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Objective: To describe a protocol of obtention of mesenchymal stem cells and to report their use as a biological adjuvant in three patients undergoing arthroscopic rotator cuff repair. Methods: Case series of patients who underwent arthroscopic repair of isolated full-thickness supraspinatus tear using mesenchymal stem cells obtained from the bone marrow as a biological adjuvant. All patients were operated on at the same institution, by a surgeon with 13 years of experience. The cells were applied at the end of the procedure, at the tendon-bone interface, at an approximate concentration of 2,000,000 mesenchymal cells/mm3 and a total volume of 5 ml. Results: All patients improved with the procedure, with one excellent and two good results. All cases overcame the minimally important clinical difference. All cases reached tendon healing, without partial or complete re-tears. We observed no complications. Conclusion: Arthroscopic rotator cuff repair with added mesenchymal cells obtained from bone marrow and submitted to a cell expansion process led to good functional results and healing in all cases in the sample, with no complications. Level of Evidence IV, Case Series.
Objetivo: Descrever o protocolo de obtenção de células mesenquimais e relatar seu uso como adjuvante biológico em três pacientes submetidos ao reparo artroscópico do manguito rotador. Métodos: Série de casos de pacientes submetidos ao reparo artroscópico de rotura transfixante do músculo supraespinal utilizando como adjuvante biológico células mesenquimais obtidas da medula óssea. Todos ospacientes foram operados na mesma instituição por um cirurgião com 13 anos de experiência. As células foram aplicadas ao final do procedimento, na interface do tendão com o osso, na concentração aproximada de 2 milhões de células mesenquimais/mm3 e volume total de 5 ml. Resultados: Todos os pacientes melhoraram após o procedimento, havendo um resultado excelente e dois bons. Todos superaram a diferença clínica minimamente importante. Em todos os casos ocorreu cicatrização tendínea, sem a presença de rerroturas parciais ou completas. Não observamos complicações. Conclusão: O reparo do manguito rotador artroscópico com adição de células mesenquimais obtidas da medula óssea e submetidas a processo de expansão celular levou a bons resultados funcionais e cicatrização, sem complicações, em todos os casos da amostra. Nível de Evidência IV, Série de Casos.
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ABSTRACT Objective: To describe a protocol of obtention of mesenchymal stem cells and to report their use as a biological adjuvant in three patients undergoing arthroscopic rotator cuff repair. Methods: Case series of patients who underwent arthroscopic repair of isolated full-thickness supraspinatus tear using mesenchymal stem cells obtained from the bone marrow as a biological adjuvant. All patients were operated on at the same institution, by a surgeon with 13 years of experience. The cells were applied at the end of the procedure, at the tendon-bone interface, at an approximate concentration of 2,000,000 mesenchymal cells/mm3 and a total volume of 5 ml. Results: All patients improved with the procedure, with one excellent and two good results. All cases overcame the minimally important clinical difference. All cases reached tendon healing, without partial or complete re-tears. We observed no complications. Conclusion: Arthroscopic rotator cuff repair with added mesenchymal cells obtained from bone marrow and submitted to a cell expansion process led to good functional results and healing in all cases in the sample, with no complications. Level of Evidence IV, Case Series.
RESUMO Objetivo: Descrever o protocolo de obtenção de células mesenquimais e relatar seu uso como adjuvante biológico em três pacientes submetidos ao reparo artroscópico do manguito rotador. Métodos: Série de casos de pacientes submetidos ao reparo artroscópico de rotura transfixante do músculo supraespinal utilizando como adjuvante biológico células mesenquimais obtidas da medula óssea. Todos ospacientes foram operados na mesma instituição por um cirurgião com 13 anos de experiência. As células foram aplicadas ao final do procedimento, na interface do tendão com o osso, na concentração aproximada de 2 milhões de células mesenquimais/mm3 e volume total de 5 ml. Resultados: Todos os pacientes melhoraram após o procedimento, havendo um resultado excelente e dois bons. Todos superaram a diferença clínica minimamente importante. Em todos os casos ocorreu cicatrização tendínea, sem a presença de rerroturas parciais ou completas. Não observamos complicações. Conclusão: O reparo do manguito rotador artroscópico com adição de células mesenquimais obtidas da medula óssea e submetidas a processo de expansão celular levou a bons resultados funcionais e cicatrização, sem complicações, em todos os casos da amostra. Nível de Evidência IV, Série de Casos.
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Objective To evaluate in vitro the viability of mesenchymal stem cells derived from adipose tissue (AD-MSCs) in different commercial solutions of hyaluronic acid (HA) before and after being sowed in collagen I/III membrane. Methods In the first stage, the interaction between AD-MSCs was analyzed with seven different commercial products of HA, phosphate buffered saline (PBS), and bovine fetal serum (BFS), performed by counting living and dead cells after 24, 48 and 72 hours. Five products with a higher number of living cells were selected and the interaction between HA with AD-MSCs and type I/III collagen membrane was evaluated by counting living and dead cells in the same time interval (24, 48 and 72 hours). Results In both situations analyzed (HA + AD-MSCs and HA + AD-MSCs + membrane), BFS presented the highest percentage of living cells after 24, 48 and 72 hours, a result higher than that of HA. Conclusion The association of HA with AD-MSCs, with or without membrane, showed no superiority in cell viability when compared with BFS.
