15 results on '"Roberta Sessa Stilhano"'
Search Results
2. Skeletal muscle healing by M1-like macrophages produced by transient expression of exogenous GM-CSF
- Author
-
Sang Won Han, Camila Congentino Gallo, Timothy J. Koh, Daniela Santoro Rosa, Tâmisa Seeko Bandeira Honda, Leonardo de Oliveira Martins, Roberta Sessa Stilhano, and Patrícia Terra Alves
- Subjects
0301 basic medicine ,Muscle tissue ,medicine.medical_specialty ,Macrophage ,Angiogenesis ,Skeletal muscle ,Medicine (miscellaneous) ,Injury ,030204 cardiovascular system & hematology ,Arteriogenesis ,Biochemistry, Genetics and Molecular Biology (miscellaneous) ,lcsh:Biochemistry ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Fibrosis ,Internal medicine ,medicine ,Animals ,lcsh:QD415-436 ,Muscle, Skeletal ,Uncategorized ,lcsh:R5-920 ,Wound Healing ,Myogenesis ,business.industry ,Macrophages ,Research ,Regeneration (biology) ,Granulocyte-Macrophage Colony-Stimulating Factor ,GM-CSF ,Contusion ,Cell Biology ,medicine.disease ,M2 Macrophage ,Mice, Inbred C57BL ,030104 developmental biology ,medicine.anatomical_structure ,Endocrinology ,Molecular Medicine ,lcsh:Medicine (General) ,business - Abstract
Background After traumatic skeletal muscle injury, muscle healing is often incomplete and produces extensive fibrosis. The sequence of M1 and M2 macrophage accumulation and the duration of each subtype in the injured area may help to direct the relative extent of fibrogenesis and myogenesis during healing. We hypothesized that increasing the number of M1 macrophages early after traumatic muscle injury would produce more cellular and molecular substrates for myogenesis and fewer substrates for fibrosis, leading to better muscle healing. Methods To test this hypothesis, we transfected skeletal muscle with a plasmid vector to transiently express GM-CSF shortly after injury to drive the polarization of macrophages towards the M1 subset. C57BL/6 mouse tibialis anterior (TA) muscles were injured by contusion and electroporated with uP-mGM, which is a plasmid vector that transiently expresses GM-CSF. Myogenesis, angiogenesis, and fibrosis were evaluated by histology, immunohistochemistry, and RT-qPCR; subpopulations of macrophages by flow cytometry; and muscle functioning by the maximum running speed on the treadmill and the recovery of muscle mass. Results Muscle injury increased the number of local M1-like macrophages and decreased the number of M2-like macrophages on day 4, and uP-mGM treatment enhanced this variation. uP-mGM treatment decreased TGF-β1 protein expression on day 4, and the Sirius Red-positive area decreased from 35.93 ± 15.45% (no treatment) to 2.9% ± 6.5% (p Hgf, Hif1α, and Mtor gene expression; arteriole density; and muscle fiber number during regeneration. The improvement in the quality of the muscle tissue after treatment with uP-mGM affected the increase in the TA muscle mass and the maximum running speed on a treadmill. Conclusion Collectively, our data show that increasing the number of M1-like macrophages immediately after traumatic muscle injury promotes muscle recovery with less fibrosis, and this can be achieved by the transient expression of GM-CSF.
