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    Scopus© Citations 11  3
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      7Scopus© Citations 8
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    Sensory neuron cultures derived from adult db/db mice as a simplified model to study type-2 diabetes-associated axonal regeneration defects
    (2021)
    DE GREGORIO CONCHA, CRISTIAN ALEJANDRO  
    ;
    EZQUER, EDUARDO FERNANDO  
    <jats:title>ABSTRACT</jats:title> <jats:p>Diabetic neuropathy (DN) is an early common complication of diabetes mellitus (DM), leading to chronic pain, sensory loss and muscle atrophy. Owing to its multifactorial etiology, neuron in vitro cultures have been proposed as simplified systems for DN studies. However, the most used models currently available do not recreate the chronic and systemic damage suffered by peripheral neurons of type-2 DM (T2DM) individuals. Here, we cultured neurons derived from dorsal root ganglia from 6-month-old diabetic db/db-mice, and evaluated their morphology by the Sholl method as an easy-to-analyze readout of neuronal function. We showed that neurons obtained from diabetic mice exhibited neuritic regeneration defects in basal culture conditions, compared to neurons from non-diabetic mice. Next, we evaluated the morphological response to common neuritogenic factors, including nerve growth factor NGF and Laminin-1 (also called Laminin-111). Neurons derived from diabetic mice exhibited reduced regenerative responses to these factors compared to neurons from non-diabetic mice. Finally, we analyzed the neuronal response to a putative DN therapy based on the secretome of mesenchymal stem cells (MSC). Neurons from diabetic mice treated with the MSC secretome displayed a significant improvement in neuritic regeneration, but still reduced when compared to neurons derived from non-diabetic mice. This in vitro model recapitulates many alterations observed in sensory neurons of T2DM individuals, suggesting the possibility of studying neuronal functions without the need of adding additional toxic factors to culture plates. This model may be useful for evaluating intrinsic neuronal responses in a cell-autonomous manner, and as a throughput screening for the pre-evaluation of new therapies for DN.</jats:p>
      2  1Scopus© Citations 3
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    Association between spontaneous internal carotid artery dissection and perivascular adipose tissue attenuation on computed tomography angiography
    (2023)
    Kevin Cheng
    ;
    Andrew Lin
    ;
    STECHER GUZMAN, XIMENA PATRICIA  
    ;
    Tomas Bernstein
    ;
    Paulo Zuñiga
    <jats:sec><jats:title>Background:</jats:title><jats:p> Spontaneous cervical artery dissection (sCAD) is a leading cause of ischemic stroke in young patients. Studies using high-resolution magnetic resonance imaging and positron emission tomography have suggested vessel wall inflammation to be a pathogenic factor in sCAD. Computed tomography (CT) attenuation of perivascular adipose tissue (PVAT) is an established non-invasive imaging biomarker of inflammation in coronary arteries, with higher attenuation values reflecting a greater degree of vascular inflammation. </jats:p></jats:sec><jats:sec><jats:title>Objectives:</jats:title><jats:p> We evaluate the CT attenuation of PVAT surrounding the internal carotid artery (PVAT<jats:sub>carotid</jats:sub>) with and without spontaneous dissection. </jats:p></jats:sec><jats:sec><jats:title>Methods:</jats:title><jats:p> Single-center prospective observational study of 56 consecutive patients with CT-verified spontaneous dissection of the internal carotid artery (ICA). Of these patients, six underwent follow-up computed tomography angiography (CTA). Twenty-two patients who underwent CTA for acute neurological symptoms but did not have dissection formed the control group. Using semi-automated research software, PVAT<jats:sub>carotid</jats:sub> was measured as the mean Hounsfield unit (HU) attenuation of adipose tissue within a defined volume of interest surrounding the ICA. </jats:p></jats:sec><jats:sec><jats:title>Results:</jats:title><jats:p> PVAT<jats:sub>carotid</jats:sub> was significantly higher around dissected ICA compared with non-dissected contralateral ICA in the same patients (−58.7 ± 10.2 vs −68.9 ± 8.1 HU, p &lt; 0.0001) and ICA of patients without dissection (−58.7 ± 10.2 vs −69.3 ± 9.3 HU, p &lt; 0.0001). After a median follow-up of 89 days, there was a significant reduction in PVAT<jats:sub>carotid</jats:sub> around dissected ICA (−57.5 ± 13.4 to −74.3 ± 10.5 HU, p &lt; 0.05), while no change was observed around non-dissected contralateral ICA (−71.0 ± 4.4 to −74.1 ± 4.1 HU, p = 0.19). ICA dissection was an independent predictor of PVAT<jats:sub>carotid</jats:sub> following multivariable adjustment for age and the presence of ICA occlusion. </jats:p></jats:sec><jats:sec><jats:title>Conclusion:</jats:title><jats:p> PVAT<jats:sub>carotid</jats:sub> is elevated in the presence of sCAD and may decrease following the acute event. </jats:p></jats:sec>
      3  1Scopus© Citations 5
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    Spheroids derived from the stromal vascular fraction of adipose tissue self-organize in complex adipose organoids and secrete leptin
    (2023)
    Fermín Robledo
    ;
    Lila González-Hodar
    ;
    Pablo Tapia
    ;
    Ana-María Figueroa
    ;
    EZQUER, EDUARDO FERNANDO  
    <jats:title>Abstract</jats:title><jats:sec> <jats:title>Background</jats:title> <jats:p>Adipose tissue-derived stromal vascular fraction (SVF) harbors multipotent cells with potential therapeutic relevance. We developed a method to form adipose spheroids (AS) from the SVF with complex organoid structure and enhanced leptin secretion upon insulin stimulation. </jats:p> </jats:sec><jats:sec> <jats:title>Methods</jats:title> <jats:p>SVF was generated from the interscapular brown adipose tissue of newborn mice. Immunophenotype and stemness of cultured SVF were determined by flow cytometry and in vitro differentiation, respectively. Spheroids were generated in hanging drops and non-adherent plates and compared by morphometric methods. The adipogenic potential was compared between preadipocyte monolayers and spheroids. Extracellular leptin was quantified by immunoassay. Lipolysis was stimulated with isoprenaline and quantified by colorimetric methods. AS viability and ultrastructure were determined by confocal and transmission electron microscopy analyses. </jats:p> </jats:sec><jats:sec> <jats:title>Results</jats:title> <jats:p>Cultured SVF contained Sca1 + CD29 + CD44 + CD11b- CD45- CD90- cells with adipogenic and chondrogenic but no osteogenic potential. Culture on non-adherent plates yielded the highest quantity and biggest size of spheroids. Differentiation of AS for 15 days in a culture medium supplemented with insulin and rosiglitazone resulted in greater <jats:italic>Pparg</jats:italic>, <jats:italic>Plin1,</jats:italic> and <jats:italic>Lep</jats:italic> expression compared to differentiated adipocytes monolayers. AS were viable and maintained leptin secretion even in the absence of adipogenic stimulation. Glycerol release after isoprenaline stimulation was higher in AS compared to adipocytes in monolayers. AS were composed of outer layers of unilocular mature adipocytes and an inner structure composed of preadipocytes, immature adipocytes and an abundant loose extracellular matrix.</jats:p> </jats:sec><jats:sec> <jats:title>Conclusion</jats:title> <jats:p>Newborn mice adipose SVF can be efficiently differentiated into leptin-secreting AS. Prolonged stimulation with insulin and rosiglitazone allows the formation of structurally complex adipose organoids able to respond to adrenergic lipolytic stimulation.</jats:p> </jats:sec>
    Scopus© Citations 9  1
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    Effect of human mesenchymal stem cell secretome administration on morphine self-administration and relapse in two animal models of opioid dependence
    (2022)
    María Elena Quintanilla
    ;
    Mauricio Quezada
    ;
    Paola Morales
    ;
    BERRIOS CARCAMO, PABLO ANDRES  
    ;
    Mario Herrera-Marschitz
