The input impedance of the peripheral vascular termination in skeletal muscle

In order to investigate the dynamic behaviour of the peripheral vessels of the arterial system, simultaneous pressure-flow measurements were made in the deep femoral artery of the rabbit at the same site. The input impedance calculated from the recorded pulses was regarded as representing the peripheral vascular termination in skeletal muscle and was expressed in terms of modulus and phase. The modulus decreases monotonously with increasing frequency while the phase angle is about –45° in the low frequency range and becomes less negative with increasing frequency. This behaviour can be simulated by a model in which the inert mass of the pulsating blood, the elastic compliance of the blood vessels, and the frictional resistance to blood flow are taken into account. An essential result is that the model is provided with a considerable degree of elastic compliance, which means that the peripheral vessels as a whole possess an important compliance and do not behave like pure frictional resistances.To Prof. Dr. Erik Wetterer on the occasion of his 75th birthday     

Reproducibility of reference tissue quantification of dynamic contrast-enhanced data: comparison with a fixed vascular input function

Reference tissues are currently used to analyse dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) data. The assessment of tumour response to treatment with anti-cancer drugs is a particularly important application of this type of analysis and requires a measure of reproducibility to define a level above which a significant change due to therapy can be inferred. This study compares the reproducibility of such quantification strategies with that found using a published, group-averaged uptake curve. It is shown that reference tissue quantification gives poorer reproducibility for most parameters than that found using a group-averaged plasma curve (a change in K(trans) of greater than 41.8% and 16.4% would be considered significant in the two approaches, respectively), but successfully incorporates some of the variability observed in plasma kinetics between visits and provides vascular input functions that, across the group, are comparable with the group-averaged curve. This study therefore provides an indirect validation of the methodology.     

一种血管张应力体外加载装置的实验研究

目的研制一种血管张应力体外加载装置,研究弹性基底(硅胶片)上的张应力、张应变分布。方法基于基底形变加载技术,研制一种接近人体血液动力学环境的血管张应力体外加载装置。利用装置中的摄像机拍摄硅胶片拉伸前后硅胶片网格点的图像并转化为数字图像,使用Matlab软件对网格点的位置特征进行计算,从而得到硅胶片的应变分布。利用万能材料试验机对硅胶片进行实验和计算得到硅胶片的力学参数,根据力学参数建立有限元模型,并对硅胶片的张应力、张应变分布进行模拟计算。将实验结果和模拟结果进行比较。结果有限元结果和实验结果基本一致,张应力、张应变的最大值均出现在加载点处,中间区域应力、应变较为均匀。硅胶片中间60%面积区域可视为均匀应变场。结论研究结果可为后期血管壁内皮细胞的动态培养以及细胞力学研究提供实验技术。     

The influence of mock circulation input impedance on valve acceleration during in vitro cardiac device testing

For a mechanical heart valve, a strong spike in pressure during closing is associated with valve wear and erythrocyte damage; thus, for valid in vitro testing, the mock circulation system should replicate the conditions, including pressure spikes, expected in vivo. To address this issue, a study was performed to investigate how mock circulation input impedance affects valve closure dynamics. A left ventricular model with polyurethane trileaflet inflow valve and tilting disc outflow valve was connected to a Louisville mock circulation system, which incorporates 2 adjustable flow resistors and 2 compliances. In the study, 116 cases matched zero frequency modulus well (982-1147 dyn x s/cm), but higher harmonics were purposely varied. Acceleration measured at the outflow valve ring (42.4-89.4 milli-Gs) was uncorrelated with impedance error (74.1-237 dyn x s/cm relative to target impedance), but was correlated with end-systolic impedance (1082-1319 dyn x s/cm) for cases with high zero frequency modulus, which exhibited just less than full ejection. These differences demonstrate that mock circulation response affects the magnitude of the closing spike, indicating that control of this parameter is necessary for authentic testing of valves. Correlation of acceleration to end-systolic impedance was weak for low zero frequency modulus, which tended toward full or hyperejection, reinforcing common laboratory observations that valve closing also depends on ventricular operating conditions.     

