Experimental Basis and New Insights for Cell Therapy in Chronic Obstructive Pulmonary Disease

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

The use of planarians as in vivo animal model to study laser biomodulation effects

A variety of effects is attributed to the photo stimulation of tissues, such as improved healing of ulcers, analgesic and anti-inflammatory effects, stimulation of the proliferation of cells of different origins and stimulation of bone repair. Some investigations that make qualitative evaluations, like wound healing and evaluation of pain and edema, can be conducted in human subjects. However, deeper investigations on the mechanisms of action of the light stimulus and other quantitative works that requires biopsies or destructive analysis has to be carried out in animal models or in cell cultures. In this work, we propose the use of planarians as a model to study laser-tissue interaction. Contrasting with cell cultures and unicellular organisms, planarians are among the simplest organism having tissue layers, central nerve system, digestive and excretory system that might have been platforms for the evolution of the complex and highly organized tissues and organs found in higher organisms. For the present study, 685 nm laser radiation was employed. Planarians were cut transversally, in a plane posterior to the auricles. The body fragments were left to regenerate and the proliferation dynamics of stem cells was studied by using histological analysis. Maximum cell count was obtained for the laser treated group at the 4th experimental day. At that experimental time, we also had the largest difference between the irradiated and the non-irradiated control group. We concluded that the studied flatworm could be an interesting animal model for in vivo studies of laser-tissue interactions.

Use of fibrin glue derived from snake venom in the repair of deep corneal ulcers - Experimental study in dogs (Canis familiaris, Linnaeus, 1758)

Fibrin glue has been researched as an alternative method for tissue synthesis and is known for its capability to promote hemostasis at the application site, good approximation of wound edges and fast healing. The current study consisted in the application of fibrin glue derived from snake venom as treatment for experimental corneal ulcers. Twenty-one dogs had their corneas experimentally prepared through lamellar keratectomy (of standardized diameter and depth). Animals were divided into seven groups of three animals each. Six experimental groups were periodically evaluated and collection was carried out on the 1st, 3rd, 7th, 15 th, 30th and 60th post-operative days, whereas one control group was evaluated throughout the experiment. Analyses consisted in the clinical evolution and in the histopathological study of all operated on eyes. Results indicated that fibrin glue was efficient in repairing keratectomy wounds in dogs and contributed to an earlier healing phenomenon, avoiding edema formation and keeping corneal clearness. The use of fibrin glue derived from snake venom showed to be easy to apply, feasible with animal models and of low cost, avoiding the lesion progress and allowing fast and appropriate corneal healing.

Importância do modelo animal para testar hipóteses sobre a fisiopatologia do binômio diabetes e incontinência urinária feminina

Aims: To discuss the importance of studying animal models to test hypotheses about the mechanisms of urinary continence and pathophysiology of diabetes and urinary incontinence. Source of Data: A literature review was conducted in PubMed and SciELO. The key words used were diabetes, urinary incontinence, urethra, human and rats. Summary of Findings: There is a strong relation between the genesis of urinary incontinence and diabetes mellitus. Due to the similarity of normal distribution of skeletal muscle and urethra anatomy between humans and rats, these animal models have been used in current research about these disorders. Conclusions: The use of rats as an animal model is suitable for experimental studies that test hypotheses about the mechanisms of continence and pathophysiology of the binomial diabetes mellitus and urinary incontinence, thus enabling solutions of great value in clinical practice.

Mild diabetes models and their maternal-fetal repercussions

The presence of diabetes in pregnancy leads to hormonal and metabolic changes making inappropriate intrauterine environment, favoring the onset of maternal and fetal complications. Human studies that explore mechanisms responsible for changes caused by diabetes are limited not only for ethical reasons but also by the many uncontrollable variables. Thus, there is a need to develop appropriate experimental models. The diabetes induced in laboratory animals can be performed by different methods depending on dose, route of administration, and the strain and age of animal used. Many of these studies are carried out in neonatal period or during pregnancy, but the results presented are controversial. So this paper, addresses the review about the different models of mild diabetes induction using streptozotocin in pregnant rats and their repercussions on the maternal and fetal organisms to propose an adequate model for each approached issue. © 2013 D. C. Damasceno et al.

