Efeito do exercício associado ao transplante de células-tronco sobre a função ventricular de ratos pós-infarto agudo do miocárdio / Effect of exercise associated with stem cell transplantation on ventricular function in rats after acute myocardial infarction

OBJETIVO: Avaliar o efeito da associação terapêutica entre o transplante autólogo de células-tronco e o exercício físico aquático, sobre a fração de ejeção do ventrículo esquerdo (FEVE) de ratos com disfunção ventricular pós-infarto agudo do miocárdio (IAM). MÉTODOS: Foram induzidos ao IAM, por ligadura da artéria coronária esquerda, 21 ratos Wistar. Os animais foram submetidos à ecocardiografia para avaliação da FEVE (%) e dos volumes diastólico e sistólico finais do ventrículo esquerdo (VDF, VSF, ml), randomizados e ao transplante das células-tronco mononucleares. Os animais foram divididos em quatro grupos: grupo sedentário sem células (n=5), sedentário com células (n=5), treinado sem células (n=5) e treinado com células (n=6). O treinamento físico foi iniciado 30 dias após o IAM e realizado em piscina adaptada durante 30 dias. No início e no final do protocolo de treinamento físico, foram realizadas dosagens de lactato. Os animais foram submetidos a nova ecocardiografia após 60 dias do IAM. RESULTADOS: Comparação dos valores de FEVE 30 dias e 60 dias pós-IAM, respectivamente: sedentário sem (35,20 ± 7,64% vs. 22,39 ± 4,56% P=0,026), com células (25,18 ± 7,73% vs. 23,85 ± 9,51% P=0,860) e no treinado sem (21,49 ± 2,70% vs. 20,71 ± 7,14% P=0,792), treinado com células (28,86 ± 6,68 vs. 38,43 ±7,56% P=0,062). Identificou-se a diminuição de fibras colágenas, nas regiões de fibrose miocárdica no grupo treinado com e sem células. CONCLUSÃO: A associação terapêutica entre exercício físico e o transplante autólogo de células-tronco foi benéfica contra as ações do remodelamento ventricular.

OBJECTIVE: To analyze the functional and anatomical-pathological effect of transplantation of bone marrow mononuclear cells associated to aquatic physical activity after myocardial infarction in rats. METHODS: Twenty-one rats were induced by myocardial infarction, through left coronary artery ligation. After a week, the animals were subjected to echocardiography for evaluation of left ventricle ejection fraction (LVEF, %) and dyastolic and end systolic volume of the left ventricle (EDV, ESV, ml), randomized and the transplantation of mononuclear stem cells. The animals were divided into four groups: sedentary group without cells (n=5), sedentary with cells (n=5), trained without cells (n=5) and trained with cells (n=6). The physical training was started 30 days after infarction and held in swimming during 30 days. At the beginning and at the end of the physical training protocol were held assay of lactate. The animals have been subjected to new echocardiography after 60 days of myocardial infarction. RESULTS: Two months after the transplant, were observed decrease in FE in the control group (35.2 to 23.54 P=0.022) and addition of LVEF and stabilization of ventricular remodeling in the group trained with cells (29.85 to 33.43% P=0.062 and 0.71 to 0.73 ml, P=0.776, respectively). Identified the reduction of collagen fibers, myocardial fibrosis regions in the group trained with and without cells. CONCLUSION: The group trained with cells improves ventricular function compared to the control group, suggesting the benefit of associated cell therapy will physical activity.

Effect of exercise associated with stem cell transplantation on ventricular function in rats after acute myocardial infarction.

OBJECTIVE: To analyze the functional and anatomical-pathological effect of transplantation of bone marrow mononuclear cells associated to aquatic physical activity after myocardial infarction in rats. METHODS: Twenty-one rats were induced by myocardial infarction, through left coronary artery ligation. After a week, the animals were subjected to echocardiography for evaluation of left ventricle ejection fraction (LVEF, %) and dyastolic and end systolic volume of the left ventricle (EDV, ESV, ml), randomized and the transplantation of mononuclear stem cells. The animals were divided into four groups: sedentary group without cells (n=5), sedentary with cells (n=5), trained without cells (n=5) and trained with cells (n=6). The physical training was started 30 days after infarction and held in swimming during 30 days. At the beginning and at the end of the physical training protocol were held assay of lactate. The animals have been subjected to new echocardiography after 60 days of myocardial infarction. RESULTS: Two months after the transplant, were observed decrease in FE in the control group (35.2 to 23.54 P=0.022) and addition of LVEF and stabilization of ventricular remodeling in the group trained with cells (29.85 to 33.43% P=0.062 and 0.71 to 0.73 ml, P=0.776, respectively). Identified the reduction of collagen fibers, myocardial fibrosis regions in the group trained with and without cells. CONCLUSION: The group trained with cells improves ventricular function compared to the control group, suggesting the benefit of associated cell therapy will physical activity.

