RESUMO
Colonization of in-dwelling catheters by microbial biofilms is a major concern in patient health eventually leading to catheter-related blood stream infections. Biofilms are less susceptible to standard antibiotic therapies that are effective against planktonic bacteria. Standard procedure for the detection of microorganisms on the catheter tip is culture. However, viable but non-culturable cells (VBNCs) may be missed. The aim of this study was to evaluate the use of fluorescence in situ hybridization (FISH) as an indicator to visualize and quantify the effect of the antibiotics daptomycin and vancomycin on biofilms in situ. We established an in vitro catheter biofilm model of Staphylococcus epidermidis biofilms on polyurethane catheters. Biofilm activity was measured by FISH and correlated to colony forming units (CFU) data. Digital image analysis was used for quantification of total biofilm mass and the area of the FISH positive biofilm cells. FISH showed a pronounced effect of both antibiotics on the biofilms, with daptomycin having a significantly stronger effect in terms of both reduction of biofilm mass and number of FISH-positive cells. This supports the anti-biofilm capacity of daptomycin. Interestingly, neither antibiotic was able to eradicate all of the FISH-positive cells. In summary, FISH succeeded in visualization, quantification, and localization of antibiotic activity on biofilms. This technique adds a new tool to the arsenal of test systems for anti-biofilm compounds. FISH is a valuable complementary technique to CFU since it can be highly standardized and provides information on biofilm architecture and quantity and localization of survivor cells.
Assuntos
Biofilmes/efeitos dos fármacos , Daptomicina/farmacologia , Hibridização in Situ Fluorescente , Staphylococcus epidermidis/efeitos dos fármacos , Staphylococcus epidermidis/fisiologia , Vancomicina/farmacologia , Antibacterianos/farmacologia , Reatores Biológicos/microbiologia , Cateteres de Demora/microbiologia , Contagem de Colônia Microbiana , Processamento de Imagem Assistida por Computador , Staphylococcus epidermidis/crescimento & desenvolvimentoRESUMO
Flow fields are one of the key factors associated with the life threatening formation of thrombi in artificial organs. Therefore, knowledge of flow field is crucial for the design and optimization of a long-term blood pump performance. The blood chamber flow of a novel counterpulsation heart assist device (CPD) has been investigated using laser Doppler velocimetry (LDV), particle image velocimetry (PIV), and near-wall PIV (wall-PIV). The wall-PIV is an in-house developed technique assessing wall shear rates (WSR). These experimental techniques analyzed complex transient three-dimensional (3D) flow fields including major and secondary structures during the whole CPD cycle (ejection, filling, and hold time). PIV measurements in the central plane investigated an evolution (development and destruction) of the blood chamber fully filling vortex as the major CPD flow structure. The wall-PIV measurements identified areas of blood stagnation (vortex center and jet impingements) and quantified WSR at the front housing. Maximal mean WSR of 2,045 ± 605 s(-1) were found at the end of the filling. The LDV, which identified helical flow structure at the outer region of the pump, was used to complete 3D flow analysis and to combine PIV and wall-PIV results. The results suggest good washing behavior of the CPD regarding thrombus formation.
Assuntos
Coração Auxiliar , Reologia/métodos , Contrapulsação , Modelos CardiovascularesRESUMO
BACKGROUND AND PURPOSE: Coil embolization procedures change the flow conditions in the cerebral aneurysm and, therefore, in the near-wall region. Knowledge of these flow changes may be helpful to optimize therapy. The goal of this study was to investigate the effect of the coil-packing attenuation on the near-wall flow and its variability due to differences in the coil structure. MATERIALS AND METHODS: An enlarged transparent model of an ACA aneurysm was fabricated on the basis of CT angiography. The near-wall flow was visualized by using a recently proposed technique called Wall-PIV. Coil-packing attenuation of 10%, 15%, and 20% were investigated and compared with an aneurysmal flow without coils. Then the flow variability due to the coil introduction was analyzed in 10 experiments by using a packing attenuation of 15%. RESULTS: A small packing attenuation of 10% already alters the near-wall flow significantly in a large part of the aneurysmal sac. These flow changes are characterized by a slow flow with short (interrupted) path lines. An increased packing attenuation expands the wall area exposed to the altered flow conditions. This area, however, depends on the coil position and/or on the 3D coil structure in the aneurysm. CONCLUSIONS: To our knowledge, this is the first time the near-wall flow changes caused by coils in an aneurysm model have been visualized. It can be concluded that future hydrodynamic studies of coil therapy should include an investigation of the coil structure in addition to the coil-packing attenuation.
