المؤلفون: Turano, Ermanna, Scambi, Ilaria, Bonafede, Roberta, Dusi, Silvia, Angelini, Gabriele, Lopez, Nicola, Marostica, Giulia, Rossi, Barbara, Furlan, Roberto, Constantin, Gabriela, Mariotti, Raffaella, Bonetti, Bruno

المساهمون: Turano, Ermanna, Scambi, Ilaria, Bonafede, Roberta, Dusi, Silvia, Angelini, Gabriele, Lopez, Nicola, Marostica, Giulia, Rossi, Barbara, Furlan, Roberto, Constantin, Gabriela, Mariotti, Raffaella, Bonetti, Bruno

مصطلحات موضوعية: EAE, adipose mesenchymal stem cells, extracellular vesicles, multiple sclerosis, stem cells, superparamagnetic iron oxide nanoparticles

الوصف: Background aims: Adipose mesenchymal stem cells (ASCs) represent a promising therapeutic approach in inflammatory neurological disorders, including multiple sclerosis (MS). Recent lines of evidence indicate that most biological activities of ASCs are mediated by the delivery of soluble factors enclosed in extracellular vesicles (EVs). Indeed, we have previously demonstrated that small EVs derived from ASCs (ASC-EVs) ameliorate experimental autoimmune encephalomyelitis (EAE), a murine model of MS. The precise mechanisms and molecular/cellular target of EVs during EAE are still unknown. Methods: To investigate the homing of ASC-EVs, we intravenously injected small EVs loaded with ultra-small superparamagnetic iron oxide nanoparticles (USPIO) at disease onset in EAE-induced C57Bl/6J mice. Histochemical analysis and transmission electron microscopy were carried out 48 h after EV treatment. Moreover, to assess the cellular target of EVs, flow cytometry on cells extracted ex vivo from EAE mouse lymph nodes was performed. Results: Histochemical and ultrastructural analysis showed the presence of labeled EVs in lymph nodes but not in lungs and spinal cord of EAE injected mice. Moreover, we identified the cellular target of EVs in EAE lymph nodes by flow cytometry: ASC-EVs were preferentially located in macrophages, with a consistent amount also noted in dendritic cells and CD4+ T lymphocytes. Conclusions: This represents the first direct evidence of the privileged localization of ASC-EVs in draining lymph nodes of EAE after systemic injection. These data provide prominent information on the distribution, uptake and retention of ASC-EVs, which may help in the development of EV-based therapy in MS.

وصف الملف: STAMPA

العلاقة: info:eu-repo/semantics/altIdentifier/pmid/38231166; volume:26; issue:3; firstpage:276; lastpage:285; numberofpages:10; journal:CYTOTHERAPY; https://hdl.handle.net/11562/1117488Test; info:eu-repo/semantics/altIdentifier/scopus/2-s2.0-85182580098; https://doi.org/10.1016/j.jcyt.2023.12.007Test

الإتاحة: https://doi.org/10.1016/j.jcyt.2023.12.007Test
https://hdl.handle.net/11562/1117488Test

المؤلفون: Bankole, Oluwamolakun, Scambi, Ilaria, Parrella, Edoardo, Muccilli, Matilde, Bonafede, Roberta, Turano, Ermanna, Pizzi, Marina, Mariotti, Raffaella

المساهمون: Bankole, Oluwamolakun, Scambi, Ilaria, Parrella, Edoardo, Muccilli, Matilde, Bonafede, Roberta, Turano, Ermanna, Pizzi, Marina, Mariotti, Raffaella

مصطلحات موضوعية: amyotrophic lateral sclerosis, epigenetics, motor neuron, resveratrol, valproate