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Abstract Objective To evaluate in vitro the viability of mesenchymal stem cells derived from adipose tissue (AD-MSCs) in different commercial solutions of hyaluronic acid (HA) before and after being sowed in collagen I/III membrane. Methods In the first stage, the interaction between AD-MSCs was analyzed with seven different commercial products of HA, phosphate buffered saline (PBS), and bovine fetal serum (BFS), performed by counting living and dead cells after 24, 48 and 72 hours. Five products with a higher number of living cells were selected and the interaction between HA with AD-MSCs and type I/III collagen membrane was evaluated by counting living and dead cells in the same time interval (24, 48 and 72 hours). Results In both situations analyzed (HA + AD-MSCs and HA + AD-MSCs + membrane), BFS presented the highest percentage of living cells after 24, 48 and 72 hours, a result higher than that of HA. Conclusion The association of HA with AD-MSCs, with or without membrane, showed no superiority in cell viability when compared with BFS.
Resumo Objetivo Avaliar in vitro a viabilidade das células-tronco mesenquimais derivadas do tecido adiposo (AD-CTMs) em diferentes soluções comerciais de ácido hialurônico (AH) antes e após serem semeadas em membrana de colágeno I/III. Métodos Na primeira etapa, analisou-se a interação entre AD-CTMs com sete diferentes produtos comerciais de AH, salina tamponada com fosfato (PBS, na sigla em inglês) e soro fetal bovino (SFB), realizada pela contagem das células vivas e mortas após 24, 48 e 72 horas. Foram selecionados cinco produtos com maior número de células vivas e avaliou-se a interação entre o AH com AD-CTMs e a membrana de colágeno tipo I/III pela contagem de células vivas e mortas no mesmo intervalo de tempo (24, 48 e 72 horas). Resultados Em ambas as situações analisadas (AH + AD-CTM e AH + AD-CTM + membrana), o SFB apresentou a maior porcentagem de células vivas após 24, 48 e 72 horas, resultado superior ao do AH. Conclusão A associação do AH com as AD-CTMs, com ou sem a membrana, não demonstrou superioridade na viabilidade celular quando comparado com SFB.
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Técnicas In Vitro , Cartilagem Articular , Colágeno Tipo I , Transplante de Células-Tronco Mesenquimais , Ácido HialurônicoRESUMO
Alzheimer's disease (AD) is a severe disabling condition with no cure currently available, which accounts for 60-70% of all dementia cases worldwide. Therefore, the investigation of possible therapeutic strategies for AD is necessary. To this end, animal models corresponding to the main aspects of AD in humans have been widely used. Similar to AD patients, the double transgenic APPswe/PS1dE9 (APP/PS1) mice show cognitive deficits, hyperlocomotion, amyloid-ß (Αß) plaques in the cortex and hippocampus, and exacerbated inflammatory responses. Recent studies have shown that these neuropathological features could be reversed by stem cell transplantation. However, the effects induced by neural (NSC) and mesenchymal (MSC) stem cells has never been compared in an AD animal model. Therefore, the present study aimed to investigate whether transplantation of NSC or MSC into the hippocampus of APP/PS1 mice reverses AD-induced pathological alterations, evaluated by the locomotor activity (open field test), short- and long-term memory (object recognition) tests, Αß plaques (6-E10), microglia distribution (Iba-1), M1 (iNOS) and M2 (ARG-1) microglial phenotype frequencies. NSC and MSC engraftment reduced the number of Αß plaques and produced an increase in M2 microglia polarization in the hippocampus of APP/PS1 mice, suggesting an anti-inflammatory effect of stem cell transplantation. NSC also reversed the hyperlocomotor activity and increased the number of microglia in the hippocampus of APP/PS1 mice. No impairment of short or long-term memory was observed in APP/PS1 mice. Overall, this study highlights the potential beneficial effects of transplanting NSC or MSC for AD treatment.
