Células tronco mesenquimais autólogas no tratamento da osteoartrite induzida por colagenase na articulação coxofemoral de coelhos (Oryctolagus cuniculus) / Autologous mesenchymal stem cells in the treatment of collagenaseinduced osteoarthritis in the coxofemoral joint of rabbits (Oryctolagus cuniculus)

Na atualidade, a pesquisa envolvendo células-tronco mesenquimais destaca-se na busca de avanços na reparação da cartilagem articular. Objetivou-se, no presente estudo, comparar a regeneração cartilaginosa da articulação coxofemoral de coelhos, com e sem o transplante de células-tronco mesenquimais autólogas, por meio de exames radiográficos e histopatológicos. Dois grupos, com 15 animais da espécie leporina cada, foram submetidos à indução química de osteoartrite com solução de colagenase 2% na articulação coxofemoral direita. No Grupo 1 (Células-tronco) realizou-se a aplicação  intra-articular de células-tronco mesenquimais autólogas. Já o Grupo 2 (Controle) foi constituído por animais submetidos à aplicação intra-articular de solução salina estéril. Foram realizadas avaliações radiográficas e histopatológicas aos 30, 60 e 90 dias após a aplicação. Os resultados histológicos indicam que células-tronco mesenquimais (Grupo 1) melhoraram discretamente a qualidade do tecido de reparo, de acordo com os critérios da escala semi-quantitativa ICRS 1 (International Cartilage Repair Society). O Grupo 1 (Células-Tronco) demonstrou superioridade em relação ao Grupo 2 nos parâmetros: Superfície articular, matriz extracelular e distribuição celular, demonstrando que as células-tronco foram benéficas no tratamento da osteoartrite.

The aim of this study was to compare cartilage regeneration of the hip in rabbits, with and without the transplantation of autologous mesenchymal stem cells. Thirty rabbits were submitted to chemical induction of osteoarthritis with a 2% colagenase in the right hip. They were divided into 2 groups of 15 animals each: Group 1 (intra-articular application of autologous mesenchymal stem cells) and Group 2 (control - intra-articular application of sterile saline solution). Radiographic and histopathological evaluations were performed at 30, 60 and 90 days after application. The mesenchymal stem cells group (Group 1) showed slight improvement of the quality of the repair tissue, according to the semi-quantitative scale criteria ICRS 1 (International Cartilage Repair Society). The Group 1 (Stem Cells) showed superiority in relation to Group 2, especially in the parameters joint surface, extracellular matrix and cellular distribution, demonstrating that stem cells were beneficial in the treatment of osteoarthritis.

Células tronco mesenquimais autólogas no tratamento da osteoartrite induzida por colagenase na articulação coxofemoral de coelhos (Oryctolagus cuniculus) / Autologous mesenchymal stem cells in the treatment of collagenaseinduced osteoarthritis in the coxofemoral joint of rabbits (Oryctolagus cuniculus)

Na atualidade, a pesquisa envolvendo células-tronco mesenquimais destaca-se na busca de avanços na reparação da cartilagem articular. Objetivou-se, no presente estudo, comparar a regeneração cartilaginosa da articulação coxofemoral de coelhos, com e sem o transplante de células-tronco mesenquimais autólogas, por meio de exames radiográficos e histopatológicos. Dois grupos, com 15 animais da espécie leporina cada, foram submetidos à indução química de osteoartrite com solução de colagenase 2% na articulação coxofemoral direita. No Grupo 1 (Células-tronco) realizou-se a aplicação  intra-articular de células-tronco mesenquimais autólogas. Já o Grupo 2 (Controle) foi constituído por animais submetidos à aplicação intra-articular de solução salina estéril. Foram realizadas avaliações radiográficas e histopatológicas aos 30, 60 e 90 dias após a aplicação. Os resultados histológicos indicam que células-tronco mesenquimais (Grupo 1) melhoraram discretamente a qualidade do tecido de reparo, de acordo com os critérios da escala semi-quantitativa ICRS 1 (International Cartilage Repair Society). O Grupo 1 (Células-Tronco) demonstrou superioridade em relação ao Grupo 2 nos parâmetros: Superfície articular, matriz extracelular e distribuição celular, demonstrando que as células-tronco foram benéficas no tratamento da osteoartrite.(AU)

