Effect of probiotic therapy on periodontal tissues and intestinal mucosa of rats with ligature-induced periodontitis / Efeito da terapia probiótica nos tecidos periodontais e mucosa intestinal de ratos com periodontite induzida por ligadura

INTRODUCTION: As periodontitis is caused by dysbiotic biofilm, it is believed that therapy with probiotics can act to control the mechanisms of adhesion and colonization, competing with invading microorganisms. OBJECTIVE: Evaluate probiotic therapy effect on periodontal tissues and intestinal mucosa of rats with ligature-induced periodontitis. METHODS: 32 Wistar rats were divided into four groups (n=8): Control Group (CG); Periodontal disease (PD); Probiotic (PROB); PD + probiotic (PDPRO). PD and PDPRO received a ligature over the first lower molars and PROB and PDPRO the probiotic Lactobacillus acidophilus based were given orally for 44 days. The animals were euthanized and the blood was collected for evaluation of triglyceride and cholesterol concentrations. The hemimandibles were collected for histomorphometric and radiographic analysis. The duodenum was removed for morphological evaluation and gingival tissue around the molars was collected for analysis of IL-17. RESULTS: The ANOVA one-way test was used followed by Tukey Test. PDPRO had a significantly lower bone loss than the PD (p<0.05) and a smaller number of osteoclasts on PDPRO when compared to the PD. As for IL-17, there was a decrease in the PDPRO when compared to the PD. The histomorphometry of the duodenum showed that there was a significant increase in the width of the villi in PROB only. CONCLUSION: The therapy with probiotics was effective to avoid the development of periodontitis by reducing alveolar bone loss and inflammation modulation and increasing the width of the duodenum villi, which may help to restabilize the balance of the gastrointestinal tract.

INTRODUÇÃO: Como a periodontite é causada por biofilme disbiótico, acredita-se que a terapia com probióticos possa atuar no controle dos mecanismos de adesão e colonização, competindo com microrganismos invasores. OBJETIVO: Avaliar o efeito da terapia probiótica nos tecidos periodontais e mucosa intestinal de ratos com periodontite induzida por ligadura. MÉTODOS: 32 ratos Wistar foram divididos em quatro grupos (n=8): Grupo Controle (GC); Doença periodontal (PD); Probiótico (PROB); PD + probiótico (PDPRO). PD e PDPRO receberam ligadura sobre os primeiros molares inferiores e PROB e PDPRO, o probiótico à base de Lactobacillus acidophilus dado via oral por 44 dias. Os animais foram sacrificados e o sangue coletado para avaliação das concentrações de triglicerídeos e colesterol total. As hemimandíbulas foram coletadas para análise histomorfométrica e radiográfica. O duodeno foi removido para avaliação morfológica e o tecido gengival ao redor dos molares foi coletado para análise de IL-17. RESULTADOS: Foi usado Teste ANOVA seguido pelo Teste de Tukey. PDPRO teve uma perda óssea significativamente menor do que o PD (p<0.05) e um menor número de osteoclastos no PDPRO quando comparado ao PD. Já para IL-17, houve diminuição do PDPRO em relação ao PD. A histomorfometria do duodeno mostrou que houve aumento significativo da largura das vilosidades no PROB somente. CONCLUSÃO: A terapia com probiótico foi eficaz para evitar o desenvolvimento de periodontite por reduzir a perda óssea alveolar e a modulação da inflamação e aumentar a largura das vilosidades duodenais, o que pode ajudar a estabilizar o equilíbrio do trato gastrointestinal.

Effects of the Association of Periodontitis and Type 1 Diabetes Mellitus Induced on Periodontal Tissues and the Duodenal Mucosa of Wistar Rats.

Forty Wistar rats were used: (1) control group (CG); (2) group of periodontal disease (PD); (3) type 1 diabetes mellitus group (T1DM); (4) type 1 diabetes mellitus + periodontal disease group (T1DM + PD). In groups T1DM and T1DM + PD, T1DM induction was performed with the administration of streptozotocin (STZ) 80 mg/kg intraperitoneal body weight. The PD and T1DM + PD groups were submitted to PD induction with ligation. After the experimental phase and euthanasia, histological, radiographic, and morphological analyses were performed. For data analysis, was used the one-way ANOVA and post-test Tukey. The T1DM + PD group had a significantly higher level of fasting blood glucose compared to the other groups. In radiographic and histomorphometric analyses, the T1DM + PD group showed greater alveolar bone loss compared to the control group. The T1DM + PD group showed greater osteoclastic activity compared to the control, T1DM, and PD groups and exhibited an intense inflammatory infiltrate, most of which were PMN, being that the amount of this group of cells (PMN) was significantly greater than the PD group. The heights of the intestinal villi were statistically higher in the PD, T1DM, T1DM + PD groups, compared to the control. Regarding the height of the crypt, only the T1DM and T1DM + PD groups were significantly higher compared to the other groups. Association of diabetes and periodontal inflammation increased the deleterious effects on bone tissue and adverse effect on the permeability of the duodenal mucosa.

Avaliações Histométrica, Radiográfica e Imunológica dos Tecidos Periodontais de Ratas Ovariectomizadas com Periodontite Induzida / Histometrical, Radiographic and Immunological Evaluations of the Periodontal Tissues of Female Ovariectomized Rats With Induced Periodontits

Objetivo: A relação da osteoporose com pós-menopausa é controversa, mas pode ser um fator de risco para doença periodontal. Assim o objetivo deste projeto foi avaliar a influência da deficiência de estrógeno nos tecidos periodontais de ratas ovariectomizadas com peridontite induzida. Material e método: Quarenta ratas com 8 semanas de vida foram divididas aleatoriamente em 4 grupos: 1) Grupo controle; 2) Grupo ligadura; 3) Grupo ovariectomia; 4) Grupo ovariectomia e ligadura; sendo que todos os animais receberam dieta padrão e água à vontade. Aos 70 dias de vida os animais foram anestesiados e foi realizada a indução da doença periodontal, sendo após este procedimento aos 79 dias de vida, as ratas também foram submetidas à anestesia e à cirurgia de ovariectomia. Aos 100 dias de vida, as ratas foram sacrificadas sendo retiradas as hemimandíbulas esquerda e direita de cada rata para análises histomorfométrica e radiográfica, bem como amostras do tecido gengival para avaliar os tecidos periodontais. Resultados: Os resultados demonstraram que na análise radiográfica e histomorfométrica do osso alveolar dos primeiros molares inferiores nos grupos onde foi feita a indução da doença periodontal por ligadura, houve maior perda óssea alveolar significativamente (p<0.05). Em relação às concentrações de citocinas, os grupos que foram ovariectomizadas apresentaram concentrações maiores estatisticamente significante em relação aos demais (p<0.05). Conclusão: Assim pode-se concluir que a deficiência de estrógeno poderia contribuir para uma evolução mais rápida da perda óssea alveolar em ratas ovarietomizadas.

