In vitro technique in estimation of passive mechanical properties of bovine heart part II. Constitutive relation and finite element analysis

A pseudo-elastic constitutive equation describing the mechanical properties of bovine myocardium was developed. The myocardium was modeled as a hyperelastic transversely isotropic material with a minimum viscoelastic loses. The material parameters for the proposed constitutive equations were determined using GA regression technique. In this work, the development of a constitutive equation based on principal stretch ratios is explained. The predictive capability of proposed model was compared against the experimental data obtained from part one. Finally, the constitutive equations were implemented into a commercial finite element program and the results of the mathematical model and FEM were compared with the experimental data.     

In vivo estimation of passive biomechanical properties of human myocardium

Identification of in vivo passive biomechanical properties of healthy human myocardium from regular clinical data is essential for subject-specific modelling of left ventricle (LV). In this work, myocardium was defined by Holzapfel-Ogden constitutive law. Therefore, the objectives of the study were (a) to estimate the ranges of the constitutive parameters for healthy human myocardium using non-invasive routine clinical data, and (b) to investigate the effect of geometry, LV end-diastolic pressure (EDP) and fibre orientations on estimated values. In order to avoid invasive measurements and additional scans, LV cavity volume, measured from routine MRI, and empirical pressure-normalised-volume relation (Klotz-curve) were used as clinical data. Finite element modelling, response surface method and genetic algorithm were used to inversely estimate the constitutive parameters. Due to the ill-posed nature of the inverse optimisation problem, the myocardial properties was extracted by identifying the ranges of the parameters, instead of finding unique values. Additional sensitivity studies were carried out to identify the effect of LV EDP, fibre orientation and geometry on estimated parameters. Although uniqueness of the solution cannot be achieved, the normal ranges of the parameters produced similar mechanical responses within the physiological ranges. These information could be used in future computational studies for designing heart failure treatments.

Open image in new window

Graphical abstract

     

In vitro hemolysis by mechanical heart valve prostheses with tilting disc.

Hemolytic and subhemolytic blood damage by mechanical heart valve prostheses have been observed in both clinical and in vitro investigations. A direct comparison between these studies is not possible. Nevertheless the transfer of some in vitro results to the behaviour of the valve in situ may be performed considering the similarity principle. This requires the use of dimensionless similarity numbers such as the plasma's hemoglobin concentration (PHb) or others, instead of dimensioned parameters. To evaluate the in vitro hemolysis of valve prosthesis a test chamber filled with human banked blood was used. An artificial ventricle ensuring an oscillatory flow through the valve was also used. The rise of PHb was evaluated in terms of a similarity number, called the lysis number. This number describes the probability of destroying a single red blood cell participating once in the hemolytic process under consideration. The lysis number, a Bj?rk-Shiley valve (TAD 29), was found to be in the order of 2 x 10(-4). From this, the survival time of erythrocytes in patients with an artificial heart valve was estimated. It was found to be in the order of 20 d of T50 Cr in agreement with clinical results.     

In vitro protective effect of bacteria-derived bovine alpha interferon I1 against selected bovine viruses.

We used bacteria-derived bovine alpha-interferon I1 (Bo IFN-alpha I1) to study its antiviral effect in a bovine turbinate cell line on bovine diarrhea virus, infectious bovine rhinotracheitis virus, parainfluenza 3 virus, and pseudorabies virus. We based our study upon replicate tests for each strain by using a block titration system with various concentrations of Bo IFN-alpha I1 against various concentrations of virus. The data were compiled in two-axis tables (replicate X concentration) and were statistically analyzed by the Spearman-K?rber method. An increase in the concentration of Bo IFN-alpha I1 enhanced its protective effect against every test virus strain. Bo IFN-alpha I1 had a marked in vitro effect on the bovine diarrhea viral strains. It demonstrated less protection against the pseudorabies and parainfluenza 3 viruses. Its effectiveness against the two infectious bovine rhinotracheitis viral strains was lesser and of a low order.     

