Caracterização, criopreservação e diferenciação in vitro de células tronco, derivadas de amniótica e gelatina de Wharton canina, visando a formação de um banco de células tronco

Pós-graduação em Biotecnologia Animal - FMVZ

Avaliação Postural em Sujeitos com Disfunção Temporomandibular Submetidos a Tratamento de Terapia Manual

Individuals with temporomandibular disorders (TMD) often have signs and symptoms such as intra-articular pain, muscle spasm, neck pain radiating from others who interfere with this balance. Over time can lead to postural changes and correction of the cervical spine, anterior head and shoulder asymmetry. The aim was to verify the effects of manual therapy on posture in subjects with TMD. Materials and methods: 30 volunteers (mean: 21.43± SD:1.43) of both sexes who had TMD classified according to axis I of the Research Diagnostic Criteria participated in this study. These were photographed in the anterior frontal and sagittal planes for analysis of the following angles (acromion clavicular joint, sternoclavicular joint, orbicular external, corners of the mouthand and protruding head). After, the volunteers were divided into groups ATM (manual therapy in the treatment of temporomandibular joint), Cervical (manual therapy in the treatment of cervical joint) and Control. The Kolmogokov-Smirnov test, followed by ANOVA, considering a significance level of 5% was used. Results: Groups ATM, Cervical and Control were considered homogeneous with respect to the values of the angles measured. These also showed no significant difference between the 1st, 5th and 10th sessions, so the manual therapy techniques applied to the TMJ and Cervical groups were not sufficient to change postural alignment. Conclusion: The postural alignment, the TMD patients evaluated in this study did not change after attending the sessions proposals with manual therapy techniques in the cervical and TMJ.

Avaliação do controle postural e do tipo do pé de pessoas com deficiência visual

The balance changing in the visually impaired is target of investigations, as balance and vision are interrelated. Also, another important factor in the maintenance of postural control is the biomechanical aspect of the feet. This study aimed to evaluate postural control and feet type of individuals with visual impairments. For then 17 adults with visual impairment participated in this study and have been evaluated by the baropodometry system. The oscillation of the center of pressure data were analyzed using the software Conformat Research 5.8, and subjected to Mann-Whitney’s statistical test. The results showed no statistically significant difference in postural control when comparing individuals with and without visual residue and which normal and foot with biomechanical alterations, which demonstrates that the visual impairment was the main factor of changes in postural control for the participants of this study.

Efeitos da Reeducação Postural Global na Hipercifose e nas Variáveis Respiratórias de Idosos Relato de Dois Casos

Examine the effects Global Postural Reeducation, (GPR) in hyperkyphosis and respiratory variables in the elderly was the objective in the research. For this, two elderly participated, one is 62 years old (voluntary 1) and another is 66 years old (voluntary 2), without lung, heart, kidney and/or skeletal-muscle diseases diagnosed. The volunteer have been assessed for Quality of Life (QOL) through the questionnaire SF-36, the degree of toracic kyphosis, the muscular respiratory strength and the thoracic-abdominal mobility. The GRP treatment consisted in eight sessions applied, once one hour each session. After the eight sessions the volunteer were reevaluated. The data concerning assessments before and after treatment were analysed describly. According to the results there was an improvement in the degree of kyphosis in both volunteers, highlighting voluntary 2. Respiratory variables also improvements after treatment. In relation to the Quality of life in most areas there has been an increase in scores indicating improved QOL. These data showed that the GPR has been effective to decrease the level of kyphosis, improves respiratory variables and the quality of life in the elderly treated. However, further work with a greater number of subjects must be carried out to analyze the effectiveness of therapy in the elderly.