The aim of this study was to compare cartilage regeneration of the hip in rabbits, with and without the transplantation of autologous mesenchymal stem cells. Thirty rabbits were submitted to chemical induction of osteoarthritis with a 2% colagenase in the right hip. They were divided into 2 groups of 15 animals each: Group 1 (intra-articular application of autologous mesenchymal stem cells) and Group 2 (control - intra-articular application of sterile saline solution). Radiographic and histopathological evaluations were performed at 30, 60 and 90 days after application. The mesenchymal stem cells group (Group 1) showed slight improvement of the quality of the repair tissue, according to the semi-quantitative scale criteria ICRS 1 (International Cartilage Repair Society). The Group 1 (Stem Cells) showed superiority in relation to Group 2, especially in the parameters joint surface, extracellular matrix and cellular distribution, demonstrating that stem cells were beneficial in the treatment of osteoarthritis.(AU)

Denervação articular aberta em cadela / Open articular denervation in bitch

A displasia coxofemoral (DCF) é uma afecção de cães e gatos definida como uma doença hereditária biomecânica ou adquirida, representada pela disparidade entre a massa muscular primária e o rápido crescimento ósseo, que pode levar à doença articular degenerativa (DAD). Com a evolução do quadro, os animais apresentarão perda da cartilagem, evoluindo para um desgaste, deformação da cabeça do fêmur e do acetábulo, levando a um quadro de dor e dificuldade de locomoção. O objetivo do presente trabalho foi relatar uma abordagem cirúrgica como tratamento de DCF e DAD de uma cadela da raça labrador. O diagnóstico foi obtido através do histórico, exame físico e exame radiológico. Na radiografia verificouse diminuição de espaço articular em ambos os membros posteriores, artrose do quadril e sub-luxação. O tratamento definitivo instituído para a correção da DCF foi a intervenção cirúrgica usando a técnica de denervação coxofemoral com abordagem aberta, com principal objetivo de amenizar a dor, prevenir a progressão da enfermidade articular degenerativa e restaurar a função normal da articulação. O tratamento se mostrou eficiente, garantindo conforto ao paciente e retorno da função articular e dos membros.

Hip dysplasia (DCF) is a condition of dogs and cats defined as a hereditary biomechanical or acquired disease, represented by the disparity between primary muscle mass and rapid bone growth, which can lead to degenerative joint disease (DAD). With the development of the condition, these animals will present loss of cartilage, evolving to a wear and tear, and deformation of the head of the femur and acetabulum, leading to a picture of pain and difficulty in locomotion. The aim of the present study was to report a surgical approach as treatment of DCF and DAD of a Labrador bitch. The diagnosis is obtained through history, physical examination and radiological examination. Radiography revealed decreased joint space in both hind limbs, hip arthrosis and sub-dislocation. The definitive treatment instituted for the correction of FDD was surgical intervention using the hip denervation technique with open approach, with the main objective of alleviating pain, preventing the progression of degenerative joint disease and restoring normal joint function. The treatment proved to be efficient, ensuring patient comfort and return of joint and limb function.

Denervação articular aberta em cadela / Open articular denervation in bitch

A displasia coxofemoral (DCF) é uma afecção de cães e gatos definida como uma doença hereditária biomecânica ou adquirida, representada pela disparidade entre a massa muscular primária e o rápido crescimento ósseo, que pode levar à doença articular degenerativa (DAD). Com a evolução do quadro, os animais apresentarão perda da cartilagem, evoluindo para um desgaste, deformação da cabeça do fêmur e do acetábulo, levando a um quadro de dor e dificuldade de locomoção. O objetivo do presente trabalho foi relatar uma abordagem cirúrgica como tratamento de DCF e DAD de uma cadela da raça labrador. O diagnóstico foi obtido através do histórico, exame físico e exame radiológico. Na radiografia verificouse diminuição de espaço articular em ambos os membros posteriores, artrose do quadril e sub-luxação. O tratamento definitivo instituído para a correção da DCF foi a intervenção cirúrgica usando a técnica de denervação coxofemoral com abordagem aberta, com principal objetivo de amenizar a dor, prevenir a progressão da enfermidade articular degenerativa e restaurar a função normal da articulação. O tratamento se mostrou eficiente, garantindo conforto ao paciente e retorno da função articular e dos membros.(AU)