Escola experimental de primatas: análise da coerência entre pressupostos e práticas empíricas

O desenvolvimento de modelos animais para o estudo de problemas no desenvolvimento cognitivo humano pressupõe como fator principal a existência de certa homologia de processos comportamentais entre os repertórios do modelo e dos humanos. A Escola Experimental de Primatas (EEP) tem utilizado o macaco-prego (Cebus ssp.) como um modelo animal para investigar a aquisição de repertórios relacionais generalizados em humanos. Este trabalho avaliou os estudos empíricos da EEP no sentido de verificar a coerência entre seus pressupostos teóricos e sua prática empírica, especialmente no que diz respeito à existência de homologias de processos comportamentais entre macacos-prego e seres humanos. Ao se analisar a 1) origem dos objetivos e dos procedimentos dos estudos (a maioria provém de estudos com animais); 2) se os resultados esperados foram alcançados ou não (de forma geral, resultados positivos em relação aos objetivos propostos); e 3) se nos estudos da EEP é discutida a questão do modelo animal e/ou a aplicabilidade dos procedimentos utilizados nos estudos para humanos (não é), verificou-se que não são apresentados dados empíricos que sustentem a homologia sugerida na proposta teórica norteadora da EEP. Esses resultados demonstram que a prática científica da EEP tem avançado enquanto pesquisa básica sobre a aprendizagem de repertórios relacionais generalizados para macacosprego, mas ainda sem evidências de transferência para o estudo desse fenômeno em humanos. Sugere-se que o caminho a seguir pode ser: 1) desenvolver estudos empíricos que validem a homologia de processos comportamentais entre alguns repertórios dos macacos-prego e humanos; e 2) dar início à aplicação dos achados da pesquisa básica da EEP no desenvolvimento de procedimentos para ensinar repertórios relacionais para pessoas com atraso no desenvolvimento cognitivo.

Eletrofisiologia da visão em modelo experimental de ratos com diabetes e hipotireoidismo

O diabetes mellitus constitui um problema de saúde pública mundial devido ao seu potencial de morbidade e mortalidade. Esta patologia é caracterizada como uma desordem metabólica com hiperglicemia crônica, resultado de uma deficiência absoluta ou relativa na secreção e/ou ação da insulina. Associado a esse quadro, problemas com a tireoide são frequentemente encontrados em pacientes acometidos por diabetes, principalmente com o avançar da idade. Além disso, há um corpo de conhecimento que descreve que tanto o diabetes quanto o hipotiroidismo provocam isoladamente perdas visuais, no entanto, não há muitas publicações acerca da fisiologia envolvendo em conjunto ambas patologias com as complicações visuais. O presente trabalho objetivou investigar questões relevantes para uma compreensão mais detalhada do quadro de evolução do comprometimento retiniano em modelos animais acometidos concomitantemente por duas doenças: diabetes e hipotireoidismo. Para isso, foram utilizados 50 ratos Wistar machos (Rattus norvegicus), com dois meses de idade, pesando entre 80 a 120 gramas, divididos em quatro grupos: um grupo controle sem procedimentos; um grupo com hipotireoidismo optando por um modelo experimental de tireoidectomia bilateral; um grupo com diabetes, com aplicação de 200 mg/Kg a 2% de Aloxana; e dois grupos com as duas patologias, adotando ambos os procedimentos experimentais, alterando a sequência das patologias em questão. Nos grupos estudados foi utilizado para avaliar as alterações visuais, um método não invasivo amplamente utilizado no meio acadêmico que é o eletrorretinograma (ERG) em diferentes períodos de 30, 45 e 60 dias. Assim, encontramos uma diminuição da amplitude média da onda-a dos animais nos grupos com diabetes e com as duas patologias em todos os registros realizados, os que apresentaram maiores diferenças estatísticas foram em relação ao registro de estocópica máxima, como também diferença no potencial oscilatório de ambos os grupos com as duas patologias. Os resultados obtidos dão suporte à hipótese que as duas patologias concomitantes (diabetes e hipotireoidismo) diminuem significativamente as respostas do ERG.

Exercício físico, ingestão de frutose e marcadores da síndrome metabólica em modelo experimental

Pós-graduação em Ciências da Motricidade - IBRC

Caracterização das alterações teciduais em dois modelos de indução de doença periodontal experimental em camundongos

Pós-graduação em Odontologia - FOAR

Clinical manifestations and experimental studies on the spine extract of the toadfish Porichthys porosissimus

Toadfish are fish from the family Batrachoididae that are found in marine and brackish environment around the world. Among the toadfish, Porichthys genus is very common, where Porichthys porosissimus, also called Atlantic Midshipman is found in Southwest Atlantic, from Rio de Janeiro, Brazil to eastern Argentina. There was no consensus about the classification of the genus Porichthys as venomous fish because so far there are no published Studies regarding human envenomations and/or toxic activities induced in animal models. Herein, we report two conclusive envenoming in human beings caused by P porosissimus spines, with clear signs and symptoms that were very important for the development of our experimental studies. We demonstrated that the P. porosissimus spine extract, now venom, can induce nociceptive and edematogenic responses in mice as well an induction of an inflammatory response elicited by intravital microscopy and leukocyte migration. Finally, we identified in the P. porosissimus spine extract, through analysis by mass spectrometry, the presence of proteins previously detected in the venoms of other fish species and other venomous animals. We believe that based on our studies we will dismiss the non-venomous nature of this fish and clarify this issue. (C) 2014 Elsevier Ltd. All rights reserved.