Previous exposure to musical auditory stimulation immediately influences the cardiac autonomic responses to the postural change maneuver in women

Background: Chronic exposure to musical auditory stimulation has been reported to improve cardiac autonomic regulation. However, it is not clear if music acutely influences it in response to autonomic tests. We evaluated the acute effects of music on heart rate variability (HRV) responses to the postural change maneuver (PCM) in women. Method: We evaluated 12 healthy women between 18 and 28 years old and HRV was analyzed in the time (SDNN, RMSSD, NN50 and pNN50) and frequency (LF, HF and LF/HF ratio) domains. In the control protocol, the women remained at seated rest for 10 minutes and quickly stood up within three seconds and remained standing still for 15 minutes. In the music protocol, the women remained at seated rest for 10 minutes, were exposed to music for 10 minutes and quickly stood up within three seconds and remained standing still for 15 minutes. HRV was recorded at the following time: rest, music (music protocol) 0–5, 5–10 and 10–15 min during standing. Results: In the control protocol the SDNN, RMSSD and pNN50 indexes were reduced at 10–15 minutes after the volunteers stood up, while the LF (nu) index was increased at the same moment compared to seated rest. In the protocol with music, the indexes were not different from control but the RMSSD, pNN50 and LF (nu) were different from the music period. Conclusion: Musical auditory stimulation attenuates the cardiac autonomic responses to the PCM.

Fractal correlation property of heart rate variability in response to the postural change maneuver in healthy women

Background: We evaluated the effects of the PCM on the fractal analysis of the HRV in healthy women Method: We evaluated healthy women between 18 and 30 years old. HRV was analyzed in the time (SDNN, RMSSD, NN50 and pNN50) and frequency (LF, HF and LF/HF ratio) domains as well as short and long-term fractal exponents (alpha-1 and alpha-2) of the detrended fluctuation analysis (DFA). HRV was recorded at rest for ten minutes at seated rest and then the women quickly stood up from a seated position in up to three seconds and remained standing for 15 minutes. HRV was recorded at the following time: rest, 0–5 min, 5–10 min and 10–15 min during standing. Results: We observed decrease (p < 0.05) in the time-domain indices of HRV between seated and 10–15 minutes after the volunteer stood up. The LF (ms2) and HF (ms2) indices were also reduced (p < 0.05) at 10–15 minutes after the volunteer stood up compared to seated while the LF (nu) was increased at 5–10 min and 10–15 min (p < 0.05). The short-term alpha-1 exponent was increased (p < 0.05) at all moments investigated compared to seated. Increase in the properties of short-term fractal correlations of heart rate dynamics accompanied by a decrease in the parasympathetic modulation and global HRV was observed in response to the postural change maneuver. Conclusion: We suggest that fractal analysis of HRV is more sensitive than frequency and time-domain analysis of HRV during the postural change maneuver.

Estudo dos mecanismos envolvidos no processo de diferenciação em linhagem osteogênica de células-tronco mesenquimais da medula óssea de ratos Wistar e ratos espontaneamente hipertensos (SHR)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Análise comparativa de diferentes metodologias para cultivo de células-tronco mesenquimais derivadas de tecido muscular

Pós-graduação em Biociências - FCLAS

Produção de anticorpos monoclonais murinos dirigidos contra células-tronco mesenquimais de coelho

Mesenchymal stem cells (MSCs) are a heterogeneous population of cells that proliferate in vitro as plastic-adherent cells, have fibroblast-like morphology and can differentiate into bone, cartilage and fat cells. Therapeutic potential of MSCs have been studied in experimental models, such as rabbit, in Laboratory of Cell Engineering of Botucatu. However, no specific markers have been reported for expanded rabbit MSCs, which hampers the isolation of pure MSC populations by immunophenotypic characterization. Thus, the objective of this study was to produce monoclonal antibodies (mAbs) to rabbit MSCs. MSCs derived from rabbit bone marrow (BM) were isolated, cultured, expanded ex vivo, and immunized into three BALB/c mices, and spleen cells subsequently harvested were used to generate hibridoma cell lines secreting antibodies against MSCs. Hybridoma cells were screened by flow cytometry and antibody-producing cells were subjected to subsequent rounds of retests. MSC1-160 obtained the best positivity for IgG expression and was cloned by limiting dilutions and micromanipulation. Ascitic fluid from ten best clones was purified by affinity chromatography in Protein A-sepharose CL-4B column and purification control was performed by electrophoresis in agarose gels. The purified IgG were tested against rabbit MSCs, obtaining high positivity by flow Cytometry. In conclusion, we developed 10 mAbs, MSC1-160 A20, A30, A41, A47, A55, A60, A63, A69, A81, and A82, that recognize rabbit MSC cell surface antigens showing potential for immunophenotypic characterization of rabbit MSC cell lines