Treatment of Philadelphia chromosome-positive acute lymphoblastic leukemia.

The presence of the Philadelphia chromosome is a poor prognosis factor in acute lymphoblastic leukemia (ALL), in both children and adults. Using molecular techniques of the gen bcr/abl, it is possible to detect the abnormality, in up to the 40% of adult patients. The unsatisfactory results with conventional chemotherapy schemes have determined the intensification of the treatments and the consideration of allogenic bone marrow transplants as the best therapeutic instance. The development of tyrosine kinase inhibitors have become a therapeutic improvement in the treatment of Philadelphia chromosome-positive ALL, being combined with chemotherapy schemes, only in a selected group of patients, even in therapeutic programs that include transplant.

Potential role of stem cell therapy in type 1 diabetes mellitus.

Type 1 diabetes mellitus is the result of the autoimmune response against pancreatic beta-cell(s). At the time of clinical diagnosis near 70% of beta-cell mass is been destroyed as a consequence of the auto-destruction that begins months or even years before the clinical diagnosis. Although marked reduction of chronic complications was seen after development and progression of insulin therapy over the years for type 1 diabetic population, associated risks of chronic end-organ damage and hypoglycemia still remain. Besides tight glucose control, beta-cell mass preservation and/or increase are known to be other important targets in management of type 1 diabetes as long as it reduces chronic microvascular complications in the eyes, kidneys and nerves. Moreover, the larger the beta-cell mass, the lower the incidence of hypoglycemic events. In this article, we discuss some insights about beta-cell regeneration, the importance of regulation of the autoimmune process and what is being employed in human type 1 diabetes in regard to stem cell repertoire to promote regeneration and/or preservation of beta-cell mass.

Potencial role of stem cell therapy in type 1 diabetes mellitus: [review]

Type 1 diabetes mellitus is the result of the autoimmune response against pancreatic beta-cell(s). At the time of clinical diagnosis near 70 percent of beta-cell mass is been destroyed as a consequence of the auto-destruction that begins months or even years before the clinical diagnosis. Although marked reduction of chronic complications was seen after development and progression of insulin therapy over the years for type 1 diabetic population, associated risks of chronic end-organ damage and hypoglycemia still remain. Besides tight glucose control, beta-cell mass preservation and/or increase are known to be other important targets in management of type 1 diabetes as long as it reduces chronic microvascular complications in the eyes, kidneys and nerves. Moreover, the larger the beta-cell mass, the lower the incidence of hypoglycemic events. In this article, we discuss some insights about beta-cell regeneration, the importance of regulation of the autoimmune process and what is being employed in human type 1 diabetes in regard to stem cell repertoire to promote regeneration and/or preservation of beta-cell mass.

O diabetes melito tipo 1 (DM1) é o resultado de uma resposta auto-imune contra as células-beta pancreáticas. Por ocasião do diagnóstico clínico do DM1, aproximadamente 70 por cento da massa de células-beta foram destruídas como conseqüência de uma autodestruição que se iniciou há anos ou meses antes dos primeiros sinais da doença. Embora a redução acentuada das complicações crônicas na população com DM1 foi observada após o desenvolvimento e evolução da insulinoterapia, os riscos associados às lesões dos órgãos-alvo e hipoglicemia persistem. Além do controle intensivo da glicemia, a preservação e/ou o aumento da massa de células-beta são reconhecidos como alvos importantes no tratamento do DM1. Isto vem associado à redução das complicações crônicas microvasculares na retina, rins e nervos e a menor incidência de eventos hipoglicêmicos. Neste artigo, discutimos alguns aspectos da regeneração das células-beta pancreáticas, a importância da regulação do processo auto-imune e o que está sendo empregado no DM1 humano com relação ao repertório das células-tronco nesse sentido.

Bone marrow cell therapy ameliorates and reverses chagasic cardiomyopathy in a mouse model.

Chronic chagasic cardiomyopathy, which is caused by the protozoan Trypanosoma cruzi, is a major cause of heart failure in Latin America. It is a disease for which effective treatment in its advanced clinical forms is lacking. We have previously shown that bone marrow mononuclear cell (BMC) transplantation is effective in reducing inflammation and fibrosis in the mouse model of Chagas disease. The present study used magnetic resonance imaging to assess changes in the cardiac morphology of infected mice after therapy with BMCs. Serial imaging of the BMC-treated mice revealed regression of the right ventricular dilatation typically observed in the chagasic mouse model.

