Efeito da terapia ultrassônica de baixa intensidade em cultura celular de fibroblastos / Efecto del ultrasonido de baja intensidad en cultivo celular de fibroblastos / Effect of low-intensity pulsed ultrasound therapy on a fibroblasts cell culture

Dentro da prática fisioterápica verifica-se a ampla utilização do ultrassom terapêutico para tratamento das diversas afecções musculoesqueléticas. O objetivo deste estudo foi avaliar o efeito da irradiação Ultrassônica de Baixa Intensidade, com diferentes regimes de pulsos e intensidade, em cultura celular de fibroblastos L929 (ATCC CCL-1 NCTC), de modo a verificar a viabilidade celular e definir parâmetros de dosimetria. Para isso, utilizou-se a aplicação de ultrassom pulsado, com frequência de 1Mhz, em cultura de células fibroblásticas, divididas em cinco grupos (controle e com intensidade instantâneas de 0,3W/cm2-10%; 0,3W/cm2 -20 %; 0,5W/cm2 -10% e US 0,5W/cm2 -20 % - 100Hz). A irradiação ocorreu com intervalos de 24, 48 e 72 horas, por dois minutos, e após 24 horas de cada irradiação foi realizado teste de MTT Brometo de [3-(4,5-dimetiltiazol)-2,5-difeniltetrazólio]. Os resultados revelaram que ao compararem-se os valores de células viáveis pelo método MTT nos cinco grupos, não foi possível encontrar diferença estatisticamente significativa em nenhum deles, nos três momentos avaliados (24, 48 e 72 horas); enquanto que, ao se realizar a análise de medida repetida nos diferentes grupos, encontrou-se diferença estatisticamente significativa apenas no grupo irradiado com ultrassom a 0,5W/ cm2com regime de pulso de 10% (p=0,003). Com base nesses resultados, conclui-se que a irradiação Ultrassônica de Baixa Intensidade em cultura celular de fibroblastos L929, somente no grupo com intensidade de 0,5W/cm2-10% obteve o crescimento numérico, com significância estatística em todos os períodos de avaliação.

En la práctica fisioterápica se utiliza bastante el ultrasonido como terapia para el tratamiento de diversos trastornos muscoloesqueléticos. Este artículo tiene por objetivo evaluar el efecto de la irradiación ultrasónica de baja intensidad, con diferentes regímenes de pulsos y de intensidades, en cultivo celular de fibroblastos L929 (ATCC CCL-1 NCTC), para verificar la viabilidad celular y establecer los parámetros de dosimetría. Se utilizó el ultrasonido pulsado, con frecuencia de 1Mhz, en un cultivo de células fibroblásticas, divididas en cinco grupos (con control y con la intensidad instantánea del 0,3W/cm2-10%; 0,3W/cm2 -20%; 0,5W/cm2 -10% y US 0,5W/cm2-20 % - 100Hz). La irradiación se llevó a cabo en intervalos de 24, 48 y 72 horas, durante dos minutos y después de las 24 horas de cada irradiación se realizó la prueba de MTT {Bromuro de [3-(4,5-dimetiltiazol)-2,5-difeniltetrazólio]}. Los resultados mostraron que en la comparación entre los valores de células viables por el método MTT en los cinco grupos evaluados no ha sido posible encontrar ninguna diferencia estadísticamente significativa en los tres momentos evaluados (24, 48 y 72 horas). En cambio, al llevar a cabo el análisis de medida repetida en los diferentes grupos, se obtuvo una diferencia estadísticamente significativa solamente en el grupo irradiado con ultrasonido a 0,5W/cm2 con el régimen de pulso del 10% (p=0,003). Basándose en estos resultados se concluyó que la irradiación ultrasónica de baja intensidad en cultivo celular de fibroblastos L929 obtuvo el aumento sólo en el grupo con intensidad de 0,5W/cm2-10%, con significancia en todos los periodos de evaluación.

Within the physiotherapy practice there is a wide use of therapeutic ultrasound for the treatment of various musculoskeletal disorders. The aim of this study was to evaluate the effect of Low Intensity Ultrasonic irradiation with differents forms of pulse and intensity in cell culture of L929 fibroblasts (ATCC CCL-1 NCTC), in order to check cell viability and define parameters of dosimetry. For this purpose, it was used the application of pulsed ultrasound with a frequency of 1MHz in cultured fibroblast cells divided into five groups (control and instantaneous intensity of 0.3W/cm2-10%, 0.3W/cm2 -20%, 0.5W/cm2-10% and US 0.5W/cm2-20% - 100Hz). Irradiation occurred at intervals of 24, 48 and 72 hours for two minutes and 24 hours after each irradiation test MTT [3-(4,5-dimethylthiazol)-2,5-diphenyltetrazolium bromide] was performed. Results showed that when comparing the values of viable cells by MTT method in the five groups, we could not find statistically significant difference in any of them, in these three conditions (24, 48 and 72 hours); while. Whereas, when performing the analysis of repeated measures in the different groups, it was found a statistically significant difference only in the group irradiated with ultrasound at 0.5W/cm2 with pulse regime of 10% (p=0.003). Based on these results, it is concluded that the Low Intensity Ultrasonic irradiation in L929 fibroblast cell culture, only in the group with an intensity of 0.5W/cm2 -10% obtained numerical growth, with statistical significance in all periods evaluation.