In vitro characterization of neurons in the ventral part of the nucleus tractus solitarius. I. Identification of neuronal types and repetitive firing properties

1. An in vitro brain stem slice preparation from adult guinea pigs was used to determine the properties of neurons located in the ventral part of the nucleus tractus solitarius (NTS), an area associated with the dorsal respiratory group. Based upon their morphology and their repetitive firing properties, three classes of ventral NTS neurons, termed types I, II, and III, were observed. 2. Type I neurons were multipolar with pyramidal-shaped cell bodies. These neurons responded to prolonged depolarizations from a resting level of -50 mV with a discrete, high-frequency burst of spikes, which rapidly adapted to a low steady-state level. When depolarized from levels more negative than -65 mV, the initial burst was diminished. 3. Type II neurons were multipolar with fusiform-shaped cell bodies. Type II neurons responded to depolarizations from -50 mV with an initial high spike frequency, which gradually adapted to a steady-state level. When depolarized from levels more negative than -60 mV, these neurons displayed a delay between the onset of the stimulus and the first spike. This delay has been termed "delayed excitation." The expression of delayed excitation was modulated by both the size and duration of hyperpolarizing prepulses that preceded depolarization. 4. Type III neurons were multipolar with spherical shaped-cell bodies. In response to depolarizations from -50 mV, these neurons displayed high-frequency firing with little adaptation. The repetitive firing properties of type III neurons were not modulated by hyperpolarization. 5. Bulbospinal neurons in the ventral NTS were identified using retrograde transport of rhodamine-labeled latex beads injected into the region of the phrenic motor nucleus at spinal cord levels C4 through C6. Only type I and type II neurons were labeled in the ventral NTS (0.2-1.0 mm rostral to the obex). Both contralateral and ipsilateral projections were observed. Contralaterally, type I and II neurons were evenly distributed. Ipsilaterally, however, type II neurons accounted for two-thirds of the labeled neurons. 6. Type I and II neurons had similar input resistances and time constants: 97.0 +/- 17.6 M omega and 14.4 +/- 2.2 ms (n = 5) for type I and 107.0 +/- 11.2 M omega and 13.7 +/- 1.6 ms for type II (n = 5).(ABSTRACT TRUNCATED AT 400 WORDS)     

Piezoelectric and mechanical properties in bovine cornea

The piezoelectric coefficient (d(31)) and Young's modulus (E) were investigated as a function of degree of hydration for bovine cornea. The piezoelectric and mechanical responses observed were anisotropic, and d(31) decreased, whereas E increased with decreasing the degree of hydration. The anisotropic mechanical and electromechanical properties observed seem to be caused by oriented crystalline collagen fibrils. In addition, the loss of water molecules appears to decrease crystallinity (of the collagen) in the cornea. With dehydration of the cornea, a reduction in crystallinity and changes in hydrogen bonding were observed by wide-angle X-ray diffraction and Fourier transform infrared measurements. The decrease of piezoelectricity in cornea during dehydration is most likely caused by the increase in modulus and the loss of order to a nonpiezoelectric phase in the collagen.     

In vitro biodegradability and mechanical properties of bioabsorbable bacterial cellulose incorporating cellulases

Bacterially produced cellulose is being actively studied as a novel scaffold material for wound care and tissue engineering applications. Bioabsorbability of the scaffold material is desired to enable improved restoration of targeted tissue. Recently, a bioabsorbable bacterial cellulose (BBC) incorporating cellulase enzymes has been demonstrated. It was revealed that some cellulases may lose up to 90% of their activity if present in a suboptimal pH environment. Therefore, a key challenge in the practical implementation of this approach rests in compensating for the variation in the wound or tissue pH, which may significantly reduce the activity of some enzymes. In this work, buffer ingredients were incorporated into the bacterial cellulose in order to create a more optimal pH microenvironment for the preferred acid cellulases, which are significantly less active at the biological pH 7.4. The results demonstrated that incorporation of buffer ingredients helped to retain the activity of the cellulases. The glucose released from degraded materials was also increased from 30% without incorporation of buffer ingredients to 97% in the presence of incorporated buffer ingredients at the suboptimal pH environment of 7.4. The use of simulated body fluid and simulated tissue padding, both mimicking the real wound environment, also demonstrated some improvements in terms of material degradation. Measurements of mechanical properties of materials revealed that BBC materials have tensile strength and extensibility similar to human skin, especially when hydrated with saline water prior to use.     

In vitro marrow culture techniques in aplastic anaemia and related disorders.

In vitro studies of marrow growth from patients with AA have generally confirmed the hypothesis that the disease is due to stem cell injury. By studies of marrow growth from AA patients (when possible) and their relatives to detect altered sensitivity of marrow stem cells to drugs or toxins, it may be possible better to identify suspected aetiological agents, and study genetic susceptibility to toxic AA. Lack of adequate stimulation has not been shown to be aetiological in AA as both ESF and CSF levels are generally higher than normal. Serum from AA patients has not been shown to be toxic to marrow and recent data suggesting that lymphocytes mediate some cases of AA have been questioned. Further studies of committed stem cell growth and the interaction between lymphocytes and stem cells are indicated in attempting to understand the complex of diseases that result in marrow aplasia.     

