A malassezíase é uma dermatopatia fúngica comum de humanos e animais, causada pela levedura lipof... more A malassezíase é uma dermatopatia fúngica comum de humanos e animais, causada pela levedura lipofílica Malassezia pachydermatis. O tratamento baseia-se na associação entre antifúngico sistêmico e tópico, contudo, recidivas são frequentes e complicadas por dermatoses intercorrentes que complicam o caso clínico. Essa recorrência comumente deve-se a erros de posologia dos fármacos, de administração dos mesmos por proprietários e por resistência da levedura. Os mecanismos de resistência antifúngica estão associados às características específicas ou adquiridas do patógeno aos fármacos. Como alternativa aos tratamentos tradicionais, o uso de produtos fitoterápicos, como óleos essenciais, na clínica veterinária vem se popularizando. Embora a possível ação dos óleos essenciais seja referida em medicina humana há cerca de 17 anos, a literatura especializada sobre seu uso na malassezíase canina e felina ainda é escassa e carecem estudos que investiguem os mecanismos de ação de óleos essenciai...
Topical treatment with bee honey (Apis mellifera), oxitetracicline or hydrocortisone on cutaneous... more Topical treatment with bee honey (Apis mellifera), oxitetracicline or hydrocortisone on cutaneous wound’s repair process by second intention in rabbits. The medical interest in anti-inflammatory and healing properties of bees honey used for the treatment of cutaneous wound’s in humans and animals has been increasing. This study investigated the therapeutics effects of bee (Apis mellifera) honey, oxitetracicline 3%, hydrocortisone 1% and oxitetracicline 3%, plus hydrocortisone 1% on cutaneous wound’s repair process by second intention, in rabbits. Six saggital dorsal lesions were created surgically, three in each antimere, which were treated using saline solution NaCl 0,9% (control), hydrocortisone 1%, oxitetracicline 3%, petrolatum (vehicle) and hydrocortisone 1% plus oxitetracicline 3%, respectively. Considering acceleration on the wound’s repair (diminishing in the size of the wounds) there weren’t differences among the treated groups comparing to the control (saline solution). Ho...
The development of ‘smart’ scaffolds has achieved notoriety among current prospects for bone repa... more The development of ‘smart’ scaffolds has achieved notoriety among current prospects for bone repair, especially for chronic osteopathy, such as osteoporosis. Millions of individuals in the world suffer from poor bone healing due to osteoporosis. The objective of this work was to produce and characterize castor polyurethane (PU) scaffolds (Ricinus communis L.) and evaluate its in vitro biocompatibility with stem cells and osteoinductive effect in vivo on bone failures in a leporid model of osteoporosis. The material was characterized using Fourier-transform infrared spectroscopy, thermogravimetric analysis, SEM, and porosity analysis. Then, the biocompatibility was assessed by adhesion using SEM and cytotoxicity in a 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H-tetrazolium assay. The osteoinductive effect in vivo was determined in bone defects in rabbit tibias (Oryctolagus cuniculus) submitted to castor PU scaffold, castor PU scaffold associated with stem cells, and negative contro...
Mesenchymal stem cells (MSCs), obtained from several anatomical sites, have already been describe... more Mesenchymal stem cells (MSCs), obtained from several anatomical sites, have already been described, characterized and used in therapeutic models for tissue repair. The umbilical cord mesenchymal stem cells, represented by cells from arteries and veins walls, as well as Wharton's jelly are easy to be obtained, highly available, require no invasive procedure, do not present risk to donors and do not present ethical limitation. The aim of this research was to analyze the plasticity of Wharton's jelly mesenchymal stem cells (WJ-MSCs) of goat, evaluating their behavior in vitro and characterizing them immunophenotypically. Thus, tests were performed on colony forming units, viability and cell growth curve, flow cytometry analysis and plasticity potential. Goat umbilical cord matrix cells exhibited fibroblastoid morphology with colony formation and self-renewal ability, always maintaining their undifferentiated state up to the eighth passage (P8). The growth curve kinetics exhibit...