Uso de células-tronco para o tratamento de tendinite em eqüinos

Brazil has the fourth largest horse herd in the world, this is due the recognition and appreciation that the different equestrian games are having within the country. Injuries of the tendon, especially in the digital flexor tendon, are the main cause of athletic life reduction among horses. The treatment of tendinitis in horses seeks full recovery of the damage tissue reestablishing the function previously lost, however conventional treatments have proven to be ineffective when considered the quality of the scar tissue and the rate of recurrence. Due to this, the use of adult stem cells to the treatment of musculoskeletal injuries of horses has been studied for some time. This method of treatment consists of aspiration of bone marrow or removal of subcutaneous fat tissue and implantation of these cells in the injured tissue. After obtaining the bone marrow the implantation can be performed with total bone marrow, with the mononuclear fraction of MSC or with cells cultured in vitro. From the fat tissue is used the stromal vascular fraction obtained by collagenase digestion, followed or not by cell culture. According to some studies, cell therapy with material obtained from bone marrow or adipose tissue has shown to be viable, given that these materials are abundant in repair components such as mesenchymal stem cells (MSC), growth factors and other components of the collagen matrix. Several studies using both types of cells have shown great potential and promising clinical results. However, knowledge of the biology and characterization of these cells remain largely unknown, and therefore is needed great care and caution when using stem cells for the treatment of musculoskeletal disorders in horses

Controle postural em pacientes com doença de Parkinson: influência da tarefa de alcance frontal e da informação sensorial

OBJETIVO: O objetivo deste estudo foi comparar o comportamento oscilatório de cada membro inferior na tarefa de alcance frontal. Além disso, verificar a influência da informação visual nestas tarefas, em pacientes com doença de Parkinson. MÉTODO: Participaram deste estudo 8 pacientes com doença de Parkinson idiopática, de ambos os sexos, classificados entre os estágios 1 a 3 da escala de Hoehn e Yahr e 8 sujeitos controle. Para avaliar o comportamento oscilatório, foi aplicada a tarefa de alcance frontal que consistiu em elevar os braços a 90º, posicionar uma mão sobre a outra, estender os dedos e tentar alcançar à frente o mais longe possível sem retirar ou mover os pés da posição inicial. Os pés foram posicionados sobre duas plataformas de força, próximas uma da outra. Para aquisição dos dados de comportamento oscilatório foi utilizado o software AMTI Net Force, com freqüência de coleta de 100 Hz. Foram realizadas 3 tentativas para cada condição (com e sem informação visual), totalizando 6 tentativas com duração de 30 segundos cada. As avaliações clínica e postural foram realizadas no estado on do medicamento. Variáveis do COP analisadas: Trajetória, Velocidades Ântero-posterior e Médio-lateral e Limite de Estabilidade. RESULTADOS: Plataforma 1- MANOVA revelou apenas efeito de momento, Wilk’s Lambda= 0,04, F(8,6)=18,74, p≤0,01. Testes univariados apontaram diferença significativa para as variáveis Trajetória (F=31,62, p<0,001), Velocidade média médio-lateral (F=83,24, p<0,001) e para Limite de estabilidade (F=27,36, p<0,001). Plataforma 2- MANOVA revelou apenas efeito de momento, Wilk’s Lambda=0,026, F(8,6)=28,23, p<0,01. Análises univariadas apontaram diferenças no Momento para trajetória F(2,26)=104,07, p<0,01, velocidade média ântero-posterior...(Resumo completo, clicar acesso eletrônico abaixo)

Transplante de medula óssea para suporte de células tronco hematopoiéticas em cães acometidos por linfoma