A method for interleaved acquisition of a vascular input function for dynamic contrast-enhanced MRI in experimental rat tumours

Dynamic contrast-enhanced MRI is widely used for the evaluation of the response of experimental rodent tumours to antitumour therapy, particularly for the newly developing antiangiogenic and antivascular agents. However, standard models require a time-course for the plasma concentration of contrast agent (usually referred to as the arterial input function) to calculate the transfer constant K(trans) from the dynamic time-course data. Ideally, the plasma concentration time-course should be measured during each experiment to obtain the most accurate measure of K(trans). This is technically difficult in rodents, so assumed values are generally used. A method is presented here using interleaved acquisitions from a tail coil to obtain the plasma concentration simultaneously with DCE-MRI data obtained from a solenoid coil around the tumour. The SNR of the resulting vascular input function data is high compared with methods using a volume coil to acquire plasma concentrations from the aorta and vena cava.     

Human embryonic stem cells as an in vitro model for human vascular development and the induction of vascular differentiation

Early embryonic blood vessels are typically composed of fragile tubes of endothelial cells encircled by vascular smooth muscle cells. Early human vasculogenesis was explored in spontaneous and directed differentiation models derived from human embryonic stem (HES) cells. In a 3-dimensional (3D) model, HES cells were studied for their potential for vascular differentiation during the spontaneous formation of embryoid bodies. Directed differentiation was investigated by means of a 2-dimensional (2D) differentiation method to promote vascular differentiation from HES cells (without the formation of embryoid bodies). Using this latter approach, up-regulation of early lineage markers of endothelial progenitors were induced. Additional culture under strict conditions and exposure to angiogenic growth factors resulted in a prolonged differentiation pathway into mature endothelial cells and up-regulation of vascular smooth muscle cell markers. The use of 3D collagen gels and Matrigel assays for the induction and inhibition of human vascular sprouting in vitro further established the vascular potential of the cells generated by the 2D differentiation system. Our study shows that HES cells can provide useful models to study early differentiation and development of blood vessels. Moreover, the 2D differentiation model facilitates both the production of vascular lineage cells from HES cells for various potential therapeutic applications and also provides a model for studying the mechanisms involved in early human embryonic blood vessel development.     

Characterization of human erythrocytes as potential carrier for pravastatin: an in vitro study

Drug delivery systems including chemical, physical and biological agents that enhance the bioavailability, improve pharmacokinetics and reduce toxicities of the drugs. Carrier erythrocytes are one of the most promising biological drug delivery systems investigated in recent decades. The bioavailability of statin drugs is low due the effects of P-glycoprotein in the gastro-intestinal tract as well as the first-pass metabolism. Therefore in this work we study the effect of time, temperature as well as concentration on the loading of pravastatin in human erythrocytes to be using them as systemic sustained release delivery system for this drug. After the loading process is performed the carriers' erythrocytes were physically and cellulary characterized. Also, the in vitro release of pravastatin from carrier erythrocytes was studied over time interval. Our results revealed that, human erythrocytes have been successfully loaded with pravastatin using endocytosis method either at 25(o)C or at 37(o)C. The loaded amount at 10 mg/ml is 0.32 mg/0.1 ml and 0.69 mg/0.1 ml. Entrapment efficiency is 34% and 94% at 25(o)C and 37(o)C respectively at drug concentration 4 mg/ml. Moreover the percent of cells recovery is 87-93%. Hematological parameters and osmotic fragility behavior of pravastatin loaded erythrocytes were similar that of native erythrocytes. Scanning electron microscopy demonstrated that the pravastatin loaded cells has no change in the morphology. Pravastatin releasing from carrier cell was 83% after 23 hours in phosphate buffer saline and decreased to 72% by treatment of carrier cells with glutaraldehyde. The releasing pattern of the drug from loaded erythrocytes obeyed first order kinetics. It concluded that pravastatin is successfully entrapped into erythrocytes with acceptable loading parameters and moderate morphological changes, this suggesting that erythrocytes can be used as prolonged release for pravastatin.     