As recentes descobertas sobre o comportamento in vitro das celulas-tronco embrionarias (CTE) e so... more As recentes descobertas sobre o comportamento in vitro das celulas-tronco embrionarias (CTE) e somaticas (CTS), assim como o aperfeicoamento metodologico das tecnicas de cultivo e aplicacao clinica experimental incentivaram o apelo popular no Brasil pela liberacao irrestrita das pesquisas com celulas-tronco (CT). Atualmente, mais um tipo de CT foi criada pela comunidade cientifica e nomeada de CT pluripotentes induzidas (iPS). Pesquisas com finalidades terapeuticas, envolvendo CTE, devem ser realizadas de forma cautelosa e, enquanto nao se desvendar toda a biologia destas in vivo, deve-se prosseguir com as pesquisas in vitro. Dentre todas as linhagens de CTS estudadas ate o presente momento, as celulas-tronco mesenquimais (CTM) apresentam maior plasticidade, originando tecidos mesodermais e nao mesodermais. No Brasil, ha um grande numero de testes clinicos em andamento com CTS, em humanos, que avaliam o uso terapeutico destas celulas em diferentes doencas e recentemente, desenvolveu...
A decada de 90 foi marcada por importantes avancos nas pesquisas com terapia celular objetivando ... more A decada de 90 foi marcada por importantes avancos nas pesquisas com terapia celular objetivando o tratamento de doencas hereditarias, auto-imunes e demais patologias com poucas perspectivas terapeuticas. Entretanto, ate entao, a terapia celular esteve restrita a utilizacao de celulas hematopoieticas no tratamento de doencas hematologicas e onco-hematologicas.
As celulas-tronco (CT) constituem grupo de celulas indiferenciadas, caracterizadas pela proprieda... more As celulas-tronco (CT) constituem grupo de celulas indiferenciadas, caracterizadas pela propriedade de originar celulas especializadas e de auto renovacao. Dentre as celulas tronco adultas, as celulas-tronco mesenquimais (CTM) compoem um grupo que vem despertando particular interesse, devido sua elevada plasticidade, com a capacidade de originar no organismo celulas de linhagem mesenquimal e nao mesenquimal. Essas celulas podem ser expandidas por mais de 40 geracoes mantendo capacidade multipotente, no entanto, os procedimentos de cultivo prolongado in vitro , podem provocar reducao nas taxas de mitose e uma grande probabilidade de acumulo de mutacoes, caracterizando uma instabilidade genetica. Este trabalho trata de uma revisao sobre estabilidade genetica das CTM em cultivo prolongado e a importância dos metodos citogeneticos. Foi realizada uma busca nas bases de dados on-line do Google Scholar, PubMed e SciELO, por meio das expressoes, “analise citogenetica”, “celulas-tronco mesen...
The present work evaluated the anesthetic association of propofol, an ultra-short acting general ... more The present work evaluated the anesthetic association of propofol, an ultra-short acting general anesthetic with a fast metabolism, with remifentanil, an opioid of great analgesic potency that is used as adjunct to general anesthetics. The main objective was to develop a safe ...
In vitro senescence of multipotent cells has been commonly associated with DNA damage induced by ... more In vitro senescence of multipotent cells has been commonly associated with DNA damage induced by oxidative stress. These changes may vary according to the sources of production and the studied lineages, which raises questions about the effect of growing time on genetic stability. This study is aimed at evaluating the evolution of genetic stability, viability, and oxidative stress of bone marrow mesenchymal stem cells (MSCBMsu) and renal progenitor cells of the renal cortex (RPCsu) of swine (Sus scrofa domesticus) in culture passages. P2, P5, and P9 were used for MSCBMsu and P1, P2, and P3 for RPCsu obtained by thawing. The experimental groups were submitted to MTT, apoptosis and necrosis assays, comet test, and reactive substance measurements of thiobarbituric acid (TBARS), nitrite, reduced glutathione (GSH), and catalase. The MTT test curve showed a mean viability of 1.14±0.62 and 1.12±0.54, respectively, for MSCBMsu and RPCsu. The percentages of MSCBMsu and RPCsu were presented, r...
