Neuroscience Institute of the Cavalieri Ottolenghi Foundation

Sant'Ambrogio di Torino, Italy

Neuroscience Institute of the Cavalieri Ottolenghi Foundation

Sant'Ambrogio di Torino, Italy
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Gartner A.,University of Porto | Pereira T.,University of Porto | Simoes M.J.,University of Porto | Armada-da-Silva P.A.S.,University of Lisbon | And 13 more authors.
Neural Regeneration Research | Year: 2012

Many studies have been dedicated to the development of scaffolds for improving post-traumatic nerve regeneration. The goal of this study was to assess the effect on nerve regeneration, associating a hybrid chitosan membrane with non-differentiated human mesenchymal stem cells isolated from Wharton's jelly of umbilical cord, in peripheral nerve reconstruction after crush injury. Chromosome analysis on human mesenchymal stem cell line from Wharton's jelly was carried out and no structural alterations were found in metaphase. Chitosan membranes were previously tested in vitro, to assess their ability in supporting human mesenchymal stem cell survival, expansion, and differentiation. For the in vivo testing, Sasco Sprague adult rats were divided in 4 groups of 6 or 7 animals each: Group 1, sciatic axonotmesis injury without any other intervention (Group 1-Crush); Group 2, the axonotmesis lesion of 3 mm was infiltrated with a suspension of 1 250 -1 500 human mesenchymal stem cells (total volume of 50 μL) (Group 2-CrushCell); Group 3, axonotmesis lesion of 3 mm was enwrapped with a chitosan type III membrane covered with a monolayer of non-differentiated human mesenchymal stem cells (Group 3-CrushChitIIICell) and Group 4, axonotmesis lesion of 3 mm was enwrapped with a chitosan type III membrane (Group 4-CrushChitIII). Motor and sensory functional recovery was evaluated throughout a healing period of 12 weeks using sciatic functional index, static sciatic index, extensor postural thrust, and withdrawal reflex latency. Stereological analysis was carried out on regenerated nerve fibers. Results showed that infiltration of human mesenchymal stem cells, or the combination of chitosan membrane enwrapment and human mesenchymal stem cell enrichment after nerve crush injury provide a slight advantage to post-traumatic nerve regeneration. Results obtained with chitosan type III membrane alone confirmed that they significantly improve post-traumatic axonal regrowth and may represent a very promising clinical tool in peripheral nerve reconstructive surgery. Yet, umbilical cord human mesenchymal stem cells, that can be expanded in culture and induced to form several different types of cells, may prove, in future experiments, to be a new source of cells for cell therapy, including targets such as peripheral nerve and muscle.


Vieira H.L.A.,Institute Biologia Experimental e Tecnologica IBET | Vieira H.L.A.,New University of Lisbon | Alves P.M.,Institute Biologia Experimental e Tecnologica IBET | Alves P.M.,New University of Lisbon | Vercelli A.,Neuroscience Institute of the Cavalieri Ottolenghi Foundation
Progress in Neurobiology | Year: 2011

Low oxygen concentrations (hypoxia) occur in several physiological and pathological cellular situations such as embryogenesis and stem cell modulation (in terms of differentiation/proliferation), or ischemic stroke and cancer. On the other side of the coin, the generation of reactive oxygen species (ROS) is tightly controlled by the cell. ROS control redox sensitive signaling pathways and thus regulate cell physiology, such as programmed cell death, inflammation and/or stem cell modulation. Herein we analyze the role of hypoxia and ROS in the modulation of neuronal differentiation focusing on: (i) in vivo neurogenesis and (ii) in vitro neuronal differentiation from neural stem/precursor cells. In vivo, hypoxia promotes neurogenesis in embryos, newborns and adults, as well as in response to noxious stimuli such as ischemia. On the other hand, oxygen and ROS also play a role in in vitro neuronal differentiation. They further impact tumor growth by influencing cell proliferation and differentiation, such as in neuroblastoma development. Therefore, manipulating hypoxia and ROS production represents a useful therapeutic tool if one needs either to enhance or to modulate neurogenesis and neuronal differentiation, such as in cell replacement or in malignant cell proliferation. © 2011 Elsevier Ltd.


Fregnan F.,University of Turin | Fregnan F.,Neuroscience Institute of the Cavalieri Ottolenghi Foundation | Muratori L.,University of Turin | Muratori L.,Neuroscience Institute of the Cavalieri Ottolenghi Foundation | And 3 more authors.
Neural Regeneration Research | Year: 2012

Inflammatory events occurring in the distal part of an injured peripheral nerve have, nowadays, a great resonance. Investigating the timing of action of the several cytokines in the important stages of Wallerian degeneration helps to understand the regenerative process and design pharmacologic intervention that promotes and expedites recovery. The complex and synergistic action of inflammatory cytokines finally promotes axonal regeneration. Cytokines can be divided into pro- and anti-inflammatory cytokines that upregulate and downregulate, respectively, the production of inflammatory mediators. While pro-inflammatory cytokines are expressed in the first phase of Wallerian degeneration and promote the recruitment of macrophages, anti-inflammatory cytokines are expressed after this recruitment and downregulate the production of all cytokines, thus determining the end of the process. In this review, we describe the major inflammatory cytokines involved in Wallerian degeneration and the early phases of nerve regeneration. In particular, we focus on interleukin-1, interleukin-2, interleukin-6, tumor necrosis factor-β, interleukin-10 and transforming growth factor-β.