الوصف: Amyotrophic lateral sclerosis (ALS) is a fatal adult-onset neurodegenerative disorder. There is no cure and current treatments fail to slow the progression of the disease. Epigenetic modulation in the acetylation state of NF-kB RelA and the histone 3 (H3) protein, involved in the development of neurodegeneration, is a drugable target for the class-I histone deacetylases (HDAC) inhibitors, entinostat or valproate, and the AMP-activated kinase (AMPK)-sirtuin 1 pathway activator, resveratrol. In this study, we demonstrated that the combination of valproate and resveratrol can restore the normal acetylation state of RelA in the SOD1(G93A) murine model of ALS, in order to obtain the neuroprotective form of NF-kB. We also investigated the sexually dimorphic development of the disease, as well as the sex-sensibility to the treatment administered. We showed that the combined drugs, which rescued AMPK activation, RelA and the histone 3 acetylation state, reduced the motor deficit and the disease pathology associated with motor neuron loss and microglial reactivity, Brain-Derived Neurotrophic Factor (BDNF) and B-cell lymphoma-extra large (Bcl-xL) level decline. Specifically, vehicle-administered males showed earlier onset and slower progression of the disease when compared to females. The treatment, administered at 50 days of life, postponed the time of onset in the male by 22 days, but not in a significant way in females. Nevertheless, in females, the drugs significantly reduced symptom severity of the later phase of the disease and prolonged the mice's survival. Only minor beneficial effects were produced in the latter stage in males. Overall, this study shows a beneficial and sexually dimorphic response to valproate and resveratrol treatment in ALS mice.

وصف الملف: ELETTRONICO

العلاقة: info:eu-repo/semantics/altIdentifier/pmid/35162978; info:eu-repo/semantics/altIdentifier/wos/WOS:000757401800001; volume:23; issue:1047; firstpage:1; lastpage:20; numberofpages:20; journal:INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES; http://hdl.handle.net/11562/1057838Test; info:eu-repo/semantics/altIdentifier/scopus/2-s2.0-85122924038; https://doi.org/10.3390/ijms23031047Test

الإتاحة: https://doi.org/10.3390/ijms23031047Test
http://hdl.handle.net/11562/1057838Test

المؤلفون: Bonafede, Roberta, Turano, Ermanna, Scambi, Ilaria, Busato, Alice, Bontempi, Pietro, Virla, Federica, Schiaffino, Lorenzo, Marzola, Pasquina, Bonetti, Bruno, Mariotti, Raffaella

المساهمون: Bonafede, Roberta, Turano, Ermanna, Scambi, Ilaria, Busato, Alice, Bontempi, Pietro, Virla, Federica, Schiaffino, Lorenzo, Marzola, Pasquina, Bonetti, Bruno, Mariotti, Raffaella

مصطلحات موضوعية: extracellular vesicles, MRI, motoneurons, neuromuscular junction, amyotrophic lateral sclerosis, stem cells, homing

الوصف: 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.

وصف الملف: ELETTRONICO

العلاقة: info:eu-repo/semantics/altIdentifier/pmid/32455791; info:eu-repo/semantics/altIdentifier/wos/WOS:000539312100247; volume:21; issue:3651; firstpage:1; lastpage:18; numberofpages:18; journal:INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES; http://hdl.handle.net/11562/1018511Test; info:eu-repo/semantics/altIdentifier/scopus/2-s2.0-85085374379; https://doi.org/10.3390/ijms21103651Test

الإتاحة: https://doi.org/10.3390/ijms21103651Test
http://hdl.handle.net/11562/1018511Test

المؤلفون: Calabria, Elisa, Scambi, Ilaria, Bonafede, Roberta, Schiaffino, Lorenzo, Peroni, Daniele, Potrich, Valentina, Capelli, Carlo, Schena, Federico, Mariotti, Raffaella

المساهمون: Calabria, Elisa, Scambi, Ilaria, Bonafede, Roberta, Schiaffino, Lorenzo, Peroni, Daniele, Potrich, Valentina, Capelli, Carlo, Schena, Federico, Mariotti, Raffaella