Associação da estimulação elétrica nervosa transcutânea (TENS), terapia manual e placas oclusais nas disfunções temporomandibulares: estudo de caso / Association of transcutaneous electrical nerve stimulation (TENS), manual therapy and occlusal splints in temporomandibular dysfunction: Case study

As disfunções temporomandibulares (DTM) apresentam etiologia multifatorial. Os pacientes com DTM apresentam características clínicas como dores de cabeça crônica, ruídos na articulação temporomandibular (ATM) como crepitações e estalidos, limitação dos movimentos mandibulares, dor e tensão na musculatura mastigatória do pescoço e da cabeça. A redução desses sinais e sintomas pode ser obtida por terapia física, medicamentosa, oclusal e psicoterapia. O objetivo deste trabalho é verificar a eficácia do tratamento conjunto da aplicação da estimulação elétrica nervosa transcutânea (TENS), da terapia manual e da utilização de placa oclusal na DTM. A terapia foi realizada em três pacientes do sexo feminino com idade entre 20 a 40 anos. Os resultados deste estudo demonstram que a associação das técnicas de TENS, terapia manual e terapia oclusal, proporcionaram aos pacientes diminuição do quadro álgico e melhora da amplitude de movimento da ATM, de acordo com os meios de avaliação utilizados. Conclui-se que a associação das técnicas pode ser indicada como método eficaz, não invasivo e de baixo custo no tratamento destes pacientes, ressaltando que ainda é preciso muitos estudos nessa área para comprovação e esclarecimento de protocolos terapêuticos, envolvendo uma equipe multiprofissional.

The temporomandibular dysfunctions (DTM) have a multifactorial etiology. Patients with DTM present clinical features as chronic headache, noises in the temporomandibular joint (ATM) as crackles and click, limitation ofmandibular movements, pain and tension in the masticatory muscles of the neck and head. The reduction of signs and symptoms can be obtained by physical therapy, medication, psychotherapy and occlusal therapy. The objective of this work is to verify the effectiveness of joint processing of the application of transcutaneous electric nerve stimulation (TENS), manual therapy and use of occlusal splints. This treatment was held in three patients females aged between 20 and 40 years. The results of this study demonstrate that the combination of the techniques of TENS, manual therapy and occlusal therapy enabled patients to decrease the pain symptoms and to improve the range of motion of the ATM, according to the methods of evaluation that were used. It is concluded that the combination of these techniques can be suggested as an effective, non invasive and cost-effective method in the treatment of these patients,emphasizing that many studies are needed in this area for evidence and clarification of treatment protocols, involving a multidisciplinary group.

Assessment of cytoskeleton and endoplasmic reticulum of fibroblast cells subjected to low-level laser therapy and low-intensity pulsed ultrasound.

OBJECTIVE: The aim of the present study was to compare the effect of low-level laser therapy (LLLT) and low-intensity pulsed ultrasound (LIPUS) on the cytoskeleton and endoplasmic reticulum of L929 cells. Thermal and non-thermal physical mechanisms such as LLLT and LIPUS induce clinically significant responses in cells, tissues, and organs. MATERIALS AND METHODS: L929 fibroblast cell cultures were irradiated with LLLT and subjected to LIPUS. Cultures irradiated with the laser (904 nm) were divided into three groups: group I, control (no irradiation); group II, irradiated at 6 J/cm(2); and group III, irradiated at 50 mJ/cm(2). Cultures subjected to ultrasound were divided into five groups: group I, control (no LIPUS); group II, LIPUS at 0.2 W/cm(2) in pulsed mode at 10% (1:9 duty cycle); group III, LIPUS at 0.6 W/cm(2) in pulsed mode at 10% (1:9 duty cycle); group IV, LIPUS at 0.2 W/cm(2) in pulsed mode at 20% (2:8 duty cycle); and group V, LIPUS at 0.6 W/cm(2) in pulsed mode at 20% (2:8 duty cycle). Each group was irradiated at 24-h intervals, with the following post-treatment incubation times: 24, 48, and 72 h. The effects of LLLT and LIPUS on the cytoskeleton and endoplasmic reticulum was evaluated by the use of fluorescent probes and with fluorescence microscopy analysis. RESULTS: The results following LLLT and LIPUS demonstrate that ultrasound was more effective than laser on fibroblast cell cultures when the endoplasmic reticulum was assessed, whereas there was a better distribution of the filaments of the cytoskeleton in the cells subjected to laser irradiation. CONCLUSION: The study demonstrated that both LLLT and LIPUS promote changes on the cellular level. However, LIPUS was more effective than LLLT at the doses used here, as assessed by fluorescence microscopy, which revealed increased reticulum activity and increased protein synthesis. However, when the organization of actin filaments was assessed, LLLT achieved a better result.