Background: There are few studies on stem cell isolation in wild animals that provide isolation a... more Background: There are few studies on stem cell isolation in wild animals that provide isolation and culture protocols of these cells in vitro. Among the wild species studied, we present the collared peccary (Tayassu tajacu) as a model with potential to obtain and use MSC in preclinical studies. These animals are phylogenetically close to the domestic pig, popularly known as peccaries and found naturally in South America, Central America and the South of the United States. The aim of the present study was to establish a protocol for the isolation, in vitro cell expansion, differentiation and assessment of the stromal MSC growth curve before and after thawing.Materials, Methods & Results: Mesenchymal stem cells (MSC) from collared peccary bone marrow (Tayassu tajacu) were isolated and expanded by centrifuge in Ficoll® solution and cultured in DMEM® High Glucose medium. The culture was assessed by assays of colony forming units CFU-F and growth curve by saturation (GCS). Cultures in th...
Introduction: Wound healing is a progressive, essential and complex physiological process that oc... more Introduction: Wound healing is a progressive, essential and complex physiological process that occurs as a restorative response after a tissue injury. It involves three phases: inflammation, proliferation and maturation. Exogenous, endogenous and pathological factors may interfere in the cicatricial process in humans and animals by altering the balance between the synthesis, degradation and remodelling of collagen and elastic fibres. Diabetes mellitus is a progressive metabolic disease that alters elastogenesis and collagenesis and induces delays in the healing process. Scientific evidence suggests that mesenchymal stem cells modulate the cicatricial response. Thus the objective of this work was to perform stereological and morphometric analysis to determine the formation of dermal fibres in cutaneous fragments of a murine model of diabetes mellitus.Materials, Methods & Results: Histological sections were obtained from the cutaneous wounds of diabetic mice. The cutaneous wounds were...
Background: Osteochondral knee failures are among the most common causes of disability among the ... more Background: Osteochondral knee failures are among the most common causes of disability among the elderly human population and animal athletes. The xenogeneic transplantation of mesenchymal stem cells is a questionable therapeutic alternative that, despite the low expression of Major Histocompatibility Complex type II by these cells, still has relevantuncertainties about the safety and clinical efficacy. The main objective of the present study was to investigate whether the xenogeneic transplantation of mesenchymal stem cells induces hyaline cartilage formation, without histopathological evidence of rejection, in osteochondralfailures of goats.Materials, Methods & Results: Five female goats were used, submitted to three surgical osteocondral failures in the right knee, treated with xenogenic mesenchymal stem cells of dental pulp, xenogenic platelet-rich plasma and hemostatic sponge of hydrolyzed collagen, respectively. The lesions were evaluated after 60 days of treatment, aiming to ...
Cell replacement through neural stem cells has been a promising alternative therapy for neurodege... more Cell replacement through neural stem cells has been a promising alternative therapy for neurodegenerative diseases. It was evaluated the possible protect and/or prevent role of neurospheres in experimental models of epilepsy by the use of biomarkers of oxidative stress and histopathological analysis. After 1 h of the epileptic inductions by pilocarpine, pentylenotetrazole and picrotoxin, rats were infused with a suspension of 2 × 10 cells/0.25 mL, marked with Qtracker® 655, via caudal vein. In the control group epilepsy was not induced, but received the cell infusion under the same conditions of other groups. After 30 days, the rats were euthanized, and the removal of the brain was proceeded to later perform the assays oxidative stress and histopathology analysis. Thiobarbituric acid and nitrite levels were elevated in epileptic groups treated with neurospheres, and the levels of reduced glutathione, superoxide dismutase and catalase were reduced when compared to non-treated groups....
To assess the efficacy of allogeneic mesenchymal stem cells and xenogenic platelet rich plasma in... more To assess the efficacy of allogeneic mesenchymal stem cells and xenogenic platelet rich plasma in the treatment of bone failure of osteoporotic rabbits secondary to estrogenic deprivation and iatrogenic hypercortisolism. Eight female rabbits underwent ovarian resection and corticoid therapy to induce clinical status of osteoporosis. Four failures were produced in the tibiae, with each failure being treated with hemostatic sponge, allogenic mesenchymal stem cells, xenogenic platelet-rich plasma and the association between both. The animals were divided into two groups, evaluated radiographically and histopathologically at 30 and 60 days post treatment. A radiographically confirmed consolidation of bone failures treated with allogeneic mesenchymal stem cells, associated with the histopathological image of mature and immature bone tissue, without evidence of osteopenia, was compared with the other groups, in which radiolucent failures with osteopenia and fibrosis were still present, de...
