Human hair follicle-derived mesenchymal stem cells promote tendon repair in a rabbit Achilles tendinopathy model / 中华医学杂志(英文版)

BACKGROUND@#Hair follicles are easily accessible and contain stem cells with different developmental origins, including mesenchymal stem cells (MSCs), that consequently reveal the potential of human hair follicle (hHF)-derived MSCs in repair and regeneration. However, the role of hHF-MSCs in Achilles tendinopathy (AT) remains unclear. The present study investigated the effects of hHF-MSCs on Achilles tendon repair in rabbits.@*METHODS@#First, we extracted and characterized hHF-MSCs. Then, a rabbit tendinopathy model was constructed to analyze the ability of hHF-MSCs to promote repair in vivo . Anatomical observation and pathological and biomechanical analyses were performed to determine the effect of hHF-MSCs on AT, and quantitative real-time polymerase chain reaction, enzyme-linked immunosorbent assay, and immunohistochemical staining were performed to explore the molecular mechanisms through which hHF-MSCs affects AT. Furthermore, statistical analyses were performed using independent sample t test, one-way analysis of variance (ANOVA), and one-way repeated measures multivariate ANOVA as appropriate.@*RESULTS@#Flow cytometry, a trilineage-induced differentiation test, confirmed that hHF-derived stem cells were derived from MSCs. The effect of hHF-MSCs on AT revealed that the Achilles tendon was anatomically healthy, as well as the maximum load carried by the Achilles tendon and hydroxyproline proteomic levels were increased. Moreover, collagen I and III were upregulated in rabbit AT treated with hHF-MSCs (compared with AT group; P  < 0.05). Analysis of the molecular mechanisms revealed that hHF-MSCs promoted collagen fiber regeneration, possibly through Tenascin-C (TNC) upregulation and matrix metalloproteinase (MMP)-9 downregulation.@*CONCLUSIONS@#hHF-MSCs can be a treatment modality to promote AT repair in rabbits by upregulating collagen I and III. Further analysis revealed that treatment of AT using hHF-MSCs promoted the regeneration of collagen fiber, possibly because of upregulation of TNC and downregulation of MMP-9, thus suggesting that hHF-MSCs are more promising for AT.

Effect of norbixin-based poly(hydroxybutyrate) membranes on the tendon repair process after tenotomy in rats

Abstract Purpose: To determine the efficacy of norbixin-based poly(hydroxybutyrate) (PHB) membranes for Achilles tendon repair. Methods: Thirty rats were submitted to total tenotomy surgery of the right Achilles tendon and divided into two groups (control and membrane; n = 15 each), which were further subdivided into three subgroups (days 7, 14, and 21; n = 5 each). Samples were analyzed histologically. Results: Histological analysis showed a significant reduction in inflammatory infiltrates on days 7, 14 (p < 0.0001 for both), and 21 (p = 0.0004) in the membrane group compared to that in the control group. There was also a significant decrease in the number of fibroblasts in the control group on days 7, 14 (p < 0.0001), and 21 (p = 0.0032). Further, an increase in type I collagen deposition was observed in the membrane group compared to that in the control group on days 7 (p = 0.0133) and 14 (p = 0.0107). Conclusion: Treatment with norbixin-based PHB membranes reduces the inflammatory response, increases fibroblast proliferation, and improves collagen production in the tendon repair region, especially between days 7 and 14.

Avaliação crítica das técnicas cirúrgicas de correção do equino / Critical evaluation of the surgical techniques to correct the equinus deformity

RESUMO A deformidade em equino leva a diversos transtornos da marcha, ao causar alterações no apoio do pé e afetar regiões anatômicas mais distantes, como o joelho, quadril e tronco. Geralmente é secundária à retração, encurtamento ou espasticidade do tríceps sural, de modo que algumas intervenções cirúrgicas podem ser necessárias para corrigi-la. Trata-se de um dos procedimentos mais antigos da Ortopedia, antes realizado apenas no tendão calcâneo e que, ao longo do tempo, evoluiu com técnicas diferentes de acordo com o grau de deformidade, doença de base e perfil do paciente. Busca-se corrigir a deformidade, com a menor interferência possível na força muscular e, com isso, diminuir a incidência de complicações, como marcha agachada, arrastada e pé calcâneo. Do ponto de vista anatômico, o tríceps sural apresenta cinco regiões que podem ser abordadas cirurgicamente para correção do equino. Em virtude da complexidade do paciente com equino, os ortopedistas devem ter experiência com ao menos uma técnica em cada zona. Neste texto são abordadas e analisadas criticamente as técnicas mais importantes para correção do equino, principalmente de modo a evitar complicações. Foi realizada uma busca sobre técnicas cirúrgicas mais comuns de correção do equino em livros clássicos e identificação e consulta aos artigos originais. Em seguida, fez-se uma busca em bases de dados nos últimos dez anos.