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Doença de Alzheimer , Células-Tronco Mesenquimais , Doença de Alzheimer/genética , Doença de Alzheimer/patologia , Doença de Alzheimer/terapia , Peptídeos beta-Amiloides/metabolismo , Animais , Hipocampo/metabolismo , Hipocampo/patologia , Humanos , Células-Tronco Mesenquimais/metabolismo , Camundongos , Camundongos Transgênicos , Placa Amiloide/patologiaRESUMO
INTRODUCTION: Sistemic Sclerosis (SSc) is a heterogeneous autoimmune disease with a high rate of progression and therapeutic failure, and treatment is a challenge, new therapeutic proposals being needed, being mesenchymal stem cells (MSCs) considered as alternative therapy for SSc for its immunomodulatory capacity. We evaluated the efficacy and safety of human MSC (hMSC) in patients with SSc through a systematic literature review (SLR). METHODS: SLR (PRISMA guideline) on MEDLINE/OVID, LILACS, EMBASE, and Cochrane/OVID bases (until July 2020, without limits). All types of clinical studies were considered: patients ≥18 years old with SSc and treatment with hMSC. EXCLUSION CRITERIA: animal models, autologous/allogenic hematopoietic stem cell transplants, narrative reviews, letters to the editor. MeSH and "Key word" terms were used. The level of evidence and the quality rating were rated [Joanna Briggs Institute (JBI) lists]. Registration in PROSPERO repository (ID CRD42020185245) The Synthesis Without Meta-analysis (SWiM) guideline was followed. RESULTS: We initially identified 508 articles, of which 11 were finally included (8 case series and 3 case reports). The 11 articles included 101 patients (85 female, age range 18-75 years). The level of evidence was mostly 4 (JBI); the quality of evidence was met (≥50% of JBI items). SWiM showed that vascular skin involvement (digital ulcers, necrosis, and gangrene) and associated pain were the predominant outcomes, while improvements were found in almost all cases. One patient died in the first month, and the frequency of complications was low. Expanded hMSCs were used in 24 patients and other cell sources in the remaining patients. CONCLUSION: There is too little reported data to reach definite conclusions about the use of hMSC in SSc. Further studies with better epidemiological designs are needed to evaluate the benefit of hMSCs in SSc patients.
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Doenças Autoimunes , Células-Tronco Mesenquimais , Escleroderma Sistêmico , Úlcera Cutânea , Adolescente , Adulto , Idoso , Animais , Feminino , Humanos , Pessoa de Meia-Idade , Escleroderma Sistêmico/terapia , Adulto JovemRESUMO
SUMMARY: Diabetic foot ulcer morbidity and mortality are dramatically increasing worldwide, reinforcing the urgency to propose more effective interventions to treat such a devastating condition. Previously, using a diabetic mouse model, we demonstrated that administration of bone marrow mesenchymal stem cells derivatives is more effective than the use of bone marrow mesenchymal stem cells alone. Here, we used the aforementioned treatments on three patients with grade 2 diabetic foot ulcers and assessed their beneficial effects, relative to the conventional approach. In the present study, two doses of cell derivatives, one dose of mesenchymal stem cells or one dose of vehicle (saline solution with 5% of human albumin), were intradermally injected around wounds. Wound healing process and changes on re-epithelialization were macroscopically evaluated until complete closure of the ulcers. All ulcers were simultaneously treated with conventional treatment (PolyMen® dressing). Patients treated with either cell derivatives or mesenchymal stem cells achieved higher percentages of wound closure in shorter times, relative to the patient treated with the conventional treatment. The cell derivative and mesenchymal stem cells approaches resulted in complete wound closure and enhanced skin regeneration at some point between days 35 and 42, although no differences between these two treatments were observed. Moreover, wounds treated with the conventional treatment healed after 161 days. Intradermal administration of cell derivatives improved wound healing to a similar extent as mesenchymal stem cells. Thus, our results suggest that mesenchymal stem cell derivatives may serve as a novel and potential therapeutic approach to treat diabetic foot ulcers. LEARNING POINTS: In diabetic mouse models, the administration of mesenchymal stem cells derivatives have been demonstrated to be more effective than the use of marrow mesenchymal stem cells alone. Mesenchymal stem cells have been explored as an attractive therapeutic option to treat non-healing ulcers. Mesenchymal stem cells derivatives accelerate the re-epithelialization on diabetic foot ulcers.
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ABSTRACT Background: Crohn's disease is a pathological condition that has different options of treatment, but there are patients who need other therapeutic approach, such as the use of adipose-derived mesenchymal stem cells. Aim: Systematic literature review to determine the different ways of adipose-derived mesenchymal stem cells administration in humans with luminal refractory and perianal fistulizing Crohn's disease. Methods: It was conducted a search for articles (from 2008 to 2018) on PubMed and ScienceDirect databases using the keywords Crohn's disease, fistulizing Crohn's disease, luminal Crohn's disease and transplantation of mesenchymal stem cells or mesenchymal stem cells or stromal cells. Thirteen publications were selected for analysis. Results: Only one study referred to the luminal Crohn´s disease. The number of cells administered was variable, occurring mainly through subcutaneous adipose tissue by liposuction. It could be highlighted the autologous transplant with exclusive infusion of mesenchymal stem cells. The procedures involved in pre-transplant were mainly curettage, setons placement and stitching with absorbable suture, and conducting tests and drug treatment for luminal Crohn´s disease. During transplant, the injection of mesenchymal stem cells across the fistula path during the transplant was mainly on the intestinal tract wall. Conclusion: Although the use of mesenchymal stem cells is promising, the transplant on the luminal region should be more investigated. The injection of mesenchymal stem cells, exclusively, is more explored when compared to treatment with other products. The preparation of the fistulizing tract and the location of cell transplantation involve standardized health care in most studies.