    <jats:title>Abstract</jats:title><jats:p>The present study investigates the possible therapeutic effects of human mesenchymal stem cell-derived secretome on morphine dependence and relapse. This was studied in a new model of chronic voluntary morphine intake in Wistar rats which shows classic signs of morphine intoxication and a severe naloxone-induced withdrawal syndrome. A single intranasal-systemic administration of MSCs secretome fully inhibited (&gt;95%; <jats:italic>p</jats:italic> &lt; 0.001) voluntary morphine intake and reduced the post-deprivation relapse intake by 50% (p &lt; 0.02). Since several studies suggest a significant genetic contribution to the chronic use of many addictive drugs, the effect of MSCs secretome on morphine self-administration was further studied in rats bred as high alcohol consumers (UChB rats). Sub-chronic intraperitoneal administration of morphine before access to increasing concentrations of morphine solutions and water were available to the animals, led UChB rats to prefer ingesting morphine solutions over water, attaining levels of oral morphine intake in the range of those in the Wistar model. Intranasally administered MSCs secretome to UChB rats dose-dependently inhibited morphine self-administration by 72% (p &lt; 0.001); while a single intranasal dose of MSC-secretome administered during a morphine deprivation period imposed on chronic morphine consumer UChB rats inhibited re-access morphine relapse intake by 80 to 85% (p &lt; 0.0001). Both in the Wistar and the UChB rat models, MSCs-secretome administration reversed the morphine-induced increases in brain oxidative stress and neuroinflammation, considered as key engines perpetuating drug relapse. Overall, present preclinical studies suggest that products secreted by human mesenchymal stem cells may be of value in the treatment of opioid addiction.</jats:p>
    Scopus© Citations 6  3
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    Herniated superficial lipomas in gluteal and thigh support areas: a special clinical ultrasound presentation
    (2022)
    WHITTLE, CAROLINA  
    ;
    FAJRE WIPE, XIMENA ELIZABETH  
    ;
    Catalina Silva
    ;
    CASTRO MENDEZ, ALEX  
    <jats:sec><jats:title>Background</jats:title><jats:p> Lipomas are the most frequent benign tumor. They have been described as soft, well-defined, slow-growing palpable masses, and classified as deep or superficial. </jats:p></jats:sec><jats:sec><jats:title>Purpose</jats:title><jats:p> To present the clinical and ultrasound findings of herniated superficial subcutaneous lipoma (HSL), located in pressure/support areas, not previously described. </jats:p></jats:sec><jats:sec><jats:title>Material and Methods</jats:title><jats:p> A seven-year retrospective review was performed. Patients with a preoperative high-resolution ultrasound (HRUS) diagnosis of HSL archived in the computational system and histological study information were selected. </jats:p></jats:sec><jats:sec><jats:title>Results</jats:title><jats:p> A total of 37 patients (mean age = 46 years) were recruited. The clinical diagnoses were “lipoma” or “suspected lipoma” in just 46% of the cases. This was three times more frequent in women. Tumors were located mainly in the gluteal (38%) and proximal thigh areas (35%). Clinically, all patients showed small, soft, rounded skin nodules. On HRUS, they appear as lipomatous tumors that protruded, compressed, and thinned the covering dermal layer. The subcutaneous portion was bigger than the herniated part, with an iceberg image. </jats:p></jats:sec><jats:sec><jats:title>Conclusion</jats:title><jats:p> HSL in support areas have a special clinical ultrasound presentation, not previously described. It is important to recognize them for a proper diagnosis and treatment, especially because they clinically may present as small superficial lesions; however, they are associated with a larger subcutaneous portion (iceberg image). It is probable that an estrogen influence on the adipose tissue can be suspected because of the female predominance. </jats:p></jats:sec>
      1  3
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    Renal peritumoral adipose tissue undergoes a browning process and stimulates the expression of epithelial-mesenchymal transition markers in human renal cells
    (2022)
    Matías Ferrando
    ;
    Flavia Alejandra Bruna
    ;
    Leonardo Rafael Romeo
    ;
    CONTADOR MARTINEZ, DAVID ERNESTO  
    ;
    Daiana Lorena Moya-Morales