Aim: The relation of osteoporosis on the post-menopause is controversial; however, it can be a risk factor for the periodontal disease. Therefore, the aim of this study was to evaluate the influence of the estrogen deficiency on the periodontal tissues of female ovariectomized rat with induced periodontitis. Materials and methods: Forty rats with 8 weeks of life were randomly divided into 4 groups: 1) Control group; 2) Ligature group; 3) Ovariectomized group; 4) Ovariectomized and ligature group; that being that all the animals were treated in a standard diet and water available all time. When reaching 70 days of life, the animals were anesthetized and a periodontal disease induction was carried, after this procedure, when reaching 79 days of life, the rats were also submitted to anesthesia and went through the ovariectomy surgery. When reaching 100 days of life, the female rats were sacrificed, at when the left and right hemimandible of each rat were taken for radiographic and histomorphometric analysis, as the samples of the gingival tissue to evaluate the periodontal tissue. Results: The results highlighted that the radiographic and histomorphometric analysis of the lower alveolar bone first molars of the groups in which the induction of the periodontal disease by ligature, there was higher alveolar bone loss significantly (p<0.05). When it comes to the relation of the cytokine concentration, the groups that were ovariectomized presented significant higher concentration when compared to the others (p<0.05). Conclusion: In this way, we could conclude that the estrogen deficiency could contribute to the quicker evolution of the alveolar bone loss on female ovariectomized rats.

Negative impact of Novaluron on the nontarget insect Bombyx mori (Lepidoptera: Bombycidae).

Due to increased use of agrochemicals and growing concerns about ecotoxicology, the development of new insecticides, moving away from those with neurotoxic and broad spectrum effects towards insecticides that are safer for the environment and nontarget beneficial species, has been a research priority. Novaluron stands out among these newer insecticides, is an insect growth regulator that is used for the control of insect pests in crops grown close to mulberry plantations. Mulberry serves as food for the silkworm Bombyx mori, which is a nontarget insect of great economic importance to silk production. We investigated the lethal and sublethal effects of Novaluron on the development of B. mori. Larvae were segregated into experimental groups: the control groups (CGs) and the treatment groups (TGs), which were treated with the Novaluron concentration of 0.15 mL/L. Following exposure, we analyzed: larval mortality, changes in the insect life cicle and cytotoxic effects on the midgut cells. This is the first report about the Novaluron's effects on B.mori. We detected rupture in the integument, complete cessation of feeding, late development, incomplete ecdysis and production of defective cocoons. After 240 h of exposure, there was 100% mortality in TG larvae exposed in the 3rd instar and 20% mortality from larvae exposed in the 5th instar. Cytotoxic effects was observed, such as dilation of cells, emission of cytoplasmic protrusions, extreme rarefaction of the cytoplasm and nuclei, dilation of the endoplasmic reticulum in addition to changes in mitochondria, the presence of large digestive vacuoles and intercellular spaces and the presence of active caspase. Novaluron exposure impairs the midgut and may affect the physiological functions of this organ. Novaluron additionally compromises several phases of insect development, indicating the importance of toxicology studies that utilize different life stages of nontarget species to evaluate the safe use of insecticides.

Morphofunctional characteristics of skeletal muscle in rats with cerebral palsy.

Knowledge of skeletal muscle adaptations is important to understand the functional deficits in cerebral palsy (CP). This study aimed to investigate the morphofunctional characteristics of skeletal muscle in a CP animal model. Initially, pregnant Wistar rats were injected intraperitoneally with saline or lipopolysaccharide over the last five days of pregnancy. The control group (n = 8) consisted of male pups born to females injected with saline. The CP group (n = 8) consisted of male pups born to females injected with lipopolysaccharide, which were submitted to perinatal anoxia [day of birth, postnatal day 0 (P0)] and sensorimotor restriction (P1-P30). The open-field test was undertaken on P29 and P45. On P48, the animals were weighed, and the plantaris muscle was collected and its weight and length were measured. Transverse sections were stained with haematoxylin-eosin, NADH-TR, Masson's trichrome and non-specific esterase reaction for analysis. and transmission electron microscopy was performed. In the CP group, reductions were observed in mobility time, number of crossings and rearing frequency, body weight, muscle weight and length, and nucleus-to-fibre and capillary-to-fibre ratios. There was a statistically significant increase in the percentage area of the muscle section occupied by collagen; reduction in the area and increase in the number of type I muscle fibres; increase in myofibrillar disorganization and Z-line disorganization and dissolution; and reduction in the area and largest and smallest diameters of neuromuscular junctions. Thus this animal model of CP produced morphofunctional alterations in skeletal muscle, that were associated with evidence of motor deficits as demonstrated by the open-field test.

Antioxidant Systems as a Response to Midgut Cellular of Bombyx mori Lineu, 1758 (Lepidoptera: Bombycidae) Infection for Baculoviruses.