RESUMO Racional: Há diferentes opções de tratamento para a doença de Crohn, porém, em alguns casos, há a necessidade de outras abordagens terapêuticas, como o uso de células-tronco mesenquimais derivadas do tecido adiposo. Objetivo: Revisar sistematicamente a literatura para determinar as diferentes formas de administração das células-tronco mesenquimais derivadas do tecido adiposo em seres humanos com doença de Crohn refratária luminal e fistulizante perianal. Método: Buscaram-se artigos publicados entre 2008 e 2018 nas bases de dados PubMed e ScienceDirect, pelos descritores: Crohn's disease, fistulizing Crohns disease, luminal Crohns disease e transplantation of mesenchymal stem cells ou mesenchymal stem cell ou stromal cells. Treze artigos foram selecionados. Resultados: Somente um trabalho se referiu à doença luminal. A quantidade de células administradas foi variável, obtendo-se principalmente do tecido adiposo subcutâneo por lipoaspiração. Destacou-se o transplante autólogo com a infusão exclusiva de células-tronco mesenquimais. Os procedimentos realizados no pré-transplante foram principalmente o de curetagem, colocação de setons e suturas com fio absorvível, e de exames e tratamento medicamentoso para a doença luminal. No transplante, ocorreu a injeção das células por todo o trajeto fistuloso, principalmente nas paredes do trato. Conclusão: Embora o uso de células-tronco mesenquimais seja promissor, o transplante na região luminal deve ser mais investigado. A injeção exclusiva de células-tronco mesenquimais é mais explorada quando comparada ao tratamento conjunto com outros produtos. A forma de preparo do trato fistuloso e o local de transplante envolvem cuidados médicos padronizados na maioria dos estudos.
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Humanos , Doença de Crohn/terapia , Tecido Adiposo/citologia , Fístula Retal/terapia , Transplante de Células-Tronco Mesenquimais/métodos , Doença de Crohn/complicações , Tecido Adiposo/transplante , Fístula Retal/etiologiaRESUMO
ABSTRACT Objective: The main purpose of this study is to evaluate, in vitro, the cytotoxicity of different commercial brands of hyaluronic acids to be used as a vehicle for injection of human adipose-derived mesenchymal stem cells (AD-MSCs). Methods: AD-MSCs were divided into seven groups: one control group where AD-MSCs were cultivated with phosphate-buffered saline (PBS) and six other groups where AD-MSCs were cultivated with different commercial brands of hyaluronic acid. AD-MSC viability analysis was performed after 4, 24, and 48 h in contact with each treatment, using the trypan staining method on a Countess automated cell counter (Thermo Fisher Scientific). Results: The results clearly demonstrated a significant difference in cell viability when AD-MSCs were exposed to different hyaluronic acids when compared with the control group. Conclusion: These data suggest that hyaluronic acid can be used as a vehicle for injection of human AD-MSCs, but caution is needed to choose the best product, aiming at its future therapeutic application.
RESUMO Objetivo: Avaliar in vitro, de forma direta, a citotoxicidade de ácidos hialurônicos como veículo de injeção para linhagens de células-tronco mesenquimais (CTMs) obtidas de tecido adiposo humano. Métodos: As CTMs foram divididas em sete grupos, os quais foram expostos ao ácido hialurônico de seis marcas comerciais, além do contato com tampão fosfato-salino PBS (grupo controle). Após quatro, 24 e 48 horas, foi feita a análise da viabilidade celular através do contador Countess pelo método de coloração com Trypan Blue (Thermo Fisher Scientific). Resultados: Os resultados demonstraram uma diferença significativa na viabilidade celular quando essas linhagens de CTMs foram expostas aos diferentes ácidos hialurônicos em comparação com o grupo controle. Conclusão: Os dados sugerem que o ácido hialurônico pode ser usado como veículo de injeção para CTMs, porém é necessária cautela na escolha do melhor produto para aplicação terapêutica futura.
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Artroscopia , Cartilagem Articular , Doenças das Cartilagens , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais , JoelhoRESUMO
OBJECTIVE: The main purpose of this study is to evaluate, in vitro, the cytotoxicity of different commercial brands of hyaluronic acids to be used as a vehicle for injection of human adipose-derived mesenchymal stem cells (AD-MSCs). METHODS: AD-MSCs were divided into seven groups: one control group where AD-MSCs were cultivated with phosphate-buffered saline (PBS) and six other groups where AD-MSCs were cultivated with different commercial brands of hyaluronic acid. AD-MSC viability analysis was performed after 4, 24, and 48 h in contact with each treatment, using the trypan staining method on a Countess automated cell counter (Thermo Fisher Scientific). RESULTS: The results clearly demonstrated a significant difference in cell viability when AD-MSCs were exposed to different hyaluronic acids when compared with the control group. CONCLUSION: These data suggest that hyaluronic acid can be used as a vehicle for injection of human AD-MSCs, but caution is needed to choose the best product, aiming at its future therapeutic application.
OBJETIVO: Avaliar in vitro, de forma direta, a citotoxicidade de ácidos hialurônicos como veículo de injeção para linhagens de células-tronco mesenquimais (CTMs) obtidas de tecido adiposo humano. MÉTODOS: As CTMs foram divididas em sete grupos, os quais foram expostos ao ácido hialurônico de seis marcas comerciais, além do contato com tampão fosfato-salino PBS (grupo controle). Após quatro, 24 e 48 horas, foi feita a análise da viabilidade celular através do contador Countess pelo método de coloração com Trypan Blue (Thermo Fisher Scientific). RESULTADOS: Os resultados demonstraram uma diferença significativa na viabilidade celular quando essas linhagens de CTMs foram expostas aos diferentes ácidos hialurônicos em comparação com o grupo controle. CONCLUSÃO: Os dados sugerem que o ácido hialurônico pode ser usado como veículo de injeção para CTMs, porém é necessária cautela na escolha do melhor produto para aplicação terapêutica futura.