- Published
- 2020
3. 17b-Estradiol, a potential ally to alleviate SARS-CoV- 2 infection
- Author
-
Carla Máximo Prado, Ana Cristina Breithaupt-Faloppa, Luiz Felipe P. Moreira, Cristiano de Jesus Correia, Roberta Sessa Stilhano, and Rodrigo Portes Ureshino
- Subjects
Male ,medicine.drug_class ,T cell ,Pneumonia, Viral ,Vasodilation ,Inflammation ,Review Article ,030204 cardiovascular system & hematology ,Nitric oxide ,Betacoronavirus ,03 medical and health sciences ,chemistry.chemical_compound ,Sex Factors ,0302 clinical medicine ,Immune system ,medicine ,Animals ,Humans ,030212 general & internal medicine ,coagulation ,Endothelial dysfunction ,Pandemics ,lcsh:R5-920 ,Coagulation ,Estradiol ,SARS-CoV-2 ,business.industry ,17β-estradiol ,COVID-19 ,General Medicine ,Hypoxia (medical) ,medicine.disease ,Immunity, Innate ,infection ,Rats ,medicine.anatomical_structure ,covid-19 ,chemistry ,inflammation ,Estrogen ,17 b -Estradio ,Immunology ,Female ,medicine.symptom ,Coronavirus Infections ,lcsh:Medicine (General) ,business ,Infection - Abstract
Considering that female sexual hormones may modulate the inflammatory response and also exhibit direct effects on the cells of the immune system, herein, we intend to discuss the sex differences and the role of estradiol in modulating the lung and systemic inflammatory response, focusing on its possible application as a treatment modality for SARS-CoV-2 patients. COVID-19 patients develop severe hypoxemia early in the course of the disease, which is silent most of the time. Small fibrinous thrombi in pulmonary arterioles and a tumefaction of endothelial were observed in the autopsies of fatal COVID-19 cases. Studies showed that the viral infection induces a vascular process in the lung, which included vasodilation and endothelial dysfunction. Further, the proportions of CD4+ T and CD8+ T lymphocytes were strongly reduced in patients with severe SARS-CoV-2 infection. Estradiol is connected with CD4+ T cell numbers and increases T-reg cell populations, affecting immune responses to infection. It is known that estradiol exerts a protective effect on endothelial function, activating the generation of nitric oxide (NO) via endothelial nitric oxide synthase. Estrogen attenuates the vasoconstrictor response to various stimuli and induces vasodilation in the pulmonary vasculature during stress situations like hypoxia. It exerts a variety of rapid actions, which are initiated after its coupling with membrane receptors, which in turn, may positively modulate vascular responses in pulmonary disease and help to maintain microvascular flow. Direct and indirect mechanisms underlying the effects of estradiol were investigated, and the results point to a possible protective effect of estradiol against COVID-19, indicating that it may be considered as an adjuvant therapeutic element for the treatment of patients affected by the novel coronavirus.
- Published
- 2020
4. SARS-CoV-2 and the Possible Connection to ERs, ACE2 and RAGE: Focus on Susceptibility Factors
- Author
-
Eduardo A. Silva, Michelle Sayuri Nishino, Cynthia Silva Bartolomeo, Roberta Sessa Stilhano, Carla Máximo Prado, Ana Cristina Breithaupt-Faloppa, Shahin Shams, Ana Lopez Ramirez, Angelica Jardim Costa, and Rodrigo Portes Ureshino
- Subjects
0301 basic medicine ,Male ,Physiology ,viruses ,Medical Physiology ,ACE2 ,Estrogen receptor ,Review ,Disease ,medicine.disease_cause ,Bioinformatics ,Biochemistry ,pharmacology_toxicology ,RAGE (receptor) ,0403 veterinary science ,0302 clinical medicine ,Receptors ,Pandemic ,estrogen ,Viral ,Lung ,Coronavirus ,0303 health sciences ,Mortality rate ,Age Factors ,04 agricultural and veterinary sciences ,RAGE ,3. Good health ,Receptors, Estrogen ,Female ,Angiotensin-Converting Enzyme 2 ,Disease Susceptibility ,Mitogen-Activated Protein Kinases ,Coronavirus Infections ,Biotechnology ,Signal Transduction ,Biochemistry & Molecular Biology ,040301 veterinary sciences ,medicine.drug_class ,Pneumonia, Viral ,Reviews ,Peptidyl-Dipeptidase A ,Betacoronavirus ,03 medical and health sciences ,Sex Factors ,Antigens, Neoplasm ,COVID‐19 ,Genetics ,medicine ,Animals ,Humans ,Antigens ,Molecular Biology ,Pandemics ,030304 developmental biology ,SARS-CoV-2 ,business.industry ,COVID-19 ,Estrogens ,Pneumonia ,030104 developmental biology ,Estrogen ,Viral Receptor ,Neoplasm ,Biochemistry and Cell Biology ,business ,030217 neurology & neurosurgery - Abstract
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has provoked major stresses on the health care systems of several countries, and caused the death of more than a quarter of a million people globally, mainly in the elderly population with pre-existing pathologies. Previous studies with coronavirus (SARS-CoV) point to gender differences in infection and disease progression with increased susceptibility in male patients, indicating that estrogens may be associated with physiological protection against the coronavirus. Therefore, the objectives of this work are threefold. First, we aim to summarize the SARS-CoV-2 infection pathway and the roles both the virus and patient play in COVID-19 (Coronavirus disease 2019) progression, clinical symptomology, and mortality. Second, we detail the effect estrogen has on viral infection and host infection response, including its role in both the regulation of key viral receptor expression and the mediation of inflammatory activity. Finally, we describe how ERs (estrogen receptors) and RAGE (receptor for advanced glycation end-products) play a critical role in metabolic pathways, which we envisage could maintain a close interplay with SARS-CoV and COVID-19 mortality rates, despite a current lack of research directly determining how. Taken together, we present the current state of the field regarding SARS-CoV-2 research and illuminate where research is needed to better define the role both estrogen and metabolic comorbidities have in the COVID-19 disease state, which can be key in screening potential therapeutic options as the search for effective treatments continue.