Assuntos
Angiografia Cerebral , Embolização Terapêutica , Aneurisma Intracraniano , Modelos Cardiovasculares , Circulação Cerebrovascular/fisiologia , Simulação por Computador , Feminino , Humanos , Técnicas In Vitro , Aneurisma Intracraniano/diagnóstico por imagem , Aneurisma Intracraniano/fisiopatologia , Aneurisma Intracraniano/terapia , Pessoa de Meia-Idade , Modelos Anatômicos , Nylons , Fluxo Sanguíneo Regional/fisiologiaRESUMO
BACKGROUND: Local wall shear stress (WSS) has an impact on local remodelling of the vessel wall. WSS in turn strongly depends on local geometry. Our aim was to characterize patterns of local wall shear stress associated with distinct types of remodelling in coronary arteries. Vessel size and flow rates are different between patients, however. To compare distribution patterns of WSS in analogy to fluid-dynamic modelling, non-dimensional WSS/area functions are calculated. METHODS: Right coronary arteries from seven controls, five patients with coronary artery disease (CAD) and five patients with aneurysmatic CAD (AnCAD) were analyzed. Flow simulations were performed in three-dimensionally reconstructed coronary vessels from biplane angiographic projections. Local WSS was normalized as percentage of maximum value in a histogram (100 classes) and corresponding area was expressed as percentage of total area. RESULTS: The normalized WSS distribution was characterized by a single peak with a large lower tie in controls, a loss of the single peak and a stochastic distribution in AnCAD and a narrowing of the lower tie in CAD. Correct classification of 16/17 coronary arteries was feasible by Fisher's discriminant functions based on median WSS, mean diameter, percentage of area with WSS
Assuntos
Aneurisma Coronário/diagnóstico por imagem , Aneurisma Coronário/fisiopatologia , Angiografia Coronária/métodos , Doença da Artéria Coronariana/diagnóstico por imagem , Doença da Artéria Coronariana/fisiopatologia , Adulto , Idoso , Angiografia Coronária/normas , Vasos Coronários/fisiologia , Feminino , Humanos , Imageamento Tridimensional , Masculino , Pessoa de Meia-Idade , Modelos Cardiovasculares , Reprodutibilidade dos Testes , Índice de Gravidade de Doença , Estresse Mecânico , Remodelação Ventricular/fisiologiaRESUMO
The assessment of flow along a vaulted wall (with two main finite radii of curvature) is of general interest; in biofluid mechanics, it is of special interest. Unlike the geometry of flows in engineering, flow geometry in nature is often determined by vaulted walls. Specifically the flow adjacent to the wall of blood vessels is particularly interesting since this is where either thrombi are formed or atherosclerosis develops. Current measurement methods have problems assessing the flow along vaulted walls. In contrast with conventional particle image velocimetry (PIV), this new method, called wall PIV, allows the investigation of a flow adjacent to transparent flexible surfaces with two finite radii of curvature. Using an optical method which allows the observation of particles up to a predefined depth enables the visualization solely of the boundary layer flow. This is accomplished by adding a specific dye to the fluid which absorbs the monochromatic light used to illuminate the region of observation. The obtained images can be analysed with the methods of conventional PIV and result in a vector field of the velocities along the wall. With wall PIV, the steady flow adjacent to the vaulted wall of a blood pump was investigated and the resulting velocity field as well as the velocity fluctuations were assessed.