ABSTRACT The equinus deformity causes changes in the foot contact and may affect more proximal anatomical regions, such as the knee, hip and trunk, potentially leading to gait disorders. The equinus is usually secondary to retraction, shortening and/or spasticity of the triceps surae, and it may require surgical correction. Surgery for the correction of equinus is one of the oldest procedures in Orthopedics, and it was initially performed only at the calcaneus tendon. The technique has evolved, so that it could be customized for each patient, depending on the degree of deformity, the underlying disease, and patient´s profile. The aim is to correct the deformity, with minimal interference in muscle strength, thus reducing the incidence of disabling complications such as crouch gait and calcaneus foot. We conducted a literature search for the most common surgical techniques to correct the equinus deformity using classic books and original articles. Further, we performed a database search for articles published in the last ten years. From the anatomical perspective, the triceps surae presents five anatomical regions that can be approached surgically for the equinus correction. Due to the complexity of the equinus, orthopedic surgeons should be experienced with at least one procedure at each region. In this text, we critically approach and analyze the most important techniques for correction of the equinus, mainly to avoid complications.

Local treatment with ascorbic acid accelerates recovery of post-sutured Achilles tendon in male Wistar rats

Tendon rupture is a very frequent accident involving average people and high-performance athletes. Clinical studies describe tendon recovery as a painful and slow process involving different biochemical and histological events. Ascorbic acid (AA) is a potent antioxidant as well as an important cofactor for collagen synthesis. In the current study, we evaluated if local treatment with AA is able to promote tendon repair in tenotomized rats. Animals were submitted to Achilles tendon rupture followed by surgical suture. Control and AA groups received in loco injection of saline solution (0.9% NaCl) and 30 mM AA, respectively. Histological and functional recovery of Achilles tendon tissue was evaluated at 7, 14, and 21 days post-surgery. Hematoxylin/eosin staining and collagen fluorescence analysis showed intense disarrangement of tendon tissue in the saline group. Tenotomized animals also showed hypercellularity in tendon tissue compared with non-tenotomized animals. The Achilles functional index (AFI) showed a significant decrease of tendon functionality in tenotomized animals at 7, 14, and 21 days post-surgery. AA accelerated tissue organization and the recovery of function of the Achilles tendons. The beneficial effect of AA treatment was also observed in the organization of the collagen network. Data presented in the current work showed that in loco treatment with AA accelerated the recovery of injured Achilles tendon post-surgery.

Effects of the essential oil of Alpinia zerumbet (Pers.) B.L. Burtt & R.M. Sm. on healing and tissue repair after partial Achilles tenotomy in rats

Abstract Purpose: To investigate the cellular response to injury, analyzing histopathologic changes associated with increased cellularity, degeneration and disorganization of collagen fibers. Methods: Thirty wistar rats were divided in two groups after partial Achilles tenotomy: the right hind paw were treated with the essential oil of Alpinia zerumbet (EOAz), diluted to 33% (0.3 mL kg-1), and the left hind paw received sunflower oil for 3, 14, 30 and 90 days. Statistical significance was determined using a Chi-square and Pearson Correlation qualitative variables test. Moreover, Mann-Whitney U-test test for comparison between different groups of the same cell, one-way ANOVA, and Tukey's test of quantitative measurement. Results: A decrease hyperemia (p < 0.001) was observed in the acute phase of inflammatory cell number (p < 0.001), whereas sub-acute phase was marked by significant correlation with macrophages in fibroblasts (r = 0.17, p = 0.03), with probable induction a dense and modeled tissue. At chronic phase, it was found an increase in the number of fibroblasts and a higher percentage of type I collagen fibers (78%) compared with control collagen fibers (55%). Conclusion: Oil of Alpinia zerumbet stimulated the process of maturation, organization and tissue repair which gave it greater resistance.