- Published
- 2020
5. PPRP and LPRP regenerative effect in 3D muscle injury celular model
- Author
-
William Filipone, Sofia Mila Cantu de Sales, Wesley Rodrigues, Beatrice Rodrigues Ranieri, and Roberta Sessa Stilhano
- Subjects
Muscle tissue ,business.industry ,Spheroid ,General Medicine ,Muscle injury ,Staining ,Calcein ,Andrology ,chemistry.chemical_compound ,medicine.anatomical_structure ,Cardiotoxin ,chemistry ,medicine ,Myocyte ,Platelet ,business - Abstract
Introduction: Extensive muscle injuries are a clinical challenge due to the formation of fibrous tissue that can lead to chronic pain and disability. Therefore, the search for alternative treatments to regenerate the injured muscle is mandatory. Platelet-Rich-Plasma is an autologous blood derivative in which platelet concentration is above peripheral blood levels. Platelets secrete growth factors and signalizing molecules and even though it is already used in clinical practice for tissue regeneration with positive outcomes, there are no concrete evidence of its efficacy on muscle tissue. The goal of this study was to evaluate the effects of PPRP (poor in leukocytes) and LPRP (rich in leukocytes) in 3D C2C12 muscle cell cultures, after injury with Cardiotoxin (CTX) and BaCl2. Methodology: On day 0 spheroids were prepared using scaffold-free technology in agarose micro-molds. Injury was induced two days later by adding CTX (1uM) or BaCl2 (0.7%). Murine PPRP and LPRP were added on day 4 and on day 6 the spheroids were collected for analysis. The volumes were calculated using brightfield images, and their viability was quantified by calcein/7AAD staining. Results: The spheroids were observed for 16 days. Regarding the spheroid volume, we could observe an exponential growth curve, reaching a plateau by day 12. On day 1, the volume was de 2,6x107 ± 3,83x106 um3 and on the last day, 8,68x107 ± 3,72x107 um3.The necrotic center volume was 5,96x106 ± 2,24x106 um3 on day one e presented a reduction tendency by the fourth day. The total spheroid volume was 2.7 X larger than the necrotic center volume after a week of growth (p
- Published
- 2021
6. PRP and BMAC for Musculoskeletal Conditions via Biomaterial Carriers
- Author
-
Fabio S.M. Yamaguchi, Shahin Shams, Roberta Sessa Stilhano, and Eduardo A. Silva
- Subjects
muscle ,Biocompatible Materials ,Review ,Regenerative Medicine ,Bioinformatics ,bone ,lcsh:Chemistry ,0302 clinical medicine ,Bone marrow aspirate ,bone marrow aspirate concentrate ,cartilage ,lcsh:QH301-705.5 ,Spectroscopy ,Clinical Trials as Topic ,030222 orthopedics ,Platelet-Rich Plasma ,Biomaterial ,Treatment options ,Platelet Rich Plasma ,General Medicine ,Computer Science Applications ,medicine.anatomical_structure ,Development of treatments and therapeutic interventions ,Biotechnology ,biomaterials ,Bone Marrow Cells ,Bioengineering ,Catalysis ,Inorganic Chemistry ,03 medical and health sciences ,Genetics ,medicine ,Humans ,bone marrow aspirate concentrate (bmac) ,Physical and Theoretical Chemistry ,Molecular Biology ,Chemical Physics ,5.2 Cellular and gene therapies ,business.industry ,Cartilage ,Regeneration (biology) ,Organic Chemistry ,Mesenchymal stem cell ,030229 sport sciences ,lcsh:Biology (General) ,lcsh:QD1-999 ,Musculoskeletal ,platelet rich plasma (prp) ,Other Biological Sciences ,Other Chemical Sciences ,business - Abstract