Assuntos
Artérias/fisiologia , Velocidade do Fluxo Sanguíneo/fisiologia , Meios de Contraste , Interpretação de Imagem Assistida por Computador/métodos , Microscopia de Fluorescência/métodos , Reologia/métodos , Tamanho da PartículaRESUMO
Platelet deposition in resting blood is well researched and understood. However, the influence of hemodynamic parameters such as wall shear rate is less clear. Clinical experience and experiments show an interaction between flow and platelet deposition. But a complete understanding of the flow influence and hence a quantification has not yet been achieved. A well defined experiment of flow dependant platelet depositions is the stagnation point flow. This kind of flow is ubiquitous in the circulatory system, to be found in every bifurcation and recirculation region. These are the areas where thrombus formations are likely to occur if other conditions are also met. First, experiments were performed in a stagnation point flow chamber. A simplified blood model, platelet rich plasma, was used as a test fluid. With a microscope the platelet deposition was observed and recorded. Platelets deposit in a characteristic pattern showing the influence of the flow. An analysis of this pattern is the objective of this study and is achieved with the help of a numerical model, which is based on a convective diffusive transport. The model results in a platelet deposition pattern, which in its shape and temporal development is very similar to experimental results. Hence it is concluded that the assumed transport processes are causal for platelet depositions and thrombus formation.
Assuntos
Simulação por Computador , Hemorreologia , Modelos Cardiovasculares , Adesividade Plaquetária/fisiologia , Animais , Cães , Método de Monte Carlo , Trombose/metabolismoRESUMO
Atherosclerosis related cardiovascular diseases are the leading cause of death in western societies. The clinical manifestations are chronic arterial obstructions or acute arterial occlusions in various vascular territories. The pathogenesis is only understood in part as yet. Arterial wall abnormalities, blood composition abnormalities and hemodynamic alterations are generally accepted to be causative (Virchow's triad). The key role is played by macrophages in the subendothelial space that are activated immunologically by oxidized LDL particles via the scavenger receptor pathway. Recently, endothelial dysfunction due to oxidative stress was identified as a priming factor in the course of the development of atherosclerotic plaques. Shear stress-induced microinjuries of the endothelium in hemodynamically compromised regions together with local coagulation activation associated with microinflammation of the plaque are currently thought to cause plaque rupture. This event is the reason for local clot formation and ultimate organ infarction. Treatment success is still insufficient, however some progress during the last decade is reflected by the improving outcome of atherosclerosis associated cardiovascular diseases. Evidence from clinical trials supports the efficacy of statins, antiplatelet agents, antihypertensive agents if necessary and omega-fatty acids in patients with overt atherosclerosis. The reduction of mortality achieved by those drugs amounts to: omega-fatty acids -21%, statins -16%, anti-platelet agents -14%, treatment of hypertension -13%. It is impossible to calculate the combined effect of these modalities since in each trial participants received co-medication containing agents tested in other trials.
Assuntos
Arteriosclerose/tratamento farmacológico , Arteriosclerose/fisiopatologia , Ensaios Clínicos como Assunto , Feminino , Humanos , Masculino , Resultado do TratamentoRESUMO
Clinical applications of ventricle assist devices continue to be problematic due to thromboembolic complications. The problem originates mainly at the valves, which are usually made of an antithrombogenic material, such as cross-linked bovine pericardium. However, wherever the blood flow is stagnant or forms a recirculation region, a thrombus is likely to form. A similar blood flow is found in the space between the housing of the valve and the leaflets, the so-called valve sinus. Consequently, thrombi are often generated in this region. The novel valve design presented in this article avoids the formation of stagnant flow in the valve sinus during systole by a purge flow. This flow is taken from the main flow through the valve and is directed into each sinus region. The effect is achieved by perforation of the valve sinus with a small orifice at the bottom of the sinus. The purge flow effect is investigated with the computational fluid dynamics (CFD) method. The simulation shows that the purge flow effectively increases flow in the valve sinuses.
