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Made available in DSpace on 2022-04-28T19:28:07Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-04-01 Objective: Acupuncture (AC) has been used for treatment of chronic and pre- operative anxiety. However, no study has evaluated the effect of AC on exam anxiety in undergraduate students. The aim of this study was to evaluate whether AC reduces pre-examination anxiety compared to placebo (PL) and non- intervention (NI). Methods: Twenty students from the first terms of physiotherapy and dentistry courses were randomized to receive AC, PL and NI in a crossover manner and subsequently completed three anatomy exams with an interval of 1 month between exams/interventions. AC was performed on acupoints PC-6 (Neiguan), HT-7 (Shenmen), LV-3 (Taichong) and Yintang one day prior to each exam. The PL was performed outside the meridians and the needles were inserted superficially without obtaining De Qi. Levels of anxiety were measured using a visual analogue scale before and after each intervention as well as before each exam. Results: Anxiety levels were reduced after AC and PL intervention compared to NI on the day before the exam, as well as on the day of the exam. Conclusion: These results suggest that AC can be used as an anxiolytic method in students under conditions of situational stress. Biomedical Bachelor Master at the State University of São Paulo Julio Mesquita Filho in General and Applied Biology Biomedical Bachelor Master at the State University of São Paulo Julio Mesquita Filho in General and Applied Biology

Cases of tendinopathy and tendon ruptures have been reported as side effects associated with statin therapy. This work assessed possible changes in the structural and biomechanical properties of the tendons after chronic treatment with statins. Wistar rats were divided into the following groups: treated with atorvastatin (A-20 and A-80), simvastatin (S-20 and S-80) and the group that received no treatment (C). The doses of statins were calculated using allometric scaling, based on the doses of 80 mg/day and 20 mg/day recommended for humans. The morphological aspect of the tendons in A-20, S-20 and S-80 presented signals consistent with degeneration. Both the groups A-80 and S-80 showed a less pronounced metachromasia in the compression region of the tendons. Measurements of birefringence showed that A-20, A-80 and S-80 groups had a lower degree of organization of the collagen fibers. In all of the groups treated with statins, the thickness of the epitenon was thinner when compared to the C group. In the biomechanical tests the tendons of the groups A-20, A-80 and S-20 were less resistant to rupture. Therefore, statins affected the organization of the collagen fibers and decreased the biomechanical strength of the tendons, making them more predisposed to ruptures.

Green tea (GT) is widely used due to its anti-inflammatory properties. Previous studies have shown beneficial effects of a glycine diet on the remodeling process in inflamed tendons. Tendinitis is commonly observed in athletes and is of concern to surgeons due to the slowness of the recovery process. Our hypothesis is that GT + a glycine diet may improve tendinitis.To analyze the effect of GT and/or glycine in the diet on tendinitis.Wistar rats were divided into seven groups (G): control group (C); G1 and G4, tendinitis; G2 and G5, tendinitis supplied with GT; and G3 and G6, tendinitis supplied with GT and a glycine diet for 7 or 21 days, respectively. We performed zymography for metalloproteinase, biochemical, morphological and biomechanics tests.G2, G3 and G5 showed high levels of hydroxyproline in relation to G1, while G4 showed high levels of glycosaminoglycans. High activity of metalloproteinase-2 was detected in G3. The organization of collagen bundles was better in G2 and G3. G5 showed similar birefringence measurements compared with C. G5 withstood a larger load compared with G4.The presence of metalloproteinase-2 indicates that a tissue is undergoing a remodeling process. High birefringence suggests a better organization of collagen bundles. After 21 days, G5 sustained a high load before rupture, unlike G4. The results suggest that GT + a glycine diet has beneficial effects that aid in the recovery process of the tendon after tendinitis.

