SUMMARY: The purpose of this study was to evaluate the chronic effects of stretching exercise on soleus muscle histomorphology and histomorphometry of young and aged rats. Thirty-eight female rats were divided into young control group (YCG, n=10;274±50 g); young stretching group (YSG, n=8;274±12 g); aged control group (ACG, n=10;335±39 g); and aged stretching group (ASG, n=10;321±32g). A mechanical apparatus was used to stretch muscle in 4 repetitions, 60 s each, 30 s interval between repetitions in each session, 3 times a week for 3 weeks. Twenty-four hours after the last stretching session, soleus muscle was removed for micromorphology and immunostaining analysis. Data analyses were performed with one-way ANOVA, post-hoc Tukey, or Kruskal-Wallis tests for parametric and nonparametric, respectively (p≤0.05). Muscle fiber cross-sectional area (MFCSA) of ACG was lower (18 %) compared to the YCG. Stretching increased MFCSA comparing YSG to YCG (5,681.15± 1,943.61 µm2 vs 5,119.84±1,857.73 µm2, p=0.00), but decreased comparing ASG to ACG (3,919.54± 1,694.65 µm2 vs 4,172.82±1,446.08 µm2, p=0.00). More serial sarcomere numbers were found in the YSG than YCG (12,062.91±1,564.68 vs 10,070.39±1,072.38, p=0.03). Collagen I and collagen III were higher in YSG than ASG (7.44±7.18 % vs 0.07±0.09 %, p=0.04) and (14.37 %± 9.54 % vs 5.51 %±5.52 %, p=0.00), respectively. TNF-a was greater in ASG than YSG (43.42 %±40.19 % vs 1.72 ± 2.02 %, p=0.00). Epimysium was larger in the YSG compared to YCG (201.83±132.07 % vs 181.09±147.04 %, p=0.00). After 3-week stretching the soleus muscles from aged rats were smaller than their younger counter-parts. Interestingly, while stretching appeared to positively affect young soleus muscle, the opposite was detected in the muscle of the aged rats.

RESUMEN: El propósito de este estudio fue evaluar los efectos crónicos del ejercicio de estiramiento sobre la histomorfología e histomorfometría del músculo sóleo de ratas jóvenes y envejecidas. Se dividieron 38 ratas hembras en un grupo control joven (YCG, n = 10; 274 ± 50 g); grupo de estiramiento joven (YSG, n = 8; 274 ± 12 g); grupo control de edad (ACG, n = 10; 335 ± 39 g); y grupo estiramiento envejecido (ASG, n = 10; 321 ± 32 g). Se usó un aparato mecánico para estirar el músculo en 4 repeticiones, 60 s cada una, intervalo de 30 s entre repeticiones en cada sesión, 3 veces por semana, durante 3 semanas. Veinticuatro horas después de la última sesión de estiramiento, se extrajo el músculo sóleo para análisis de micromorfología e inmunotinción. Los análisis de datos se realizaron con pruebas ANOVA de una vía, Tukey post-hoc o Kruskal-Wallis para pruebas paramétricas y no paramétricas, respectivamente (p≤0,05). El área de la sección transversal de fibra muscular (MFCSA) de GCE fue menor (18 %) en comparación con el GCJ. El estiramiento aumentó ASTFM comparando GEJ con GCJ (5.681,15 ± 1.943,61 µm2 vs 5.119,84 ± 1.857,73 µm2, p = 0,00), pero disminuyó comparando GEE con GCE (3.919,54 ± 1.694,65 µm2 vs 4.172,82 ± 1.446,08 µm2, p = 0,00). Se encontraron más sarcómeros en serie en el GEJ que en el GCJ (12.062,91 ± 1.564,68 vs 10.070,39 ± 1,072.38, p = 0,03). El colágeno I y el colágeno III fueron más numerosos en GEJ que en GEE (7,44 ± 7.18 % vs 0,07 ± 0,09 %, p = 0,04) y (14,37 % ± 9,54 % vs 5,51 % ± 5,52 %, p = 0,00), respectivamente. TNF-α fue mayor en GEE que GEJ (43,42 % ± 40,19 % vs 1,72 ± 2,02 %, p = 0,00). El epimisio fue mayor en el GEJ en comparación con el GCJ (201,83 ± 132,07 % vs 181,09 ± 147,04 %, p = 0,00). Después de 3 semanas de estiramiento, los músculos sóleo de las ratas envejecidas eran más pequeños que sus contrapartes más jóvenes. Curiosamente, si bien el estiramiento pareció afectar positivamente el músculo sóleo joven, se detectó lo contrario en el músculo de las ratas envejecidas.