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Introdução: hepatopatias crônicas figuram entre as principais causas de morte no mundo e o transplante hepático permanece como a única alternativa terapêutica curativa disponível. A terapia celular utilizando células tronco tem sido demonstrada como uma alternativa potencial, porém os mecanismos envolvidos na resolução da doença permanecem em debate. Metodologia: trata-se de uma revisão de literatura, na qual foi realizada uma consulta por artigos científicos selecionados através de busca no banco de dados do PubMed e Scielo no período de agosto de 2016 a junho de 2017. Objetivo: este estudo tem como objetivo descrever os mecanismos envolvidos na regeneração/reparo da cirrose hepática através do tratamento com células tronco mesenquimais derivadas do tecido adiposo (AD-MSCs). Resultados e discussão: os estudos utilizando terapia celular apontam como principais mecanismos a fibrinólise, imunomodulação e atividade antioxidante. Com relação a atividade fibrinolítica, o aumento proeminente na expressão de enzimas como MMP2, MMP3 e MMP9 explica a redução da fibrose normalmente observada. A atividade imunomodulatória das ADMSCs parece estar relacionada com a secreção de agentes imunossupressores e fatores que promovem a hematopoiese. Finalmente, as AD-MSCS são capazes de aumentar a atividade do NF-E2-related fator (Nrf2), um fator de transcrição crucial na expressão de diversas enzimas relacionadas à resolução do estresse oxidativo. Conclusão: os estudos utilizando a terapia celular evidenciaram que fibrinólise, imunomodulação e efeito antioxidante podem, de forma associada, contribuir para os bons resultados apresentados. Apesar dos principais mecanismos propostos terem sido apresentados neste presente trabalho, mais estudos são necessários a fim de determinar a precisa contribuição de cada um deles.
Introduction: chronic liver diseases are among the leading causes of death in the world and liver transplantation remains the only curative alternative available. Cell therapy using stem cells has been demonstrated as a potential alternative, but the mechanisms involved in this proccess remain unclear. Methodology: we performed a review of the literature, in which a query was made for scientific articles selected through a search in the PubMed and Scielo database from August 2016 to June 2017. Objectives: this study aims to describe the mechanisms involved in the regeneration/repair of liver cirrhosis by treatment with adipose tissue-derived mesenchymal stem cells (AT-MSCs). Results and discussion: studies using cellular therapy point out as main mechanisms fibrinolysis, immunomodulation and antioxidant activity. With regard to fibrinolytic activity, the prominent increase in the expression of enzymes such as MMP2, MMP3 and MMP9 explains the reduction in fibrosis normally observed. The immunomodulatory activity of AT-MSCs appears to be related to the secretion of immunosuppressive agents and factors that promote hematopoiesis. Finally, AT-MSCS are able to increase the activity of the NF-E2-related factor (Nrf2), a crucial transcription factor in the expression of several enzymes related to the resolution of oxidative stress. Conclusion: studies using cell therapy have shown that fibrinolysis, immunomodulation and antioxidant effect may, in an associated way, contribute to the good results presented. Although the main proposed mechanisms have been presented in this present study, more studies are necessary in order to determine the precise contribution of each of them.
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Tecido Adiposo , Transplante de Células-Tronco Mesenquimais , Imunomodulação , Fibrinólise , Cirrose Hepática , Antioxidantes , Estudos de Avaliação como AssuntoRESUMO
Abstract Purpose: To evaluate the effects of mesenchymal stem cells on liver regeneration in rats following a 70% hepatectomy. Methods: Forty rats were subjected to 70% hepatectomy and then ~106 mesenchymal stem cells (test group), or saline solution (control group), were infused into their livers via the portal vein. Each treatment group was divided into early and late subgroups (euthanized 3 d and 5 d following the operation, respectively). Group comparisons of Albumin, aminotransaminases (AST, ALT), and Alcaline Phosphatase (AP) levels, proliferative index (ki-67+ straining), and mitotic cell counts were conducted. Results: No significant differences in liver regeneration rate, number of mitoses, proliferative index, or serum levels of albumin, AST, or AP were observed. ALT levels were higher in the test group than in the control group (p<.05). Conclusions: Mesenchymal stem-cell therapy did not improve liver regeneration rate 3 d or 5 d after 70% hepatectomy in rats. Likewise, the therapy appeared not to affect liver function, proliferative index, or number of mitoses significantly.