مصطلحات موضوعية: ALS, NSC-34 cell line, complex I, coupling efficiency, exosome, high resolution respirometry, membrane potential, mitochondria

الوصف: The amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder characterized by motoneurons death. Mutations in the superoxide dismutase 1 (SOD1) protein have been identified to be related to the disease. Beyond the different altered pathways, the mitochondrial dysfunction is one of the major features that leads to the selective death of motoneurons in ALS. The NSC-34 cell line, overexpressing human SOD1(G93A) mutant protein [NSC-34(G93A)], is considered an optimal in vitro model to study ALS. Here we investigated the energy metabolism in NSC-34(G93A) cells and in particular the effect of the mutated SOD1(G93A) protein on the mitochondrial respiratory capacity (complexes I-IV) by high resolution respirometry (HRR) and cytofluorimetry. We demonstrated that NSC-34(G93A) cells show a reduced mitochondrial oxidative capacity. In particular, we found significant impairment of the complex I-linked oxidative phosphorylation, reduced efficiency of the electron transfer system (ETS) associated with a higher rate of dissipative respiration, and a lower membrane potential. In order to rescue the effect of the mutated SOD1 gene on mitochondria impairment, we evaluated the efficacy of the exosomes, isolated from adipose-derived stem cells, administrated on the NSC-34(G93A) cells. These data show that ASCs-exosomes are able to restore complex I activity, coupling efficiency and mitochondrial membrane potential. Our results improve the knowledge about mitochondrial bioenergetic defects directly associated with the SOD1(G93A) mutation, and prove the efficacy of adipose-derived stem cells exosomes to rescue the function of mitochondria, indicating that these vesicles could represent a valuable approach to target mitochondrial dysfunction in ALS.

وصف الملف: ELETTRONICO

العلاقة: info:eu-repo/semantics/altIdentifier/pmid/31680811; info:eu-repo/semantics/altIdentifier/wos/WOS:000497587300001; volume:13; issue:1070; firstpage:1; lastpage:13; numberofpages:13; journal:FRONTIERS IN NEUROSCIENCE; http://hdl.handle.net/11562/1002257Test; info:eu-repo/semantics/altIdentifier/scopus/2-s2.0-85074996399; https://doi.org/10.3389/fnins.2019.01070Test

الإتاحة: https://doi.org/10.3389/fnins.2019.01070Test
http://hdl.handle.net/11562/1002257Test

المؤلفون: Kassa, Roman M., Bonafede, Roberta, Boschi, Federico, Malatesta, Manuela, Mariotti, Raffaella

المساهمون: Kassa, Roman M., Bonafede, Roberta, Boschi, Federico, Malatesta, Manuela, Mariotti, Raffaella

مصطلحات موضوعية: amyotrophic lateral sclerosi, SOD1(G93A) mice, motoneuron, synaptophysin, major histocompatibility complex I, facial axotomy

الوصف: Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease characterized by motoneuron death. Several cellular pathways have been described to be involved in ALS pathogenesis; however, the involvement of presynaptic stripping and the related MHC class I molecules in mutant SOD1 motoneurons remains to be clarified. To this purpose, we here investigated, for the first time, the motoneurons behavior, di per seand after facial axonal injury, in terms of synaptic stripping and MHC class I expression in wild-type (Wt) mice and in a murine model of ALS, the SOD1(G93A) mice, at the presymptomatic and symptomatic stage of the disease. Concerning Wt animals, we found a reduction in synaptophysin immunoreactivity and an increase of MHC class I molecules in facial motoneurons after axotomy. In uninjured motoneurons of SOD1(G93A) mice, an altered presynaptic framework was evident, and this phenomenon increased during the disease course. The alteration in the presynaptic input is related to excitatory fibers. Moreover, after injury, a further decrease of excitatory input was not associated to an upregulation of MHC class I molecules in motoneuron soma. This study demonstrates, for the first time, that the presence of mutated SOD1 protein affects the MHC class I molecules expression, altering the presynaptic input in motoneurons. Nevertheless, a positive MHC class I immunolabeling was evident in glial cells around facial injured motoneurons, underlying an involvement of these cells in synaptic stripping. This study contributes to better understand the involvement of the mutated SOD1 protein in the vulnerability of motoneurons after damage.