Bombyx mori nucleopolyhedrovirus (BmNPV) is a DNA virus that infects different tissues in Bombyx mori at immature stage. Caterpillars become infected after ingesting polyhedral occlusion bodies (POB) present in contaminated mulberry leaves and spread through the body after passing the epithelium of the midgut. As this organ is responsible for digestion, most absorption of nutrients requires an intact epithelium to maintain gastrointestinal physiology. Considering the importance of this organ in the feeding of caterpillars and in the production of quality silk threads, and because it is also the first barrier faced by the BmNPV, the study analyzed details of cytopathological events in the intestinal cells as well as evaluated the action of the antioxidant systems as a response to cellular infection. For this purpose, B. mori hybrid caterpillars of fifth instar were inoculated with a suspension of 7.8 × 107 POB ml-1 and, from the first to the eighth day post-inoculation (dpi), segments of the midgut were collected and processed for light and electronic microscopy. The nuclei of columnar cells showed polyhedric occlusion bodies in the seventh dpi and fragmentation of those cells, with peritrophic matrix disorganization. Analysis of antioxidant systems shows some moments of changes of the catalase enzymes and superoxide dismutase. Analysis of the cholinergic system revealed changes only at the beginning of the infection. Thus, the article acknowledges the antioxidant system as a barrier to stop viral infection, albeit it cannot stop infection from occurring, once a coevolutionary bond is maintained between virus and host.

Sericin silk protein in peripheral nervous repair associated with the physical exercise of swimming in Wistar rats.

OBJECTIVE: To verify the action of sericin associated to swim exercise with overload, on sciatic nerve repair in Wistar rats, after 22 days of nerve compression. METHODS: Forty animals been composed of five groups: control, injury, injury-sericin, injury-swimming and injury-sericin-swimming. During the lesion procedure, sericin, in hydrolyzed form, applied directly to the injury in the injury-sericin and injury-sericin-swimming groups. Injury-swimming and injury-sericin-swimming groups underwent to 5 days per week for 3 weeks, with a 10% overload of the animal's body weight, and a weekly progressive evolution of swimming time, lasting 15, 20 and 25 min/day. Pre and throughout the treatment period the animals performed evaluation of sciatic functional index and pressure pain threshold with digital von Frey filament. Euthanasia was performed on the 22nd postoperative day, and two fragments of the nerve were collected and prepared for descriptive and quantitative analysis. RESULTS: The sciatic functional index assessment showed significant differences in the motor function of the control until the 14th day. Regarding the allodynia, there was revealed a significant improvement in injury-swimming performance relative to injury, injury-sericin and injury-sericin-swimming, and the number of viable and non-viable nerve fibers smaller than 4 µm in diameter was significantly higher in the injury-sericin-swimming. CONCLUSION: swimming showed a better evolution of the nociceptive threshold and allodynia. Sericin treatment had exacerbated pro-inflammatory characteristics. On the other hand, the association of sericine and swimming showed a possible regulatory effect by resting swimming exercise, with a significant increase of fibers of smaller diameter.

Morphological and nociceptive effects of mechanical vibration on the sciatic nerve of oophorectomized Wistar rats

Aim: To evaluate the effects of whole-body vibration in the sciatic nerve of oophorectomized Wistar rats, on nociceptive and morphological parameters, such as fiber, axon, and myelin sheath diameters, G ratio, number of nerve fiber and nuclei of Schwann cells, and percentage of connective tissue. Method: Sixty-four rats were used in the groups sham-operate and oophorectomy (n = 32/group); after surgical procedures, each group was subdivided into four: euthanized in the 12th week, untreated and treated for four weeks; and euthanized in the 16th week, untreated and treated for eight weeks. The treatment with vibration was performed with a 60 Hz frequencies, for 10 minutes, three days a week, with duration of 4 or 8 weeks. Nociception was evaluated later, in the right paw, by means of a digital analgesimeter, prior to surgery, at the beginning and at the end of the protocol. After the trial period, the sciatic nerve was dissected for examination of the general morphology of the tissue and morphometric analysis; later, the animals were euthanized. Results: Regarding nociception and the morphometry of the sciatic nerve, independent of oophorectomy and treatment time, there was no statistically significant difference within and between groups. Also, the general morphology of the tissue in all groups had characteristics that were preserved. Conclusion: The mechanical vibration did not alter the nociceptive threshold and the morphological aspects of nerve fibers in oophorectomized Wistar rats.(AU)

Avaliações funcionais e morfométrica de modelo de lesão muscular por agulha de biópsia em gastrocnêmio de ratos / Functional and morphometric evaluations of model of muscle lesion with biopsy needle in gastrocnemius of rats

Introdução: Dentre os modelos experimentais, o uso de agulha de biópsia mostrou ser eficaz para reproduzir as lesões em atletas. Objetivo: Avaliar a força de preensão, a nocicepção e as alterações morfológicas do músculo gastrocnêmio de animais submetidos à lesão muscular. Métodos: Foram utilizados seis ratos Wistar. A lesão foi realizada na junção miotendínea do músculo gastrocnêmio direito, utilizando agulha de biópsia. Para avaliação funcional utilizou-se o teste de força de preensão e avaliação nociceptiva em três momentos: 24 horas antes da lesão (AV1); no segundo dia pós-lesão (AV2) e no sexto dia pós-lesão (AV3). Ao final do experimento o músculo foi retirado para análise morfológica. Resultados: Houve redução da força de preensão entre AV1 e AV2 (p=0,0031). Para a nocicepção os valores de AV2 (p<0,001) e AV3 (p=0,0109) foram maiores que AV1. Na região das fibras musculares observou-se desarranjo do padrão fascicular, aumento do calibre de vasos sanguíneos, da quantidade de tecido conjuntivo, de fibroblastos e de células inflamatórias. Conclusão: A lesão muscular por agulha de biópsia provocou redução da força de preensão e da nocicepção do membro lesado, como também evidenciou as etapas características do processo de degeneração muscular e início do processo de regeneração.

