From isolation of adult adipose tissue derived stem cells ADAS to labelling with superparamagnetic iron oxide nanoparticles: first approaches to unleash the potential

The use of adult adipose-derived stem cells (ADAS) as a treatment for neurological diseases is in promising development. Extracellular vesicles such as exosomes (EXO), which impact surrounding cells, are the main biological agents of ADAS. Exosome localization and tracking techniques need to be effective and non-invasive in the current development of exosome therapies. Exosomes must be labeled with contrast agents, such as ferrous superparamagnetic nanoparticles (NPs). The current research project aims to validate the therapeutic efficacy of ADAS-derived EXOs labelled with different NPs in models of neurodegenerative diseases, capable of providing an imaging and cell therapy approach

Chemical shift imaging at 4.7 tesla of brown adipose tissue.

In vivo distinction between small deposits of brown adipose tissue (BAT) and surrounding tissues may be difficult. In this article, we propose an experiment paradigm, based on techniques of chemical shift magnetic resonance imaging (CSI), which can improve the methods presently available for the study of BAT. Male rats were examined in an imager-spectrometer equipped with a 4.7 T magnet. Proton spectra of isolated BAT deposits showed that both fat and water protons contributed significantly to the genesis of the magnetic resonance signal. An equivocal definition of BAT deposits was obtained by three (respectively, spin-echo, water-selective, and fat-selective) images. The spin-echo (SE), T1-weighted image provided the best anatomical description of the structures. The images selective for fat-protons displayed the degree of lipid accumulation in each area. The images selective for water-protons provided an internal control of adipose tissue localization. The proposed paradigm allows an unequivocal definition of BAT deposits and appears particularly useful in studies where experimental manipulation (i.e., cold acclimation or drug treatment) produces changes in this issue

In vivo quantitative lipidic map of brown adipose tissue by chemical shift imaging at 4.7 tesla

In the present paper, chemical shift imaging techniques are applied to quantitative in vivo evaluation of fat and water content in interscapular brown adipose tissue (BAT). The experiments have been carried out on five female Sprague-Dawley rats after calibration and testing with suitable phantoms containing known amounts of water and oil. We found that, in the interscapular BAT, the fat is about 50% at the surface (mainly unilocular) region, but its percentage drops to 20–30% in the deepest (mainly multilocular) portion. The perirenal deposits of white adipose tissue (WAT) contained significantly higher amount of fat with large areas ranging from 70 to 90%. Later the rats were killed and the same procedure was repeated with dead animals. Experiments performed in dead rats show a modification of the hydro-lipidic ratio more evident in the multilocular portions of the deposit. The present work demonstrates that MRI-based methods allow a non-invasive, in vivo quantitative mapping of the lipid content which can be applied to investigation of brown adipose tissue deposits in small experiment animals.—Lunati, E., P. Marzola, E. Nicolato, M. Fedrigo, M. Villa, and A. Sbarbati. In vivo quantitative lipidic map of brown adipose tissue by chemical shift imaging at 4.7 tesla. J. Lipid Res. 1999. 40: 1395–1400

Heterogeneous enhancement pattern in DCE-MRI reveals the morphology of normal lymph nodes: an experimental study

Purpose: To investigate the heterogeneous enhancement pattern in normal lymph nodes of healthy mice by different albumin-binding contrast agents. Methods: The enhancement of normal lymph nodes was assessed in mice by dynamic contrast-enhanced MRI (DCE-MRI) after the administration of two contrast agents characterized by different albumin-binding properties: gadopentetate dimeglumine (Gd-DTPA) and gadobenate dimeglumine (Gd-BOPTA). To take into account potential heterogeneities of the contrast uptake in the lymph nodes, k-means cluster analysis was performed on DCE-MRI data. Cluster spatial distribution was visually assessed. Statistical comparison among clusters and contrast agents was performed on semiquantitative parameters (AUC, wash-in rate, and wash-out rate) and on the relative size of the segmented clusters. Results: Cluster analysis of DCE-MRI data revealed at least two main clusters, localized in the outer portion and in the inner portion of each lymph node. With both contrast agents, AUC (p < 0.01) and wash-in (p < 0.05) rates were greater in the inner cluster, which also showed a steeper wash-out rate than the outer cluster (Gd-BOPTA, p < 0.01; Gd-DTPA, p=0.056). The size of the outer cluster was greater than that of the inner cluster by Gd-DTPA (p < 0.05) and Gd-BOPTA (p < 0.01). The enhancement pattern of Gd-DTPA was not significantly different from the enhancement pattern of Gd-BOPTA. Conclusion: DCE-MRI in normal lymph nodes shows a characteristic heterogeneous pattern, discriminating the periphery and the central portion of the lymph nodes. Such a pattern deserves to be investigated as a diagnostic marker for lymph node staging