A malassezíase é uma dermatopatia fúngica comum de humanos e animais, causada pela levedura lipof... more A malassezíase é uma dermatopatia fúngica comum de humanos e animais, causada pela levedura lipofílica Malassezia pachydermatis. O tratamento baseia-se na associação entre antifúngico sistêmico e tópico, contudo, recidivas são frequentes e complicadas por dermatoses intercorrentes que complicam o caso clínico. Essa recorrência comumente deve-se a erros de posologia dos fármacos, de administração dos mesmos por proprietários e por resistência da levedura. Os mecanismos de resistência antifúngica estão associados às características específicas ou adquiridas do patógeno aos fármacos. Como alternativa aos tratamentos tradicionais, o uso de produtos fitoterápicos, como óleos essenciais, na clínica veterinária vem se popularizando. Embora a possível ação dos óleos essenciais seja referida em medicina humana há cerca de 17 anos, a literatura especializada sobre seu uso na malassezíase canina e felina ainda é escassa e carecem estudos que investiguem os mecanismos de ação de óleos essenciai...
Topical treatment with bee honey (Apis mellifera), oxitetracicline or hydrocortisone on cutaneous... more Topical treatment with bee honey (Apis mellifera), oxitetracicline or hydrocortisone on cutaneous wound’s repair process by second intention in rabbits. The medical interest in anti-inflammatory and healing properties of bees honey used for the treatment of cutaneous wound’s in humans and animals has been increasing. This study investigated the therapeutics effects of bee (Apis mellifera) honey, oxitetracicline 3%, hydrocortisone 1% and oxitetracicline 3%, plus hydrocortisone 1% on cutaneous wound’s repair process by second intention, in rabbits. Six saggital dorsal lesions were created surgically, three in each antimere, which were treated using saline solution NaCl 0,9% (control), hydrocortisone 1%, oxitetracicline 3%, petrolatum (vehicle) and hydrocortisone 1% plus oxitetracicline 3%, respectively. Considering acceleration on the wound’s repair (diminishing in the size of the wounds) there weren’t differences among the treated groups comparing to the control (saline solution). Ho...
The development of ‘smart’ scaffolds has achieved notoriety among current prospects for bone repa... more The development of ‘smart’ scaffolds has achieved notoriety among current prospects for bone repair, especially for chronic osteopathy, such as osteoporosis. Millions of individuals in the world suffer from poor bone healing due to osteoporosis. The objective of this work was to produce and characterize castor polyurethane (PU) scaffolds (Ricinus communis L.) and evaluate its in vitro biocompatibility with stem cells and osteoinductive effect in vivo on bone failures in a leporid model of osteoporosis. The material was characterized using Fourier-transform infrared spectroscopy, thermogravimetric analysis, SEM, and porosity analysis. Then, the biocompatibility was assessed by adhesion using SEM and cytotoxicity in a 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H-tetrazolium assay. The osteoinductive effect in vivo was determined in bone defects in rabbit tibias (Oryctolagus cuniculus) submitted to castor PU scaffold, castor PU scaffold associated with stem cells, and negative contro...
Mesenchymal stem cells (MSCs), obtained from several anatomical sites, have already been describe... more Mesenchymal stem cells (MSCs), obtained from several anatomical sites, have already been described, characterized and used in therapeutic models for tissue repair. The umbilical cord mesenchymal stem cells, represented by cells from arteries and veins walls, as well as Wharton's jelly are easy to be obtained, highly available, require no invasive procedure, do not present risk to donors and do not present ethical limitation. The aim of this research was to analyze the plasticity of Wharton's jelly mesenchymal stem cells (WJ-MSCs) of goat, evaluating their behavior in vitro and characterizing them immunophenotypically. Thus, tests were performed on colony forming units, viability and cell growth curve, flow cytometry analysis and plasticity potential. Goat umbilical cord matrix cells exhibited fibroblastoid morphology with colony formation and self-renewal ability, always maintaining their undifferentiated state up to the eighth passage (P8). The growth curve kinetics exhibit...