Introduction: Among the experimental models, the use of a biopsy needle was shown to be effective to reproduce lesions that occur with athletes. Objective: To evaluate the grip strength, nociception and morphological alterations of the gastrocnemius muscle of animals submitted to muscle injury. Methods: Six Wistar rats were used. The lesion was performed at the myotendinous junction of the right gastrocnemius muscle using a biopsy needle. For functional evaluation, the grip strength test and nociceptive evaluation were used at three moments: 24 hours before the injury (AV1); on the second postoperative day (AV2) and on the sixth postoperative day (AV3). At the end of the experiment the muscle was removed for morphological analysis. Results: There was a reduction in grip strength between AV1 and AV2 (p = 0.0031). For the nociception the values of AV2 (p < 0.001) and AV3 (p = 0.0109) were higher than AV1. In the muscle fibers region, there was disarrangement of the fascicular pattern, increased blood vessel size, the amount of connective tissue, fibroblasts and inflammatory cells. Conclusion: Muscle lesion by biopsy needle caused reduction of grip strength and nociception of the damaged limb, as well as the characteristic stages of the muscular degeneration process and the beginning of the regeneration process.

Analysis of the muscle tissue of Wistar rats submitted to the sciatic nerve compression model and cryotherapy.

OBJECTIVE: To evaluate the effects of right sciatic nerve compression and cryotherapy on muscle tissue. METHODS: We used 42 male Wistar rats, subdivided in the following Groups Control, Injury 3, Injury 8 and Injury 15 submitted to nerve compression and euthanized in the 3rd, 8th and 15th day after surgery. The Cryotherapy Injury 3 was entailed treatment with cryotherapy by immersion of the animal in recipient for 20 minutes during 1 day, then animals were euthanized at the 3rd day after surgery, and the Cryotherapy Injury 8 and the Cryotherapy Injury 15 was treated for 6 days, and euthanized at the 8th and 15th day after surgery. Functional evaluation was performed by the grasping strength of the right pelvic limb. The right tibialis anterior muscles were evaluated for mass, smaller diameter and cross-sectional area. In the Cryotherapy Injury 8 and the Cryotherapy Injury 15 groups, the hydroxyproline was dosed in the right soles. RESULTS: In the compression there was a significant difference in the Injury Groups compared with the Control Group (p<0.05). In the smaller diameter, the compression in Control Group was higher than Injury 8 (p=0.0094), Injury 15 (p=0.002) and Cryotherapy Injury 15 (p<0.001) groups. The comparison between groups with euthanasia in the same post-operative period, a significant difference (p=0.0363) was seen in day 8th after surgery, and this result in Cryotherapy Injury Group was greater than Injury Group. In the fiber area, Control Group was also higher than the Injury 8 (p=0.0018), the Injury 15 (p<0.001) and the Cryotherapy Injury 15 (p<0.001). In hydroxyproline, no significant difference was seen between groups. CONCLUSION: Nerve damage resulted in decreased muscle strength and trophism, the cryotherapy delayed hypotrophy, but this effect did not persist after cessation of treatment.

Neurotrophin expression and histomorphometric evaluation in Wistar rats subjected to neural mobilization after compression of the median nerve.

OBJECTIVE: To evaluate the neurotrophin mRNA expression and axon count in the median nerve of Wistar rats submitted to neural mobilization (NM) after nerve compression. METHODS: Eighteen animals were randomly divided into G1 (nerve compression only), G2 (NM for 1 min), and G3 (NM for 3 min). For NM, the animals were anesthetized and the right scapula received the mobilization, adapted as indicated for humans, on alternate days, from the third to the 13th postoperative (PO) day, totaling six days of therapy. On the 14th PO day, animals were anesthetized and euthanized. Fragments of the median nerve, distal to the compression procedure, were removed for histomorphometric analysis and expression of neurotrophins, nerve growth factor (NGF), and brain-derived neurotrophic factor (BDNF) by RT-PCR. RESULTS: Histomorphometric analysis revealed differences in the number of axons in the injured side, which was significantly lower in the injured limb nerve compared to the control limb, whereas the RT-PCR analysis showed no significant differences in the expression of NGF or BDNF. CONCLUSION: NM treatment did not affect median nerve regeneration, which maintained normal recovery rates.

OBJETIVO: Avaliar a expressão de RNAm de neurotrofinas e a contagem de axônios no nervo mediano de ratos Wistar submetidos à mobilização neural (MN) após compressão nervosa. MÉTODOS: Foram divididos aleatoriamente 18 animais em G1 (apenas compressão nervosa), G2 (MN por 1 minuto) e G3 (MN por 3 minutos). Para a MN, os animais foram anestesiados e o membro escapular direito recebeu a mobilização, adaptada da forma indicada para humanos, em dias alternados, do terceiro ao 13° dia de pós-operatório (PO), em seis dias de terapia. No 14° dia PO, os animais foram anestesiados e eutanasiados. Fragmentos do nervo mediano, distais ao procedimento de compressão, foram retirados para análise histomorfométrica e de expressão das neutrotrofinas, fator de crescimento do nervo (NGF) e fator de crescimento derivado do cérebro (BNDF) por RT-PCR. RESULTADOS: A análise histomorfométrica evidenciou diferenças no número de axônios nos lados lesionados, que foi significativamente menor no nervo do membro lesado comparado com o membro controle; por sua vez, a análise por RT-PCR não apontou diferenças significativas na expressão de NGF e nem de BNDF. CONCLUSÃO: O tratamento de MN não afetou a regeneração do nervo mediano, que manteve índices normais de recuperação.

Neurotrophin expression and histomorphometric evaluation in Wistar rats subjected to neural mobilization after compression of the median nerve / Expressão de neurotrofinas e avaliação histomorfométrica em ratos Wistar submetidos à mobilização neural após compressão do nervo mediano

ABSTRACT Objective: To evaluate the neurotrophin mRNA expression and axon count in the median nerve of Wistar rats submitted to neural mobilization (NM) after nerve compression. Methods: Eighteen animals were randomly divided into G1 (nerve compression only), G2 (NM for 1 min), and G3 (NM for 3 min). For NM, the animals were anesthetized and the right scapula received the mobilization, adapted as indicated for humans, on alternate days, from the third to the 13th postoperative (PO) day, totaling six days of therapy. On the 14th PO day, animals were anesthetized and euthanized. Fragments of the median nerve, distal to the compression procedure, were removed for histomorphometric analysis and expression of neurotrophins, nerve growth factor (NGF), and brain-derived neurotrophic factor (BDNF) by RT-PCR. Results: Histomorphometric analysis revealed differences in the number of axons in the injured side, which was significantly lower in the injured limb nerve compared to the control limb, whereas the RT-PCR analysis showed no significant differences in the expression of NGF or BDNF. Conclusion: NM treatment did not affect median nerve regeneration, which maintained normal recovery rates.