Amniotic membrane as an option for treatment of acute Achilles tendon injury in rats

Abstract Purpose: To evaluate the effect of human amniotic membrane (hAM) fragment on inflammatory response, proliferation of fibroblast and organization of collagen fibers in injured tendon. Methods: Sixty rats were divided into 3 groups: C - surgical procedures without tendon lesion and with simulation of hAM application; I - surgical procedures, tendon injury and simulation of hAM application; T - surgical procedures, tendon injury and hAM application. These groups were subdivided into four experimental times (3, 7, 14 and 28 days). The samples underwent histological analysis and ATR-FTIR spectroscopy. Results: Histological analysis at 14 days, the T group showed collagen fibers with better alignment. At 28 days, the I group presented the characteristics described for the T group at 14 days, while this group presented aspects of a mature connective tissue. FT-IR analysis showed a clear distinction among the three groups at all experimental times and groups T and I presented more similarities to each other than to group C. Conclusion: Acute injury of tendon treated with human amniotic membrane fragment showed a faster healing process, reduction in inflammatory response, intense proliferation of fibroblasts and organization of collagen fibers.

Histomorphometric analysis of the Achilles tendon of Wistar rats treated with laser therapy and eccentric exercise / Análise histomorfométrica do tendão calcanear comum de ratos Wistar tratado com laserterapia e exercício excêntrico

Low-level laser therapy is recommended for the treatment of tendinopathies despite the contradictory results related to the ideal dose of energy, wavelength and time of application. This study aimed to assess the effects of laser therapy and eccentric exercise on tendinopathy of the Achilles tendon of Wistar rats. Forty-eight adult male rats were randomly distributed into four groups (L= laser; E= eccentric exercise; LE = laser and eccentric exercise; and R= rest). Laser therapy (904nm/3J/cm2) and/or eccentric exercise (downhill walking; 15o incline treadmill; 12m/min; 50min/day) was started 24h after induction of unilateral tendinopathy and remained for 20 days. At 3, 7, 14 and 21 days after lesion induction, three rats from each group were euthanized and the tendons were collected for histological and morphometric analyses. There was no difference among groups or among times for the characteristics hemorrhage (p=0.4154), fibrinous adhesion formation (p=0.0712), and organization of collagen fibers (p=0.2583) and of the connective tissue (p=0.1046). For these groups, regardless of the time, eccentric exercise led to epitenon thickening (p=0.0204), which was lower in the group treated with laser therapy. Histological analysis revealed differences (p=0.0032) in the number of inflammatory cells over time. They were more numerous in the group that only exercised. This result was confirmed by morphometric analysis, which showed a significant interaction (groups x time) for this characteristic. Eccentric exercise increased (p=0.0014) the inflammatory infiltrate over time (3 and 21 days). However, association with laser therapy reduced inflammatory reaction. On the other hand, the combination of the treatments increased angiogenesis in morphometric (p=0.0000) and histological (p=0.0006) analyses compared with the other groups, while the isolated application of low-level laser reduced this characteristic over time. Animals maintained at rest presented the lowest amount (p=0.0000) of fibroblasts, according to the morphometric analysis. However, histological evaluation showed a significant group x time interaction (p=0.0024). Greater amounts of fibroblasts were observed in groups E, L and LE on the 7th, 14th and 21st days, respectively. The animals that received laser therapy and were exercised showed a greater (p=0.0000) amount of collagen fibers over time. Laser therapy at a dose of 3J and at a wavelength of 904nm, starting 24h after surgical induction of tendinopathy in Wistar rats, is suitable for angiogenesis and prevention of tendon thickening, which can be associated with the intensity of inflammatory process. When associated with eccentric exercise, the therapy has the advantage of increasing the amount of collagen fibers, reducing fibrinous adhesions and inflammatory infiltrate, despite prolonging angiogenesis. Therefore, eccentric exercise performed concomitantly with laser therapy improves the histological properties of the injured tendon.(AU)