Platelet-rich plasma (PRP) and bone marrow aspirate concentrate (BMAC) are orthobiologic therapies considered as an alternative to the current therapies for muscle, bone and cartilage. Different formulations of biomaterials have been used as carriers for PRP and BMAC in order to increase regenerative processes. The most common biomaterials utilized in conjunction with PRP and BMAC clinical trials are organic scaffolds and natural or synthetic polymers. This review will cover the combinatorial strategies of biomaterial carriers with PRP and BMAC for musculoskeletal conditions (MsCs) repair and regeneration in clinical trials. The main objective is to review the therapeutic use of PRP and BMAC as a treatment option for muscle, bone and cartilage injuries.
- Published
- 2019
7. Platelet-Rich Plasma in a Murine Model
- Author
-
Roberta Sessa Stilhano, Sang Won Han, Priscila Martins Andrade Denapoli, Rene Jorge Abdalla, and Sheila Jean McNeill Ingham
- Subjects
Male ,Vascular Endothelial Growth Factor A ,medicine.medical_specialty ,Platelet-derived growth factor ,Contusions ,medicine.medical_treatment ,Enzyme-Linked Immunosorbent Assay ,Physical Therapy, Sports Therapy and Rehabilitation ,Transforming Growth Factor beta1 ,Mice ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Epidermal growth factor ,Internal medicine ,Leukocytes ,medicine ,Animals ,Humans ,Orthopedics and Sports Medicine ,Platelet-Derived Growth Factor ,Wound Healing ,030222 orthopedics ,Vascular Endothelial Growth Factor Receptor-1 ,biology ,Platelet-Rich Plasma ,business.industry ,Muscles ,Growth factor ,030229 sport sciences ,Mice, Inbred C57BL ,Vascular endothelial growth factor ,Disease Models, Animal ,Vascular endothelial growth factor A ,Endocrinology ,chemistry ,Platelet-rich plasma ,biology.protein ,Hepatocyte growth factor ,business ,Platelet-derived growth factor receptor ,medicine.drug - Abstract
Background: It is well known that platelet-rich plasma (PRP) preparations are not the same and that not all preparations include white blood cells, but the part that leukocytes play on the healing role of PRP is still unknown. Purpose: The primary aim of this study was to evaluate the influence of leukocytes in different PRP preparations with a special emphasis on growth factor concentrations. The secondary aim was to evaluate the influence of PRP on muscle healing. Study Design: Controlled laboratory study. Methods: Two PRP preparation procedures were evaluated. Blood fractions were stained with Rapid Panoptic, and growth factors (transforming growth factor beta 1 [TGF-β1], vascular endothelial growth factor [VEGF], insulin-like growth factor [IGF], epidermal growth factor [EGF], hepatocyte growth factor [HGF], and platelet-derived growth factor [PDGF]) were quantified by enzyme-linked immunosorbent assay. Western blotting analysis was performed for Fms-related tyrosine kinase 1 (Flt-1). A muscle contusion injury was created and treated with PRP at different time points. Results: Leukocytes were the main source of VEGF, and all other growth factors measured had a higher concentration in the preparations that included the buffy coat and consequently had a higher concentration of white blood cells. Flt-1 was also found in platelet-poor plasma (PPP). There were higher concentrations of PDGF and HGF in the preparations that encompassed the buffy coat. A PRP injection 7 days after the injury provided significantly increased exercise performance and decreased the fibrotic area when compared with other PRP-treated groups. Conclusion: VEGF is only present in PRP′s buffy coat, while Flt-1 is present in PPP. A PRP injection 7 days after an injury resulted in improved exercise performance. Clinical Relevance: The presence of Flt-1 in PRP provides yet another explanation for results described in the literature after a PRP injection. This information is relevant for selecting the best PRP for each type of injury.
