Drug-based mobilisation of mesenchymal stem/stromal cells improves cardiac function post myocardial infarction

There is an unmet need for treatments that prevent the progressive cardiac dysfunction following myocardial infarction. Mesenchymal stem/stromal cells (MSCs) are under investigation for cardiac repair; however, culture expansion prior to transplantation is hindering their homing and reparative abilities. Pharmacological mobilisation could be an alternative to MSC transplantation. Here, we report that endogenous MSCs mobilise into the circulation at day 5 post myocardial infarction in male Lewis rats. This mobilisation can be significantly increased by using a combination of the FDA-approved drugs mirabegron (β3-adrenoceptor agonist) and AMD3100 (CXCR4 antagonist). Blinded cardiac magnetic resonance imaging analysis showed the treated group to have increased left ventricular ejection fraction and decreased end systolic volume at 5 weeks post myocardial infarction. The mobilised group had a significant decrease in plasma IL-6 and TNF-α levels, a decrease in interstitial fibrosis, and an increase in the border zone blood vessel density. Conditioned medium from blood-derived MSCs supported angiogenesis in vitro, as shown by tube formation and wound healing assays. Our data suggest a novel pharmacological strategy that enhances myocardial infarction-induced MSC mobilisation and improves cardiac function after myocardial infarction

Ventricular fibrillation mechanism and global fibrillatory organisation are determined by gap junction coupling and fibrosis pattern

Aims Conflicting data exist supporting differing mechanisms for sustaining ventricular fibrillation (VF), ranging from disorganised multiple-wavelet activation to organised rotational activities (RAs). Abnormal gap junction (GJ) coupling and fibrosis are important in initiation and maintenance of VF. We investigated whether differing ventricular fibrosis patterns and the degree of GJ coupling affected the underlying VF mechanism. Methods and Results Optical mapping of 65 Langendorff-perfused rat hearts was performed to study VF mechanisms in control hearts with acute GJ modulation, and separately in three differing chronic ventricular fibrosis models; compact (CF), diffuse (DiF) and patchy (PF). VF dynamics were quantified with phase mapping and frequency dominance index (FDI) analysis, a power ratio of the highest amplitude dominant frequency in the cardiac frequency spectrum. Enhanced GJ coupling with rotigaptide (n = 10) progressively organised fibrillation in a concentration-dependent manner; increasing FDI (0nM: 0.53±0.04, 80nM: 0.78±0.03, p < 0.001), increasing RA sustained VF time (0nM:44±6%, 80nM: 94±2%, p < 0.001) and stabilised RAs (maximum rotations for a RA; 0nM:5.4±0.5, 80nM: 48.2±12.3, p < 0.001). GJ uncoupling with carbenoxolone progressively disorganised VF; the FDI decreased (0µM: 0.60±0.05, 50µM: 0.17±0.03, p < 0.001) and RA-sustained VF time decreased (0µM: 61±9%, 50µM: 3±2%, p < 0.001). In CF, VF activity was disorganised and the RA-sustained VF time was the lowest (CF: 27±7% versus PF: 75±5%, p < 0.001). Global fibrillatory organisation measured by FDI was highest in PF (PF: 0.67±0.05 versus CF: 0.33±0.03, p < 0.001). PF harboured the longest duration and most spatially stable RAs (patchy: 1411±266ms versus compact: 354±38ms, p < 0.001). DiF (n = 11) exhibited an intermediately organised VF pattern, sustained by a combination of multiple-wavelets and short-lived RAs. Conclusion The degree of GJ coupling and pattern of fibrosis influences the mechanism sustaining VF. There is a continuous spectrum of organisation in VF, ranging between globally organised fibrillation sustained by stable RAs and disorganised, possibly multiple-wavelet driven fibrillation with no RAs. Translational perspective Multiple competing mechanisms have been proposed for sustaining VF. We reframed conflicting mechanisms reported in sustaining fibrillation and defined them as part of a continuum of varying global organisation, with some sustained by stable rotationalactivities. The underlying cardiac electroarchitecture, namely gap junction coupling and fibrosis, were important determinants of the VF mechanism. Characterising the VF mechanism and its relationship to the cardiac electroarchitecture may facilitate a patient-tailored treatment approach towards VF prevention in VF survivors. Organised fibrillation sustained by stable rotational activities could be considered for targeted ablation. Disorganised fibrillation dynamics may be better suited for conventional pharmacotherapy

