The objective of this study was to evaluate the effect of full-thickness chondral defects on intratissue deformation patterns and matrix constituents in an experimental model mimicking in vivo ...cartilage-on-cartilage contact conditions.
Pairs of bovine osteochondral explants, in a unique cartilage-on-cartilage model system, were compressed uniaxially by 350 N during 2 s loading and 1.4 s unloading cycles (≈1700 repetitions). Tissue deformations under quasi-steady state load deformation response were measured with displacement encoded imaging with stimulated echoes (DENSE) in a 9.4 T magnetic resonance imaging (MRI) scanner. Pre- and post-loading, T1, T2 and T1ρ relaxation time maps were measured. We analyzed differences in strain patterns and relaxation times between intact cartilage (n = 8) and cartilage in which a full-thickness and critical sized defect was created (n = 8).
Under compressive loading, strain magnitudes were elevated at the defect rim, with elevated tensile and compressive principal strains (Δϵmax = 4.2%, P = 0.02; Δϵmin = −4.3%, P = 0.02) and maximum shear strain at the defect rim (Δγmax = 4.4%, P = 0.007). The opposing cartilage showed minimal increase in strain patterns at contact with the defect rim but decreased strains opposing the defect. After defect creation, T1, T2 and T1ρ relaxation times were elevated at the defect rim only. Following loading, the overall relaxations times of the defect tissue and especially at the rim, increased compared to intact cartilage.
This study demonstrates that the local biomechanical changes occurring after defect creation may induce tissue damage by increasing shear strains and depletion of cartilage constituents at the defect rim under compressive loading.
With the growing prevalence of psychotropic drug prescriptions among children and adolescents, the need for studies on lasting effects of drug exposure on the developing brain rises. Fluoxetine is ...the only selective serotonin reuptake inhibitor (SSRI) officially registered to treat major depressive disorder in children. Although various (pre)clinical studies have assessed the (long-term) effects of fluoxetine exposure in the perinatal period and in adulthood, limited data is available on its effects on the developing brain later in life, i.e. during adolescence.
The present study aimed at investigating the effects of age following chronic SSRI treatment on the central serotonin (5-HT) system. To this end, pharmacological MRI (phMRI) was performed in chronic fluoxetine-treated (5mg/kg, oral gavage for 3weeks) juvenile (PND25) and adult rats (PND65) after a 1-week washout period, using an acute fluoxetine challenge (5mg/kg, i.v.) to trigger the 5-HT system.
We observed a diminished brain response to the acute challenge in adult treated animals when compared to control animals, whereas this response was increased in juvenile treated rats. As a result, a significant age by treatment interaction effect was seen in several (subcortical) 5-HT related brain regions.
An opposite effect of chronic fluoxetine treatment was seen in the developing brain compared to that in matured brain, as assessed non-invasively using phMRI. These findings most likely reflect neuronal imprinting effects of juvenile SSRI treatment and may underlie emotional disturbances seen in animals and children treated with this drug. Also, our findings suggest that phMRI might be ideally suited to study this important issue in the pediatric population.
► We performed 5-HT phMRI in chronic fluoxetine-treated juvenile and adult rats. ► Juvenile treatment caused an opposite phMRI response compared to adult treatment. ► An age by treatment interaction effect was seen in 5-HT related brain regions. ► Our findings probably reflect neuronal imprinting effects of early SSRI treatment. ► phMRI is suitable to non-invasively assess age-related differences in 5-HT response.
Abstract
Magnetic nanoparticles are versatile materials that have boosted the development of different biomedical applications, being superparamagnetic magnetite nanoparticles a milestone in the ...field, after achieving clinical approval as contrast agents in magnetic resonance imaging (Feridex
®
), magnetic hyperthermia agents for oncological treatments (NanoTherm
®
), or iron deficiency supplement (Feraheme
®
). However, its potential as theragnostic agent could be further expanded by its encapsulation within a biodegradable hydrogel, capable of enhancing the biocompatibility and loading abilities, to simultaneously carry drugs, radiotracers, or biomolecules. Gelatin, is a natural biopolymer with optimal
in vivo
feature and gelling capacity that has been extensively used for decades in pharmaceuticals. In this work, we have addressed the preparation of gelatin nanoparticles, bare and loaded with magnetite nanoparticles, with controlled size to be used as contrast agents in magnetic resonance imaging. The main formulation parameters influencing the preparation of gelatin nanoparticles with controlled size by single-step desolvation method, were studied and optimized, to produce small gelatin nanoparticles (97nm) and highly loaded (38% w/w) Fe
3
O
4
@citrate gelatin nanoparticles (150 nm) with high magnetic response (65emus/g). The viability assays of the magnetic gelatin nanoparticles, tested with mesenchymal stem cells, showed negligible toxicity and
in vitro
magnetic resonance imaging tests, performed in agar phantoms, revealed a good contrast for T2 weighting MRI, r2 = 265.5(mM
−1
s
−1
), superior to commercial products, such as Resovist or Endorem.