As recentes descobertas sobre o comportamento in vitro das celulas-tronco embrionarias (CTE) e so... more As recentes descobertas sobre o comportamento in vitro das celulas-tronco embrionarias (CTE) e somaticas (CTS), assim como o aperfeicoamento metodologico das tecnicas de cultivo e aplicacao clinica experimental incentivaram o apelo popular no Brasil pela liberacao irrestrita das pesquisas com celulas-tronco (CT). Atualmente, mais um tipo de CT foi criada pela comunidade cientifica e nomeada de CT pluripotentes induzidas (iPS). Pesquisas com finalidades terapeuticas, envolvendo CTE, devem ser realizadas de forma cautelosa e, enquanto nao se desvendar toda a biologia destas in vivo, deve-se prosseguir com as pesquisas in vitro. Dentre todas as linhagens de CTS estudadas ate o presente momento, as celulas-tronco mesenquimais (CTM) apresentam maior plasticidade, originando tecidos mesodermais e nao mesodermais. No Brasil, ha um grande numero de testes clinicos em andamento com CTS, em humanos, que avaliam o uso terapeutico destas celulas em diferentes doencas e recentemente, desenvolveu...
A decada de 90 foi marcada por importantes avancos nas pesquisas com terapia celular objetivando ... more A decada de 90 foi marcada por importantes avancos nas pesquisas com terapia celular objetivando o tratamento de doencas hereditarias, auto-imunes e demais patologias com poucas perspectivas terapeuticas. Entretanto, ate entao, a terapia celular esteve restrita a utilizacao de celulas hematopoieticas no tratamento de doencas hematologicas e onco-hematologicas.
As celulas-tronco (CT) constituem grupo de celulas indiferenciadas, caracterizadas pela proprieda... more As celulas-tronco (CT) constituem grupo de celulas indiferenciadas, caracterizadas pela propriedade de originar celulas especializadas e de auto renovacao. Dentre as celulas tronco adultas, as celulas-tronco mesenquimais (CTM) compoem um grupo que vem despertando particular interesse, devido sua elevada plasticidade, com a capacidade de originar no organismo celulas de linhagem mesenquimal e nao mesenquimal. Essas celulas podem ser expandidas por mais de 40 geracoes mantendo capacidade multipotente, no entanto, os procedimentos de cultivo prolongado in vitro , podem provocar reducao nas taxas de mitose e uma grande probabilidade de acumulo de mutacoes, caracterizando uma instabilidade genetica. Este trabalho trata de uma revisao sobre estabilidade genetica das CTM em cultivo prolongado e a importância dos metodos citogeneticos. Foi realizada uma busca nas bases de dados on-line do Google Scholar, PubMed e SciELO, por meio das expressoes, “analise citogenetica”, “celulas-tronco mesen...
The present work evaluated the anesthetic association of propofol, an ultra-short acting general ... more The present work evaluated the anesthetic association of propofol, an ultra-short acting general anesthetic with a fast metabolism, with remifentanil, an opioid of great analgesic potency that is used as adjunct to general anesthetics. The main objective was to develop a safe ...
In vitro senescence of multipotent cells has been commonly associated with DNA damage induced by ... more In vitro senescence of multipotent cells has been commonly associated with DNA damage induced by oxidative stress. These changes may vary according to the sources of production and the studied lineages, which raises questions about the effect of growing time on genetic stability. This study is aimed at evaluating the evolution of genetic stability, viability, and oxidative stress of bone marrow mesenchymal stem cells (MSCBMsu) and renal progenitor cells of the renal cortex (RPCsu) of swine (Sus scrofa domesticus) in culture passages. P2, P5, and P9 were used for MSCBMsu and P1, P2, and P3 for RPCsu obtained by thawing. The experimental groups were submitted to MTT, apoptosis and necrosis assays, comet test, and reactive substance measurements of thiobarbituric acid (TBARS), nitrite, reduced glutathione (GSH), and catalase. The MTT test curve showed a mean viability of 1.14±0.62 and 1.12±0.54, respectively, for MSCBMsu and RPCsu. The percentages of MSCBMsu and RPCsu were presented, r...