Tendinopathy of the Achilles tendon is a clinical problem that motivates the scientific community to search for treatments that assist in restoring its functional properties. Glycine has broad biological effects, acting as a modulator of the inflammatory cascade, and is the predominant amino acid in collagen. A 5% glycine diet provided beneficial effects against toxicity and inflammation since glycine may restructure the collagen molecules faster due to its broad anti-inflammatory effects. The purpose was analyze the effects of a 5% glycine diet in rats as a treatment for the inflammatory process. The experimental groups were as follows: C (control group), G1 and G3 (inflammatory group), and G2 and G4 (glycine+inflammatory group). G1 and G2 were euthanized 8 days following injury, and G3 and G4 were euthanized 22 days following injury. The concentrations of hydroxyproline, non-collagenous proteins, and glycosaminoglycans, as well as the activity of MMP-2 and -9 were analyzed. Biomechanical and morphological tests were employed. Higher concentrations of hydroxyproline and glycosaminoglycans were found in G4 and an increased activity of MMP-2 was found in G2. Higher birefringence was noted in group G2. The biomechanical results indicated that the tendon was more resistant to loading to rupture upon treatment with a glycine diet in group G4. Glycine induced the synthesis of important components of the tendon. A rapid remodeling was noted when compared with the inflamed-only groups. These data suggest that glycine may be a beneficial supplement for individuals with inflammation of the Achilles tendon.

A previous study demonstrated that acupuncture increases the synthesis and reorganisation of collagen molecules in rat tendons after injury. Clinical studies have shown that acupuncture improves pain and functional activity in patients with tendinopathy. However, the molecular mechanisms underlying these effects are unknown. Recent studies have shown that acupuncture can modulate both anti-inflammatory (AI) and mechanotransduction (MT) molecular pathways. Moreover, the modulation of these pathways can increase type I collagen synthesis, which is the main factor that influences tendon biomechanical properties. Our hypothesis is that acupuncture increases synthesis and subsequent reorganisation of type I collagen during tendon healing by concomitant modulation of the Toll-like receptor-nuclear factor-κB AI pathway, the mitogen-activated protein kinases pathway and the Rho/Rac-F-actin MT pathway. Increased collagen synthesis and reorganisation requires that at least one acupoint is anatomically connected with the site of the injury because of the local tenoblast MT mechanism. Confirmation of this hypothesis will increase the knowledge of acupuncture modulation of the previously mentioned molecular pathways, and such confirmation may also help to establish the relationships between the different types of acupuncture needle stimulation and the influence of acupuncture stimuli on pathway activity levels. In addition, the downstream therapeutic effects of acupuncture therapy may be established. This hypothesis can be verified in a rat tendon healing model, and subsequent clinical protocols for tendon healing can be developed and evaluated as standalone therapies or as a component of a combination therapy.

Statins have been widely prescribed as lipid-lowering drugs and are associated with tendon rupture. Therefore, this study aimed to evaluate the possible biochemical changes in the Achilles tendon of rats after chronic treatment with statins. Dosages of statins were calculated using allometric scaling with reference to the 80mg/day and 20mg/day, doses recommended for humans. The rats were divided into the following groups: treated with simvastatin (S-20 and S-80), treated with atorvastatin (A-20 and A-80), and the control group that received no treatment (C). Measurements of low-density lipoprotein (LDL) in the plasma were performed. The levels of non-collagenous proteins, glycosaminoglycans (GAGs) and hydroxyproline were quantified. Western blotting for collagen I was performed, and the presence of metalloproteinases (MMPs)-2 and -9 was investigated through zymography. The concentration of non-collagenous proteins in S-20 was less than the C group. There was a significant increase in pro-MMP-2 activity in A-80 group and in active MMP-2 in S-20 group compared to the C group. A significant increase in latent MMP-9 activity was observed in both the A-80 and S-20 groups when compared to C group. In the A-20 group, there was a lower amount of collagen I in relation to C group. In addition, a higher concentration of hydroxyproline was found in the S-20 group than the C group. The analysis of GAGs showed a significant increase in the A-20 group when compared to C group. The treatment induced remarkable alterations in the Achilles tendon and the response of the tissue seems to depend of the used statin dosage. The presence of MMP-2 and MMP-9 is evidence of the degradation and remodeling processes in the extracellular matrix of the tendons. Our results show that statins induce imbalance of extracellular matrix components and possibly induce microdamage in tendons.