Competitive interactions between C. albicans, C. glabrata and C. krusei during biofilm formation and development of experimental Candidiasis

In this study, we evaluated the interactions between Candida albicans, Candida krusei and Candida glabrata in mixed infections. Initially, these interactions were studied in biofilms formed in vitro. CFU/mL values of C. albicans were lower in mixed biofilms when compared to the single biofilms, verifying 77% and 89% of C. albicans reduction when this species was associated with C. glabrata and C. krusei, respectively. After that, we expanded this study for in vivo host models of experimental candidiasis. G. mellonella larvae were inoculated with monotypic and heterotypic Candida suspensions for analysis of survival rate and quantification of fungal cells in the haemolymph. In the groups with single infections, 100% of the larvae died within 18 h after infection with C. albicans. However, interaction groups achieved 100% mortality after 72 h of infection by C. albicans-C. glabrata and 96 h of infection by C. albicans-C. krusei. C. albicans CFU/mL values from larvae hemolymph were lower in the interacting groups compared with the monoespecies group after 12 h of infection. In addition, immunosuppressed mice were also inoculated with monotypic and heterotypic microbial suspensions to induce oral candidiasis. C. albicans CFU/mL values recovered from oral cavity of mice were higher in the group with single infection by C. albicans than the groups with mixed infections by C. albicans-C. glabrata and C. albicans-C. krusei. Moreover, the group with single infection by C. albicans had a higher degree of hyphae and epithelial changes in the tongue dorsum than the groups with mixed infections. We concluded that single infections by C. albicans were more harmful for animal models than mixed infections with non-albicans species, suggesting that C. albicans establish competitive interactions with C. krusei and C. glabrata during biofilm formation and development of experimental candidiasis.

Experimental evidence of heparanase, Hsp70 and NF-κB gene expression on the response of anti-inflammatory drugs in TNBS-induced colonic inflammation

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Corrositex®, BCOP and HET-CAM as alternative methods to animal experimentation

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Experimental Basis and New Insights for Cell Therapy in Chronic Obstructive Pulmonary Disease

Chronic Obstructive Pulmonary Disease (COPD) can be briefly described as air flow limitation and chronic dyspnea associated to an inflammatory response of the respiratory tract to noxious particles and gases. Its main feature is the obstruction of airflow and consequent chronic dyspnea. Despite recent advances, and the development of new therapeutic, medical and clinical approaches, a curative therapy is yet to be achieved. Therapies involving the use of tissue-specific or donor derived cells present a promising alternative in the treatment of degenerative diseases and injuries. Recent studies demonstrate that mesenchymal stem cells have the capacity to modulate immune responses in acute lung injury and pulmonary fibrosis in animal models, as well as in human patients. Due to these aspects, different groups raised the possibility that the stem cells from different sources, such as those found in bone marrow or adipose tissue, could act preventing the emphysematous lesion progression. In this paper, it is proposed a review of the current state of the art and future perspectives on the use of cell therapy in obstructive lung diseases.

Effects of Positive End-expiratory Pressure Titration and Recruitment Maneuver on Lung Inflammation and Hyperinflation in Experimental Acid Aspiration-induced Lung Injury

Background: In acute lung injury positive end-expiratory pressure (PEEP) and recruitment maneuver are proposed to optimize arterial oxygenation. The aim of the study was to evaluate the impact of such a strategy on lung histological inflammation and hyperinflation in pigs with acid aspiration-induced lung injury. Methods: Forty-seven pigs were randomly allocated in seven groups: (1) controls spontaneously breathing; (2) without lung injury, PEEP 5 cm H2O; (3) without lung injury, PEEP titration; (4) without lung injury, PEEP titration + recruitment maneuver; (5) with lung injury, PEEP 5 cm H2O; (6) with lung injury, PEEP titration; and (7) with lung injury, PEEP titration + recruitment maneuver. Acute lung injury was induced by intratracheal instillation of hydrochloric acid. PEEP titration was performed by incremental and decremental PEEP from 5 to 20 cm H2O for optimizing arterial oxygenation. Three recruitment maneuvers (pressure of 40 cm H2O maintained for 20 s) were applied to the assigned groups at each PEEP level. Proportion of lung inflammation, hemorrhage, edema, and alveolar wall disruption were recorded on each histological field. Mean alveolar area was measured in the aerated lung regions. Results: Acid aspiration increased mean alveolar area and produced alveolar wall disruption, lung edema, alveolar hemorrhage, and lung inflammation. PEEP titration significantly improved arterial oxygenation but simultaneously increased lung inflammation in juxta-diaphragmatic lung regions. Recruitment maneuver during PEEP titration did not induce additional increase in lung inflammation and alveolar hyperinflation. Conclusion: In a porcine model of acid aspiration-induced lung injury, PEEP titration aimed at optimizing arterial oxygenation, substantially increased lung inflammation. Recruitment maneuvers further improved arterial oxygenation without additional effects on inflammation and hyperinflation.