minutes after medium exchange a redevelopment of the slightly disturbed microfilament structure with a condensation and narrowing of the marginal filament band was seen. 12âh later a further consolidation of the microfilament structure occurred. In addition, a perturbation of the cultured HUVEC occurred after cell culture medium exchange. The prostacyclin concentration in the supernatant increased significantly after 1.5âmin to 466 ± 543 pg·mL-1 (pâ< â0.001) and after 5âmin to 408 ± 458 pg·mL-1 (pâ< â0.001), while in control cells the prostacyclin concentration did not change remaining in the range of 50 ± 48.9 pg·mL-1. CONCLUSION: This study revealed that the exchange of the cell culture medium led to a rapid disturbance of the HUVEC with stress fiber formation, disconnection of cell-cell contacts and an altered prostacyclin secretion, which had regressed nearly completely after 12 hours. Therefore, the evaluation of HUVEC on body foreign materials should be performed not earlier than 12 hours after cell culture medium exchange to avoid a misinterpretation of the endothelial cell morphological state. This procedure minimizes the risk of a misinterpretation of the endothelial cell morphology - caused by the culture medium exchange and not by the interaction between biomaterials and HUVEC.
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Técnicas de Cultivo de Célula , Medios de Cultivo , Células Endoteliales de la Vena Umbilical Humana/citología , Células Cultivadas , Matriz Extracelular , HumanosRESUMEN
The proper morphology and function of the vascular endothelium are prerequisites for a sufficient supply of the tissues. Endothelial cell (EC) dysfunction can lead to circulatory disorders and the development of cardiovascular diseases. The endothelialization of cardiovascular implants is a sophisticated task since EC miss their natural environment and physiological stimuli in vitro. In addition, different studies revealed that the EC behavior and morphology depended on the substrate and the passage number of the EC. Therefore, the comparison of endothelialization studies is very difficult, when passage and substrate are unknown.The aim of this study was to investigate the growth potential and cell morphology of human venous endothelial cells (HUVEC) as a function of different cell passages and different substrates (pristine polystyrene, tissue-typical ECM-coated polystyrene). The study revealed that HUVEC morphology and growth potential were significantly different on pristine polystyrene compared to the basal lamina-like ECM-coated polystyrene surface. Furthermore, it became obvious that the passage of the cells affected the endothelialization of the polystyrene surface significantly. In conclusion, this study emphasized the need for a critical consideration of EC data whereas a simple comparison of results is not possible if EC age and passage is unknown.
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Materiales Biocompatibles Revestidos/metabolismo , Matriz Extracelular/metabolismo , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Poliestirenos/farmacología , Técnicas de Cultivo de Célula , Células Cultivadas , Endotelio Vascular/citología , Células Endoteliales de la Vena Umbilical Humana/citología , Humanos , Poliestirenos/químicaRESUMEN
Radiographic contrast media can lead to drastic changes of the morphology of erythrocytes. The change of the erythrocyte morphology is associated with a decreased deformability possibly resulting from distinctions in the loss of constituents of the membrane cytoskeleton. However, it is unclear whether there is an intravascular hemolysis as a consequence of the disintegration of the erythrocyte membrane. The results of this study showed, that free haemoglobin increased from 16.8 ± 10.0 mg/dl to 21.6 ± 12.6 mg/dl after Iopromide application (p = 0.240), while it slightly decreased from 20.5 ± 10.3 mg/dl to 19.5 ± 12.2 mg/dl after Iodixanol application (p = 0.547). The slight decrease of free haemoglobin after application of Iodixanol differed significantly compared to the increase of free haemoglobin after Iopromide application (p < 0.05). This different response is thought to give evidence to the assumption that the erythrocyte membrane integrity was compromised leading to the release of free haemoglobin as an indicator of hemolysis as well.
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Medios de Contraste/química , Hemólisis/efectos de los fármacos , Yohexol/análogos & derivados , Radiografía , Ácidos Triyodobenzoicos/química , Anciano , Angiografía Coronaria , Membrana Eritrocítica , Eritrocitos/efectos de los fármacos , Femenino , Tasa de Filtración Glomerular , Hemoglobinas/química , Humanos , Yohexol/química , Masculino , Persona de Mediana EdadRESUMEN
Different radiographic contrast media (RCM) were shown to induce morphological changes of blood cells (e.g. erythrocytes or thrombocytes) and endothelial cells. The echinocytic shape change of erythrocytes, particularly, affords alterations of the membrane cytoskeleton. The cytoskeleton plays a crucial role for the shape and deformability of the red blood cell. Disruption of the interaction between components of the red blood cell membrane cytoskeleton may cause a loss of structural and functional integrity of the membrane. In this study band4.9 and actin as components of the cytoskeletal junctional complex were examined in human erythrocytes after suspension in autologous plasma or in plasma RCM mixtures (30% v/v Iodixanol-320 or Iopromide-370) followed by a successive double staining with TRITC-/FITC-coupled monoclonal antibodies. After adding Iopromide-370 to the plasma in practically none of the cells the rounded conformation of the membrane cytoskeleton - as it appeared in cells suspended in autologous plasma - was found. In addition, Iopromide-370 induced thin lines and coarse knob-like structures of band4.9 at the cell periphery while most cell centers were devoid of band4.9, and a box-like arrangement of bands of band4.9. A dissociation between colours red (actin) and green (band4.9) occurred as well. In contrast, erythrocytes suspended in a plasma/Iodixanol-320 mixture showed a membrane cytoskeleton comparable to cells suspended in autologous plasma, Similar results were found with respect to the distribution of actin. This study revealed for the first time RCM-dependent differences in band4.9 activities as possible pathophysiological mechanism for the chemotoxicity of radiographic contrast media.