OBJECTIVES: To determine the effects of three sessions of a passive stretching exercise protocol on the muscles of elderly female rats. METHODS: The effects of the stretching exercises on the soleus muscle were analyzed using immunohistochemistry [tissue inhibitors of matrix metalloproteinases (TIMP), the tumor necrosis factor-alpha (TNF-α), and the gene expression levels using real-time PCR of the transforming growth factor-beta 1 (TGF-β1), collagen type 1 (COL1), and collagen type 3 (COL3)]. Fifteen 26-month-old female Wistar rats were randomly divided into two groups, namely, Stretching (SG, n=8) and Control (CG, n=7). The passive mechanical stretching protocol consisted of a set of 4 1-minute repetitions, with 30 seconds between each repetition (total treatment of 4 minutes), three times a week for 1 week. RESULTS: Immunohistochemical analysis revealed an increase of 71.4% in the TNF-α (p=0.04) gene expression levels for the SG and a 58% decrease in the TGF-β1 gene expression levels (p=0.005) in the SG compared to that in the CG. No significant differences were observed between the groups for the immunostaining of TIMP-1 or the gene expression levels of COL1 and COL3. CONCLUSION: Three sessions of static stretching reduced the gene expression level of TGF-β1, which, owing to its anti-fibrotic role, might contribute to the remodeling of the intramuscular connective tissue of the aging muscle. In addition, immunostaining revealed that TNF-α levels increased in the aging muscle tissue in response to stretching, indicating its effect on stimulating extracellular matrix degradation. These outcomes have important clinical implications in reinforcing the use of stretching exercises in the elderly, considering that the aging muscle presents an infiltration of connective tissue.

Resumo Contexto Pacientes com úlceras venosas reportam múltiplas comorbidades e são mais propensos a ser fisicamente inativos. A sarcopenia e a fragilidade aumentam a vulnerabilidade de um indivíduo para maior dependência e/ou morte. Objetivos Verificar presença da sarcopenia e fragilidade em pacientes portadores de úlceras venosas crônicas. Métodos Estudo observacional e transversal, realizado com 9 pacientes com idade média de 67,4 ± 8,42 anos e portadores de úlcera venosa nos membros inferiores classificadas no Consenso Internacional de Doenças Venosas Crônicas (CEAP) em estágio 6. Para identificação e classificação da sarcopenia, foi avaliada a força (dinamometria manual), a velocidade da marcha (teste de caminhada de 10 metros) e a massa muscular (circunferência da panturrilha). Para triagem de fragilidade, foram utilizados os critérios de Fried: perda de peso não intencional, fadiga, redução da força e da velocidade da caminhada e baixa atividade física. Resultados O fenótipo de fragilidade foi mais frequente (n = 9; 100%) em relação à sarcopenia (n = 1; 11,1%). Entre os critérios de Fried, os mais frequentes foram a exaustão (n = 9; 100%), seguida pela baixa atividade física (n = 8; 88,8%) e fraqueza muscular (n = 5; 55%). Por fim, o critério menos frequente foi a diminuição da velocidade da marcha (n = 2; 22,2%). No diagnóstico de sarcopenia, foi observada redução da força associada à redução da massa muscular (n = 1; 11,1%). Conclusões Pacientes com úlceras venosas crônicas apresentam condição de fragilidade ou pré-fragilidade, enquanto a condição de sarcopenia foi pouco frequente.

Abstract Background Patients with venous ulcers report multiple comorbidities and are more likely to be physically inactive. Sarcopenia and frailty increase vulnerability to dependence and/or death. Objectives To investigate the occurrence of sarcopenia and frailty in patients with chronic venous ulcers. Methods Observational study with cross-sectional design. Nine patients (67.4 ± 8.42 years) with lower limb venous ulcers classified as CEAP 6 according to International Consensus on Chronic Venous Diseases criteria (open and active ulcer) were evaluated. Sarcopenia was assessed and classified by assessment of strength (manual dynamometry), gait speed (10-meter walk test), and muscle mass (calf circumference). Frailty screening was based on the Fried criteria, consisting of five components: unintentional weight loss; exhaustion; weakness; slow gait speed; and low physical activity. Results Frailty was more frequent (n=9; 100%) than sarcopenia (n=1; 11,1%). The most common Fried criterion was exhaustion (n=9; 100%), followed by low physical activity (n=8; 88,8%), muscle weakness (n=5; 55%), and unintentional weight loss. Finally, the least frequent criterion was slow walking speed (n=2; 22.2%). In the subject diagnosed with sarcopenia, both weakness and reduced muscle mass were observed (n=1; 11,1%). Conclusions Patients with chronic venous ulcers exhibit frailty or pre-frailty and the components that comprise the condition of frailty in this population are exhaustion, low physical activity, and muscle weakness. Sarcopenia was identified in a small proportion of the patients.