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Animais , Masculino , Ratos , Transplante de Células-Tronco Mesenquimais/métodos , Hepatectomia/métodos , Regeneração Hepática , Fatores de Tempo , Ratos Wistar , Modelos Animais , Proliferação de Células , Injeções IntravenosasRESUMO
Purpose: To evaluate the effects of mesenchymal stem cells on liver regeneration in rats following a 70% hepatectomy. Methods: Forty rats were subjected to 70% hepatectomy and then ~106 mesenchymal stem cells (test group), or saline solution (control group), were infused into their livers via the portal vein. Each treatment group was divided into early and late subgroups (euthanized 3 d and 5 d following the operation, respectively). Group comparisons of Albumin, aminotransaminases (AST, ALT), and Alcaline Phosphatase (AP) levels, proliferative index (ki-67+ straining), and mitotic cell counts were conducted. Results: No significant differences in liver regeneration rate, number of mitoses, proliferative index, or serum levels of albumin, AST, or AP were observed. ALT levels were higher in the test group than in the control group (p .05). Conclusions: Mesenchymal stem-cell therapy did not improve liver regeneration rate 3 d or 5 d after 70% hepatectomy in rats. Likewise, the therapy appeared not to affect liver function, proliferative index, or number of mitoses significantly.(AU)
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Animais , Células-Tronco Embrionárias Murinas/transplante , Regeneração Hepática , HepatectomiaRESUMO
Keratoconjunctivitis sicca (KCS) is a dysfunction in tear production associated with clinical signs, which include conjunctival hyperemia, ocular discharge, discomfort, pain, and, eventually, corneal vascularization and pigmentation. Immunosuppressive drugs are routinely administrated for long periods to treat KCS but with side effects and limited results. Evaluation of the clinical benefits of intralacrimal transplantation of allogeneic mesenchymal stem cells (MSCs) in dogs with mild-moderate and severe KCS was done. A total of 24 eyes with KCS from 15 dogs of different breeds were enrolled in the present study. A single transplantation of MSCs (1 × 106) directly into lacrimal glands (dorsal and third eyelid) was performed. The Schirmer tear tests (STTs) and ocular surface improvements were used to assess short- and long-term effects of these cells. The STTs were carried out on day 0 (before MSCs transplantation) and on days 7, 14, 21, and 28, as well as 6 and 12 months after MSC transplantation. Our data demonstrate that allogeneic MSC transplantation in KCS dogs is safe since no adverse effects were observed immediately after transplantation and in short- and long-term follow-ups. A statistically significant increase in the STT and ocular surface improvements was found in all eyes studied. In all the eyes with mild-moderate KCS, STT values reverted to those of healthy eyes, while in eyes with severe KCS, although complete reversion was not found, there was improvement in tear production and in other clinical signs. Our study shows that a single dose of a low number of MSCs can be used to treat KCS in dogs. In contrast to immunosuppressive drug use, MSC transplantation has an effect over a long period (up to 12 months), even after a single administration, and does not require daily drug administration.
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AbstractPulmonary hypertension is a devastating and refractory disease and there is no cure for this disease. Recently, microRNAs and mesenchymal stem cells emerged as novel methods to treat pulmonary hypertension. More than 20 kinds of microRNAs may participate in the process of pulmonary hypertension. It seems microRNAs or mesenchymal stem cells can ameliorate some symptoms of pulmonary hypertension in animals and even improve heart and lung function during pulmonary hypertension. Nevertheless, the relationship between mesenchymal stem cells, microRNAs and pulmonary hypertension is not clear. And the mechanisms underlying their function still need to be investigated. In this study we review the recent findings in mesenchymal stem cells - and microRNAs-based pulmonary hypertension treatment, focusing on the potential role of microRNAs regulated mesenchymal stem cells in pulmonary hypertension and the role of exosomes between mesenchymal stem cells and pulmonary hypertension.
ResumoA hipertensão pulmonar é uma doença devastadora e refratária, para a qual não existe cura. Recentemente, microRNAs e células-tronco mesenquimais emergiram como novos métodos para tratar a hipertensão pulmonar. Mais de 20 tipos de microRNAs podem participar no processo de hipertensão pulmonar. Ao que parece, microRNAs ou células-tronco mesenquimais podem atenuar alguns sintomas de hipertensão pulmonar em animais de e até mesmo melhorar a função cardíaca e do pulmão durante a hipertensão pulmonar. No entanto, a relação entre células-tronco mesenquimais, microRNAs e hipertensão pulmonar não é clara. E os mecanismos subjacentes a sua função ainda precisam ser investigados. Neste estudo, revisamos as descobertas recentes no tratamento da hipertensão pulmonar baseado em células-tronco mesenquimais e microRNAs, enfocando o papel potencial dos microRNAs para regular as células-tronco mesenquimais na hipertensão pulmonar e o papel dos exossomos entre células-tronco mesenquimais e hipertensão pulmonar.
Assuntos
Animais , Humanos , Hipertensão Pulmonar/terapia , Transplante de Células-Tronco Mesenquimais/métodos , Células-Tronco Mesenquimais/fisiologia , MicroRNAs/uso terapêutico , Exossomos/fisiologia , Hipertensão Pulmonar/fisiopatologia , Pulmão/fisiopatologiaRESUMO
OBJECTIVE: To evaluate the importance of stem cells derived from adipose tissue in reducing graft inflammation in a murine model of allogeneic heterotopic tracheal transplant. METHODS:We performed a heterotopic tracheal allografting in dorsal subcutaneous pouch and systemically injected 5x105 mesenchymal stem cells derived from adipose tissue. The animals were divided into two groups according to the time of sacrifice: T7 and T21. We also carried out histological analysis and digital morphometry. RESULTS:The T7 animals treated with cell therapy had median obstructed graft area of 0 versus 0.54 of controls (p = 0.635). The treated T21 subjects had median obstructed graft area of 0.25 versus 0 in controls (p = 0.041). CONCLUSION:The systemically injected cell therapy in experimental murine model of bronchiolitis obliterans did not reduce the severity of the allograft inflammation in a statistically significant way in seven days; Conversely, in 21 days, it increased the allograft inflammatory process.