Matrix superhighways configurations: new concepts for complex organ regeneration.

New ideas and experimental models for tissue and organ regeneration are urgently needed. There are several exciting challenges in the field of organogenesis that need to be defined. The integrated signals and molecular repertoires that shape the particular architecture of specific organs like the kidney or the liver are not completely understood yet. To develop a new scientific platform to be able to build up complex organs we have established a research program using basically Acellular Xenogeneic Isomorphic Matrices (AXIMs) and mesenchymal stem cells (MSCs) generating the necessary concepts for the definition, production, and application of the specific configurations of these matrices for organ regeneration. New and interesting pathways for MSC differentiation were identified. We believe that all extracellular matrices were created fundamentally equal or at least very similar in nature. We also believe that there are true "matrix superhighway configurations" with different three-dimensional geometrical architectures as well as biochemical, electrical, and molecular properties that are tissue and organ specific that influence cell differentiation and organogenesis and will be fundamental for the in vitro regeneration of complex organs for transplantation.

Feasibility of a cost-effective approach to evaluate short tandem repeat markers suitable for chimerism follow-up.

BACKGROUND: Precise chimerism monitoring is important for the prediction of the success of allogeneic bone marrow transplantation (BMT). Most of the current procedures employed for chimerism follow-up with short tandem repeat (STR) markers are either time-consuming, labor-intensive, or use expensive assays, making it burdensome to perform large-scale studies of transplanted patients. AIM: To set-up a simple nonradioactive method to investigate a set of STR markers that could be used in the evaluation of chimerism status after allogeneic BMT. METHOD: Six dinucleotide STRs (D2S123, D5S107, CRTL1, D7S500, D11S1356, and TP53) were analyzed by touchdown (TD)-PCR followed by medium size non-denaturing polyacrylamide gel electrophoresis and silver staining. The sensitivity of the approach was evaluated by dilution competition assays. Peripheral blood samples were taken from a group of 50 healthy Argentinean donors, two transplanted patients, and their respective bone marrow donors. Buccal mucosa samples were also obtained from the BMT recipients. RESULTS: Four markers, D2S123, D7S500, D11S1356, and TP53, presented the highest heterozygosities (0.67-0.88) under our experimental system. A sensitivity of 0.8-1.6% for chimerism detection was consistently found for the different STR. The usefulness of these STR in chimerism analysis was illustrated with the screening of related siblings analyzing two transplanted patients with persistent mixed chimerism, which were previously studied by fluorescence in situ hybridization (FISH). Similar proportions of mixed chimerism were obtained with STR analysis compared with those estimated by FISH. DISCUSSION: To our knowledge, this was the first study of mixed chimerism using TD-PCR to achieve a highly specific STR amplification. This approach allows simple and accurate chimerism quantification because it avoids slippage of Taq polymerase on repeat stretches and prevents the differential amplification of the shorter allele. STR heterozygosities and the high level of sensitivity of this method demonstrated that this approach is not only very informative in this population, but is also rapid (taking less than 14 hours) and cost-efficient. CONCLUSION: The data confirms that this method is a useful tool applicable to routine large-scale STR genotyping and mixed chimerism analysis in low-complexity laboratories worldwide.

Effect of bone marrow transplantation on the immunolocalization of p53, hMSH2, and hMLH1 proteins on oral mucosa.

OBJECTIVE: Considering that hMSH2, hMLH1 and p53 are important in maintaining genomic stability of the oral mucosa epithelium, the purpose of the present study was to investigate the immunolocalization of these proteins in the epithelium of the oral mucosa of patients submitted to bone marrow transplantation (BMT) compared with controls. MATERIALS AND METHODS: Twenty-one samples of lip biopsies from BMT recipients were retrieved. Twenty samples of normal lower labial mucosa associated with mucocele in non-transplanted patients were included as control group. The streptavidin-biotin complex stain was used to detect the human DNA mismatch repair proteins hMSH2, hMLH1 and p53 protein. RESULTS: The main findings demonstrated that the mean number of suprabasal epithelial cells positive for MSH2 was statistically higher than the control group. The immunostaining of hMLH1 and p53 at the basal and suprabasal epithelial layers were statistically higher in the oral labial mucosa of the BMT patients compared with controls. CONCLUSION: The present study shows that oral epithelial cells of BMT patients show increased immunolocalization of the DNA repair related proteins.