Lymphoma represents the most prevalent hematopoietic malignancy in small animal medicine. It is highly responsible to chemotherapy and therefore several protocols are used as therapeutic tools. For that reason, the bone marrow transplantation, enshrined in human medicine through initial trial in canine patients, has increasingly become the focus of studies in order to make it a reality also in veterinary medicine. First, the treatment with the chosen chemotherapy protocol is made. As complete remission of lymphoma is observed, it must be initiated the bone marrow harvesting. The obtained material is subjected to the processes of erythrocyte depletion, plasma depletion, cryoprotectants addition, total nuclear cells counting, hematopoietic progenitor quantification, analysis of cell viability and freezing. Following that, with radiotherapy or application of cyclophosphamide, the conditioning phase of the patient who is receiving the transplantation is carried out. The bags containing hematopoietic stem cells are then thawed and transplanted into the receptor organism. Support with hematopoietic stem cells allows the use of lethal doses of chemotherapy or radiotherapy and has been shown to considerably raise the disease remission time and survival rate of the canine patients

Papel reparador e imunomodulador das células-tronco mesenquimais (CTMs) deficientes de receptor para interferon-gamma (IFNy) em modelos renais agudos

A insuficiência renal aguda (IRA) é uma patologia que apresenta alta incidência na população e elevada morbimortalidade. Apesar de todos os avanços terapêuticos já obtidos, essas taxas ainda continuam elevadas. Uma possível alternativa, atualmente sugerida, seria o transplante de células-tronco. O processo regenerativo das células-tronco mesenquimais (CTMs) já foi demonstrado em diversos modelos experimentais e em alguns ensaios clínicos. O mecanismo de ação mais sugerido é a ação parácrina das CTMs na área lesada. Ainda, sabe-se que nesse ambiente, citocinas pró-inflamatórias, como TNF-α e IFNγ, ativam as CTMs para seu papel reparador. O presente estudo busca analisar o papel do IFNγ na ativação das CTMs em modelos renais. As CTMs de animais nocautes para receptor de IFNγ (IFNγR KO) e de animais selvagens (controle/ C57/Bl6) foram isoladas do tecido adiposo. Essas células foram caracterizadas por imunofenotipagem e diferenciação em adipócitos e osteócitos. A lesão renal aguda foi obtida através do clampeamento dos pedículos renais de camundongos machos C57/Bl6, por 45 min. Após 4hs da lesão isquêmica, as CTMs IFNγR KO e CTMs controles foram administradas intraperitonealmente, e 24hs após a cirurgia os animais foram sacrificados. O tratamento com CTMs selvagens apresentou significativa redução dos níveis de uréia e creatinina sérica. No entanto, a redução desses níveis séricos com CTMs IFNγR KO foi menos intensa. Com relação à análise da resposta inflamatória do rim, os dados demonstram que a expressão de RNAm de IL-6 é maior nos animais tratados com CTMs IFNγR KO quando comparada ao tratamento com CTMs selvagens; porém, os dois tratamentos apresentam expressão reduzida em comparação aos animais não tratados. Já a expressão de RNAm de IL-10 é maior em animais tratados com CTMs em comparação aos não tratados... (Resumo completo, clicar acesso eletrônico abaixo)

Marcadores de superfície de Células-tronco: estudo de reação cruzada inter-espécies

Mesenchymal stem cells (MSCs) are adult multipotent cells with fibroblastoid morphology and adherent to plastic. Furthermore, they can be obtained from different sources. Besides bone marrow, these cells are taken from umbilical cord blood, umbilical vein, saphenous vein, peripheral blood, arteries, liver and fetal pancreas, placenta, dental pulp and adipose tissue. MSCs derived from adipose tissue are important because of the abundant number of cells that can be obtained from this tissue, easy access and little discomfort to the patient. This study compared two techniques for obtaining MSCs from adipose tissue: mechanical dissociation (MD) and enzymatic digestion (ED). We also analyzed the inter-species cross-reactions using commercial monoclonal antibodies directed against surface antigens of stem cells from different species: mouse, horse, rabbit, monkey and human. We found that MD technique is favorable in relation to ED within 15 days of culture, and ED is more efficient in the first days of culture. The data also showed that MD causes less damage to cellular DNA. About inter-species cross-reactions, the monoclonal antibody A69 directed against stem cells from rabbits, which can be used in veterinary medicine, particularly in research involving horses