O laser de baixa potência é recomendado para o tratamento das tendinopatias apesar dos resultados contraditórios no que se refere a dose de energia ideal, comprimento de onda e tempo de aplicação. O objetivo desse estudo foi avaliar os efeitos da laserterapia e do exercício excêntrico na tendinopatia do tendão calcanear comum de ratos Wistar. Foram utilizados 48 ratos machos adultos. Os animais foram distribuídos aleatoriamente em quatro grupos (L=laser; E=exercício excêntrico; LE=laser e exercício excêntrico, e R=repouso). Após 24h da indução de tendinopatia unilateral, foi iniciada laserterapia (904nm/3J/cm2) e/ou exercício excêntrico (caminhada em declive; esteira com 15o de inclinação; 12m/min; 50min/dia) que permaneceu por 20 dias. Aos 3, 7, 14 e 21 dias após indução da lesão, três ratos de cada grupo foram submetidos à eutanásia, e os tendões foram obtidos para análises histológica e morfométrica. Não houve diferença entre grupos nem entre tempos nas características hemorragia (p=0,4154), formação de aderência fibrinosa (p=0,0712) e organização das fibras colágenas (p=0,2583) e do tecido conjuntivo (p=0,1046). Considerando os grupos, independentemente do tempo, foi observado que o exercício excêntrico ocasionou espessamento do epitendão (p=0,0204), que foi menor no grupo submetido à laserterapia. A análise histológica revelou diferença (p=0,0032) na quantidade de células inflamatórias ao longo do tempo, sendo observado em maior quantidade no grupo apenas exercitado. Esse resultado foi confirmado pela análise morfométrica, que demonstrou haver interação significativa (grupos x tempo) nessa característica, na qual o exercício excêntrico aumentou (p=0,0014) o infiltrado inflamatório ao longo do tempo (3 e 21 dias), porém, quando associado à laserterapia, ocorreu redução da reação inflamatória. Por outro lado, a associação dos tratamentos ocasionou maior angiogênese observada nas análises morfométrica (p=0,0000) e histológica (p=0,0006), quando comparada com os demais grupos, enquanto a aplicação isolada do laser de baixa potência reduziu essa característica ao longo do tempo. Os animais mantidos em repouso foram os que apresentaram menor (p=0,0000) quantidade de fibroblastos, na análise morfométrica, porém na avaliação histológica foi observada interação significativa entre grupo e tempo (p=0,0024). Maior quantidade de fibroblastos foi observada nos grupos E, L e LE, no 7o, 14o e 21o dias, respectivamente. Os animais que receberam laserterapia e foram exercitados apresentaram maior (p=0,0000) quantidade de fibras colágenas ao longo do tempo. A laserterapia na dose de 3J e comprimento de onda de 904nm, iniciada 24h após indução cirúrgica de tendinopatia em ratos Wistar é adequada para angiogênese e para evitar o espessamento do tendão, que pode estar relacionado com a intensidade do processo inflamatório. Quando associada ao exercício excêntrico, a terapia apresenta a vantagem de aumentar a quantidade de fibras colágenas, reduzir a aderência fibrinosa e infiltrado inflamatório, apesar de prolongar a angiogênese. Portanto, o exercício excêntrico realizado concomitantemente com a laserterapia melhora as propriedades histológicas do tendão lesionado.(AU)

Ecografía del tendón de Aquiles: de la imagen a la patología / Achilles tendon ultrasound: from image to the pathology

El tendón de Aquiles o calcáneo constituye la extensión tendinosa en la pantorrilla del gemelo, soleo y plantar delgado con inserción distal en la tuberosidad del calcáneo siendo el tendón más grueso y fuerte de todo el organismo. La ecografía contribuye a discernir el tipo de lesiones de los tendones, lo que permite establecer el tratamiento más adecuado. Tiene cuatro funciones principales: confirmar la existencia real de la lesión tendinosa, precisar la ubicación exacta de la misma, determinar la gravedad de la lesión y evaluar su carácter "activo" o de secuela. En este artículo trataremos de definir los aspectos ecográficos normales del tendón de Aquiles y sus lesiones más frecuentes: rupturas completas e incompletas, tendinitis, tendinosis y bursitis

Effect of reversed polarity microcurrent electrical stimulation on an experimentally induced Achilles tendon injury in male albino rats: a histological and immunohistochemical study