RESUMO Laser e alongamento são usados para tratar lesões musculares. Este estudo objetivou avaliar os efeitos do laser GaAs e alongamento na morfologia do músculo Tibial anterior (TA) após contusão. Trinta e seis ratos (349±23 g) foram divididos em seis grupos (n=6): grupo controle (GC); grupo lesão e laser (GLL); grupo lesão e alongamento (GLA); grupo lesão, laser e alongamento (GLLA) e grupo alongamento (GA). Foi realizada lesão no TA por meio de um aparato de contusão. O tratamento com laser GAAS foi usado com dose de 4,5 J/cm2 durante 32 s, iniciando 48 h após lesão, por 7 dias. Alongamento passivo manual consistiu de 10 repetições de 1 minuto de duração, iniciando no 8º dia, 3 vezes por semana, durante 3 semanas. Após 4 semanas, os ratos foram eutanasiados para retirada do TA para análise de: peso e comprimento musculares, área de secção transversa das fibras musculares (ASTFM), número de sarcômeros em série (NSS), comprimento dos sarcômeros e porcentagem de tecido conjuntivo. A comparação entre os grupos deu-se por meio da ANOVA e post hoc Tukey, com nível de significância ≤ 0,05. O número de sarcômeros em série do GLLS foi maior que o GLS. O comprimento dos sarcômeros no GLA foi superior ao GLL, GLLA e GA. No GA houve aumento do NSS comparado com o GC, enquanto a porcentagem de tecido conjuntivo do GA diminuiu em comparação com o GLLA. Assim, a sarcomerogênese dos músculos lesionados foi aumentada pelo uso do laser, alongamento e pela associação destes. O alongamento foi suficiente para aumentar o NSS em músculos intactos.

ABSTRACT Laser and stretching are used to treat skeletal muscle injuries. This study aimed to evaluate the effects of GaAs laser and stretching in the morphology of the tibialis anterior (TA) muscle after contusion. Thirty-six male rats (349±23g) were divided into six groups (n=6): control group (CG); lesion group (LG); lesion and laser group (LLG); lesion and stretching group (LSG); lesion, laser and stretching group (LLSG); and stretching group (SG). TA was wounded by a contusion apparatus. We used GaAs laser 4.5 J/cm2 dose for 32 s each, beginning 48 h after lesion, for 7 days, once a day. Manual passive stretching was applied by 10 repetitions for 1 minute, initiating on the 8th day, once a day, 3 times a week, during 3 weeks. After 4 weeks, rats were euthanized and we analyzed: muscle weight and length, cross sectional area of muscle fibers (CSAMF), serial sarcomere number (SSN), sarcomere length, and percentage of connective tissue. Comparisons among groups were made by ANOVA and post hoc Tukey tests, with the significance level set at ≤ 0.05. The serial sarcomere number of LLSG was higher than LSG. The sarcomere length of LSG was superior to LLG, LLSG, and SG. SG increased SSN compared to CG, while the percentage of connective tissue of SG decreased in comparison to LLSG. Thus, the sarcomerogenesis of injured muscles was enhanced by laser therapy, stretching, and association of both. The stretching protocol was enough to increase SSN of intact muscles.

RESUMEN Láser y estiramiento son utilizados para sanar lesiones musculares. Este estudio tuvo como objetivo evaluar los efectos del láser GaAs y del estiramiento en la morfología del músculo tibial anterior (TA) después de contusión. Treinta y seis ratones (349±23 g) fueron divididos en seis grupos (n=6): grupo control (GC); grupo lesión y láser (GLL); grupo lesión y estiramiento (GLE); grupo lesión, laser y estiramiento (GLLA) y grupo estiramiento (GA). Se realizó lesión en el TA mediante un aparato de contusión. Se utilizó el tratamiento con láser GaAs con dosis de 4,5 J/cm2 durante 32 s, iniciado 48 h después de la lesión, durante 7 días. El estiramiento pasivo manual consistió de 10 repeticiones de 1 minuto de duración, iniciado en el 8º día, 3 veces por semana, durante 3 semanas. Después de 4 semanas, los ratones sufrieron eutanasia para la retirada del TA para análisis de: peso y longitud musculares, área de sección trasversa de las fibras musculares (ASTFM), número de sarcómeros en serie (NSS), longitud de los sarcómeros y porcentaje de tejido conjuntivo. La comparación entre los grupos ocurrió mediante la Anova y post hoc Tukey, con nivel de significancia de ≤0,05. El número de sarcómeros en serie del GLLS fue mayor que el GLS. La longitud de los sarcómeros en el GLA fue superior al GLL, GLLA y GA. En el GA hubo aumento del NSS en comparación con el GC, mientras el porcentaje del tejido conjuntivo del GA disminuyó en comparación al del GLLA. Así, la sarcomerogénesis de los músculos lesionados fue aumentada por el uso del láser, estiramiento y por su asociación. El estiramiento fue suficiente para aumentar el NSS en músculos intactos.