Assuntos
Valvas Cardíacas , Coração Auxiliar , Valvas Cardíacas/fisiopatologia , Humanos , Modelos Cardiovasculares , Modelos Teóricos , Trombose/prevenção & controleRESUMO
BACKGROUND: Hydrodynamic performance of a decellularized pulmonary porcine valve was evaluated with a computer versatile pulse duplicator and compared to glutaraldehyde fixated stentless porcine bioprosthesis and a polyurethane heart valve. METHODS: Decellularized pulmonary porcine matrices (Group I, n = 5) were treated chemically to become cell-free collagen matrices. The findings of this heart valve were compared with aortic glutaraldehyde treated porcine prostheses (Group II, n = 5) and polyurethane three leaflet valve prostheses (Group III, n = 1). Measurements were performed in 0.9% saline test fluid at room temperature. Measurements compared were closing time, closing volume, systemic pressure difference and energy losses. Each valve was measured 6 times with 70 beats/minute, a stroke volume of 70 ml corresponds to a cardiac output of 4.9 L/minute. RESULTS: Group I and group III showed no significant differences between parameters. The measured closing time was significantly different (p < 0.001) between group I and II, respectively 24.333 and 53.600 ms and group II and III respectively 53.600 and 24.000. Difference in closing volume was significant (p < 0.05) between groups II and I respectively 3.67 and 0.68 ms and group II and III respectively 3.67 and 0.71. Systolic mean pressure gradient was 18.25 +/- 1.04 mm Hg in group II which was significantly different (p < 0.001) from groups I and III, respectively 10.65 +/- 0.29 mm Hg and 7.70 +/- 0.30 mm Hg. CONCLUSIONS: Decellularized pulmonary porcine valves showed the same excellent performance as polyurethane valve prosthesis, which are superior to the investigated glutaraldehyde fixed xenograft.
Assuntos
Materiais Biocompatíveis , Bioprótese , Prótese Vascular , Glutaral , Próteses Valvulares Cardíacas/normas , Engenharia Tecidual , Animais , Próteses Valvulares Cardíacas/tendências , Teste de Materiais , Probabilidade , Desenho de Prótese , Valva Pulmonar , Fluxo Pulsátil , Sensibilidade e Especificidade , SuínosRESUMO
In patients that are in rehabilitation the physical ability is measured with stationary cycle ergometers. A new ergometer, that can be mounted easily to a bicycle, allows the mobile measurement independent of the location. The principle is based on the simultaneous measurement of chain force and chain velocity. One component of the chain force is measured, that is generated by the change of the chain's direction by pulleys. The velocity of the chain is measured by the rotational speed of one of the pulleys. The new ergometer is easy to attach and remove. The measured values are digitized, numerically processed and recorded. Through a GSM-Modem these data are transmitted to a remote central host computer and allow the control/evaluation of the patient's performance.
Assuntos
Teste de Esforço/instrumentação , Aptidão Física/fisiologia , Reabilitação/instrumentação , Processamento de Sinais Assistido por Computador/instrumentação , Desenho de Equipamento , HumanosRESUMO
The walls in blood pumps are made of artificial material and thus are thrombogenic to a lesser or larger degree. Also the flow plays a role: a blood flow with no flow separations and stagnation zones is required to avoid the generation of thrombi. A precondition for solving this problem is the assessment of the wall shear rate. However this parameter is difficult to assess because of the deformability of the walls and the pulsation of the flow. Two methods are proposed to estimate the wall shear stress in bloodpumps. The paint erosion method allows a characterisation of the flow near the wall. The second method is a special development of standard Particle Image Velocimetry (PIV). A vector field of the flow close to the wall results. Both methods should permit the assessment of the wall shear stress in bloodpumps.
Assuntos
Velocidade do Fluxo Sanguíneo/fisiologia , Coração Auxiliar , Modelos Cardiovasculares , Trombose/fisiopatologia , Humanos , Resistência ao CisalhamentoRESUMO
Objective of the present study is the hemodynamic investigation of the fluidmechanics a ball valve developed for ventricular assist devices with improved parameters. The flow was investigated using Digital Particle Image Velocimetry with an enlarged model of the valve (2.8:1). The flow was recorded using a high speed video camera with 250 fps and analyzed by a cross-correlation method implemented in the software DaVis by LaVision. The results confirmed the improvement of the ball valve performed by a numerical study. The flow separations on the ball valve housing observed in the first design version was eliminated in the final design of the novel valve. Hence a valve of this design applied in ventricular assist devices can be expected to have a lower rate of thromboembolic complications.