In the last decades, the tendon injuries have increased substantially. Previous results suggested that low-level laser treatment (LLLT) promotes synthesis of extracellular matrix and improves the functional properties of the tendon. The aim of this study was to evaluate the effects of different protocols of LLLT on partially tenotomized tendons. Adult male rats were divided into the following: G1-intact, G2-injured, G3-injured + LLLT (4 J/cm(2) continuous), G4-injured + LLLT (4 J/cm(2) at 20 Hz). G2, G3, and G4 were euthanized 8 days after injury. G5-injured, G6-injured + LLLT (4 J/cm(2) continuous), and G7-injured + LLL (4 J/cm(2) at 20 Hz until the seventh day and 2 kHz from 8 to 14 days). G5, G6, and G7 were euthanized on the 15th day. Glycosaminoglycan (GAG) level was quantified by dimethylmethylene blue method and analyzed on agarose gel. Toluidine blue (TB) stain was used to observe metachromasy. CatWalk system was used to evaluate gait recovery. Collagen organization was analyzed by polarization microscopy. The GAG level increased in all transected groups, except G5. In G6 and G7, there was a significant increase in GAG in relation to G5. In G3 and G4, the presence of dermatan sulfate band was more prominent than G2. TB stains showed intense metachromasy in the treated groups. Birefringence analysis showed improvement in collagen organization in G7. The gait was significantly improved in G7. In conclusion, pulsed LLLT leads to increased organization of collagen bundles and improved gait recovery.

Tendinopathy is a pathology found mainly in the rotator cuff, patellar, Achilles and flexor tendons. Tendinopathy is a significant impediment to performance in athletes and in workers in the labor market. Some studies have indicated that inflammation in adjacent tissues may affect the rotator cuff and Achilles tendon. In this study alterations were verified in the extracellular matrix (ECM) of the deep digital flexor tendon after two periods (12 and 24 hr) of induction inflammation in rat paw. Wistar rats were divided into three groups: those that received injection of 1% carrageenan; those that received 0.9% NaCl; and those that received no application. The tendon was divided into distal (d), proximal (p), and intermediate (i) regions. Biochemical analyses were performed and included non-collagenous proteins (NCP), glycosaminoglycans (GAGs), hydroxyproline (HoPro) and metalloproteinases 2 and 9. Tissue sections were stained with toluidine blue, hematoxylin-eosin, and Ponceau SS and observed under polarization microscopy. Remarkable results were detected that included the presence of MMP-9, degradation of NCP and GAG and the presence of cellular infiltrate closer to digits in d region. The different concentrations of HoPro, as well as alterations in the organization of the collagen fibers showed the collagenous matrix undergoing some alterations. The results indicated that the induced inflammation in rat paw exhibited characteristics similar to the typical acute inflammatory process observed in tendons. Anat Rec, 2013. © 2013 Wiley Periodicals, Inc.

The Achilles tendon has a high incidence of rupture, and the healing process leads to a disorganized extracellular matrix (ECM) with a high rate of injury recurrence. To evaluate the effects of different conditions of low-level laser (LLL) application on partially tenotomized tendons, adult male rats were divided into the following groups: G1, intact; G2, injured; G3, injured + LLL therapy (LLLT; 4 J/cm(2) continuous); G4, injured + LLLT (4 J/cm(2), 20 Hz); G5, injured; G6, injured + LLLT (4 J/cm(2) continuous); and G7, injured + LLLT (4 J/cm(2), 20 Hz until the 7th day and 2 kHz from 8 to 14 days). G2, G3, and G4 were euthanized 8 days after injury, and G5, G6, and G7 were euthanized on the 15th day. The quantification of hydroxyproline (HOPro) and non-collagenous protein (NCP), zymography for matrix metalloproteinase (MMP)-2 and MMP-9, and Western blotting (WB) for collagen types I and III were performed. HOPro levels showed a significant decrease in all groups (except G7) when compared with G1. The NCP level increased in all transected groups. WB for collagen type I showed an increase in G4 and G7. For collagen type III, G4 presented a higher value than G2. Zymography for MMP-2 indicated high values in G4 and G7. MMP-9 increased in both treatment groups euthanized at 8 days, especially in G4. Our results indicate that the pulsed LLLT improved the remodeling of the ECM during the healing process in tendons through activation of MMP-2 and stimulation of collagen synthesis.