Wave transmission and input impedance of a model of skeletal muscle microvasculature

We analyzed wave transmission properties and input impedance of a microvascular network model. The model, derived from rat spinotrapezius muscle and previously described and validated by other investigators for steady pressure-flow relations, was expanded to include pulsatile phenomena. Microvessels are considered purely elastic, with compliances a function of vessel type; viscous dissipation follows Poiseuille's law. Linear and nonlinear results are presented. In the nonlinear case, shear rate-dependent viscosity of blood and transmural pressure-dependent vascular diameters were calculated and small signal perturbations were imposed around several working points. We investigated effects on input impedance of physiological variability of network parameters and structure: distribution of capillary diameters, capillary segment length, and presence or absence of cross-connecting capillaries. Results show that although wave transmission properties are complex, input impedance is simple. Apparent wave speeds differ substantially from phase velocities and change markedly from branch to branch; pressure and flow waves appear to travel at different speeds. These features result from the mesh-like structure of the network and the prominence of reflection at branchpoints. Input impedance displays a similar form under all conditions: Magnitude is a monotonically decreasing function of frequency, and phase decreases from 0 to approximately −45°. Consideration of the characteristic impedance of a microvessel leads to modification of the three-element Windkessel as a reduced model of the observed input impedance.     

In vitro study of human vascular endothelial cell function on materials with various surface roughness

In recent years, creating a biodegradable polymer scaffold with an endothelialized surface has become an attractive concept for replacement of small-diameter blood vessels. Toward this end, a better understanding of the interaction between endothelial cells and biodegradable polymer substrates is particularly important. Surface roughness of biomaterials is one of the important parameters that affect cell behavior. In this study, human vascular endothelial cells were cultured on electrospun and solvent-cast poly(L-lactic acid) substrates with different surface roughness. Cell responses were evaluated via both qualitative examinations of cell morphology changes as well as quantitative assessment of cell adhesion and proliferation rate on the different substrates. The results proved that endothelial cell function was enhanced on the smooth solvent-cast surface rather than on the rough electrospun surface of poly(L-lactic acid). Together with our previous findings that electrospun substrates favor vascular smooth muscle cell behavior, it is possible to design a unique three-dimensional scaffold for application of tissue-engineered small-diameter vessel replacement by combining the fabrication technique of solvent casting and electrospinning.     

Function of a new internal bioartificial liver: an in vitro study

The goal of this study was to determine whether a new internal bioartificial liver utilizing porcine hepatocytes can perform detoxification and other metabolic functions. Such a system might aid in treating patients with moderate to severe liver failure and prolong patient survival until a matching organ is found for transplantation. Porcine hepatocytes were attached to a microcarrier and an internal artificial liver was constructed by perfusing the hepatocytes into a polysulfon hollow fiber. The 4 experimental groups were: (a) control group, (b) microcarrier group, (c) hollow fiber group, and (d) internal bioartificial liver group. Viability of hepatocytes, alanine aminotransferase (ALT) and lactate dehydrogenase (LD) activities in the medium, urea production, diazepam transformation, protein synthesis, and glucose-6-phosphatase activity of cells were monitored during a 7-day culture period. Viability of porcine hepatocytes in the internal bioartificial liver group was maintained at >80% during the culture period, and alanine aminotransferase and lactate dehydrogenase activities did not fluctuate significantly. These enzyme activities were significantly lower in the internal bioartificial liver group than in the control or microcarrier groups. Urea production, diazepam transformation, [3H]-leucine incorporation, and glucose-6-phosphatase activity were significantly higher in the internal bioartificial liver group than in the control and hollow fiber groups. These results show that the new internal bioartificial liver produces small amounts of ALT and LD and exhibits detoxification and protein synthetic functions.     

Quantitative assessment of cerebral autoregulation from transcranial Doppler pulsatility: a computer simulation study