Comparison between the effect of low-level laser therapy and low-intensity pulsed ultrasonic irradiation in vitro.

OBJECTIVE: The objective of this study was to compare the effect of low-level laser therapy (LLLT) and low-intensity pulsed ultrasound (LIPUS) on fibroblast cell culture. Several methods, including ultrasound treatment and LLLT, are being used to facilitate tissue repair and healing processes. MATERIALS AND METHODS: L929 fibroblast cell cultures were irradiated with low-level laser energy and LIPUS. Cultures irradiated with ultrasound were divided into five groups: group 1: control (did not receive irradiation); group 2: 0.2 W/cm(2) in pulsed mode at 10% (1:9 duty cycle); group 3: 0.6 W/cm(2) in pulsed mode at 10% (1:9 duty cycle); group 4: 0.2 W/cm(2) in pulsed mode at 20% (2:8 duty cycle); and group 5: 0.6 W/cm(2) in pulsed mode at 20% (2:8 duty cycle). Cultures irradiated with laser energy were divided into three groups: group 1: control (did not receive irradiation); group 2: 6 J/cm(2); and group 3: 50 mJ/cm(2). Each group was irradiated at 24-h intervals, with the following incubation periods post-irradiation: 24, 48, and 72 h; after each irradiation cycle the cultures were analyzed using MTT [3-(4.5-dimethylthiazol-2-yl)-2.5 diphenyltetrazolium bromide]. RESULTS: Analysis of results after LLLT and LIPUS demonstrated that the effect of laser therapy on fibroblast cell culture was greater than that of LIPUS (p < 0.05). CONCLUSION: Results demonstrated that LLLT significantly increased fibroblastic activity more than LIPUS. Therefore, in the first and second phases of tissue repair, laser treatment may be more effective than ultrasound treatment.

Estudo da resposta motora do músculo vasto lateral e dos componentes longo e oblíquo do músculo vasto medial, em contração isométrica máxima, duranteextensão do joelho / Study of the vastus lateralis muscle and long and oblique components of the vastus medialis muscle motor response in maximum isometric contraction during knee extension

A proposta deste trabalho foi analisar o comportamento eletromiográfico do músculo vasto lateral e dos componentes longo e oblíquo do músculo vasto medial em contração isométrica máxima, durante extensão da articulação do joelho, tomando-se as medidas de ângulo de 150º, 165º e 180º de extensão do joelho. Foram analisados 26 voluntários, do sexo feminino, normais, utilizando-se um módulo de aquisição de sinais biológicos, eletrodos de superfície para aquisição e processamento dos sinais eletromiográficos, utilizou-se o software AqDados. O método estatístico empregado foi Análise de Variância e o teste de Tukey. Os resultados mostraram que o músculo vasto lateral e o componente oblíquo do músculo vasto medial, apresentaram atividade semelhante e maior que aquela desenvolvida pelo componente longo do músculo vasto medial e quando avaliado o comportamento de cada músculo em relação aos três ângulos estudados, verifica-se que todos os músculos não apresentaram diferença significante entre os três ângulos analisados. Portanto, para desenvolver-se um protocolo de fortalecimento muscular pelas contrações isométricas diante desse grupo muscular, podemos utilizar qualquer ângulo (150º, 165º e 180º), na postura utilizada na pesquisa, pois os mesmos não apresentaram diferenças significantes.

The aim of this investigation was to analyze the electromyographic behavior of the vastus lateralis muscle and long and oblique components of the vastus medialis muscle in a maximum isometric contraction, during extention of the knee articulation, using angles of 150º, 165º, and 180º of knee extention. A total of 26 healthy female volunteers were analyzed, using a biological signal acquisition module and surface electrodes; electromyographic signals were collected and processed by an AqDados software. The statistical methods employed were Analysis of Variance and the Tukey test. The results of this investigation showed that the vastus lateralis muscle and the oblique component of the vastus medialis muscle have similar activity, and that both of them have higher activity than the long component of the vastus medialis muscle; when the behavior of each muscle in the three studied angles had been evaluated, we verified that they did not show significant difference between them. Thus, for developing a muscular strenghtening protocol by isometric contractions of this muscular group, we can apply any angle (150º, 165º and 180º), in the posture used in our experiment, since the latter did not show significant differences.