Hip dysplasia (DCF) is a condition of dogs and cats defined as a hereditary biomechanical or acquired disease, represented by the disparity between primary muscle mass and rapid bone growth, which can lead to degenerative joint disease (DAD). With the development of the condition, these animals will present loss of cartilage, evolving to a wear and tear, and deformation of the head of the femur and acetabulum, leading to a picture of pain and difficulty in locomotion. The aim of the present study was to report a surgical approach as treatment of DCF and DAD of a Labrador bitch. The diagnosis is obtained through history, physical examination and radiological examination. Radiography revealed decreased joint space in both hind limbs, hip arthrosis and sub-dislocation. The definitive treatment instituted for the correction of FDD was surgical intervention using the hip denervation technique with open approach, with the main objective of alleviating pain, preventing the progression of degenerative joint disease and restoring normal joint function. The treatment proved to be efficient, ensuring patient comfort and return of joint and limb function.(AU)

Cartilagem articular, patogênese e tratamento da osteoartrite / Articular cartilage, and pathogenesis and treatment of the osteoarthritis / Cartílago articular, patogénesis y tratamiento de la osteoartritis

A osteoartrite caracteriza-se por dor e claudicação, sendo considerada processo não específico, mais sim uma sequela comum a várias formas de lesão articular, com envolvimento de fatores biológicos e mecânicos. Desta forma, o objetivo da presente revisão foi discorrer sobre alguns aspectos da cartilagem articular (generalidades, histogênese), patogênese da osteoartrite, bem como algumas possibilidades de tratamentos intra-articulares para a osteoatrite. A cartilagem articular, juntamente com o líquido sinovial, tem papel mecânico importante nas articulações diartrodiais. Os condrócitos estão entre os principais componentes da cartilagem articular e são apontados como células altamente diferenciadas com capacidade limitada de proliferação e migração. O outro componente é a matriz extracelular constituída de colágeno tipo II, proteoglicanos, proteínas não colagenosas, glicoproteínas, água e eletrólitos. A osteoartrite é classificada como doença articular não inflamatória, com múltiplas interações bioquímicas e biomecânicas. Todos os componentes da articulação podem ser comprometidos, porém a cartilagem é considerada o tecido alvo. A afecção pode ser classificada como primária, que ocorre predominantemente em animais idosos, e secundária, decorrente de doenças que afetam a articulação e as estruturas de suporte. Entre os tratamentos intra-articulares para a osteoartrite estão a viscossuplementação com ácido hialurônico, o uso de corticosteroides, e a combinação de ácido hialurônico e corticosteroides. Contudo, há ainda várias controvérsias quanto à eficácia.

Osteoarthritis is characterized by pain and lameness, being considered a non-specific process, but rather a common sequel to various forms of joint lesion, with involvement of biological and mechanical factors. Therefore, the purpose of the present review is to discuss some aspects of articular cartilage (generalities and histogenesis), pathogenesis of osteoarthritis, as well as some possibilities of intra-articular treatments for osteoarthritis. The articular cartilage, along with the synovial fluid, have an important mechanical role in diarthrodial joints. Chondrocytes are one of principal components of articular cartilage and are designated as highly differentiated cells with limited proliferation and migration capacity. The other component is the extracellular matrix composed of collagen type II, proteoglycans, non-collagenous proteins, glycoproteins, water and electrolytes. Osteoarthritis is classified as a non-inflammatory joint disease with multiple biochemical and biomechanical interactions. All components of the joint can be compromised, but cartilage is considered the target tissue. The osteoarthritis can be classified as primary, which occurs predominantly in elderly animals, and secondary, due to diseases that affect the joint and supporting structures. Among the intra-articular treatments for osteoarthritis are viscosupplementation with hyaluronic acid, the use of corticosteroids, and the combination of hyaluronic acid and corticosteroids. However, there are still several controversies regarding efficacy.