The aim of this study was to investigate the effect of electroacupuncture (EA) on the composition and organization of the extracellular matrix of the rat Achilles tendon after a partial transection during the proliferative phase of healing. Wistar rats were divided into three groups: rats that were not tenotomized (G1), tenotomized rats (G2), and rats that were tenotomized and submitted to EA (G3). EA was applied 15 days after injury at the ST36 and BL57 acupoints for 20 min, three times per week on alternate days for a total of six sessions. Biochemical analyses were performed using non-collagenous proteins, glycosaminoglycans, and hydroxyproline quantifications. An analysis of metalloproteinase-2 was carried out by zymography. The general organization of the extracellular matrix and the metachromasy of the tendons were analyzed under light microscopy. The organization of the bundles of collagen fibers was analyzed by birefringence analysis. The results showed that EA did not alter the concentration of non-collagenous proteins or glycosaminoglycans or the enzymatic activity of metalloproteinase-2 in the transected tendons. However, the concentration of hydroxyproline was significantly increased when these tendons were treated by EA. The analysis of birefringence showed a higher organization of collagen fibers in the group treated by EA. These results indicate, for the first time, that EA may offer therapeutic benefits for the treatment of tendon injuries by increasing the concentration of collagen and by inducing a better molecular organization of the collagen fibers, which may improve the mechanical strength of the tendon after injury.

OBJETIVO: Analisar as características de tendões de Aquiles de ratos após indução de processo inflamatório localizado na pata. MÉTODOS: Foram utilizados três grupos experimentais: grupo inflamado com carragenina na pata de rato (G1); grupo salina (G2) e grupo controle (G3). Após 4 horas os animais foram eutanaziados e o tendão de Aquiles foi removido. RESULTADOS:Não foram observadas diferenças significativas nas análises de proteínas não colagênicas, glicosaminoglicanos e hidroxiprolina, mas uma tendência a diminuição foi verificada em G1. Em organização de moléculas de colágeno não foram observadas diferenças entre os grupos. Com respeito à atividade de MMPs, foi observada uma presença maior da isoforma ativa da MMP-2 em G1, sugerindo que a remodelação do tecido está ocorrendo. CONCLUSÃO: Desta forma, nós concluímos que o processo inflamatório desencadeado em pata de rato pode afetar o remodelamento de tendões situados próximo ao local inflamado. Nível de Evidência I, Estudos Prognósticos - Investigação do Efeito de Característica de um Paciente Sobre o Desfecho da Doença. OBJECTIVE: To analyze the characteristics of the Achilles tendon of rats after induction of localized inflammation in the rat paw. METHODS: In our study three groups were used: inflamed group with carrageenan in rat paw (G1); saline group (G2) and control group (G3). After 4 hours the animals were euthanized and the Achilles tendon removed. RESULTS: No significant differences were observed in the analysis of non-collagenous proteins, glycosaminoglycans and hydroxyproline in the groups but a tendency of reduction was verified in G1. About the organization of collagen molecules, no differences were observed between groups. With respect to MMPs activity, a stronger presence of the active isoform of MMP-2 in G1 was observed, suggesting that the remodeling was occurring. CONCLUSION: Thus, we conclude that the inflammatory process in rat paw may affect the remodeling of tendons located near the inflamed site. Level of Evidence I, Prognostic Studies - Investigating the Effect of a Patient Characteristic on the Outcome of Disease.

Tendinopathy of the Achilles tendon is a clinical problem that motivates the scientific community to search for treatments that assist in restoring its functional properties. Glycine has broad biological effects, acting as a modulator of the inflammatory cascade, and is the predominant amino acid in collagen. A 5% glycine diet provided beneficial effects against toxicity and inflammation since glycine may restructure the collagen molecules faster due to its broad anti-inflammatory effects. The purpose was analyze the effects of a 5% glycine diet in rats as a treatment for the inflammatory process. The experimental groups were as follows: C (control group), G1 and G3 (inflammatory group), and G2 and G4 (glycine+inflammatory group). G1 and G2 were euthanized 8 days following injury, and G3 and G4 were euthanized 22 days following injury. The concentrations of hydroxyproline, non-collagenous proteins, and glycosaminoglycans, as well as the activity of MMP-2 and -9 were analyzed. Biomechanical and morphological tests were employed. Higher concentrations of hydroxyproline and glycosaminoglycans were found in G4 and an increased activity of MMP-2 was found in G2. Higher birefringence was noted in group G2. The biomechanical results indicated that the tendon was more resistant to loading to rupture upon treatment with a glycine diet in group G4. Glycine induced the synthesis of important components of the tendon. A rapid remodeling was noted when compared with the inflamed-only groups. These data suggest that glycine may be a beneficial supplement for individuals with inflammation of the Achilles tendon.