Transcranial Doppler (TCD) ultrasonography is largely used today to achieve non-invasive assessment of cerebral autoregulation and cerebrovascular reactivity in neurosurgical patients. Recent experimental and clinical studies suggest that not only the pattern of mean velocity, but also velocity pulse amplitude alterations during changes in cerebral perfusion pressure (CPP) contain information on autoregulation status. The aim of this work is to investigate the relationship between cerebral autoregulation and TCD pulsatility by means of a comprehensive mathematical model of intracranial dynamics and cerebrovascular regulation. Simulation results, performed using different values of the most important clinical parameters of the model (autoregulation strength, cerebrospinal fluid (CSF) outflow resistance and intracranial elastance coefficient) show that velocity pulse amplitude increases with a reduction in CPP in patients with intact autoregulation, whereas changes in velocity pulsatility are modest in patients with weak autoregulation. Finally, velocity pulse amplitude decreases during a CPP reduction in patients with impaired autoregulation. Moreover, the relationship between the velocity pulse amplitude changes and autoregulation strength is almost linear in a wide range of CPP values, and is scarcely affected by changes in CSF circulation and intracranial elasticity. Starting from these results, we suggest a new quantitative index to assess autoregulation strength, i.e. G(aut)% = (s-b)/a, where G(aut)% is autoregulation strength (100% means intact autoregulation, 0% means impaired autoregulation), a approximately -0.03; b approximately 1.5 and s is the slope of the relationship ' percentage changes of velocity pulse amplitude to arterial pressure pulse amplitude vs. CPP changes'.     

Quantification of bacterial mass recovery as a function of pore-water velocity and ionic strength

Transport of bacteria in aquifer systems plays an important role in bioaugmentation, which relies upon successful bacterial delivery to a target area. In the present study, we conducted a set of laboratory column experiments under various conditions of pore-water velocity (upsilon(omega)) and ionic strength (IS) of culture medium for Pseudomonas aeruginosa, known to be a benzene-degrading bacteria, in order to investigate their relationship to mass recovery in saturated quartz sands. The column experiments revealed that both peak concentrations and mass recoveries of bacteria were lower than those of a conservative tracer KCl when deionized water was used as leaching water for all ranges of pore-water velocity (0.18-6.23 cm/min). Thus, the parameter responsible for transport of P. aeruginosa was only the deposition coefficient. Bacterial cells could not be attached to the mineral surfaces by predominance of electrostatic charge or repulsive forces over hydrophobicity or attractive forces due to the very low ionic strength ( approximately 0 mM) of deionized water. The loss of bacterial mass was attributed to the deposition in the crevice formed on the quartz surfaces, as evidenced by SEM images. For a given pore-water velocity, the ionic strength markedly influenced bacterial deposition, showing decreased peak concentrations and mass recoveries with increasing ionic strength of column leaching water. An optimum range of upsilon(omega) and IS for achieving bacterial mass recovery higher than 70% in the studied quartz sand was found such that: (i) at low IS ( approximately 0 mM), a pore-water velocity higher than 0.30 cm/min, and (ii) at pore-water velocity of 0.52 cm/min, an IS lower than 290 mM, were required, respectively.     

Bone-particle-impregnated bone cement: an in vitro study

Bone-particle-impregnated bone cement specimens (up to 30% by weight) were characterized by various test methods. The experimental bone cement showed decreased crack propagation rates and increased Young's modulus, while the ultimate tensile strength and impact strength were decreased. The viscosity could be adjusted by adding initiators lost when substituting the PMMA powder with bone particles. The present study warranted further in vivo experiments on the possibility of tissue ingrowth for which the new bone cement was developed.     

Cell swelling, seizures and spreading depression: an impedance study

The cellular processes that take place during the transition from pre-seizure state to seizure remain to be defined. In this study in awake, paralyzed rats, we used an electrical impedance measure of changes in extra-cellular intracranial volume to estimate changes in cell size in acute models of epilepsy. Animals were prepared with extradural electroencephalographic (EEG)/impedance electrodes and a venous catheter. On a subsequent day, animals were paralyzed, ventilated and treated with picrotoxin, kainic acid or fluorocitrate in doses that usually induce epileptiform discharges. We now report that increases in baseline impedance were induced by kainic acid and smaller increases by picrotoxin. We also demonstrated that epileptiform discharges were preceded by small, accelerated increases in impedance. Increases in baseline impedance were highly correlated with increases in power of non-ictal high frequency EEG activity. Seizures were accompanied by increases in impedance and all treatments induced transient, relatively large, increases in impedance often associated with unilateral reductions in low frequency EEG, likely periods of spreading depression. We conclude: cerebral cells swell in convulsant models of epilepsy, that there are pre-ictal accelerations in cell swelling, and that spreading depression-like events are frequently associated with seizures.