La osteoartritis se caracteriza por dolor e claudicación, considerándose como un proceso no especifico, y si, una secuela común a varias formas de lesión articular, involucrando factores biológicos e mecánicos. De esta forma, el objetivo de la presente revisión es mencionar algunos aspectos del cartílago articular (generalidades, histogénesis), patogénesis de la osteoartritis, así como algunas posibilidades de tratamientos intra-articulares para la osteoartritis. El cartílago articular, junto con el líquido sinovial, tienen un papel mecánico importante en las articulaciones diartrodiales. Los condrocitos están entre los principales componentes del cartílago articular y son fijados como células altamente diferenciadas con capacidad limitada de proliferación y migración. El otro componente es la matriz extracelular constituida de colágeno tipo II, proteoglicanos, proteínas no colagenosas, glicoproteínas, agua y electrólitos. La osteoartritis es clasificada como una enfermedad articular no inflamatoria, con múltiples interacciones bioquímicas y biomecánicas. Todos los componentes de la articulación pueden ser comprometidos, sin embargo, el cartílago articular es considerado el tejido de interés. La afección puede ser clasificada como primaria, que ocurre predominantemente en animales de edad avanzada, y secundaria, debido a enfermedades que afectan la articulación y las estructuras de soporte. Entre los tratamientos intra-articulares para la osteoartritis esta la viscosuplementación con ácido hialurônico, el uso de corticosteroides, e la combinación de ácido hialurônico e corticosteroides. Sin embargo, hay aun varias controversias en cuanto la eficacia.

Cartilagem articular, patogênese e tratamento da osteoartrite / Articular cartilage, and pathogenesis and treatment of the osteoarthritis / Cartílago articular, patogénesis y tratamiento de la osteoartritis

A osteoartrite caracteriza-se por dor e claudicação, sendo considerada processo não específico, mais sim uma sequela comum a várias formas de lesão articular, com envolvimento de fatores biológicos e mecânicos. Desta forma, o objetivo da presente revisão foi discorrer sobre alguns aspectos da cartilagem articular (generalidades, histogênese), patogênese da osteoartrite, bem como algumas possibilidades de tratamentos intra-articulares para a osteoatrite. A cartilagem articular, juntamente com o líquido sinovial, tem papel mecânico importante nas articulações diartrodiais. Os condrócitos estão entre os principais componentes da cartilagem articular e são apontados como células altamente diferenciadas com capacidade limitada de proliferação e migração. O outro componente é a matriz extracelular constituída de colágeno tipo II, proteoglicanos, proteínas não colagenosas, glicoproteínas, água e eletrólitos. A osteoartrite é classificada como doença articular não inflamatória, com múltiplas interações bioquímicas e biomecânicas. Todos os componentes da articulação podem ser comprometidos, porém a cartilagem é considerada o tecido alvo. A afecção pode ser classificada como primária, que ocorre predominantemente em animais idosos, e secundária, decorrente de doenças que afetam a articulação e as estruturas de suporte. Entre os tratamentos intra-articulares para a osteoartrite estão a viscossuplementação com ácido hialurônico, o uso de corticosteroides, e a combinação de ácido hialurônico e corticosteroides. Contudo, há ainda várias controvérsias quanto à eficácia.(AU)

Osteoarthritis is characterized by pain and lameness, being considered a non-specific process, but rather a common sequel to various forms of joint lesion, with involvement of biological and mechanical factors. Therefore, the purpose of the present review is to discuss some aspects of articular cartilage (generalities and histogenesis), pathogenesis of osteoarthritis, as well as some possibilities of intra-articular treatments for osteoarthritis. The articular cartilage, along with the synovial fluid, have an important mechanical role in diarthrodial joints. Chondrocytes are one of principal components of articular cartilage and are designated as highly differentiated cells with limited proliferation and migration capacity. The other component is the extracellular matrix composed of collagen type II, proteoglycans, non-collagenous proteins, glycoproteins, water and electrolytes. Osteoarthritis is classified as a non-inflammatory joint disease with multiple biochemical and biomechanical interactions. All components of the joint can be compromised, but cartilage is considered the target tissue. The osteoarthritis can be classified as primary, which occurs predominantly in elderly animals, and secondary, due to diseases that affect the joint and supporting structures. Among the intra-articular treatments for osteoarthritis are viscosupplementation with hyaluronic acid, the use of corticosteroids, and the combination of hyaluronic acid and corticosteroids. However, there are still several controversies regarding efficacy.(AU)