The application of electrical stimulation can lead to a greater and faster increase in the rate of wound healing, especially when applying the cathedral [negative] stimulation for the first week, followed by the anodal [positive] polarity for the rest of the treatment period. The present work aimed to study the effect of polarity reversal of microelectrical current stimulation [MES] on the healing process in an experimentally induced Achilles tendon injury in rats. Forty three male albino rats were used in this study; they were classified into group I [control group] and group II [experimentally injured group], which was further classified into subgroups, I, II, III and IV. Subgroup I represents the injured tendon without MES treatment, whereas subgroups II, III and IV represent the MES-treated ones. The obtained tendon sections were subjected to H and E staining. Masson's trichrome stain, and immunohistochemical staining for alpha smooth muscle actin [alpha SMA], followed by morphometric study and statistical analysis. Subgroup I showed signs of inflammation, a few thin irregularly arranged collagen, active fibroblasts that start to align in rows on the regenerating collagen bundles, and alpha SMA immunoreactivity. In anodal-treated tendons, signs of inflammation had started to disappear; collagen fibers appeared thin and irregularly arranged, active fibroblasts were obviously observed and minimal alpha SMA immunoreactivity were recorded only in subgroup II. Cathodal-treated tendons showed rapid disappearance of cellular infiltration; most collagen fibers appeared regularly arranged with mature fibrocytes in between and multiple avoid alpha SMA immunoreactive myofibroblasts were maximally observed in subgroup II. alpha SMA was suggested to play a role in wound healing due to its high immunoreactivity in myofibroblasts during wound healing. Moreover, the application of electrical stimulation by applying cathodal [negative] stimulation for the first week, followed by anodal [positive] polarity for the rest of the treatment period may lead to better repaired tissue due to myofibroblast directional attraction to the cathode, especially when applied for 4-week duration

Xantomatosis cerebrotendínea: una causa de aumento de volumen bilateral del tendón de Aquiles: caso clínico / Cerebrotendinous xanthomatosis: report of one case

Cerebrotendinous xanthomatosis is an inherited autosomal recessive disease caused by a mutation in the gene for the sterol 27-hydroxylase enzyme, which determines the accumulation of plasmatic cholestanol in various tissues. The natural history of this disease is characterized by chronic diarrhea beginning in childhood, cataract in youth, tendinous xanthomas in adulthood and later progressive neurological dysfunction manifested as dementia, psychiatric disorders, cerebellar, pyramidal or extra pyramidal signs or seizures. We report a 39 year-old male with a history of diarrhea during childhood and bilateral cataracts requiring surgery at 20 years of age, who evolves later with psychiatric disorders and bilateral increased volume in Achules tendons. High levels of plasmatic cholestanol and magnetic resonance imaging confirmed the diagnosis of this disease.

Cambios histopatologicos en las roturas espontaneas del tendon de Aquiles / Histological changes in spontaneous ruptures of the the Achilles tendon

Introducción: La rotura del tendón de Aquiles es una patología que se observa con relativa frecuencia en nuestro medio. Distintos factores se han asociado con la rotura espontánea de ese tendón. El objetivo del presente trabajo es determinar la existencia de lesiones degenerativas tendinosas previas predisponentes a la rotura del Aquiles. Materiales y métodos: Se analizaron 31 muestras de roturas tendinosas aquilianas, de 29 hombres y 2 mujeres, con edad promedio de 42,5 años. Las biopsias fueron tomadas dentro de los 6 días de producida la lesión. Se utilizaron criterios de inclusión y de exclusión. Se realizaron las técnicas de tinción de hematoxilina-eosina, azul alciano a pH 2,5, tricrómico de Mallory, PAS y técnica de Perls, para su estudio microscópico. Se analizaron los datos con métodos estadísticos de Mann-Whitney y de Spearman.Resultados: Todos los pacientes presentaron alteración de la estructura fibrilar, disminución de la tinción del colágeno y desorganización fibrilar. Conclusiones: Los cambios degenerativos encontrados fueron independientes de la edad del paciente y del tiempo de evolución de la rotura. La degeneración mucoide se encontró con mayor frecuencia (67,7 por ciento). Los cambios reparativos variaron de acuerdo con el tiempo de rotura del tendón.