Background: There are few studies on stem cell isolation in wild animals that provide isolation a... more Background: There are few studies on stem cell isolation in wild animals that provide isolation and culture protocols of these cells in vitro. Among the wild species studied, we present the collared peccary (Tayassu tajacu) as a model with potential to obtain and use MSC in preclinical studies. These animals are phylogenetically close to the domestic pig, popularly known as peccaries and found naturally in South America, Central America and the South of the United States. The aim of the present study was to establish a protocol for the isolation, in vitro cell expansion, differentiation and assessment of the stromal MSC growth curve before and after thawing.Materials, Methods & Results: Mesenchymal stem cells (MSC) from collared peccary bone marrow (Tayassu tajacu) were isolated and expanded by centrifuge in Ficoll® solution and cultured in DMEM® High Glucose medium. The culture was assessed by assays of colony forming units CFU-F and growth curve by saturation (GCS). Cultures in th...
Introduction: Wound healing is a progressive, essential and complex physiological process that oc... more Introduction: Wound healing is a progressive, essential and complex physiological process that occurs as a restorative response after a tissue injury. It involves three phases: inflammation, proliferation and maturation. Exogenous, endogenous and pathological factors may interfere in the cicatricial process in humans and animals by altering the balance between the synthesis, degradation and remodelling of collagen and elastic fibres. Diabetes mellitus is a progressive metabolic disease that alters elastogenesis and collagenesis and induces delays in the healing process. Scientific evidence suggests that mesenchymal stem cells modulate the cicatricial response. Thus the objective of this work was to perform stereological and morphometric analysis to determine the formation of dermal fibres in cutaneous fragments of a murine model of diabetes mellitus.Materials, Methods & Results: Histological sections were obtained from the cutaneous wounds of diabetic mice. The cutaneous wounds were...
Background: Osteochondral knee failures are among the most common causes of disability among the ... more Background: Osteochondral knee failures are among the most common causes of disability among the elderly human population and animal athletes. The xenogeneic transplantation of mesenchymal stem cells is a questionable therapeutic alternative that, despite the low expression of Major Histocompatibility Complex type II by these cells, still has relevantuncertainties about the safety and clinical efficacy. The main objective of the present study was to investigate whether the xenogeneic transplantation of mesenchymal stem cells induces hyaline cartilage formation, without histopathological evidence of rejection, in osteochondralfailures of goats.Materials, Methods & Results: Five female goats were used, submitted to three surgical osteocondral failures in the right knee, treated with xenogenic mesenchymal stem cells of dental pulp, xenogenic platelet-rich plasma and hemostatic sponge of hydrolyzed collagen, respectively. The lesions were evaluated after 60 days of treatment, aiming to ...
Cell replacement through neural stem cells has been a promising alternative therapy for neurodege... more Cell replacement through neural stem cells has been a promising alternative therapy for neurodegenerative diseases. It was evaluated the possible protect and/or prevent role of neurospheres in experimental models of epilepsy by the use of biomarkers of oxidative stress and histopathological analysis. After 1 h of the epileptic inductions by pilocarpine, pentylenotetrazole and picrotoxin, rats were infused with a suspension of 2 × 10 cells/0.25 mL, marked with Qtracker® 655, via caudal vein. In the control group epilepsy was not induced, but received the cell infusion under the same conditions of other groups. After 30 days, the rats were euthanized, and the removal of the brain was proceeded to later perform the assays oxidative stress and histopathology analysis. Thiobarbituric acid and nitrite levels were elevated in epileptic groups treated with neurospheres, and the levels of reduced glutathione, superoxide dismutase and catalase were reduced when compared to non-treated groups....
To assess the efficacy of allogeneic mesenchymal stem cells and xenogenic platelet rich plasma in... more To assess the efficacy of allogeneic mesenchymal stem cells and xenogenic platelet rich plasma in the treatment of bone failure of osteoporotic rabbits secondary to estrogenic deprivation and iatrogenic hypercortisolism. Eight female rabbits underwent ovarian resection and corticoid therapy to induce clinical status of osteoporosis. Four failures were produced in the tibiae, with each failure being treated with hemostatic sponge, allogenic mesenchymal stem cells, xenogenic platelet-rich plasma and the association between both. The animals were divided into two groups, evaluated radiographically and histopathologically at 30 and 60 days post treatment. A radiographically confirmed consolidation of bone failures treated with allogeneic mesenchymal stem cells, associated with the histopathological image of mature and immature bone tissue, without evidence of osteopenia, was compared with the other groups, in which radiolucent failures with osteopenia and fibrosis were still present, de...
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