OBJETIVO: Avaliar a importância das células-tronco derivadas de tecido adiposo na redução do processo inflamatório no enxerto em modelo murino de transplante traqueal heterotópico alogênico. MÉTODOS:Foi realizado alotransplante traqueal heterotópico em bolsa dorsal subcutânea e injetado 5x105 células-tronco mesenquimais, derivadas de tecido adiposo, sistemicamente. Os animais foram distribuídos em dois grupos, conforme o tempo de sacrifício: T7 e T21. Procedida a análise em HE e morfometria digital. RESULTADOS:Os T7 tratados com terapia celular apresentaram mediana de área obstruída do enxerto de 0 contra 0,54 dos controles (p=0,635). Os T21 tratados apresentaram mediana de área obstruída da luz do enxerto de 0,25 nos tratados e 0 nos controles (p=0,041). CONCLUSÃO: A terapia celular injetada sistemicamente em modelo experimental murino de bronquiolite obliterante não reduziu a gravidade do processo inflamatório no aloenxerto de forma estatisticamente significativa em sete dias; de modo contrário, em 21 dias, aumentou o processo inflamatório no aloenxerto.
Assuntos
Humanos , Bronquiolite Obliterante , Terapia Baseada em Transplante de Células e Tecidos , Transplante de Células-Tronco Mesenquimais , Células-Tronco , Transplante HeterotópicoRESUMO
As células-tronco mesenquimais (CTM) constituem uma ferramenta promissora para o campo de terapia celular. Além de seu potencial de diferenciação em diferentes tipos celulares, as CTM apresentam a habilidade de secretar moléculas bioativas e, assim, exercer múltiplos efeitos biológicos, tais como indução da regeneração de tecidos lesionados, redução de fibrose e modulação do sistema imune. A superexpressão dos fatores de crescimento G-CSF e IGF-1, conhecidos por seus efeitos sobre os processos de imunomodulação, sobrevivência celular e reparo tecidual, pode ampliar as ações terapêuticas das CTM. O objetivo deste trabalho consiste em gerar e caracterizar linhagens de CTM de camundongo superexpressando hGCSF ou hIGF-1. Um sistema lentiviral de segunda geração foi utilizado para modificação de CTM para expressão ectópica dos genes de interesse. As sequências codificantes de hG-CSF e hIGF-1 foram amplificadas por PCR e subclonadas em um vetor lentiviral de transferência, contendo um promotor constitutivo. As partículas lentivirais foram produzidas a partir da cotransfecção de células da linhagem HEK293FT...
Mesenchymal stem cells (MSCs) are a promising tool for the cell therapy field. In addition to their potential for differentiation into different cell types, MSCs have the ability to secrete bioactive molecules and thus exert multiple biological effects such as induction of the injured tissue regeneration, fibrosis reduction and modulation of the immune system. The overexpression of the growth factors G-CSF and IGF-1, known for their effects on immune modulation processes, cell survival and tissue repair, can result in a magnification of MSCs' therapeutic actions. The objective of this work is to generate and characterize mouse MSCs lines overexpressing hG-CSF or hIGF-1. A second generation lentiviral system was used to modify MSCs derived from mice for the ectopic expression of the genes of interest. The coding sequences of hG-CSF and hIGF-1 were amplified by PCR and subcloned into a lentiviral transfer vector containing a constitutive promoter. The lentiviral particles were produced from the co-transfection of HEK293FT...
Assuntos
Animais , Células-Tronco/classificação , Células-Tronco/imunologia , Células-Tronco/patologia , Insulina , Insulina/análise , Insulina/genética , Insulina/sangue , Insulina/urinaRESUMO
PURPOSE: To investigate the experimental model for obtaining adipose tissue, isolation, characterization of mesenchymal stem cells and evaluation of their distribution in the tram flap in rats. METHODS: Five rats of Wistar were randomly assigned to two groups. In group I, three animals underwent removal of adipose tissue in the groin procedure to establish the experimental model and obtain a cell lineage. The animals of group II (n = 2) underwent surgical flap procedure, and satisfaction injection of mesenchymal stem cells pretreated with marker fluorescente. RESULTS: obtaining adipose tissue of the inguinal region of the rat proved to be possible. The isolated cells were characterized as mesenchymal stem cells and fluorescence microscopy showed the presence of multiple cells arranged around blood vessels and capillaries. CONCLUSION: It was possible to establish an experimental model for obtaining adipose tissue for isolation of mesenchymal stem cells and their distribution in the TRAM flap in rats.(AU)
Assuntos
Animais , Tecido Adiposo/anatomia & histologia , Células-Tronco , Retalhos Cirúrgicos , Ratos/classificaçãoRESUMO
PURPOSE: To investigate the experimental model for obtaining adipose tissue, isolation, characterization of mesenchymal stem cells and evaluation of their distribution in the tram flap in rats. METHODS: Five rats of Wistar were randomly assigned to two groups. In group I, three animals underwent removal of adipose tissue in the groin procedure to establish the experimental model and obtain a cell lineage. The animals of group II (n = 2) underwent surgical flap procedure, and satisfaction injection of mesenchymal stem cells pretreated with marker fluorescente. RESULTS: obtaining adipose tissue of the inguinal region of the rat proved to be possible. The isolated cells were characterized as mesenchymal stem cells and fluorescence microscopy showed the presence of multiple cells arranged around blood vessels and capillaries. CONCLUSION: It was possible to establish an experimental model for obtaining adipose tissue for isolation of mesenchymal stem cells and their distribution in the TRAM flap in rats. .