وصف الملف: ELETTRONICO

العلاقة: info:eu-repo/semantics/altIdentifier/pmid/29943955; info:eu-repo/semantics/altIdentifier/wos/WOS:000454458500008; volume:62; issue:2; firstpage:158; lastpage:165; numberofpages:8; journal:EUROPEAN JOURNAL OF HISTOCHEMISTRY; http://hdl.handle.net/11562/979848Test; info:eu-repo/semantics/altIdentifier/scopus/2-s2.0-85051057145; https://doi.org/10.4081/ejh.2018.2904Test

الإتاحة: https://doi.org/10.4081/ejh.2018.2904Test
http://hdl.handle.net/11562/979848Test

المؤلفون: Farinazzo, Alessia, Angiari, Stefano, Turano, Ermanna, Bistaffa, Edoardo, Dusi, Silvia, Ruggieri, Serena, Bonafede, Roberta, Mariotti, Raffaella, Constantin, Gabriela, Bonetti, Bruno

المساهمون: Farinazzo, Alessia, Angiari, Stefano, Turano, Ermanna, Bistaffa, Edoardo, Dusi, Silvia, Ruggieri, Serena, Bonafede, Roberta, Mariotti, Raffaella, Constantin, Gabriela, Bonetti, Bruno

مصطلحات موضوعية: multiple sclerosi, neuroimmunology

الوصف: Cell based-therapies represent promising strategies for the treatment of neurological diseases. We have previously shown that adipose stem cells (ASC) ameliorate chronic experimental autoimmune encephalomyelitis (EAE). Recent evidence indicates that most ASC paracrine effects are mediated by extracellular vesicles, i.e. micro- and nanovesicles (MVs and NVs). We show that preventive intravenous administration of NVs isolated from ASC (ASC-NVs) before disease onset significantly reduces the severity of EAE and decreases spinal cord inflammation and demyelination, whereas therapeutic treatment with ASC-NVs does not ameliorate established EAE. This treatment marginally inhibits antigen-specific T cell activation, while reducing microglial activation and demyelination in the spinal cord. Importantly, ASC-NVs inhibited integrin-dependent adhesion of encephalitogenic T cells in vitro, with no effect on adhesion molecule expression. In addition, intravital microscopy showed that encephalitogenic T cells treated with ASC NVs display a significantly reduced rolling and firm adhesion in inflamed spinal cord vessels compared to untreated cells. Our results show that ASC-NVs ameliorate EAE pathogenesis mainly by inhibiting T cell extravasation in the inflamed CNS, suggesting that NVs may represent a novel therapeutic approach in neuro-inflammatory diseases, enabling the safe administration of ASC effector factors.

وصف الملف: ELETTRONICO

العلاقة: info:eu-repo/semantics/altIdentifier/pmid/29748664; info:eu-repo/semantics/altIdentifier/wos/WOS:000431763700006; volume:8; issue:7473; firstpage:1; lastpage:11; numberofpages:11; journal:SCIENTIFIC REPORTS; http://hdl.handle.net/11562/979791Test; info:eu-repo/semantics/altIdentifier/scopus/2-s2.0-85046950823; https://doi.org/10.1038/s41598-018-25676-2Test

الإتاحة: https://doi.org/10.1038/s41598-018-25676-2Test
http://hdl.handle.net/11562/979791Test

المؤلفون: Schiaffino, Lorenzo, Bonafede, Roberta, Scambi, Ilaria, Parrella, Edoardo, Pizzi, Marina, Mariotti, Raffaella