In the last decade an important role for the progression of neuronal cell death in Alzheimer's disease (AD) has been ascribed to oxidative stress.
trans-4-Hydroxy-2-nonenal, a product of lipid ...peroxidation, forms conjugates with a variety of nucleophilic groups such as thiols or amino moieties. Here we report for the first time the quantitation of glutathione conjugates of trans-4-hydroxy-2-nonenal (HNEGSH) in the human postmortem brain using the specific and very sensitive method of electrospray ionization triple quadrupole mass spectrometry (ESI-MS-MS). Levels of HNEGSH conjugates calculated as the sum of three chromatographically separated diastereomers were determined in hippocampus, entorhinal cortex, substantia innominata, frontal and temporal cortex, as well as cerebellum from patients with AD and controls matched for age, gender, postmortem delay and storage time. Neither age, nor postmortem delay, nor storage time did correlate with levels of HNEGSH conjugates which ranged between 1 and 500
pmol/g fresh weight in the brain areas examined. The brain specimen from patients with clinically and neuropathologically probable AD diagnosed according to criteria of the consortium to establish a registry for AD (CERAD) show increased levels of HNEGSH in the temporal and frontal cortex, as well as in the substantia innominata. Classification of disease severity according to Braak and Braak, which takes into consideration the amount of neurofibrillary tangles and neuritic plaques, revealed highest levels of HNEGSH in the substantia innominata and the hippocampus, two brain regions known to be preferentially affected in AD. These results substantiate the link between conjugates of glutathione with a product of lipid peroxidation and Alzheimer's disease and justify further studies to evaluate the role of HNE metabolites as potential biomarkers for disease progression in AD.
Background
A reproducible Roux-en-Y gastric bypass (RYGB) model in mice is needed to study the physiological alterations after surgery.
Methods
Male C57BL6 mice weighing 29.0 ± 0.8 g underwent either ...RYGB (
n
= 14) or sham operations (
n
= 6). RYGB surgery consisted of a small gastric pouch (~2 % of the initial stomach size), a biliopancreatic and alimentary limb of 10 cm each and a common channel of 15 cm. Animals had free access to standard chow in the postoperative period. Body mass and food intake were recorded for 60 days. Bomb calorimetry was used for faecal analysis. Anatomical rearrangement was assessed using planar X-ray fluoroscopy and computed tomography (CT) after oral Gastrografin® injection.
Results
RYGB surgery led to a sustained reduction in body weight compared to sham-operated mice (postoperative week 1: sham 27.8 ± 0.7 g vs. RYGB 26.5 ± 1.0 g,
p
= 0.008; postoperative week 8: sham 30.7 ± 0.8 g vs. RYGB 28.4 ± 1.1 g,
p
= 0.003). RYGB mice ate less compared to shams (sham 4.6 ± 0.2 g/day vs. RYGB 4.3 ± 0.4 g/day,
p
< 0.001). There were no differences in faecal mass (
p
= 0.13) and faecal energy content (
p
= 0.44) between RYGB and shams. CT scan demonstrated the expected anatomical rearrangement without leakage or stenosis. Fluoroscopy revealed rapid pouch emptying.
Conclusions
RYGB with a small gastric pouch is technically feasible in mice. With this model in place, genetically manipulated mouse models could be used to study the physiological mechanisms involved with metabolic changes after gastric bypass.
Abstract
Introduction
The acute inflammatory response contributes substantially to functional recovery and remodelling of the left ventricle after acute ischemic injury. Previously, we have shown ...that the C-Type Lectin Receptor CLEC4E plays a role in early leukocyte recruitment during the acute inflammatory response of ischemia-reperfusion injury (I/R). However, the role of CLEC4E signalling in functional recovery of the left ventricle after I/R remains unknown. Therefore, we studied the chronic inflammatory response and left-ventricular remodelling in murine gene deletion model of Clec4e, subjected to I/R.