RESUMO Objetivo: Avaliar a expressão de RNAm de neurotrofinas e a contagem de axônios no nervo mediano de ratos Wistar submetidos à mobilização neural (MN) após compressão nervosa. Métodos: Foram divididos aleatoriamente 18 animais em G1 (apenas compressão nervosa), G2 (MN por 1 minuto) e G3 (MN por 3 minutos). Para a MN, os animais foram anestesiados e o membro escapular direito recebeu a mobilização, adaptada da forma indicada para humanos, em dias alternados, do terceiro ao 13° dia de pós-operatório (PO), em seis dias de terapia. No 14° dia PO, os animais foram anestesiados e eutanasiados. Fragmentos do nervo mediano, distais ao procedimento de compressão, foram retirados para análise histomorfométrica e de expressão das neutrotrofinas, fator de crescimento do nervo (NGF) e fator de crescimento derivado do cérebro (BNDF) por RT-PCR. Resultados: A análise histomorfométrica evidenciou diferenças no número de axônios nos lados lesionados, que foi significativamente menor no nervo do membro lesado comparado com o membro controle; por sua vez, a análise por RT-PCR não apontou diferenças significativas na expressão de NGF e nem de BNDF. Conclusão: O tratamento de MN não afetou a regeneração do nervo mediano, que manteve índices normais de recuperação.

Sericin and swimming on histomorphometric parameters of denervated plantar muscle in Wistar rats.

Objective To analyze the combined effects of the silk protein sericin and swimming exercise on histomorphometry of the plantar muscle in Wistar rats. Methods Forty adult rats were randomly allocated into 5 groups comprising 8 animals each, as follows: Control, Injury, Sericin, Swim, and Swim plus Sericin. Three days after crushing of the sciatic nerve the rats in the Swim and Swim plus Sericin Groups were submitted to swimming exercise for 21 days. Rats were then euthanized and the plantar muscle harvested and processed. Results Cross-sectional area, peripheral nuclei and muscle fiber counts, nucleus/fiber ratio and smallest muscle fiber width did not differ significantly between groups. Morphological analysis revealed hypertrophic fibers in the Swim Group and evident muscle damage in the Swim plus Sericin and Injury Groups. The percentage of intramuscular collagen was apparently maintained in the Swim Group compared to remaining groups. Conclusion Combined treatment with sericin and swimming exercise did not improve muscle properties. However, physical exercise alone was effective in maintaining intramuscular connective tissue and preventing progression of deleterious effects of peripheral nerve injury.

Association of sericin and swimming on the phenotype, motor plate, and functionality of the denervated plantar muscle of Wistar rats.

Physical exercise may help maintain muscle properties and functional improvement after peripheral nerve lesion, which may be enhanced by using biocompatible substances, such as sericin. The aim of this study was analyse the effect of sericin associated with swimming exercise on the phenotype, innervation, and functionality of the plantar muscle of Wistar rats. Forty randomly divided adult rats were used in five groups of eight animals: control, injury, sericin, exercise, exercise and sericin. The application of sericin was done on the spot, 100 µL, shortly after nerve compression. Three days after sciatic nerve compression, the swimming and swimming and sericin groups were submitted to physical swimming exercise for 21 days. Afterwards, the animals were euthanised and the plantar muscle was dissected and submitted to histochemical and histoenzymological techniques. The grip strength test did not show alterations in muscular functionality, and the control presented greater muscle mass in relation to the other groups, the same did not occur for muscle length. Polymorphic neuromuscular junctions were detected in the groups, although without significant morphometric alterations of the area, major and minor diameters. The percentage of type I fibres was lower in the lesion group and there was no difference in fibres IIa and IIb between groups. The area of fibres I, IIa and IIb remained constant between groups. Sericin biopolymer combined with swimming exercise did not affect plantar muscle function, submitted to experimental axonotmosis, whose contractile properties were altered by nerve injury.

Crioterapia em modelo de compressão do nervo isquiático: análise funcional e morfológica / Cryotherapy in sciatic nerve compression model: functional and morphological analysis / Crioterapia en modelo de compresión del nervio ciático: análisis funcional y morfológico

RESUMO Introdução: A crioterapia é uma modalidade terapêutica que visa reduzir processos álgicos e inflamatórios, sendo que a imersão é considerada a forma mais eficaz; no entanto, a literatura apresenta possíveis efeitos deletérios com relação à aplicação da crioterapia em nervos superficiais. Objetivo: Avaliar o efeito da crioterapia em modelo experimental de compressão do nervo isquiático em ratos Wistar, por meio de análise funcional e morfológica. Métodos: Foram utilizados 42 ratos, sendo seis animais por grupo: G1 - controle, submetido à eutanásia no 15º dia de pós-operatório (PO); G2, G3 e G4 - submetidos à compressão do nervo isquiático, submetidos à eutanásia no 3º, 8º e 15º dias de PO, respectivamente; G5, G6 e G7 - submetidos à compressão do nervo isquiático e tratados com crioterapia, submetidos à eutanásia no 3º, 8º e 15º dias de PO, respectivamente. As avaliações do índice funcional do isquiático (IFC) e do teste de incapacidade funcional aconteceram nos momentos pré-lesão, no 2º de PO e no dia da eutanásia em cada grupo com lesão. Após o período de intervenção, os animais foram devidamente anestesiados e o nervo isquiático distal ao procedimento de compressão foi dissecado e coletado para análise morfológica. A análise estatística foi realizada pelo teste de ANOVA mista, com nível de significância de 5%. Resultados: Houve diminuição do IFC após a lesão e o teste de incapacidade funcional mostrou aumento do tempo de elevação da pata. Com relação à análise morfológica, o G1 apresentou fibras nervosas com aspecto normal e nos grupos com lesão houve degeneração nervosa, sendo que o G6 teve uma discreta recuperação das fibras nervosas, além de leve regeneração no G4 e G7. Conclusão: A crioterapia não foi eficaz para recuperar os parâmetros funcionais analisados, entretanto, houve discreta melhora dos aspectos morfológicos do grupo submetido à eutanásia no 8º dia de PO. Nível de Evidência II; Estudos terapêuticos - Investigação dos resultados do tratamento.