Минеральные воды и их рациональное использование в гастроэнтерологии и гепатологии

Department of Pharmacology and Clinical Pharmacology, Nicolae Testemitanu State Medical and Pharmaceutical University, Congresul III al Medicilor de Familie din Republica Moldova, 17–18 mai, 2012, Chişinău, Republica Moldova, Conferinţa Naţională „Maladii bronhoobstructive la copii”, consacrată profesorului universitar, doctor habilitat Victor Gheţeul, 27 aprilie, Chişinău, Republica MoldovaThe rational utilization of mineral water in gastroenterology and hepatology is based on the study of the medicinal and prophylactic properties depending on the composition, mineral degree, and the effects of its components. Type, volume, and temperature of the mineral water determine the dosage regime in the stomach, gut, liver, and biliary duct’s pathologies.Рациональное применение минеральных вод в гастроэнтерологии и гепатологии основано на изучении лечебных и профилактических эффектов в зависимости от состава, степени минерализации и эффектов компонентов. Тип, объём, и температура минеральной воды определяют режим дозирования при соответствующей патологии желудка, кишечника, печени и желчевыводящих путей

From biomechanics to pathology: predicting axonal injury from patterns of strain after traumatic brain injury

The relationship between biomechanical forces and neuropathology is key to understanding traumatic brain injury. White matter tracts are damaged by high shear forces during impact, resulting in axonal injury, a key determinant of long-term clinical outcomes. However, the relationship between biomechanical forces and patterns of white matter injuries, associated with persistent diffusion MRI abnormalities, is poorly understood. This limits the ability to predict the severity of head injuries and the design of appropriate protection. Our previously developed human finite element model of head injury predicted the location of post-traumatic neurodegeneration. A similar rat model now allows us to experimentally test whether strain patterns calculated by the model predicts in vivo MRI and histology changes. Using a Controlled Cortical Impact, mild and moderate injuries (1 and 2 mm) were performed. Focal and axonal injuries were quantified with volumetric and diffusion 9.4T MRI two weeks post injury. Detailed analysis of the corpus callosum was conducted using multi-shell diffusion MRI and histopathology. Microglia and astrocyte density, including process parameters, along with white matter structural integrity and neurofilament expression were determined by quantitative immunohistochemistry. Linear mixed effects regression analyses for strain and strain rate with the employed outcome measures were used to ascertain how well immediate biomechanics could explain MRI and histology changes. The spatial pattern of mechanical strain and strain rate in the injured cortex shows good agreement with the probability maps of focal lesions derived from volumetric MRI. Diffusion metrics showed abnormalities in segments of the corpus callosum predicted to have a high strain, indicating white matter changes. The same segments also exhibited a severity-dependent increase in glia cell density, white matter thinning and reduced neurofilament expression. Linear mixed effects regression analyses showed that mechanical strain and strain rate were significant predictors of in vivo MRI and histology changes. Specifically, strain and strain rate respectively explained 33% and 28% of the reduction in fractional anisotropy, 51% and 29% of the change in neurofilament expression and 51% and 30% of microglia density changes. The work provides evidence that strain and strain rate in the first milliseconds after injury are important factors in determining patterns of glial and axonal injury and serve as experimental validators of our computational model of TBI. Our results provide support for the use of this model in understanding the relationship of biomechanics and neuropathology and can guide the development of head protection systems, such as airbags and helmets

The feasibility and superiority of high frame rate strain imaging compared to ejection fraction in a rat model of ischamia-reperfusion myocardial infarction using cardiac MRI