Assuntos
Coração Auxiliar , Hemodinâmica/fisiologia , Modelos Cardiovasculares , Fenômenos Biomecânicos , Velocidade do Fluxo Sanguíneo/fisiologia , Viscosidade Sanguínea/fisiologia , Desenho de Equipamento , HumanosRESUMO
Saccular aneurysm of the cerebral arteries occur in 1 to 5% of the population; the major risk of this disease is aneurysm rupture causing subarachnoid hemorrhage associated with a mortality rate of 50 to 60%. Two methods exist to treat cerebral aneurysms: neurosurgical clipping at the base of the aneurysm and endovascular introduction of a platin coils. Both methods have advantages and disadvantages. With the knowledge of the flow in an aneurysm it is easier to choose the appropriate method. In this study we simulated the flow in four different aneurysms. The shape of the aneurysms are gained from computertomographic data. The simulated flow in four aneurysms shows the great variability of possible flow patterns.
Assuntos
Angiografia Cerebral/métodos , Hemodinâmica/fisiologia , Processamento de Imagem Assistida por Computador/métodos , Imageamento Tridimensional/métodos , Aneurisma Intracraniano/diagnóstico por imagem , Tomografia Computadorizada por Raios X/métodos , Análise de Elementos Finitos , Humanos , Aneurisma Intracraniano/cirurgia , Fluxo Sanguíneo Regional/fisiologiaRESUMO
The carotid artery is of special interest for the pathologist because of its frequent depositions, and for the fluidmechanician because of its complex flow properties. However, there is a distinct lack in current knowledge of its geometry. Therefore, a vessel cast study was undertaken. At post mortem, the arteries are excised and filled with a special resin at the proper transmural pressure. Eighty-six vessel casts of the carotid artery were performed, and some etiological factors of atherosclerosis, such as age, sex and disease, were collected. The following selected geometric parameters of these vessel casts were measured in this study: the diameters of the main branches of carotid bifurcation (common, internal and external arteries), and the angles between internal, external and common carotid arteries. The averaged geometric parameters and their variability over 86 vessel casts of the carotid artery were investigated. Furthermore, the relationship between these measured parameters and the etiological factors age, sex and disease was analyzed. The geometric parameters varied considerably, presumably contributing to a corresponding variability in the local hemodynamic and distribution of the atherosclerotic lesions.
Assuntos
Artéria Carótida Primitiva/anatomia & histologia , Idoso , Idoso de 80 Anos ou mais , Doenças das Artérias Carótidas/patologia , Artéria Carótida Primitiva/patologia , Molde por Corrosão , Feminino , Humanos , Hipertensão/patologia , Masculino , Matemática , Pessoa de Meia-IdadeRESUMO
The blood flow immediately adjacent to the wall of a blood vessel or an artificial surface is of great interest. This flow defines the shear stress at the wall and is known to have a great physiological importance. The use of models is a viable method to investigate this flow. However, even in models the shear stress at the wall is difficult to assess. A new optical method is based on transparent models and uses particles in the model fluid, which are only visible near the wall. This is achieved with a model fluid having a defined opacity. This fluid obscures particles in the center of the models, but permits the observation and recording of particles close to the wall. The method has been applied for Hagen-Poiseuille flow and for the likewise well researched flow in a tube with a sudden expansion.
Assuntos
Artérias/fisiopatologia , Endotélio Vascular/fisiopatologia , Modelos Cardiovasculares , Hemorreologia , Humanos , Fluxo Sanguíneo Regional , Estresse MecânicoRESUMO
The influence of blood flow on the depositions and development of atherosclerotic lesions have been observed and described since the 19th century. Observations have shown that depositions correlate with regions of low wall shear stress. However, the exact correlations between depositions, vessel geometry and flow parameters are not yet known. The purpose of this study was the quantification of atherosclerosis risk factors in carotid bifurcation. This artery has attracted particular interest because lesions are often found in this bifurcation. Post mortem, the arteries are excised and vessel casts are produced. Afterwards, the arteries are analyzed morphometrically. The vessel casts are used for the assessment of some geometrical parameters. 31 carotid bifurcations were analyzed in this study. Eight vessel casts were digitized and rendered three-dimensional mathematical models of the arteries. These data were imported by the computational fluid dynamics program FLUENT. Further, the blood flow was reconstructed in a computer model based on the individual vessel geometry. The flow parameters, such as velocity, pressure and wall shear stress were computed. At the same time the geometrical parameters and wall alterations are known. This permits the comparison of the anatomical shape and its flow with the distribution and level of the wall alterations.