المساهمون: Schiaffino, Lorenzo, Bonafede, Roberta, Scambi, Ilaria, Parrella, Edoardo, Pizzi, Marina, Mariotti, Raffaella

مصطلحات موضوعية: Amyotrophic lateral sclerosis (ALS), epigenetic drug, behaviour

الوصف: Dysregulation in acetylation homeostasis has been implicated in the pathogenesis of the amyotrophic lateral sclerosis (ALS), a fatal neurodegenerative disorder. It is known that the acetylation of transcriptional factors regulates their activity. The acetylation state of NF-kB RelA has been found to dictate the neuroprotective versus the neurotoxic effect of p50/RelA. Here we showed that the pro-apoptotic acetylation mode of RelA, involving a general lysine deacetylation of the subunit with the exclusion of the lysine 310, is evident in the lumbar spinal cord of SOD1(G93A) mice, a murine model of ALS. The administration of the HDAC inhibitor MS-275 and the AMPK/sirtuin 1 activator resveratrol restored the normal RelA acetylation in SOD1(G93A) mice. The SOD1(G93A) mice displayed a 3 weeks delay of the disease onset, associated with improvement of motor performance, and 2 weeks increase of lifespan. The epigenetic treatment rescued the lumbar motor neurons affected in SOD1(G93A) mice, accompanied by increased levels of protein products of NF-kB-target genes, Bcl-xL and brain-derived neurotrophic factor. In conclusion, we here demonstrate that MS-275 and resveratrol restore the acetylation state of RelA in the spinal cord, delaying the onset and increasing the lifespan of SOD1(G93A) mice.

وصف الملف: ELETTRONICO

العلاقة: info:eu-repo/semantics/altIdentifier/pmid/30150770; info:eu-repo/semantics/altIdentifier/wos/WOS:000442870300053; volume:8; issue:12875; firstpage:1; lastpage:13; numberofpages:13; journal:SCIENTIFIC REPORTS; http://hdl.handle.net/11562/984460Test; info:eu-repo/semantics/altIdentifier/scopus/2-s2.0-85052313837; https://doi.org/10.1038/s41598-018-30659-4Test

الإتاحة: https://doi.org/10.1038/s41598-018-30659-4Test
http://hdl.handle.net/11562/984460Test

المؤلفون: Bonafede, Roberta, MARIOTTI, Raffaella

المساهمون: Bonafede, Roberta, Mariotti, Raffaella

مصطلحات موضوعية: amyotrophic lateral sclerosi, pathogenesi, therapy, mesenchymal stem cell, extracellular vesicle

الوصف: Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by progressive muscle paralysis determined by the degeneration of motoneurons in the motor cortex brainstem and spinal cord. The ALS pathogenetic mechanisms are still unclear, despite the wealth of studies demonstrating the involvement of several altered signaling pathways, such as mitochondrial dysfunction, glutamate excitotoxicity, oxidative stress and neuroinflammation. To date, the proposed therapeutic strategies are targeted to one or a few of these alterations, resulting in only a minimal effect on disease course and survival of ALS patients. The involvement of different mechanisms in ALS pathogenesis underlines the need for a therapeutic approach targeted to multiple aspects. Mesenchymal stem cells (MSC) can support motoneurons and surrounding cells, reduce inflammation, stimulate tissue regeneration and release growth factors. On this basis, MSC have been proposed as promising candidates to treat ALS. However, due to the drawbacks of cell therapy, the possible therapeutic use of extracellular vesicles (EVs) released by stem cells is raising increasing interest. The present review summarizes the main pathological mechanisms involved in ALS and the related therapeutic approaches proposed to date, focusing on MSC therapy and their preclinical and clinical applications. Moreover, the nature and characteristics of EVs and their role in recapitulating the effect of stem cells are discussed, elucidating how and why these vesicles could provide novel opportunities for ALS treatment.