Resaz R.,G Gaslini Institute | Emionite L.,Italian National Cancer Institute | Vanni C.,G Gaslini Institute | Astigiano S.,Italian National Cancer Institute | And 16 more authors.
Journal of Hepatology | Year: 2011

Background & Aims: Several studies have shown that bone marrow-derived committed myelomonocytic cells can repopulate diseased livers by fusing with host hepatocytes and can restore normal liver function. These data suggest that myelomonocyte transplantation could be a promising approach for targeted and well-tolerated cell therapy aimed at liver regeneration. We sought to determine whether bone marrow-derived myelomonocytic cells could be effective for liver reconstitution in newborn mice knock-out for glucose-6-phosphatase-α. Methods: Bone marrow-derived myelomonocytic cells obtained from adult wild type mice were transplanted in newborn knock-out mice. Tissues of control and treated mice were frozen for histochemical analysis, or paraffin-embedded and stained with hematoxylin and eosin for histological examination or analyzed by immunohistochemistry or fluorescent in situ hybridization. Results: Histological sections of livers of treated knock-out mice revealed areas of regenerating tissue consisting of hepatocytes of normal appearance and partial recovery of normal architecture as early as 1 week after myelomonocytic cells transplant. FISH analysis with X and Y chromosome paints indicated fusion between infused cells and host hepatocytes. Glucose-6-phosphatase activity was detected in treated mice with improved profiles of liver functional parameters. Conclusions: Our data indicate that bone marrow-derived myelomonocytic cell transplant may represent an effective way to achieve liver reconstitution of highly degenerated livers in newborn animals. © 2011 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.


Gartner A.,Abel Salazar Biomedical Sciences Institute | Gartner A.,Institute Ciencias e Tecnologias Agrarias e Agro Alimentares ICETA | Pereira T.,Abel Salazar Biomedical Sciences Institute | Pereira T.,Institute Ciencias e Tecnologias Agrarias e Agro Alimentares ICETA | And 19 more authors.
Journal of Stem Cells and Regenerative Medicine | Year: 2014

Peripheral nerves have the intrinsic capacity of self-regeneration after traumatic injury but the extent of the regeneration is often very poor. Increasing evidence demonstrates that mesenchymal stem/stromal cells (MSCs) may play an important role in tissue regeneration through the secretion of soluble trophic factors that enhance and assist in repair by paracrine activation of surrounding cells. In the present study, the therapeutic value of a population of umbilical cord tissue-derived MSCs, obtained by a proprietary method (UCX®), was evaluated on end-to-end rat sciatic nerve repair. Furthermore, in order to promote both, end-to-end nerve fiber contacts and MSC cell-cell interaction, as well as reduce the flush away effect of the cells after administration, a commercially available haemostatic sealant, Floseal®, was used as vehicle. Both, functional and morphologic recoveries were evaluated along the healing period using extensor postural thrust (EPT), withdrawal reflex latency (WRL), ankle kinematics analysis, and either histological analysis or stereology, in the hyper-acute, acute and chronic phases of healing. The histological analysis of the hyper-acute and acute phase studies revealed that in the group treated with UCX® alone the Wallerian degeneration was improved for the subsequent process of regeneration, the fiber organization was higher, and the extent of fibrosis was lower. The chronic phase experimental groups revealed that treatment with UCX® induced an increased number of regenerated fibers and thickening of the myelin sheet. Kinematics analysis showed that the ankle joint angle determined for untreated animals was significantly different from any of the treated groups at the instant of initial contact (IC). At opposite toe off (OT) and heel rise (HR), differences were found between untreated animals and the groups treated with either UCX® alone or UCX® administered with Floseal®. Overall, the UCX® application presented positive effects in functional and morphologic recovery, in both the acute and chronic phases of the regeneration process. Kinematics analysis has revealed positive synergistic effects brought by Floseal® as vehicle for MSCs. © Journal of Stem Cells and Regenerative Medicine. All rights reserved.