Longitudinal evaluation of cardiopulmonary performance during exercise after bone marrow transplantation in children.

OBJECTIVE: Abnormalities in cardiopulmonary performance during exercise have been reported in children after bone marrow transplantation (BMT). We sought to study changes in exercise performance over time in pediatric BMT survivors. STUDY DESIGN: We retrospectively reviewed the results of serial cardiopulmonary exercise tests performed by patients who had undergone BMT at our institution. Four measurements of cardiopulmonary function are reported: maximum cardiac index (MCI), maximal oxygen consumption (Max VO(2)), oxygen consumption at ventilatory threshold (VO(2) at VT), and maximum work (Max Work) performed. A linear mixed-effects model was fitted to assess changes in these parameters over time. RESULTS: Thirty-three patients performed 96 cardiopulmonary exercise tests. MCI and VO(2) at VT were depressed at initial testing and did not change over time. Max VO(2) increased by 4% per year to 69% predicted, and Max Work increased to 77% predicted at 6 years after BMT. CONCLUSIONS: In spite of an impaired cardiovascular response to exercise as indicated by the persistently low MCI, aerobic and physical working capacity increase. Improved Max VO(2) suggests that oxygen extraction at the musculoskeletal level becomes more efficient with recovery from BMT. This may represent a compensatory response to an impaired ability to increase cardiac output.

Reconstitution by bone marrow grafting of the defective protective capacity at the migratory phase but not at the intestinal phase of Nippostrongylus brasiliensis infection in W/Wv mice.

After a primary infection by subcutaneous inoculation with the infective larvae (L3) of Nippostrongylus brasiliensis, the intestinal worm burden was higher and expulsion was slower in W/Wv mice than in +/+ mice. When the course of infection was segregated into the migratory and intestinal phases, protection during the migratory phase examined by the larval recovery from the lungs and that during the intestinal phase measured by worm burden after intraduodenal implantation with adult worms were both defective in W/Wv mice. The higher susceptibility of W/Wv mice during the migratory phase was normalized by bone marrow reconstitution. On the other hand, higher susceptibility of W/Wv mice during the intestinal phase, which was measured by worm burden 24 h after intraduodenal implantation of the larvae recovered from the lungs of rats, was not normalized by bone marrow grafting. Furthermore, slower expulsion seen in W/Wv mice after intraduodenal implantation with adult worms was not hastened by bone marrow reconstitution. These results indicate that the protective mechanisms against N. brasiliensis operating during the migratory phase and those during the intestinal phase were different in terms of bone marrow dependency and that non-myeloid cells utilizing c-kit ligand/receptor system to express their functions are involved in the mucosal defence against N. brasiliensis.

Trasplante de medula osea: un tratamiento para los anos 90 / Bone marrow transplantation: a treatment for the 90th years

El trasplante de medula osea es un tratamiento ampliamente aceptado para las enfermedades congenitas y adquiridas de la medula osea, tumores solidos y alteraciones metabolicas congenitas. Actualmente, mas de 240 grupos de TMO trabajan activamente en el mundo y se estima que cada ano se efectuan alrededor de 5.000 procedimientos tanto en la modalidad alogenica (donante genticamente no identico) como autologa (utilizando la medula del mismo paciente). Los resultados mas satisfactorios se han obtenido en el tratamiento de las leucemias agudas, la leucemia mieloide cronica y la anemia aplastica adquirida severa con sobrevidas libres de enfermdead de 60, 70 y 85%, respectivamente. En nuestro medio el primer TMO se efectuo hace 14 anos, y desde 1986 el grupo de TMO de la Universidad de Antioquia en el Hospital San Vicente de Paul, de Medellin, ha realizado 5 procedimientos en pacientes con Sindrome de Wiskott-Aldrich, purpura amegacariocitica y leucemia mieloide cronica

Transplante de médula ósea de trilliza idéntica a paciente con hemoglobinuria paroxística nocturna (HPN) y anemia aplástica / Transplantation of bone marrow from triplet of patient with paroxysmal nocturnal hemoglobin (PNH) and aplastic anemia

Una paciente con hemoglobinuria paroxística nocturna y aplasia severa de la médula ósea fue tratada con transplante de la médula ósea de su hermana trilliza idéntica. La paciente fue preparada con ciclofosfamida y ha tenido recuperación medular completa y constante. Las pruebas que identifican la hemoglobinuria paroxística nocturna han sido persistentemente negativas. La paciente ha estado clínica y hematológicamente normal durante más de 8 años sin ninguna medicación