وصف الملف: STAMPA

العلاقة: info:eu-repo/semantics/altIdentifier/pmid/28377696; info:eu-repo/semantics/altIdentifier/wos/WOS:000396985400002; volume:11; firstpage:1; lastpage:16; numberofpages:16; journal:FRONTIERS IN CELLULAR NEUROSCIENCE; http://hdl.handle.net/11562/959541Test; info:eu-repo/semantics/altIdentifier/scopus/2-s2.0-85018345046; https://doi.org/10.3389/fncel.2017.00080Test

الإتاحة: https://doi.org/10.3389/fncel.2017.00080Test
http://hdl.handle.net/11562/959541Test

المؤلفون: Baglio, S. Rubina, Lagerweij, Tonny, Pérez-Lanzón, Maria, Ho, Xuan Dung, Léveillé, Nicolas, Melo, Sonia A., Cleton-Jansen, Anne-Marie, Jordanova, Ekaterina S., Roncuzzi, Laura, Greco, Michelina, van Eijndhoven, Monique A. J., Grisendi, Giulia, Dominici, Massimo, Bonafede, Roberta, Lougheed, Sinead M., de Gruijl, Tanja D., Zini, Nicoletta, Cervo, Silvia, Steffan, Agostino, Canzonieri, Vincenzo, Martson, Aare, Maasalu, Katre, Köks, Sulev, Wurdinger, Tom, Baldini, Nicola, Pegtel, D. Michiel

المساهمون: Baglio, S. Rubina, Lagerweij, Tonny, Pérez-Lanzón, Maria, Ho, Xuan Dung, Léveillé, Nicola, Melo, Sonia A., Cleton-Jansen, Anne-Marie, Jordanova, Ekaterina S., Roncuzzi, Laura, Greco, Michelina, van Eijndhoven, Monique A. J., Grisendi, Giulia, Dominici, Massimo, Bonafede, Roberta, Lougheed, Sinead M., de Gruijl, Tanja D., Zini, Nicoletta, Cervo, Silvia, Steffan, Agostino, Canzonieri, Vincenzo, Martson, Aare, Maasalu, Katre, Köks, Sulev, Wurdinger, Tom, Baldini, Nicola, Pegtel, D. Michiel

مصطلحات موضوعية: Exosome, Cell, Recipient cells

الوصف: Purpose: Human osteosarcoma is a genetically heterogeneous bone malignancy with poor prognosis despite the employment of aggressive chemotherapy regimens. Because druggable driver mutations have not been established, dissecting the interactions between osteosarcoma cells and supporting stroma may provide insights into novel therapeutic targets.Experimental Design: By using a bioluminescent orthotopic xenograft mouse model of osteosarcoma, we evaluated the effect of tumor extracellular vesicle (EV)-educated mesenchymal stem cells (TEMSC) on osteosarcoma progression. Characterization and functional studies were designed to assess the mechanisms underlying MSC education. Independent series of tissue specimens were analyzed to corroborate the preclinical findings, and the composition of patient serum EVs was analyzed after isolation with size-exclusion chromatography.Results: We show that EVs secreted by highly malignant osteosarcoma cells selectively incorporate a membrane-associated form of TGF beta, which induces proinflammatory IL6 production by MSCs. TEMSCs promote tumor growth, accompanied with intratumor STAT3 activation and lung metastasis formation, which was not observed with control MSCs. Importantly, intravenous administration of the anti-IL6 receptor antibody tocilizumab abrogated the tumor-promoting effects of TEMSCs. RNA-seq analysis of human osteosarcoma tissues revealed a distinct TGF beta-induced prometastatic gene signature. Tissue microarray immunostaining indicated active STAT3 signaling in human osteosarcoma, consistent with the observations in TEMSC-treated mice. Finally, we isolated pure populations of EVs from serum and demonstrated that circulating levels of EV-associated TGF beta are increased in osteosarcoma patients.Conclusions: Collectively, our findings suggest that TEMSCs promote osteosarcoma progression and provide the basis for testing IL6- and TGF beta-blocking agents as new therapeutic options for osteosarcoma patients. (C) 2017 AACR.