- Published
- 2016
8. Epidemiology and Clinical Features of Muscle Injuries
- Author
-
Roberta Sessa Stilhano, Rogério Teixeira de Carvalho, Rene Jorge Abdalla, Sheila Jean McNeill Ingham, and Leonardo Addêo Ramos
- Subjects
Hamstring injury ,Amateur sports ,medicine.medical_specialty ,business.industry ,Muscle strains ,Football ,Muscle damage ,medicine.disease ,Muscle injury ,film.subject ,Physical medicine and rehabilitation ,film ,Epidemiology ,medicine ,Eccentric training ,business ,human activities - Abstract
Muscle injuries are extremely frequent in professional and amateur sports. Muscle strains, particularly of the hamstrings, are the most frequent injuries in football (soccer) and Australian football. Muscle injuries can be classified in relation to the time of absence into minor, moderate or SEVERE. One of the main problems in muscle injuries is the recurrence rate and this is especially common in the hamstrings group. The clinical presentation of muscle injuries is highly variable and depends on many factors related the mechanism, intensity and type of trauma. Understanding the injury as well as its cause is crucial for the development of specific treatment and prevention strategies.
- Published
- 2017
9. Treatment of Muscle Injury
- Author
-
Roberta Sessa Stilhano, Rogério Teixeira de Carvalho, Leonardo Addêo Ramos, Rene Jorge Abdalla, and Sheila Jean McNeill Ingham
- Subjects
Novel gene ,Muscle regeneration ,medicine.anatomical_structure ,business.industry ,Eccentric exercise ,medicine.medical_treatment ,Anesthesia ,Cell ,Medicine ,Stem-cell therapy ,business ,Muscle injury ,Return to play - Abstract
Muscle injuries are one of the most frequent lesions in sports. They can be treated conservatively or operatively. Return to play should be based on the patient’s ability to stretch the injured muscle as much as the contralateral healthy muscle, pain-free use of the injured muscle in sports-specific movements (mainly in eccentric exercise), comparable strength between injured and healthy muscles and functional tests. Strategies can be adopted to minimize the recurrence rate and to enhance performance. Novel gene and cell therapies including stem cell therapy are emerging but need more research before they are ready for routine clinical use.
- Published
- 2017
10. Molecular Pathology
- Author
-
Edna Sadayo Miazato Iwamura, Roberta Sessa Stilhano, Denise Barcelos, Ricardo Artigiani Neto, and Sang Won Han
- Subjects
Oncology ,medicine.medical_specialty ,Histology ,Stromal cell ,GiST ,business.industry ,Internal medicine ,medicine ,Brazilian population ,Sample (statistics) ,General Medicine ,business ,Pathology and Forensic Medicine - Published
- 2012
11. Gene and cell therapy for muscle regeneration
- Author
-
Roberta Sessa Stilhano, Johnny Huard, João Bosco Pesquero, Leonardo de Oliveira Martins, and Sheila Jean McNeill Ingham
- Subjects
business.industry ,Genetic enhancement ,Regeneration (biology) ,Growth factor ,medicine.medical_treatment ,Skeletal muscle ,Inflammation ,Bioinformatics ,medicine.disease ,Cell therapy ,Muscle Injuries (SJ McNeill Ingham, Section Editor) ,medicine.anatomical_structure ,Fibrosis ,medicine ,Orthopedics and Sports Medicine ,medicine.symptom ,Stem cell ,business - Abstract
Skeletal muscle injury and healing are multifactorial processes, involving three steps of healing: (1) degeneration and inflammation, (2) regeneration, and (3) fibrosis. Fibrous tissue hinders the muscle’s complete recovery and current therapies fail in achieving total muscle recovery. Gene and cell therapy (or both) are potential future treatments for severe muscular injuries. Stem cells’ properties associated with growth factors or/and cytokines can improve muscle healing and permit long-term recovery.