In vitro mechanical properties comparsion of four room temperature curing denture base resin:

The room temperature curing denture base resin has low mechanical properties,so itwas limited in clinical application.Ithad previously disscused thatreinfored ma-chinical properties with metal fiber,glass fiber,plastic fiberand by adding the frac-ture resistance substances in powder　　The aim of this study was to determined the mechanical properitiesof fourroomtemperature curing denture base resin which had been modified performance withhigh boil pointmethacrylate.MADTERIALSAND METHOD…     

生物型人工韧带的体外检测和体内组织相容性的实验研究

目的:应用最新生化处理技术研制了一种以动物组织为原料的去抗原生物型人工韧带,检测其结构、力学性能及组织相容性,探讨应用去抗原生物型人工韧带重建膝关节叉韧带的可行性。方法:对动物肌腱/韧带组织进行环氧交联固定、多方位去抗原、力学增强改性以及表面活性修饰处理制得生物型人工韧带,应用组织学检查、扫描电镜、细胞培养及力学测试等多种方法分析生物型人工韧带的组织结构、生物力学特性和体内/体外组织相容性。结果:生物型人工韧带的外观与正常肌腱外观相似。组织学检查显示韧带为无细胞的胶原纤维。电镜检查显示韧带为走向一致的发状纤维样物紧密平行排列。异种骨髓基质细胞于韧带表面培养3周,可见细胞黏附并可见细胞分泌的基质沉积于细胞周围,韧带内部未见细胞。人工韧带平均直径约5 mm。在速度100 mm/m in条件下进行力学测试,平均最大拉力为927.19 N,抗拉强度47.22 N/mm2,与正常山羊的前交叉韧带(ACL)相应参数相近。人工韧带置换羊ACL术后12周,韧带表面可见滑膜组织覆盖并有小血管长入,术后52周人工韧带材料被新生的韧带替代,近关节面骨隧道内的人工韧带材料纤维周边可见韧带-纤维软骨-钙化软骨-骨的直接连接模式。结论:应用新的生化处理技术研制的生物型人工韧带有效地去除了猪肌腱的异种蛋白的免疫原性,具有良好的生物力学特性和组织相容性,作为膝关节叉韧带重建的韧带替换材料具有良好的应用前景。     

Decreased mechanical properties of heart valve tissue constructs cultured in platelet lysate as compared to fetal bovine serum

In autologous heart valve tissue engineering, there is an ongoing search for alternatives of fetal bovine serum (FBS). Human platelet-lysate (PL) might be a promising substitute. In the present article, we aimed to examine the tissue formation, functionality, and mechanical properties of engineered three-dimensional tissue constructs cultured in PL as a substitute for FBS. Our results show that tissue constructs that were cultured in PL and FBS produce similar amounts of collagen, glycosoaminoglycans, and collagen crosslinks, and that the cellular phenotype remains unchanged. Nevertheless, mechanical testing showed that the ultimate tensile strength in PL constructs was on average approximately three times lower as compared to FBS (0.25 vs. 0.74 MPa, respectively, p<0.01), and also the elastic modulus was almost three times lower (1.33 MPa of PL constructs vs. 3.94 MPa of FBS constructs, p<0.01). Additional tests indicated that this difference might be explained by different collagen fiber architecture possibly due to increased production of matrix-degrading proteases by cells cultured in PL. In summary, our results indicate that PL is not preferred for the culture of strong heart valve tissue constructs.     

In vitro and in vivo genotoxicity evaluation of hormonal drugs. I. Hydrocortisone

Genotoxic evaluation of a widely used glucocorticoid, hydrocortisone, was undertaken using a battery of in vitro and in vivo test systems. Human lymphocyte cultures and mouse bone marrow studies (micronuclei and sister chromatid exchange analyses) showed the drug to be very potent clastogen. However, the Ames/Salmonella assay both with and without S9 did not show an increase in the His+ revertants.     

In vitro IgE-secreting cells in man I. Mitogen-independent and -dependent subpopulations

In vitro IgE secretion by atopic and normal peripheral-blood lymphocytes was examined in culture with pokeweed mitogen or Staphylococcus aureus strain Cowan-I (StaCw) or without mitogen. IgE secreted in culture supernatants was measured with double antibody radioimmunoassay. Enumeration of IgE-secreting cells was made by a protein-A plaque assay. IgE was detected in increasing quantities in supernatants of cultured lymphocytes without mitogen up to the 12th day. IgE-plaque-forming cells were formed by the lymphocytes in large numbers on days 4–7 in cultures with mitogen. These results suggest that not only mitogen-independent but also mitogen-dependent subpopulations may exist in the IgE-secreting cells.     