ABSTRACT Introduction: Cryotherapy is a therapeutic modality that aims to reduce inflammatory and painful processes, with immersion being considered the most effective form; however, the literature has possible deleterious effects related to the application of cryotherapy to superficial nerves. Objective: To evaluate the effect of cryotherapy in experimental model of sciatic nerve compression in Wistar rats, through morphologic and functional analysis. Methods: Forty-two rats were used, six animals per group: G1 - control euthanized on the 15th postoperative day (PO); G2, G3 and G4 - submitted to sciatic nerve compression, euthanized at 3rd, 8th and 15th PO days, respectively; G5, G6 and G7 - submitted to sciatic nerve compression and treated with cryotherapy, euthanized at 3rd, 8th and 15th PO days, respectively. The assessments of sciatic functional index (SFI) and the functional disability test took place at the pre-injury, 2nd PO and on the day of euthanasia in each group with injury. After the intervention period, the animals were anesthetized properly and the sciatic nerve distal to the compression procedure was dissected and collected for morphological analysis. Statistical analysis was by the mixed ANOVA test with a significance level of 5%. Results: There was a decrease of SFI after injury and the functional disability test showed an increase in paw elevation time. Regarding the morphological analysis, the G1 showed normal nerve fibers and in the groups with lesion, there was nerve degeneration, G6 had a slight recovery of the nerve fibers, besides mild regeneration in G4 and G7. Conclusion: Cryotherapy was not effective to recover the functional parameters analyzed; however, there was a slight improvement in the morphological aspects of the group euthanized on the 8th PO day. Level of Evidence II; Therapeutic studies - Investigating the results of treatment.

RESUMEN Introducción: La crioterapia es una modalidad terapéutica que busca reducir procesos de dolor e inflamatorios, siendo que la inmersión es considerada la forma más eficaz; sin embargo, la literatura presenta posibles efectos deletéreos con relación a la aplicación de la crioterapia en nervios superficiales. Objetivo: Evaluar el efecto de la crioterapia en modelo experimental de compresión del nervio ciático en ratas Wistar, por medio de análisis funcional y morfológico. Métodos: Se utilizaron 42 ratas, siendo seis animales por grupo: G1 - control, sometido a la eutanasia en el 15º día de postoperatorio (PO); G2, G3 y G4 - sometidos a la compresión del nervio ciático, sometidos a la eutanasia en los 3º, 8º y 15º días de PO, respectivamente; G5, G6 y G7 - sometidos a la compresión del nervio ciático y tratados con crioterapia, sometidos a la eutanasia en los 3º, 8º y 15º días de PO, respectivamente. Las evaluaciones del índice funcional del ciático (IFC) y la prueba de incapacidad funcional ocurrieron en los momentos pre-lesión, en el 2º día de PO y el día de la eutanasia en cada grupo con lesión. Después del período de intervención, los animales fueron debidamente anestesiados y el nervio ciático distal al procedimiento de compresión fue disecado y recogido para análisis morfológico. El análisis estadístico fue realizado por la prueba de ANOVA mixta, con un nivel de significancia del 5%. Resultados: Hubo disminución del IFC después de la lesión y la prueba de incapacidad funcional mostró aumento del tiempo de elevación de la pata. En cuanto al análisis morfológico, el G1 presentó fibras nerviosas con aspecto normal y en los grupos lesionados hubo degeneración nerviosa, siendo que el G6 tuvo una discreta recuperación de las fibras nerviosas, además de ligera regeneración en el G4 y G7. Conclusión: La crioterapia no fue eficaz para recuperar los parámetros funcionales analizados, sin embargo, hubo discreta mejora de los aspectos morfológicos del grupo sometido a la eutanasia en el 8º día de PO. Nivel de Evidencia II; Estudios terapéuticos - Investigación de los resultados del tratamento.

Analysis of the muscle tissue of Wistar rats submitted to the sciatic nerve compression model and cryotherapy / Análise do tecido muscular de ratos Wistar submetidos ao modelo de compressão do nervo isquiático e à crioterapia

ABSTRACT Objective: To evaluate the effects of right sciatic nerve compression and cryotherapy on muscle tissue. Methods: We used 42 male Wistar rats, subdivided in the following Groups Control, Injury 3, Injury 8 and Injury 15 submitted to nerve compression and euthanized in the 3rd, 8th and 15th day after surgery. The Cryotherapy Injury 3 was entailed treatment with cryotherapy by immersion of the animal in recipient for 20 minutes during 1 day, then animals were euthanized at the 3rd day after surgery, and the Cryotherapy Injury 8 and the Cryotherapy Injury 15 was treated for 6 days, and euthanized at the 8th and 15th day after surgery. Functional evaluation was performed by the grasping strength of the right pelvic limb. The right tibialis anterior muscles were evaluated for mass, smaller diameter and cross-sectional area. In the Cryotherapy Injury 8 and the Cryotherapy Injury 15 groups, the hydroxyproline was dosed in the right soles. Results: In the compression there was a significant difference in the Injury Groups compared with the Control Group (p<0.05). In the smaller diameter, the compression in Control Group was higher than Injury 8 (p=0.0094), Injury 15 (p=0.002) and Cryotherapy Injury 15 (p<0.001) groups. The comparison between groups with euthanasia in the same post-operative period, a significant difference (p=0.0363) was seen in day 8th after surgery, and this result in Cryotherapy Injury Group was greater than Injury Group. In the fiber area, Control Group was also higher than the Injury 8 (p=0.0018), the Injury 15 (p<0.001) and the Cryotherapy Injury 15 (p<0.001). In hydroxyproline, no significant difference was seen between groups. Conclusion: Nerve damage resulted in decreased muscle strength and trophism, the cryotherapy delayed hypotrophy, but this effect did not persist after cessation of treatment.