Tendinopathy is a pathology found mainly in the rotator cuff, patellar, Achilles and flexor tendons. Tendinopathy is a significant impediment to performance in athletes and in workers in the labor market. Some studies have indicated that inflammation in adjacent tissues may affect the rotator cuff and Achilles tendon. In this study alterations were verified in the extracellular matrix (ECM) of the deep digital flexor tendon after two periods (12 and 24 hr) of induction inflammation in rat paw. Wistar rats were divided into three groups: those that received injection of 1% carrageenan; those that received 0.9% NaCl; and those that received no application. The tendon was divided into distal (d), proximal (p), and intermediate (i) regions. Biochemical analyses were performed and included non-collagenous proteins (NCP), glycosaminoglycans (GAGs), hydroxyproline (HoPro) and metalloproteinases 2 and 9. Tissue sections were stained with toluidine blue, hematoxylin-eosin, and Ponceau SS and observed under polarization microscopy. Remarkable results were detected that included the presence of MMP-9, degradation of NCP and GAG and the presence of cellular infiltrate closer to digits in d region. The different concentrations of HoPro, as well as alterations in the organization of the collagen fibers showed the collagenous matrix undergoing some alterations. The results indicated that the induced inflammation in rat paw exhibited characteristics similar to the typical acute inflammatory process observed in tendons.

The tendon is commonly affected by inflammation, and in such situations, the tissue undergoes a process of reorganization of the extracellular matrix to improve and regenerate the affected region. Little is known about the mechanisms that trigger inflammation in the tissues surrounding the affected area. The objective of this study was to biochemically and morphologically analyze the deep digital flexor tendon at the peak of acute inflammation in the rat paw. Wistar rats were divided into the following three groups: those that received injection of 1% carrageenan, those that received 0.9% NaCl, and those that received nothing. The deep digital flexor tendon was divided into the distal, proximal, and intermediate regions. For biochemical analysis, the tendons were treated with guanidine hydrochloride and analyzed by sodium dodecyl sulfate-polyacrilamide gel electrophoresis. Proteins, glycosaminoglycans (GAGs), and hydroxyproline were quantified, and metalloproteinases were analyzed. The GAGs were analyzed by agarose gel electrophoresis. Tissue sections were stained with hematoxylin-eosin, toluidine blue, and Ponceau SS. The content of proteins and GAGs was smaller in the group receiving the application of carrageenan. The concentration of hydroxyproline in the two tendon regions that respond to tension forces was higher in the inflammation group. The metalloproteinase-9 was detected in the distal region, and a thicker epitenon with cellular infiltrate was observed in the groups with inflamed paws. Meanwhile, a better organization of collagen bundles was observed in the two tension regions of that same group. Our results show that although the tendon was not directly inflamed, changes in the surrounding structural and biochemical parameters were observed.