- Published
- 2015
12. Changes in gene expression and methylation in the blood of patients with first-episode psychosis
- Author
-
Eduardo Gouvea, Sang Won Han, Sintia Iole Belangero, Cristiano Noto, Rodrigo A. Bressan, Vanessa Kiyomi Ota, Patricia Natalia Silva, Ary Gadelha, Roberta Sessa Stilhano, Marcos L. Santoro, Leticia Spindola, Quirino Cordeiro, Bruno Bertolucci Ortiz, and João Ricardo Sato
- Subjects
Oncology ,Adult ,Male ,Psychosis ,medicine.medical_specialty ,Adolescent ,Bisulfite sequencing ,Young Adult ,Internal medicine ,Tachykinin receptor 2 ,Medicine ,Humans ,Serial analysis of gene expression ,RNA, Messenger ,GTP Cyclohydrolase ,Biological Psychiatry ,Genetics ,Psychiatric Status Rating Scales ,Risperidone ,Positive and Negative Syndrome Scale ,business.industry ,Receptors, Neurokinin-2 ,DNA Methylation ,medicine.disease ,Psychiatry and Mental health ,Gene Expression Regulation ,Psychotic Disorders ,Schizophrenia ,Case-Control Studies ,DNA methylation ,Female ,business ,medicine.drug ,Antipsychotic Agents - Abstract
Schizophrenia is a severe mental health disorder with high heritability. The investigation of individuals during their first-episode psychosis (FEP), before the progression of psychotic disorders and especially before treatment with antipsychotic medications, is particularly helpful for understanding this complex disease and for the identification of potential biomarkers. In this study, we compared the expression of genes that are involved in neurotransmission and neurodevelopment of antipsychotic-naive FEP in the peripheral blood of patients (n = 51) and healthy controls (n = 51). In addition, we investigated the differentially expressed genes with respect to a) DNA methylation, b) the correlation between gene expression and clinical variables (PANSS), and c) gene expression changes after risperidone treatment. Expression levels of 11 genes were quantified with SYBR Green. For methylation analysis, bisulfite sequencing was performed. A significant decrease in GCH1 mRNA levels was observed in FEP patients relative to controls. Also, when we compare the FEP patients after risperidone treatment with controls, this difference remains significant, and no significant differences were observed in GCH1 mRNA levels when comparing patients before and after risperidone treatment. Additionally, although the differences were non-significant after Bonferroni correction, the expression of GCH1 seemed to be correlated with PANSS scores, and the GCH1 promoter region was more methylated in FEP than in controls, thus corroborating the results obtained at the mRNA level. Few studies have been conducted on GCH1, and future studies are needed to clarify its potential role in the progression of schizophrenia.
- Published
- 2014
13. Mesenchymal stem cells do not prevent antibody responses against human α-L-iduronidase when used to treat mucopolysaccharidosis type I
- Author
-
Vanessa Gonçalves Pereira, Vivian Yochiko Samoto, Christina Maeda Takiya, Sang Won Han, Fábio Marques, Andréia Hanada Otake, Priscila Keiko Matsumoto Martin, Roberta Sessa Stilhano, Roger Chammas, Giovani B. Peres, Yara M. Michelacci, Vânia D'Almeida, and Flávia Helena da Silva
- Subjects
Mucopolysaccharidosis I ,Iduronidase ,Mice ,Animal Cells ,Molecular Cell Biology ,Medicine and Health Sciences ,Medicine ,Tissue Distribution ,Immune Response ,Cells, Cultured ,Mice, Knockout ,Multidisciplinary ,biology ,Stem Cells ,Antibody titer ,Enzyme replacement therapy ,Gene Therapy ,Animal Models ,Combined Modality Therapy ,Cytokines ,Antibody ,Cellular Types ,Genetic Dominance ,Injections, Intraperitoneal ,Research Article ,Science ,Immunology ,Mouse Models ,Mesenchymal Stem Cell Transplantation ,Research and Analysis Methods ,Mucopolysaccharidosis type I ,Model Organisms ,Autosomal Recessive Diseases ,Genetics ,Animals ,Humans ,Enzyme Replacement Therapy ,Molecular Biology Techniques ,Molecular Biology ,Autoantibodies ,Clinical Genetics ,Autosomal Recessive Traits ,business.industry ,Mesenchymal stem cell ,Autoantibody ,Biology and Life Sciences ,Mesenchymal Stem Cells ,Human Genetics ,Cell Biology ,Mucopolysaccharidoses ,Mice, Inbred C57BL ,Genetics of Disease ,biology.protein ,business ,Developmental Biology - Abstract