RESUMO Objetivo: Avaliar os efeitos da compressão nervosa do isquiático direito e da crioterapia no tecido muscular. Métodos: Foram utilizados 42 ratos Wistar machos, subdivididos nos Grupos Controle, Lesão 3, Lesão 8 e Lesão 15, submetidos a compressão nervosa e eutanasiados, respectivamente, no 3°, 8° e 15° dias pós-operatório; Lesão Crioterapia 3, tratado com crioterapia, por imersão durante 20 minutos, por 1 dia, e eutanasiados no 3° dia pós-operatório; e Lesão Crioterapia 8 e Lesão Crioterapia 15, tratados durante 6 dias e eutanasiados no 8° e 15° dias pós-operatório. A avaliação funcional foi realizada pela força de preensão do membro pélvico direito. Os músculos tibiais anteriores direitos foram avaliados quanto a massa, menor diâmetro e área de secção transversa. Em Lesão Crioterapia 8 e Lesão Crioterapia 15, foi dosada a hidroxiprolina nos sóleos direitos. Resultados: Na preensão, houve diferença significativa nos Grupos Lesão quando comparados ao Grupo Controle (p<0,05). No menor diâmetro, o Grupo Controle foi maior que Lesão 8 (p=0,0094), Lesão 15 (p = 0,002) e Lesão Crioterapia 15 (p<0,001). Na comparação entre os grupos com eutanásia no mesmo pós-operatório, houve diferença significativa (p=0,0363) no 8° pós-operatório, sendo Lesão Crioterapia maior que Lesão. Na área das fibras, o Grupo Controle também foi maior que Lesão 8 (p=0,0018), Lesão 15 (p<0,001) e Lesão Crioterapia 15 (p<0,001). Na hidroxiprolina, não houve diferença significativa entre os grupos. Conclusão: A lesão nervosa resultou na diminuição da força e em trofismo muscular, e a crioterapia retardou a hipotrofia, porém este efeito não se manteve após o tratamento cessar.

Sericin and swimming on histomorphometric parameters of denervated plantar muscle in Wistar rats / Sericina e natação sobre parâmetros histomorfométricos de músculo plantar desnervado de ratos Wistar

Abstract Objective To analyze the combined effects of the silk protein sericin and swimming exercise on histomorphometry of the plantar muscle in Wistar rats. Methods Forty adult rats were randomly allocated into 5 groups comprising 8 animals each, as follows: Control, Injury, Sericin, Swim, and Swim plus Sericin. Three days after crushing of the sciatic nerve the rats in the Swim and Swim plus Sericin Groups were submitted to swimming exercise for 21 days. Rats were then euthanized and the plantar muscle harvested and processed. Results Cross-sectional area, peripheral nuclei and muscle fiber counts, nucleus/fiber ratio and smallest muscle fiber width did not differ significantly between groups. Morphological analysis revealed hypertrophic fibers in the Swim Group and evident muscle damage in the Swim plus Sericin and Injury Groups. The percentage of intramuscular collagen was apparently maintained in the Swim Group compared to remaining groups. Conclusion Combined treatment with sericin and swimming exercise did not improve muscle properties. However, physical exercise alone was effective in maintaining intramuscular connective tissue and preventing progression of deleterious effects of peripheral nerve injury.

RESUMO Objetivo Analisar o efeito da proteína sericina associada ao exercício físico de natação na histomorfometria do músculo plantar de ratos Wistar. Métodos Foram utilizados 40 ratos adultos divididos aleatoriamente em 5 grupos, com 8 animais cada: Controle, Lesão, Sericina, Natação, Natação e Sericina. Três dias após a compressão do nervo isquiático, os Grupos Natação e Exercício e Sericina foram submetidos ao exercício físico de natação durante 21 dias. Após, os animais foram sacrificados, e o músculo plantar foi processado. Resultados Não houve diferença da área da secção transversa entre os grupos, quantidade de núcleos periféricos, quantidade de fibra, relação núcleo/fibra e diâmetro menor. A análise morfológica revelou que no Grupo Natação ocorreu hipertrofia das fibras, assim como nos Grupos Exercício e Sericina e Lesão, o dano muscular foi evidente. O percentual de conjuntivo intramuscular parece ter sido mantido no Grupo Exercício em relação aos demais grupos. Conclusão A associação da proteína sericina e exercício físico de natação não foi eficiente na melhora das propriedades musculares, embora a aplicação do exercício físico tenha sido eficiente na manutenção do conjuntivo intramuscular, e no não agravamento dos efeitos deletérios consequentes da lesão nervosa periférica.

Changes in the soleus muscular tissue of rats with experimental periodontitis under physical exercise influences / Cambios en el tejido muscular del sóleo de las ratas con periodontitis experimental bajo influencia del ejercicio físico

Until the moment, no study explored conjunctively the physical activity relation, using the inflammatory biomarkers, with the periodontitis. This way, the objective of this experiment was to evaluate the muscular tissue behavior of rats submitted to physical exercise in aquatic environment with experimental disease. Twenty-four male Wistar rats were divided in 4 groups: 1) control and sedentary (CS); 2) control and active (CA); 3) with the periodontal disease and sedentary (PDS); with the periodontal disease and active (PDA). On the group that the periodontitis was induced, it was for ligature and the groups with active swimming activity have practiced it for 4 weeks. At the end of 30 days the animals were euthanized and a portion of the gingival tissue and the soleus muscle were removed and underwent analysis by ELISA and morphological and morphometrical analysis of the muscle. Data drawn from the analysis was analyzed through ANOVA and Tukey. Results have shown that there is a higher expression of TNF-α in the gingival tissue and on the muscular tissue of the rats that underwent the induced periodontitis independently of the physical activity (PDS and PDA), as a meaningful decrease on the conjunctive tissue on the groups with induced periodontitis, that have or have not undergone active swimming activity which could suggest a predisposition to muscular injury or difficulty of muscular recovering on these groups. Therefore, it was possible to highlight a correlation between the periodontal disease and the muscle morphological changes, and, moreover, the physical swimming activity promoting an acceleration of the regeneration of the muscle tissue