Orientador: Edson Rosa Pimentel Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia Resumo: A eletroacupuntura (EA) tem sido utilizada amplamente para o tratamento de uma variedade de doenças inflamatórias e para o alivio da dor. No entanto, não há relatos na literatura sobre o efeito da EA na composição e organização da matriz extracelular (MEC) de tendões em processo de cicatrização. Objetivo: Investigar através de análises bioquímicas e morfológicas, o efeito da EA durante o pico de síntese dos componentes da MEC, 15 dias após transecção parcial do tendão calcâneo de ratos. Materiais e Métodos: Ratos Wistar machos com 60 dias de idade foram divididos em 3 grupos: Não tenotomizados (G1), tenotomizados (G2) e tenotomizados e submetidos a EA (G3). Agulhas de acupuntura (AC) foram inseridas nos pontos E-36 e B-57 e uma corrente elétrica com onda farádica bipolar assimétrica na freqüência de 2 Hz e intensidade de 2-4 Volts foi aplicada nas agulhas por 20 minutos, 3 vezes por semana em dias alternados, em um total de 6 sessões. Análises bioquímicas foram feitas através da SDS-PAGE, dosagem de proteínas não colagênicas (PNCs), glicosaminoglicanos (GAGs) e hidroxiprolina. Análise da metaloproteinase-2 (MMP-2) foi realizada por zimografia. Para análise morfológica em microscopia de luz comum, os cortes foram corados com hematoxilina-eosina (HE) e azul de toluidina (AT). Para análise em microscopia de polarização os cortes foram corados com Ponceau SS e a birrefringência foi avaliada em termos de valor de média de cinza em pixels utilizando um analisador de imagem. Resultados: EA nos pontos utilizados não alterou a concentração de PNCs, GAGs ou a atividade enzimática da MMP-2 nos tendões transeccionados. Já a concentração de hidroxiprolina foi significativamente aumentada nestes tendões quando tratados com EA. Na análise morfológica nos cortes corados com HE e AT, aparentemente não houve alterações na composição nem na organização da MEC. No entanto, na análise da birrefringência houve uma maior reorganização das fibras de colágeno no grupo tratado com EA. Conclusão: Nossos resultados indicam que a EA pode oferecer benefícios terapêuticos para o tratamento de lesões tendíneas através do aumento do conteúdo do colágeno e da reorganização molecular de suas fibras Abstract: Electroacupuncture (EA) has been used to treat a variety of inflammatory diseases and pain relief. Therefore, there are no reports in the literature about the effect of EA on the composition and organization of the extracellular matrix (ECM) of tendons in the healing process. Objective: To investigate by morphological and biochemical analysis, the effect of EA during the peak of synthesis of ECM components, 15 days after partial transection of the Achilles tendon of rats. Methods: Male Wistar rats with 60 days old were divided into three groups: no tenotomized (G1), tenotomized (G2) and tenotomized and submitted to EA (G3). Acupuncture (AC) needles were inserted in the E-36 and B-57 points and an electrical current with asymmetrical bipolar faradic wave at frequency of 2 Hz and intensity of 2-4 Volts was applied to the needles for 20 minutes, 3 times per week on alternate days, for a total of 6 sessions. Biochemical analyses were done by SDS-PAGE, dosage of non collagenous proteins (NCPs), glycosaminoglycans (GAGs) and hydroxyproline. Analysis of metalloproteinase-2 (MMP-2) was carried out by zymography. For morphological analyses, the sections were stained with hematoxilin-eosin (HE) and toluidine blue. For polarization microscopy analysis the sections were stained with Ponceau SS and birefringence was evaluated in terms of gray average values in pixels using an image analyzer. Results: EA at points used did not alter the concentration of NCPs, GAGs or the enzymatic activity of MMP-2 in transected tendons. However, the concentration of hydroxyproline was significantly increased when these tendons were treated with EA. In the morphological analysis, apparently there were no changes in the composition and organization of the ECM. However, analysis of the birefringence showed higher reorganization of collagen fibers in the group treated with EA. Conclusion: Our results indicate that EA may offer therapeutic benefits for treatment of tendon injuries by increasing the content of collagen and the molecular reorganization of its fibers Mestrado Anatomia Mestre em Biologia Celular e Estrutural

Objective To analyze the characteristics of the Achilles tendon of rats after induction of localized inflammation in the rat paw. Methods In our study three groups were used: inflamed group with carrageenan in rat paw (G1); saline group (G2) and control group (G3). After 4 hours the animals were euthanized and the Achilles tendon removed. Results No significant differences were observed in the analysis of non-collagenous proteins, glycosaminoglycans and hydroxyproline in the groups but a tendency of reduction was verified in G1. As regards the organization of collagen molecules, no differences were observed between groups. With respect to MMPs activity, a stronger presence of the active isoform of MMP-2 in G1 was observed, suggesting that the remodeling was occurring. Conclusion Thus, we conclude that the inflammatory process in rat paw may affect the remodeling of tendons located near the inflamed site. Level of Evidence I, Prognostic Studies - Investigating the Effect of a Patient Characteristic on the Outcome of Disease