    <jats:title>Abstract</jats:title><jats:p>Tumor cells can interact with neighboring adipose cells and adipocyte dedifferentiation appears to be an important aspect of tumorigenesis. We evaluated the size of adipocytes in human adipose explants from normal (hRAN) and kidney cancer (hRAT); changes in the expression of WAT and BAT/beige markers in hRAN and hRAT; the expression of epithelial-mesenchymal transition (EMT) cell markers in human kidney tumor (786-O, ACHN and Caki-1); and non-tumor (HK-2) epithelial cell lines incubated with the conditioned media (CMs) of hRAN and hRAT. We observed that hRAT adipocytes showed a significantly minor size compared to hRAN adipocytes. Also, we observed that both Prdm16 and Tbx1 mRNA and the expression of UCP1, TBX1, PPARγ, PCG1α, c/EBPα LAP and c/EBPα LIP was significantly higher in hRAT than hRAN. Finally, we found an increase in vimentin and N-cadherin expression in HK-2 cells incubated for 24 h with hRAT-CMs compared to hRAN- and control-CMs. Furthermore, desmin and N-cadherin expression also increased significantly in 786-O when these cells were incubated with hRAT-CMs compared to the value observed with hRAN- and control-CMs. We observed a significant decrease in E-cadherin expression in the ACHN cell line incubated with hRAT-CMs versus hRAN- and control-CMs. However, we did not observe changes in E-cadherin expression in HK-2, 786-O or Caki-1. The results obtained, together with the results previously published by our group, allow us to conclude that perirenal white adipose tissue browning contributes to tumor development in kidney cancer. In addition, hRAT-CMs increases the expression of mesenchymal markers in renal epithelial cells, which could indicate a regulation of EMT due to this adipose tissue. </jats:p>
      1Scopus© Citations 11  1
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    Microencapsulation of cellular aggregates composed of differentiated insulin and glucagon-producing cells from human mesenchymal stem cells derived from adipose tissue
    (2020)
    Claudia Jara
    ;
    Felipe Oyarzun-Ampuero
    ;
    Flavio Carrión
    ;
    Esteban González-Echeverría
    ;
    Claudio Cappelli
    <jats:title>Abstract</jats:title><jats:sec> <jats:title>Background</jats:title> <jats:p>In type I diabetes mellitus (T1DM) pancreatic β cells are destroyed. Treatment entails exogenous insulin administration and strict diet control, yet optimal glycemic control is hardly attainable. Islet transplant could be an alternative in patients with poor glycemic control, but inefficient islet purification and autoimmune response of patients is still a challenge. For these reasons, it is necessary to explore new cellular sources and immunological isolation methods oriented to develop T1DM cell-based therapies.</jats:p> </jats:sec><jats:sec> <jats:title>Aims</jats:title> <jats:p>We postulate human adipose-derived stem cell (hASC) as an adequate source to generate pancreatic islet cells in vitro, and to produce islet-like structures. Furthermore, we propose microencapsulation of these aggregates as an immunological isolation strategy.</jats:p> </jats:sec><jats:sec> <jats:title>Methods</jats:title> <jats:p>hASC obtained from lipoaspirated fat tissue from human donors were differentiated in vitro to insulin (Ins) and glucagon (Gcg) producing cells. Then, insulin producing cells (IPC) and glucagon producing cells (GPC) were cocultured in low adhesion conditions to form cellular aggregates, and later encapsulated in a sodium alginate polymer. Expression of pancreatic lineage markers and secretion of insulin or glucagon in vitro were analyzed.</jats:p> </jats:sec><jats:sec> <jats:title>Results</jats:title> <jats:p>The results show that multipotent hASC efficiently differentiate to IPC and GPC, and express pancreatic markers, including insulin or glucagon hormones which they secrete upon stimulation (fivefold for insulin in IPC, and fourfold for glucagon, compared to undifferentiated cells). In turn, calculation of the Feret diameter and area of cellular aggregates revealed mean diameters of ~ 80 µm, and 65% of the aggregates reached 4000 µm<jats:sup>2</jats:sup> at 72 h of formation. IPC/GPC aggregates were then microencapsulated in sodium-alginate polymer microgels, which were found to be more stable when stabilized with Ba<jats:sup>2+</jats:sup>, yielding average diameters of ~ 300 µm. Interestingly, Ba<jats:sup>2+</jats:sup>-microencapsulated aggregates respond to high external glucose with insulin secretion.</jats:p> </jats:sec><jats:sec> <jats:title>Conclusions</jats:title> <jats:p>The IPC/GPC differentiation process from hASC, followed by the generation of cellular aggregates that are later microencapsulated, could represent a possible treatment for T1DM.</jats:p> </jats:sec>
      16Scopus© Citations 9
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      21Scopus© Citations 6