Background: In the fields of cardiac regeneration and cardioprotection, robust quantification of the cardiac function is paramount for the assessment of the effects of experimental interventions on the heart. We hypothesized the feasibility of performing cardiac MRI strain imaging and assessed its superiority to ejection fraction (EF) in I/R MI rats. Methods: Lewis rats (normal versus 24–48-hour post I/R MI, each n = 6) were imaged using cine FLASH and high-frame rate IntraGate cine (Bruker BioSpec 9.4T MRI, Ettlingen) conforming to AHA's planes recommendations. MI was confirmed using multi-slice IR LGE. Segment (Medviso AB, Lund) and 2D CPA MR (Tomtec) were used respectively for volumes measurements and strain analysis. Results: Normal rat versus MI rats: EF (mean 64.0%, SD 2.3% versus mean 59.5%, SD 5.1%), p = 0.089; endo peak GLS (global longitudinal strain) (mean -21.9.0%, SD 1.1% vs. -14.1%, SD 5.5%), p = 0.017; endo peak GCS (global circumferential strain) (mean -34.0%, SD 1.4% versus mean -26.1%, SD 6.4%), p = 0.028; myo peak GCS (mean -23.5%, SD 1.7% vs -17.0%, SD 5.4%), p = 0.032; GRS (global radial strain) (mean 45.8%, SD 5.3% versus mean 33.7%, SD 9.8%), p = 0.029. Conclusion: Strain imaging indices show statistically significant changes after MI where EF does not. A small change in EF was observed despite large infarct sizes. Increase in heart rate and alterations of remote regional strain and tissue velocities (data not shown) are suggested to be compensatory mechanisms which preserve EF

Sensitivity of intervertebral disc finite element models to internal geometric and non-geometric parameters

Finite element models are useful for investigating internal intervertebral disc (IVD) behaviours without using disruptive experimental techniques. Simplified geometries are commonly used to reduce computational time or because internal geometries cannot be acquired from CT scans. This study aimed to 1) investigate the effect of altered geometries both at endplates and the nucleus-anulus boundary on model response, and 2) to investigate model sensitivity to material and geometric inputs, and different modelling approaches (graduated or consistent fibre bundle angles and glued or cohesive interlamellar contact). Six models were developed from 9.4T MRIs of bovine IVDs. Models had two variations of endplate geometry (a simple curved profile from the centre of the disc to the periphery, and precise geometry segmented from MRIs), and three variations of NP-AF boundary (linear, curved, and segmented). Models were subjected to axial compressive loading (to 0.86mm at a strain rate of 0.1/sec) and the effect on stiffness and strain distributions, and the sensitivity to modelling approaches was investigated. The model with the most complex geometry (segmented endplates, curved NP-AF boundary) was 3.1 times stiffer than the model with the simplest geometry (curved endplates, linear NP-AF boundary). Peak strains were close to the endplates at locations of high curvature in the segmented endplate models which were not captured in the curved endplate models. Differences were also seen in sensitivity to material properties, graduated fibre angles, cohesive rather than glued interlamellar contact, and NP:AF ratios. These results show that FE modellers must take care to ensure geometries are realistic so that load is distributed and passes through IVDs accurately

The Effect of Degeneration on Internal Strains and the Mechanism of Failure in Human Intervertebral Discs Analyzed Using Digital Volume Correlation (DVC) and Ultra-High Field MRI

The intervertebral disc (IVD) plays a main role in absorbing and transmitting loads within the spinal column. Degeneration alters the structural integrity of the IVDs and causes pain, especially in the lumbar region. The objective of this study was to investigate non-invasively the effect of degeneration on human 3D lumbar IVD strains (n = 8) and the mechanism of spinal failure (n = 10) under pure axial compression using digital volume correlation (DVC) and 9.4 Tesla magnetic resonance imaging (MRI). Degenerate IVDs had higher (p < 0.05) axial strains (58% higher), maximum 3D compressive strains (43% higher), and maximum 3D shear strains (41% higher), in comparison to the non-degenerate IVDs, particularly in the lateral and posterior annulus. In both degenerate and non-degenerate IVDs, peak tensile and shear strains were observed close to the endplates. Inward bulging of the inner annulus was observed in all degenerate IVDs causing an increase in the AF compressive, tensile, and shear strains at the site of inward bulge, which may predispose it to circumferential tears (delamination). The endplate is the spine's “weak link” in pure axial compression, and the mechanism of human vertebral fracture is associated with disc degeneration. In non-degenerate IVDs the locations of failure were close to the endplate centroid, whereas in degenerate IVDs they were in peripheral regions. These findings advance the state of knowledge on mechanical changes during degeneration of the IVD, which help reduce the risk of injury, optimize treatments, and improve spinal implant designs. Additionally, these new data can be used to validate computational models