Hasta este momento, ningún estudio ha explorado conjuntamente la relación de la actividad física con la periodontitis utilizando biomarcadores de inflamación. Siendo así, el objetivo de este estudio fue evaluar el comportamiento del tejido muscular de ratas con periodontitis experimental al realizar ejercicio físico en medio acuático. Veinticuatro ratas Wistar machos fueron divididas en cuatro grupos: 1) control y sedentario (CS); 2) control y ejercicio (CA); 3) con enfermedad periodontal y sedentario (PDS); 4) con enfermedad periodontal y ejercicio (PDA). En los grupos con periodontitis, la enfermedad periodontal fue inducida por ligadura y los grupos con ejercicio realizaron natación durante cuatro semanas. A los treinta días, los animales fueron sacrificados y una parte del tejido de las encías y del músculo soleo se resecaron y utilizaron para análisis con ELISA y para análisis morfológicos y morfométricos. Los datos obtenidos fueron analizados y evaluados a través de los tests ANOVA y TUKEY. Los datos mostraron una mayor expresión de TNF-α tanto en el tejido de las encías como en el tejido muscular de los ratones sometidos a periodontitis inducida independiente del ejercicio físico (PDS y PDA). Se percibió también una disminución significativa en el tejido conjuntivo en los grupos con periodontitis inducida sometidos o no al ejercicio de natación, lo que podría sugerir una predisposición a lesión muscular o una dificultad en la reparación de las lesiones musculares de esos grupos. Por lo tanto, fue posible destacar una correlación entre la enfermedad periodontal y los cambios morfológicos musculares y, además, que la actividad física de natación favoreció una aceleración de la regeneración del tejido muscular

Action of vanillin (Vanilla planifolia) on the morphology of tibialis anterior and soleus muscles after nerve injury.

OBJECTIVE: To evaluate the action of vanillin (Vanilla planifolia) on the morphology of tibialis anterior and soleus muscles after peripheral nerve injury. METHODS: Wistar rats were divided into four groups, with seven animals each: Control Group, Vanillin Group, Injury Group, and Injury + Vanillin Group. The Injury Group and the Injury + Vanillin Group animals were submitted to nerve injury by compression of the sciatic nerve; the Vanillin Group and Injury + Vanillin Group, were treated daily with oral doses of vanillin (150mg/kg) from the 3rd to the 21st day after induction of nerve injury. At the end of the experiment, the tibialis anterior and soleus muscles were dissected and processed for light microscopy and submitted to morphological analysis. RESULTS: The nerve compression promoted morphological changes, typical of denervation, and the treatment with vanillin was responsible for different responses in the studied muscles. For the tibialis anterior, there was an increase in the number of satellite cells, central nuclei and fiber atrophy, as well as fascicular disorganization. In the soleus, only increased vascularization was observed, with no exacerbation of the morphological alterations in the fibers. CONCLUSION: The treatment with vanillin promoted increase in intramuscular vascularization for the muscles studied, with pro-inflammatory potential for tibialis anterior, but not for soleus muscle. OBJETIVO: Avaliar a ação da vanilina (Vanilla planifolia) sobre a morfologia dos músculos tibial anterior e sóleo após lesão nervosa periférica. MÉTODOS: Ratos Wistar foram divididos em quatro grupos, com sete animais cada, sendo Grupo Controle, Grupo Vanilina, Grupo Lesão e Grupo Lesão + Vanilina. Os animais dos Grupos Lesão e Grupo Lesão + Vanilina foram submetidos à lesão nervosa por meio da compressão do nervo isquiático, e os Grupos Vanilina e Grupo Lesão + Vanilina foram tratados diariamente com doses orais de vanilina (150mg/kg) do 3o ao 21o dia após a indução da lesão nervosa. Ao término do experimento, os músculos tibial anterior e sóleo foram dissecados e seguiram o processamento de rotina em microscopia de luz, para posterior análise morfológica. RESULTADOS: A compressão nervosa promoveu alterações morfológicas características de denervação, sendo que o tratamento com vanilina foi responsável por respostas distintas nos músculos estudados. Para o tibial anterior, houve aumento do número de células satélites, núcleos centrais e atrofia das fibras, bem como desorganização fascicular. Já no sóleo, houve apenas aumento da vascularização, sem exacerbação das alterações morfológicas nas fibras. CONCLUSÃO: O tratamento com vanilina promoveu o aumento da vascularização intramuscular para os músculos estudados, com potencial pró-inflamatório para o tibial anterior, o que não ocorreu no músculo sóleo.

Experimental Periodontitis in the Potentialization of the Effects of Immobilism in the Skeletal Striated Muscle.

This study aims to evaluate if ligature-induced periodontitis can potentiates the deleterious effects of immobilization in the skeletal striated muscle, contributing to the development of muscle atrophy due to disuse. Forty Wistar rats were divided into four groups: (1) Control Group (CG), (2) Periodontal Disease (PDG), (3) Immobilized (IG), and (4) Immobilized with Periodontal Disease (IPDG). Periodontal disease was induced for 30 days, with ligature method, and the immobilization was performed with cast bandage for 15 days. Prior to euthanasia, nociceptive threshold and muscular grasping force were evaluated. Afterwards, the soleus muscle was dissected and processed for sarcomere counting and morphological/morphometric analysis. For data analysis, was used the one-way ANOVA and post-test Tukey (p < 0.05). The IG and IPDG presented lower muscle weight, lower muscular grip strength, and less number of sarcomeres compared to CG. The PDG showed reduction of muscle strength and nociceptive threshold after 15 days of periodontal disease and increased connective tissue compared to CG. The IPDG presented lower muscle length and nociceptive threshold. The IG presented reduction in cross-sectional area and smaller diameter, increase in the number of nuclei and a nucleus/fiber ratio, decrease in the number of capillaries and capillary/fiber ratio, with increase in connective tissue. The IPDG had increased nucleus/fiber ratio, decreased capillaries, and increased connective tissue when compared to the IG. The IPDG presented greater muscle tissue degeneration and increased inflammatory cells compared to the other groups. Ligature-induced periodontitis potentiated the deleterious effects of immobilization of the skeletal striated muscle.