Preclinical In vivo Imaging for Fat Tissue Identification, Quantification, and Functional Characterization

Localization, differentiation, and quantitative assessment of fat tissues have always collected the interest of researchers. Nowadays, these topics are even more relevant as obesity (the excess of fat tissue) is considered a real pathology requiring in some cases pharmacological and surgical approaches. Several weight loss medications, acting either on the metabolism or on the central nervous system, are currently under preclinical or clinical investigation. Animal models of obesity have been developed and are widely used in pharmaceutical research. The assessment of candidate drugs in animal models requires non-invasive methods for longitudinal assessment of efficacy, the main outcome being the amount of body fat. Fat tissues can be either quantified in the entire animal or localized and measured in selected organs/regions of the body. Fat tissues are characterized by peculiar contrast in several imaging modalities as for example Magnetic Resonance Imaging (MRI) that can distinguish between fat and water protons thank to their different magnetic resonance properties. Since fat tissues have higher carbon/hydrogen content than other soft tissues and bones, they can be easily assessed by Computed Tomography (CT) as well. Interestingly, MRI also discriminates between white and brown adipose tissue (BAT); the latter has long been regarded as a potential target for anti-obesity drugs because of its ability to enhance energy consumption through increased thermogenesis. Positron Emission Tomography (PET) performed with (18)F-FDG as glucose analog radiotracer reflects well the metabolic rate in body tissues and consequently is the technique of choice for studies of BAT metabolism. This review will focus on the main, non-invasive imaging techniques (MRI, CT, and PET) that are fundamental for the assessment, quantification and functional characterization of fat deposits in small laboratory animals. The contribution of optical techniques, which are currently regarded with increasing interest, will be also briefly described. For each technique the physical principles of signal detection will be overviewed and some relevant studies will be summarized. Far from being exhaustive, this review has the purpose to highlight some strategies that can be adopted for the in vivo identification, quantification, and functional characterization of adipose tissues mainly from the point of view of biophysics and physiology

Magnetic resonance imaging of adipose-derived adult stem cells labelled with superparamagnetic iron oxide nanoparticles

The application of mesenchymal stem cells (MSCs) represents a new promising approach for treating neurodegenerative diseases. Recently, considerable attention has been paid to adipose-derived adult MSC (ADAS), thanks to the easy availability of adipose tissue and to the possibility of autologous cells transplantation. Any possible application of therapies based on ADAS in the clinics cannot occur without elucidation of their homing. Superparamagnetic iron-oxide nanoparticles (NPs) can be used to label and track cells in vivo via Magnetic Resonance Imaging (MRI

Co-Transplantation of endothelial progenitor cells and pancreatic islets to induce long-lasting normoglycemia in streptozotocin-treated diabetic rats

Graft vascularization is a crucial step to obtain stable normoglycemia in pancreatic islet transplantation. Endothelial progenitor cells (EPCs) contribute to neoangiogenesis and to the revascularization process during ischaemic events and play a key role in the response to pancreatic islet injury. In this work we co-transplanted EPCs and islets in the portal vein of chemically-induced diabetic rats to restore islet vascularization and to improve graft survival. Syngenic islets were transplanted, either alone or with EPCs derived from green fluorescent protein (GFP) transgenic rats, into the portal vein of streptozotocin-induced diabetic rats. Blood glucose levels were monitored and intraperitoneal glucose tolerance tests were performed. Real time-PCR was carried out to evaluate the gene expression of angiogenic factors. Diabetic-induced rats showed long-lasting (6 months) normoglycemia upon co-transplantation of syngenic islets and EPCs. After 3–5 days from transplantation, hyperglycaemic levels dropped to normal values and lasted unmodified as long as they were checked. Further, glucose tolerance tests revealed the animals' ability to produce insulin on-demand as indexed by a prompt response in blood glucose clearance. Graft neovascularization was evaluated by immunohistochemistry: for the first time the measure of endothelial thickness revealed a donor-EPC-related neovascularization supporting viable islets up to six months after transplant. Our results highlight the importance of a newly formed viable vascular network together with pancreatic islets to provide de novo adequate supply in order to obtain enduring normoglycemia and prevent diabetes-related long-term health hazards