Boido M.,Neuroscience Institute of the Cavalieri Ottolenghi Foundation | Garbossa D.,University of Turin | Vercelli A.,Neuroscience Institute of the Cavalieri Ottolenghi Foundation
Journal of Neurosurgery: Spine | Year: 2011

Object. Spinal cord injury (SCI) often results in irreversible and permanent neurological deficits below the injury site and is considered a pathological state of functional damage to local neurons and axon fibers. There are several experimental treatments to minimize tissue damage, and recently cell transplantation has emerged as a promising approach in spinal cord repair. The authors undertook this study to evaluate grafting of neural tube precursors as a possible therapeutic strategy in a model of spinal cord compression in the mouse. Methods. Compression SCI was induced at the T-13 level in adult male mice. Immediately after injury, neural precursor cells (NPs) were transplanted into the SCI lesion cavity in 18 mice; the remaining 19 mice received saline injections into the lesion cavity and were used as controls. Spinal cords were examined 12, 19, and 26 days postinjury to investigate the survival of the NPs and their effects on the cellular environment, glial scar and glial cyst formation, astrogliosis, and microglial activation. Results. Grafted NPs survived well and integrated into the host spinal cord tissue. Some NPs had differentiated into cells expressing glial and neuronal markers at all 3 end points. Analysis of glial cyst volume showed a lesion volume reduction of 63.2% in the NP-treated mice compared with volume in the injured but untreated mice. There appeared to be no difference in astroglial and microglial activation between untreated mice and treated ones. Sensory and motor tests demonstrated that transplantation of NPs promoted improvement in injured and treated animals compared with controls. Conclusions. These results support the therapeutic potential of NPs, demonstrating that they can survive for a long time, differentiate, integrate into the injured spinal cord, and promote functional recovery after SCI.


Piras A.,Neuroscience Institute of the Cavalieri Ottolenghi Foundation | Piras A.,University of Turin | Gianetto D.,Neuroscience Institute of the Cavalieri Ottolenghi Foundation | Gianetto D.,University of Turin | And 5 more authors.
PLoS ONE | Year: 2011

Acute primary open angle glaucoma is an optic neuropathy characterized by the elevation of intraocular pressure, which causes retinal ischemia and neuronal death. Rat ischemia/reperfusion enhances endocytosis of both horseradish peroxidase (HRP) or fluorescent dextran into ganglion cell layer (GCL) neurons 24 h after the insult. We investigated the activation of autophagy in GCL-neurons following ischemia/reperfusion, using acid phosphatase (AP) histochemistry and immunofluorescence against LC3 and LAMP1. Retinal I/R lead to the appearance of AP-positive granules and LAMP1-positive vesicles 12 and 24 h after the insult, and LC3 labelling at 24 h, and induced a consistent retinal neuron death. At 48 h the retina was negative for autophagic markers. In addition, Western Blot analysis revealed an increase of LC3 levels after damage: the increase in the conjugated, LC3-II isoform is suggestive of autophagic activity. Inhibition of autophagy by 3-methyladenine partially prevented death of neurons and reduces apoptotic markers, 24 h post-lesion. The number of neurons in the GCL decreased significantly following I/R (I/R 12.21±1.13 vs controls 19.23±1.12 cells/500 μm); this decrease was partially prevented by 3-methyladenine (17.08±1.42 cells/500 μm), which potently inhibits maturation of autophagosomes. Treatment also prevented the increase in glial fibrillary acid protein immunoreactivity elicited by I/R. Therefore, targeting autophagy could represent a novel and promising treatment for glaucoma and retinal ischemia. © 2011 Piras et al.


PubMed | Neuroscience Institute of the Cavalieri Ottolenghi Foundation
Type: Journal Article | Journal: PloS one | Year: 2011

Acute primary open angle glaucoma is an optic neuropathy characterized by the elevation of intraocular pressure, which causes retinal ischemia and neuronal death. Rat ischemia/reperfusion enhances endocytosis of both horseradish peroxidase (HRP) or fluorescent dextran into ganglion cell layer (GCL) neurons 24 h after the insult. We investigated the activation of autophagy in GCL-neurons following ischemia/reperfusion, using acid phosphatase (AP) histochemistry and immunofluorescence against LC3 and LAMP1. Retinal I/R lead to the appearance of AP-positive granules and LAMP1-positive vesicles 12 and 24 h after the insult, and LC3 labelling at 24 h, and induced a consistent retinal neuron death. At 48 h the retina was negative for autophagic markers. In addition, Western Blot analysis revealed an increase of LC3 levels after damage: the increase in the conjugated, LC3-II isoform is suggestive of autophagic activity. Inhibition of autophagy by 3-methyladenine partially prevented death of neurons and reduces apoptotic markers, 24 h post-lesion. The number of neurons in the GCL decreased significantly following I/R (I/R 12.211.13 vs controls 19.231.12 cells/500 m); this decrease was partially prevented by 3-methyladenine (17.081.42 cells/500 m), which potently inhibits maturation of autophagosomes. Treatment also prevented the increase in glial fibrillary acid protein immunoreactivity elicited by I/R. Therefore, targeting autophagy could represent a novel and promising treatment for glaucoma and retinal ischemia.

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