Methods
In anesthetized C57Bl6/J wild-type (n=14) and Clec4e−/− (n=13) mice, we transiently occluded the left-descending artery for 60 min, followed by 4 weeks reperfusion (I/R). A blood sample was collected at 90 minutes reperfusion to measure high-sensitivity troponin I (TnI) levels, as a surrogate marker of cardiac damage. At 4 weeks, mice underwent MRI (7T) to investigate the effect of Clec4e-gene deletion on LV-remodelling.
Results
Plasma TnI-levels showed no statistical difference between both groups, indicating that the initial insult was comparable. In wild-type mice, plasma TnI-levels negatively correlated with ejection fraction (EF, R2=0.92 p<0.0001) at 4 weeks I/R, while Clec4e−/− mice showed preserved EF, irrespective of 90 minutes TnI-levels. MRI-analysis at 4 weeks after I/R showed significantly smaller end-diastolic and end-systolic volumes in Clec4e−/− mice, together with a trend towards a higher ejection fraction, suggesting better preserved structural and functional LV-remodelling (Fig.1).
Conclusion
The inflammatory leukocyte-associated Clec4e signalling pathway impairs functional recovery of the left ventricle after myocardial I/R injury. Inhibition of the Clec4e receptor may be a promising strategy in the treatment of ischemic injury.
Figure 1
Funding Acknowledgement
Type of funding source: Private grant(s) and/or Sponsorship. Main funding source(s): Scholarship Ir. Jozef en Mevr. Reinhilde De Swerts 2018-2022 by the Royal Academy of Medicine of Belgium
Magnetoliposomes (MLs) have shown great potential as magnetic resonance imaging contrast agents and as delivery vehicles for cancer therapy. Targeting the MLs towards the tumor cells or ...neovascularization could ensure delivery of drugs at the tumor site. In this study, we evaluated the potential of MLs targeting the αvβ3 integrin overexpressed on tumor neovascularization and different tumor cell types, including glioma and ovarian cancer.
MLs functionalized with a Texas Red fluorophore (anionic MLs), and with the fluorophore and the cyclic Arginine-Glycine-Aspartate (cRGD; cRGD-MLs) targeting the αvβ3 integrin, were produced in-house. Swiss nude mice were subcutaneously injected with 10
human ovarian cancer SKOV-3 cells. Tumors were allowed to grow for 3 weeks before injection of anionic or cRGD-MLs. Biodistribution of MLs was followed up with a 7T preclinical magnetic resonance imaging (MRI) scanner and fluorescence imaging (FLI) right after injection, 2h, 4h, 24h and 48h post injection. Ex vivo intratumoral ML uptake was confirmed using FLI, electron paramagnetic resonance spectroscopy (EPR) and histology at different time points post injection.
In vivo, we visualized a higher uptake of cRGD-MLs in SKOV-3 xenografts compared to control, anionic MLs with both MRI and FLI. Highest ML uptake was seen after 4h using MRI, but only after 24h using FLI indicating the lower sensitivity of this technique. Furthermore, ex vivo EPR and FLI confirmed the highest tumoral ML uptake at 4 h. Last, a Perl's stain supported the presence of our iron-based particles in SKOV-3 xenografts.
Uptake of cRGD-MLs can be visualized using both MRI and FLI, even though the latter was less sensitive due to lower depth penetration. Furthermore, our results indicate that cRGD-MLs can be used to target SKOV-3 xenograft in Swiss nude mice. Therefore, the further development of this particles into theranostics would be of interest.
Human immunodeficiency virus infection (HIV) at late stages of the disease is accompanied by neurological complications, including motor, behavioral and cognitive impairment. Using simian ...immunodeficiency virus (SIV)-infected rhesus monkeys, an animal model of HIV infection, we found that during the asymptomatic SIV infection dopamine (DA) deficits are early components of central nervous system (CNS) dysfunction. To investigate the role of the DA system in SIV infection and to restore the DA deficiency, we administered selegiline, an agent with DAergic and neuroprotective properties, to SIV-infected monkeys. Selegiline increased DA availability but induced CNS vacuolization, SIV encephalitic lesions, and enhanced CNS viral replication during early SIV infection. The pathological changes seem to be mediated by DA, as treatment with L-DOPA, the precursor of DA, had similar effects. We propose that any natural or induced DAergic dysregulation which results in increased DA availability may potentiate HIV-associated neurological disease (ND). Our findings raise new questions regarding the pathogenesis of HIV-ND and generate concerns about the safety of dopaminergic drugs in the clinical management of HIV-infected patients.
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