العلاقة: info:eu-repo/semantics/altIdentifier/pmid/28053020; info:eu-repo/semantics/altIdentifier/wos/WOS:000405678400029; volume:23; issue:14; firstpage:3721; lastpage:3733; numberofpages:13; journal:CLINICAL CANCER RESEARCH; http://hdl.handle.net/11368/2967826Test; info:eu-repo/semantics/altIdentifier/scopus/2-s2.0-85024115637; https://clincancerres.aacrjournals.org/content/23/14/3721.longTest

الإتاحة: https://doi.org/10.1158/1078-0432.CCR-16-2726Test
http://hdl.handle.net/11368/2967826Test
https://clincancerres.aacrjournals.org/content/23/14/3721.longTest

المؤلفون: BUSATO, ALICE, Bonafede, Roberta, Bontempi, Pietro, SCAMBI, Ilaria, Schiaffino, Lorenzo, BENATI, Donatella, MALATESTA, Manuela, SBARBATI, Andrea, MARZOLA, Pasquina, MARIOTTI, Raffaella

المساهمون: Busato, Alice, Bonafede, Roberta, Bontempi, Pietro, Scambi, Ilaria, Schiaffino, Lorenzo, Benati, Donatella, Malatesta, Manuela, Sbarbati, Andrea, Marzola, Pasquina, Mariotti, Raffaella

مصطلحات موضوعية: MRI, superparamagnetic iron oxide nanoparticles, cellular imaging, stem cells labeling, exosome labeling

الوصف: Purpose: Recent findings indicate that the beneficial effects of adipose stem cells (ASCs), reported in several neurodegenerative experimental models, could be due to their paracrine activity mediated by the release of exosomes. The aim of this study was the development and validation of an innovative exosome-labeling protocol that allows to visualize them with magnetic resonance imaging (MRI).Materials and methods: At first, ASCs were labeled using ultrasmall superparamagnetic iron oxide nanoparticles (USPIO, 4–6 nm), and optimal parameters to label ASCs in terms of cell viability, labeling efficiency, iron content, and magnetic resonance (MR) image contrast were investigated. Exosomes were then isolated from labeled ASCs using a standard isolation protocol. The efficiency of exosome labeling was assessed by acquiring MR images in vitro and in vivo as well as by determining their iron content. Transmission electron microscopy images and histological analysis were performed to validate the results obtained.Results: By using optimized experimental parameters for ASC labeling (200 μg Fe/mL of USPIO and 72 hours of incubation), it was possible to label 100% of the cells, while their viability remained comparable to unlabeled cells; the detection limit of MR images was of 102 and 2.5×103 ASCs in vitro and in vivo, respectively. Exosomes isolated from previously labeled ASCs retain nanoparticles, as demonstrated by transmission electron microscopy images. The detection limit by MRI was 3 μg and 5 μg of exosomes in vitro and in vivo, respectively.Conclusion: We report a new approach for labeling of exosomes by USPIO that allows detection by MRI while preserving their morphology and physiological characteristics.

وصف الملف: ELETTRONICO

العلاقة: info:eu-repo/semantics/altIdentifier/pmid/27330291; info:eu-repo/semantics/altIdentifier/wos/WOS:000377086000001; volume:11; firstpage:2481; lastpage:2490; numberofpages:10; journal:INTERNATIONAL JOURNAL OF NANOMEDICINE; http://hdl.handle.net/11562/944325Test; info:eu-repo/semantics/altIdentifier/scopus/2-s2.0-84971517018; http://dx.doi.org/10.2147/IJN.S104152Test

الإتاحة: https://doi.org/10.2147/IJN.S104152Test
http://hdl.handle.net/11562/944325Test