In vivo mechanical properties of thoracic aortic aneurysmal wall estimated from in vitro biaxial tensile test

To investigate the mechanism of aneurysm rupture, it is necessary to examine the mechanical properties of aneurysm tissues in vivo. A new approach to evaluate in vivo mechanical properties of aortic aneurysmal tissues has been proposed in this study. The shape of the aneurysm was modeled as a sphere, and equi-biaxial stress in the in vivo state was estimated from the diameter and the wall thickness of each aneurysm and mean blood pressure of each patient. The mechanical properties of the aneurysm at the in vivo stress were estimated from its in vitro biaxial tensile properties. There were no significant correlations among maximum diameter D, wall thickness t, and mean infinitesimal strain in the in vivo state epsilon(m). This indicates the wall deformation during aneurysm development was not elastic but plastic. The mean incremental elastic modulus H(m), an index of tissue stiffness, had a significant positive correlation with elastic modulus anisotropy index K(H). This indicates the aneurysmal wall got more anisotropic in vivo as it becomes stiffer.     

In vitro exposure of preimplantation bovine embryos to vesicular stomatitis virus.

Vesicular stomatitis virus New Jersey serotype (VSV-NJ) adhered to 14 of 20 zonae pellucidae intact (ZP-I) bovine embryos exposed in vitro. The VSV-NJ-exposed ZP-I bovine embryos were washed by a single- or multiple-pipette procedure. The multiple-pipette washing procedure was more efficient in removing unattached virus than the single-pipette procedure, but neither washing procedure was effective in consistently removing attached virus from ZP-I embryos. The virus plaque assay with Vero-MARU cells was more sensitive than was the suckling mouse intracerebral inoculation procedure for detection of VSV-NJ from the sonic extracts of bovine embryos. A maximum of 15 infective VSV-NJ particles were detected adhering to one virus-exposed, washed ZP-I bovine embryo.     

Analysis of passive and active electrophysiologic properties of neurons in mammalian nodose ganglia maintained in vitro.

1. We studied the passive and active electrical properties of the soma membrane of neurons in nodose ganglia removed from cats and rabbits and maintained in vitro. The ganglia were superfused at 37 degrees C with a solution formulated to approximate the extracellular fluid of each species. The solution was buffered to pH 7.34, continuously equilibrated with 95% O2 and 5% CO2, and contained dialyzed calf serum and glucose. We also examined these properties in nodose ganglion neurons in vivo. Intracellular recordings were obtained with glass micropipettes filled with either 3 M KCl or 5 M K acetate. 2. We determined mean values for a variety of passive and active electrophysiologic properties. Values obtained in vitro did not differ significantly from those obtained in vivo. Based on the passive electrical properties of the soma membrane, neurons in the nodose ganglion appear to be a uniform population, despite the different sensory modalities conveyed by the afferent fibers. 3. Cell bodies of neurons generated action potentials in response to impulses in their afferent fibers. Somatic spikes could be evoked by stimulation of either the supranodose or infranodose vagus nerve, and an inflection point could be seen on their rising phase. When the vagus nerve was stimulated at frequencies greater than 10-20 Hz, the generation of somatic spikes often became progressively delayed and then failed completely, leaving a smaller potential (IS spike) which was apparently generated in the initial complex. The afterhyperpolarization was associated only with the somatic spike. 4. Many neurons, both in vitro and in vivo, developed a persistent hyperpolarization when repetitive action potentials occurred in the soma. This hyperpolarization was apparent at frequencies as low as 1-2 Hz, persisted for up to 5 s after the occurrence of the last somatic spike, and sometimes caused failure of somatic spikes to be generated. 5. Neurons in both species differed in their responses to suprathreshold depolarization applied through the recording electrode. Some neurons produced a train of action potentials which lasted for the duration of the depolarizing pulse, the frequency of the train being related to the magnitude of depolarization. The trains were characterized by gradually decreasing spike amplitudes and increasing interspike intervals. Other neurons responded with only a single spike or brief burst of action potentials at the beginning of depolarization to threshold. 6. It is suggested that the adaptive properties of the soma membrane of a peripheral sensory neuron are similar to those of its sensory ending, and that electrophysiological studies of the soma membrane may provide an opportunity to examine mechanisms of receptor adaptation.