Mucopolysaccharidosis type I (MPSI) is an autosomal recessive disease that leads to systemic lysosomal storage, which is caused by the absence of α-L-iduronidase (IDUA). Enzyme replacement therapy is recognized as the best therapeutic option for MPSI; however, high titers of anti-IDUA antibody have frequently been observed. Due to the immunosuppressant properties of MSC, we hypothesized that MSC modified with the IDUA gene would be able to produce IDUA for a long period of time. Sleeping Beauty transposon vectors were used to modify MSC because these are basically less-immunogenic plasmids. For cell transplantation, 4×10(6) MSC-KO-IDUA cells (MSC from KO mice modified with IDUA) were injected into the peritoneum of KO-mice three times over intervals of more than one month. The total IDUA activities from MSC-KO-IDUA before cell transplantation were 9.6, 120 and 179 U for the first, second and third injections, respectively. Only after the second cell transplantation, more than one unit of IDUA activity was detected in the blood of 3 mice for 2 days. After the third cell transplantation, a high titer of anti-IDUA antibody was detected in all of the treated mice. Anti-IDUA antibody response was also detected in C57Bl/6 mice treated with MSC-WT-IDUA. The antibody titers were high and comparable to mice that were immunized by electroporation. MSC-transplanted mice had high levels of TNF-alpha and infiltrates in the renal glomeruli. The spreading of the transplanted MSC into the peritoneum of other organs was confirmed after injection of 111In-labeled MSC. In conclusion, the antibody response against IDUA could not be avoided by MSC. On the contrary, these cells worked as an adjuvant that favored IDUA immunization. Therefore, the humoral immunosuppressant property of MSC is questionable and indicates the danger of using MSC as a source for the production of exogenous proteins to treat monogenic diseases.
- Published
- 2014
14. Comparison of treatments of peripheral arterial disease with mesenchymal stromal cells and mesenchymal stromal cells modified with granulocyte and macrophage colony-stimulating factor
- Author
-
Flavia Franco da Cunha, Roberta Sessa Stilhano, Leonardo de Oliveira Martins, Edgar Julian Paredes Gamero, Sang Won Han, and Priscila Keiko Matsumoto Martin
- Subjects
Macrophage colony-stimulating factor ,Cancer Research ,Pathology ,medicine.medical_specialty ,Stromal cell ,Necrosis ,Immunology ,Ischemia ,Bone Marrow Cells ,Mesenchymal Stem Cell Transplantation ,Mice ,Peripheral Arterial Disease ,Fibrosis ,medicine ,Immunology and Allergy ,Animals ,Microvessel ,Genetics (clinical) ,Transplantation ,business.industry ,Mesenchymal stem cell ,Granulocyte-Macrophage Colony-Stimulating Factor ,Extremities ,Mesenchymal Stem Cells ,Cell Biology ,medicine.disease ,Recombinant Proteins ,medicine.anatomical_structure ,Oncology ,Bone marrow ,medicine.symptom ,business ,Signal Transduction - Abstract
Background aims Granulocyte macrophage-colony stimulating factor (GM-CSF) promotes vessel formation through several molecular signaling pathways. Mesenchymal stromal cells (MSCs) have an important role in neovasculogenesis during ischemia because they release pro-angiogenic paracrine factors, pro-survival and immunomodulatory substances and can differentiate into endothelial cells. The objective of this study was to evaluate whether there is synergy between GM-CSF and MSCs in recovering ischemic limbs. Methods MSCs from mouse bone marrow were transduced with a lentiviral vector expressing GM-CSF and injected into animals with surgically induced limb ischemia, with unmodified MCSs used as control. The evolution of limb necrosis was evaluated for 1 month. Muscle strength was assessed on the 30th day, and the animals were euthanized to determine the muscle mass and to perform histological analyses to determine the degree of cellular infiltration, capillary and microvessel densities, fibrosis, necrosis and tissue regeneration. Results Both treatments were able to ameliorate ischemia, decrease the areas of fibrosis, necrosis, adipocytes and leukocyte infiltrates and increase the number of capillaries. The addition of GM-CSF promoted the formation of larger vessels, but it also resulted in more fibrosis and less muscle mass without affecting muscle force. Conclusions Both treatments resulted in a remarkable amelioration of ischemia. More fibrosis and less muscle mass produced by the overexpression of GM-CSF did not affect muscle functionality significantly. Importantly, MSCs overexpressing GM-CSF produced larger vessels, which is an important long-term advantage because larger vessels are more efficient in the reperfusion of ischemic tissues physiologically.