Multiscale modelling of cerebrovascular injury reveals the role of vascular anatomy and parenchymal shear stresses

Neurovascular injury is often observed in traumatic brain injury (TBI). However, the relationship between mechanical forces and vascular injury is still unclear. A key question is whether the complex anatomy of vasculature plays a role in increasing forces in cerebral vessels and producing damage. We developed a high-fidelity multiscale finite element model of the rat brain featuring a detailed definition of the angioarchitecture. Controlled cortical impacts were performed experimentally and in-silico. The model was able to predict the pattern of blood–brain barrier damage. We found strong correlation between the area of fibrinogen extravasation and the brain area where axial strain in vessels exceeds 0.14. Our results showed that adjacent vessels can sustain profoundly different axial stresses depending on their alignment with the principal direction of stress in parenchyma, with a better alignment leading to larger stresses in vessels. We also found a strong correlation between axial stress in vessels and the shearing component of the stress wave in parenchyma. Our multiscale computational approach explains the unrecognised role of the vascular anatomy and shear stresses in producing distinct distribution of large forces in vasculature. This new understanding can contribute to improving TBI diagnosis and prevention

Characterization of acute TLR-7 agonist-induced hemorrhagic myocarditis in mice by multi-parametric quantitative cardiac MRI

Hemorrhagic myocarditis is a potentially fatal complication of excessive levels of systemic inflammation. It has been reported in viral infection, but is also possible in systemic autoimmunity. Epicutaneous treatment of mice with the TLR-7 agonist Resiquimod induces auto-antibodies and systemic tissue damage including in the heart, and is used as an inducible mouse model of Systemic Lupus Erythematosus (SLE). Here, we show that over-activation of the TLR-7 pathway of viral recognition by Resiquimod-treatment of CFN mice induces severe thrombocytopenia and internal bleeding which manifests most prominently as hemorrhagic myocarditis. We optimized a cardiac magnetic resonance (CMR) tissue mapping approach for the in vivo detection of diffuse infiltration, fibrosis and hemorrhages using a combination of T1, T2 and T2* relaxation times, and compared results to ex vivo histopathology of cardiac sections corresponding to CMR tissue maps. This allowed a detailed correlation between in vivo CMR parameters and ex vivo histopathology, and confirmed the need to include T2* measurements to detect tissue iron for accurate interpretation of pathology associated with CMR parameter changes. In summary, we provide detailed histological and in vivo imaging-based characterization of acute hemorrhagic myocarditis as acute cardiac complication in the mouse model of Resiquimod-induced SLE, and a refined CMR protocol to allow non-invasive longitudinal in vivo studies of heart involvement in acute inflammation. We propose that adding T2* mapping to CMR protocols for myocarditis diagnosis will improve interpretation of disease mechanisms and diagnostic sensitivity

Myocardial damage induced by a single high dose of isoproterenol in C57BL/6J mice triggers a persistent adaptive immune response against the heart

Heart failure is the common final pathway of several cardiovascular conditions and a major cause of morbidity and mortality worldwide. Aberrant activation of the adaptive immune system in response to myocardial necrosis has recently been implicated in the development of heart failure. The ß-adrenergic agonist isoproterenol-hydrochloride isused for its cardiac effects in a variety of different dosing regimens with high doses causing acute cardiomyocyte necrosis.To assess if isoproterenol-induced cardiomyocyte necrosistriggersan adaptive immune response against the heart, we treated C57BL/6J mice with a single intraperitoneal injection of 160mg/kg isoproterenol. We confirmed tissue damage reminiscent of human type 2 myocardial infarction. This is followed by an adaptive immune response targeting the heart as demonstrated by the activation of T cells, the presence of anti-heart auto-antibodies in the serum, as late as 12 weeks after initial challenge and IgG deposition in the myocardium. All of these are hallmark signs of an established autoimmune response. Adoptive transfer of splenocytes from isoproterenol-treated mice induces left ventricular dilation and impairs cardiac function in healthy recipients. In summary, a single administration of a high dose of isoproterenol is a suitable high-throughput model for future studies of the pathological mechanisms of anti-heart autoimmunity and to test potential immunomodulatory therapeutic approaches