Calcinose peritendínea do tendão calcâneo associada a dermatomiosite: correlacão entre radiografia convencional, ultra-sonografia, ressonância magnética e macroscopia cirúrgica / Peritendinous calcinosis of calcaneus tendon associated with dermatomyositis: correlation between conventional radiograph, ultrasound, magnetic resonance imaging and gross surgical pathology

A calcinose intersticial é uma afeccão incomum, na qual existe deposicão de cálcio localizada ou disseminada na pele, tecido celular subcutâneo, músculos e tendões. Freqüentemente a calcinose está associada com doencas do tecido conjuntivo, como esclerodermia e dermatomiosite. Os autores relatam um caso de calcinose intersticial associada a dermatomiosite, estudada com radiografia convencional, ultra-sonografia, ressonância magnética, e com correlacão com a macroscopia cirúrgica.

Reconstrucción tardía con aloinjertos de ruptura del mecanismo extensor después de artroplastia total de rodilla: más frustaciones / Late reconstruction with aloinjertos of rupture of the extending mechanism after total artroplastia of knee: more frustaciones

Entre mayo 2003 y mayo 2006 cuatro pacientes que habian sufrido ruptura del mecanismo extensor después de una artroplastia total de rodilla fueron sometidos a reconstrucción tardía usando aloinjertos liofilizados de tendón de Aquiles. El promedio de edad era de 70,75 (70-72) años. Por lo menos 3 cirugías anteriores en promedio. Todos los pacientes alcanzaron un alivio del dolor con un promedio de seguimiento de 16,75 (3-38) meses. Dos pacientes presentaron en el post operatorio retardo para la extensión de 10 (50 por ciento) y dos de -40° (50 por ciento), comparando al promedio preoperatorio de -65° por ciento. La deambulación apoyando todo el peso fue posible en todos los pacientes; sin embargo dos pacientes usaban bastón (50 por ciento) y dos andadera (50 por ciento).

Xantoma y ateroma, hipercolesterolemia familiar homo y heterocigota: riesgo de infarto precoz / Xanthoma and atheroma, homo and heterozygous familiar hypercholesterolemia: risk of early myocardiac infarct

Se presentan dos pacientes con hipercolesterolemia familiar y xantomas en cada una de sus variantes genéticas: homocigota y heterocigota. Se hace revisión y actualización de ambos cuadros: clínica, dermatológica, histopatológica, de laboratorio, por ultrasonografía, genética y terapéutica, destacando la importancia del dignóstico precoz a través de xantoma; para la prevención y el tratamiento oportuno de enfermedades cardiovasculares graves

Valor de la resonancia magnetica en la evaluacion de la patologia del tendon de Aquiles / Magnetic resonance imaging value in evaluation of the achilles tendon pathology

Si bien el tendon de Aquiles es una estructura anatomica accesible al examen fisico, su estudio a traves de imagenes es a veces necesario para el analisis de decisiones terapeuticas. La resonancia magnetica es una excelente tecnica para la evaluacion no invasiva de los tejidos blandos y ha demostrado ser mas sensible y exacto que la evaluacion clinica en la localizacion de lesiones en el tendon de Aquiles (resumen truncado)

Xantomas tendinosos: caso clínico / Tendinous xanthomas: clinical case

Se presenta el caso de una mujer de 36 años portadora de xantomas tendinosos de ambos tendornes aquilianos, sin antecedentes familiares aparentes ni evidencias de hipercolesterolemia (colesterol total: 140 mg/dl). Se aprovecha para revisar la relativamente escasa literatura existente en relación con los xantomas tendinosos, destacando algunas conclusiones. La mayoría de los xantomas están asociados a hipercolesterolemia o hiperlipidemia. Sin embargo, hay casos excepcionales, como el que motiva esta presentación, que cursan con valores normales de lípidos sanguíneos. Por los general, la severidad de los xantomas es proporcional a la alteración lipídica. El diagnóstico diferencial se plantea con lesiones traumáticas y tumorales y se resuelve con el estudio histológico. El tratamiento de los xantomas tendinoso es quirúrgico y consiste en la resección parcial o total del tendón afectado. Ocasionalmente puede recurrirse al tratamiento médico con hipolipemiantes. como generalmente persiste el trastorno metabólico, la recurrencia es frecuente