- Published
- 2012
15. Treatment of adult MPSI mouse brains with IDUA-expressing mesenchymal stem cells decreases GAG deposition and improves exploratory behavior
- Author
-
Vivian Yochiko Samoto, Roberta Sessa Stilhano, Leonardo Carvalho, Priscila Keiko Matsumoto, Eduardo Gallatti Yasumura, Leny Toma, Bianca Cristina Garcia Lisboa, Sang Won Han, Letícia de Campos Brandão, Marimelia Porcionatto, Vânia D'Almeida, Vanessa Gonçalves Pereira, Bruno Frederico Aguilar Calegare, Flávia Helena da Silva, Valderez Bastos Valero, Helena B. Nader, Melissa Camassola, Nance Beyer Nardi, Thais R. M. Filippo, and Ligia Zacchi Tenório
- Subjects
Retroviral vectors ,Lysosomal storage disorder ,IDUA ,Genetic enhancement ,Immunology ,Neuroprotection ,Glycosaminoglycan ,Mucopolysaccharidosis type I ,chemistry.chemical_compound ,Gene therapy ,Murine leukemia virus ,Immunology and Allergy ,Medicine ,Mesenchymal stem cell ,biology ,business.industry ,Research ,Heparan sulfate ,biology.organism_classification ,Cell biology ,chemistry ,MPSI ,Molecular Medicine ,Iduronidase ,business ,Biotechnology - Abstract
Background Mucopolysaccharidosis type I (MPSI) is caused by a deficiency in alpha-L iduronidase (IDUA), which leads to lysosomal accumulation of the glycosaminoglycans (GAGs) dermatan and heparan sulfate. While the currently available therapies have good systemic effects, they only minimally affect the neurodegenerative process. Based on the neuroprotective and tissue regenerative properties of mesenchymal stem cells (MSCs), we hypothesized that the administration of MSCs transduced with a murine leukemia virus (MLV) vector expressing IDUA to IDUA KO mouse brains could reduce GAG deposition in the brain and, as a result, improve neurofunctionality, as measured by exploratory activity. Methods MSCs infected with an MLV vector encoding IDUA were injected into the left ventricle of the brain of 12- or 25-month-old IDUA KO mice. The behavior of the treated mice in the elevated plus maze and open field tests was observed for 1 to 2 months. Following these observations, the brains were removed for biochemical and histological analyses. Results After 1 or 2 months of observation, the presence of the transgene in the brain tissue of almost all of the treated mice was confirmed using PCR, and a significant reduction in GAG deposition was observed. This reduction was directly reflected in an improvement in exploratory activity in the open field and the elevated plus maze tests. Despite these behavioral improvements and the reduction in GAG deposition, IDUA activity was undetectable in these samples. Overall, these results indicate that while the initial level of IDUA was not sustainable for a month, it was enough to reduce and maintain low GAG deposition and improve the exploratory activity for months. Conclusions These data show that gene therapy, via the direct injection of IDUA-expressing MSCs into the brain, is an effective way to treat neurodegeneration in MPSI mice.
- Published
- 2012
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.