Assuntos
Animais , Masculino , Tecido Adiposo/citologia , Separação Celular/métodos , Células-Tronco Mesenquimais , Modelos Animais , Retalhos Cirúrgicos , Diferenciação Celular , Células Cultivadas , Técnicas de Cultura de Células/métodos , Imunofenotipagem , Microscopia de Fluorescência , Distribuição Aleatória , Ratos Wistar , Reprodutibilidade dos TestesRESUMO
Introdução: Pacientes com câncer na região de cabeça e pescoço normalmente são tratados por combinação de cirurgia, radioterapia (RT) e quimioterapia. Em muitos casos, a reabilitação oral com próteses implanto-suportadas representa a melhor opção para uma recuperação funcional adequada. Entretanto, em pacientes irradiados procedimentos como exodontias e instalações de implantes dentários são fatores de risco ao desenvolvimento de osteorradionecrose. Diversos estudos experimentais têm demonstrado que o uso de células-tronco mesenquimais (CTMs) associadas a fatores de crescimento como plasma rico em plaquetas (PRP) proporciona melhora no reparo ósseo e na osseointegração, podendo ser considerada uma alternativa viável para defeitos ósseos ou injúria. Objetivos: Isolar e caracterizar as CTMs da medula óssea (MO) de miniporcos brasileiros (Minipigs BR-1), avaliar a interferência da RT e o efeito da associação de CTMs-MO+PRP no processo de osseointegração de implantes instalados em alvéolos frescos em mandíbulas de miniporcos, por análise histológica e histomorfométrica da interface osso-implante. Métodos: CTMs-MO de 12 miniporcos adultos machos foram isoladas da crista ilíaca. Após 21 dias de cultura, o potencial de diferenciação celular foi avaliado por meio de coloração e RT-PCR. O perfil imunofenotípico foi caracterizado por citometria de fluxo. Os animais foram divididos em três grupos: Grupo A (grupo controle, sem RT), Grupo B (implantes instalados 15 dias antes da RT) e Grupo C (implantes instalados três meses após a RT). A dose total de radiação para cada lado da mandíbula foi de 24 Gy, divididos em três doses de 8 Gy com intervalo de 7 dias entre as doses, a qual equivale biologicamente a aproximadamente 56 Gy, com 28 exposições de 2 Gy cada. Quatro implantes de titânio foram instalados nos alvéolos frescos, imediatamente após as extrações dos terceiro e quarto pré-molares, totalizando 48 implantes controles e 48 experimentais (uso de CTM-MO+PRP)...
Introduction: Patients with head and neck cancer are usually treated by a combination of surgery, radiotherapy (RT) and chemotherapy. In many cases, oral rehabilitation with implant-supported prostheses is the best option for a proper functional recover. However, in irradiated patients procedures as dental extractions and implants are risk factors for developing osteoradionecrosis. Several experimental studies have shown that the use of mesenchymal stem cells (MSCs) associated with growth factors such as platelet-rich plasma (PRP) provides improvement in bone regeneration and osseointegration, being considered an alternative to bone defects or injury. Objectives: To isolate and characterize MSCs from bone marrow (BM) of Brazilian minipigs (Minipigs BR-1), to evaluate the effect of RT and of BM-MSCs+PRP in the osseointegration of implants placed in fresh sockets, by histological and histomorphometric analysis of the bone-implant interface. Methods: BM-MSCs from 12 adult male minipigs were isolated from the iliac crest. After 21 days of culture, cell differentiation potential was assessed by staining and RT-PCR. The immunophenotypic profile was characterized by flow cytometry. The animals were divided into three groups: Group A (control group, no RT), Group B (implants placement 15 days before RT) and Group C (implants placement three months after RT). The total radiation dose for each side of the mandible was 24 Gy, divided into 3 doses of 8 Gy with a 7 dayinterval for each dose, which is biologically equivalent to approximately 56 Gy, with 28 sections of 2 Gy each. Four titanium implants were installed in the alveoli fresh, immediately after the extraction of the third and fourth premolars of each hemimandible, totalling 48 implants on the control side and 48 on the experimental side (using BM-MSCs+PRP). The animals were euthanized 90 days post-implantation....