An in vivo study of quantum dots tissue accumulation

Nanotechnology represents a new frontier for the science progress and there are great expectations in relation to diagnostic and therapeutic envelopes [1]. Living organisms are built of cells that are typically 10 \u3bcm across. However, the cell parts are much smaller and are in the sub-micron size domain, for example a red blood cell is approximately 7,000 nm wide. Even smaller are the proteins with a typical size of just 5 nm, which is comparable with the dimensions of smallest manmade nanoparticles. This simple size comparison gives an idea of using nanoparticles as very small probes that would allow us to spy at the cellular machinery without introducing too much interference. Understanding of biological processes on the nanoscale level is a strong driving force behind development of nanotechnologyOur current knowledge of the toxicology of nanoparticles in vivo is poor [2] but suggests that nanoparticles may able to have adverse effects at their portal of entry , for example, the lungs, but that some nanoparticles may also escape the normal defences and translocate from their portal of entry to have diverse effects in other target organs [3].There is no cut-off below witch particles suddenly become harmful, in the lung at least. This is because harmful particles have their effects as a consequence of two factors that act together to determine their potential to cause harm: their large surface area, and the reactivity or intrinsic toxicity of the surface. It is self evident that the smaller particles are, the more surface area they have per unit mass; therefore any intrinsic toxicity of the particles surface will be emphasised. Some of the most complex nanoparticles are likely to be produced for therapeutic purposes, furthermore nanoparticles binding to protein may result in a series of consequences not expected to occur when proteins bind to large particles. Very small particles may be not detected by the normal phagocytic defences, allowing them to gain access to the blood or nervous system [4]. Very small particles are smaller than some molecules and could act like haptens to modify protein structures, either altering their function or rendering them antigenic, raising the potential for autoimmune effects.Tracers that we have used are nanoparticles with optical properties, fluorescent semiconductors, that absorb photons of light and re-emit photons at a different wavelength They are known as quantum dots (QDs), nanocrystals that are nanometres-scale (10-20nm, roughly protein-sized) atom clusters, containing from a few hundred to a few thousand atoms of a semiconductor material (cadmium mixed with selenium), which has been coated with an additional semiconductor shell (zinc sulfide) to improve the optical properties of the material. These nanoparticles fluoresce in a different way than do traditional fluorophores, they exhibit some important differences as compared to organic fluorescent dyes and naturally fluorescent proteins: they have an extinction coefficient 10-50 times bigger than them. These nanoparticles projected around their optical properties: stable , bright and photo-stable fluorescence, observed and measured for hours, and that persists also into isolated tissues. Nanoparticles like QDs, could be targeted and not targeted and provided several unique features and capabilities[5, 6]: the size-effect does the QDs cancer biomarkers and there is the possibility to functionalize their surface area with a several numbers of functional groups that can be linked with multiple diagnostic (e.g. radio-isotopic or magnetic) and therapeutic agents. The aim of the study is to monitor nanoparticles behaviour into blood system: kinetic, T1/2, bio distribution, and tissues accumulation. We would extrapolate from optics parameters physiological ones, in specific districts so as liver and lungs that are the most probably targets of toxicity

ASC-exosomes ameliorate the disease progression in SOD1(G93A) murine model underlining their potential therapeutic use in human ALS

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by progressive degeneration of motoneurons. To date, there is no effective treatment available. Exosomes are extracellular vesicles that play important roles in intercellular communication, recapitulating the effect of origin cells. In this study, we tested the potential neuroprotective effect of exosomes isolated from adipose-derived stem cells (ASC-exosomes) on the in vivo model most widely used to study ALS, the human SOD1 gene with a G93A mutation (SOD1(G93A)) mouse. Moreover, we compared the effect of two different routes of exosomes administration, intravenous and intranasal. The effect of exosomes administration on disease progression was monitored by motor tests and analysis of lumbar motoneurons and glial cells, neuromuscular junction, and muscle. Our results demonstrated that repeated administration of ASC-exosomes improved the motor performance; protected lumbar motoneurons, the neuromuscular junction, and muscle; and decreased the glial cells activation in treated SOD1(G93A) mice. Moreover, exosomes have the ability to home to lesioned ALS regions of the animal brain. These data contribute by providing additional knowledge for the promising use of ASC-exosomes as a therapy in human ALS

Learning approach to analyze tumour heterogeneity in DCE-MRI data during anti-cancer treatment

The paper proposes a learning approach to support medical researchers in the context of in-vivo cancer imaging, and specifically in the analysis of Dynamic Contrast-Enhanced MRI (DCE-MRI) data. Tumour heterogeneity is characterized by identifying regions with different vascular perfusion. The overall aim is to measure volume differences of such regions for two experimental groups: the treated group, to which an anticancer therapy is administered, and a control group. The proposed approach is based on a three-steps procedure: (i) robust features extraction from raw time-intensity curves, (ii) sample-regions identification manually traced by medical researchers on a small portion of input data, and (iii) overall segmentation by training a Support Vector Machine (SVM) to classify the MRI voxels according to the previously identified cancer areas. In this way a non-invasive method for the analysis of the treatment efficacy is obtained as shown by the promising results reported in our experiments. © 2009 Springer Berlin Heidelberg