La osteoartritis se caracteriza por dolor e claudicación, considerándose como un proceso no especifico, y si, una secuela común a varias formas de lesión articular, involucrando factores biológicos e mecánicos. De esta forma, el objetivo de la presente revisión es mencionar algunos aspectos del cartílago articular (generalidades, histogénesis), patogénesis de la osteoartritis, así como algunas posibilidades de tratamientos intra-articulares para la osteoartritis. El cartílago articular, junto con el líquido sinovial, tienen un papel mecánico importante en las articulaciones diartrodiales. Los condrocitos están entre los principales componentes del cartílago articular y son fijados como células altamente diferenciadas con capacidad limitada de proliferación y migración. El otro componente es la matriz extracelular constituida de colágeno tipo II, proteoglicanos, proteínas no colagenosas, glicoproteínas, agua y electrólitos. La osteoartritis es clasificada como una enfermedad articular no inflamatoria, con múltiples interacciones bioquímicas y biomecánicas. Todos los componentes de la articulación pueden ser comprometidos, sin embargo, el cartílago articular es considerado el tejido de interés. La afección puede ser clasificada como primaria, que ocurre predominantemente en animales de edad avanzada, y secundaria, debido a enfermedades que afectan la articulación y las estructuras de soporte. Entre los tratamientos intra-articulares para la osteoartritis esta la viscosuplementación con ácido hialurônico, el uso de corticosteroides, e la combinación de ácido hialurônico e corticosteroides. Sin embargo, hay aun varias controversias en cuanto la eficacia.(AU)

The correlation between score-based protocol for equine joint assessment and subsequent arthroscopic intervention outcomes / Protocolo de avaliação baseado em escore das afecções articulares em equinos e correlação com o resultado do tratamento por artroscopia

This study aimed to grade joint abnormalities in horses submitted to therapeutic arthroscopy using score-based protocols for equine joint assessment, correlated with arthroscopic treatment outcomes and owner satisfaction. In this prospective study, we evaluated 126 joints of athletic horses referred for arthroscopy. The joints were scored according to findings of medical history and physical, radiographic, ultrasonographic and arthroscopic examination. Lameness, positive response to flexion test and decreased maximum joint flexion angle were detected in more than 50% of joints. Soft tissue swelling, sclerosis, subchondral bone osteolysis and single osteochondral fragments were the most common radiographic findings. Ultrasonographic examination revealed changes in synovial fluid volume and appearance, and subchondral bone irregularities. Increased vascularity of the synovial villi, chondral fibrillation, chondral fissures and superficial cartilage erosions were the most significant arthroscopic findings. The approaches that demonstrated greater sensitivity and correlation with treatment outcome and owner satisfaction were medical history, ultrasonographic and arthroscopic examination. The scoring protocol employed to grade joint abnormalities enabled the creation of a joint score system for the objective assessment, thus determining the most frequent findings and establishing an

O objetivo desse estudo foi avaliar as anormalidades articulares observadas em equinos submetidos à artroscopia terapêutica, utilizando protocolos baseados em pontuações e correlacionando os resultados do tratamento cirúrgico com a satisfação do proprietário. Neste estudo prospectivo, foram avaliadas 126 articulações de cavalos atletas, encaminhados para artroscopia. As articulações foram pontuadas de acordo com os achados da anamnese e exame físico, exame radiográfico, exame ultrassonográfico e avaliação artroscópica. A claudicação, resposta positiva ao teste de flexão e diminuição do ângulo máximo de flexão articular foram detectados em mais de 50% das articulações. Aumento de volume dos tecidos moles, esclerose subcondral, osteólise subcondral e fragmentos osteocondrais únicos foram os achados radiográficos mais comuns. O exame ultrassonográfico revelou, com frequência, as alterações no volume e na aparência do líquido sinovial, além de irregularidades osteocondrais. O aumento da vascularização das vilosidades sinoviais, fibrilação condral, fissuras condrais e erosões superficiais da cartilagem foram os achados artroscópicos mais significativos. As abordagens diagnósticas que demonstraram maior sensibilidade e melhor correlação entre o resultado do tratamento e satisfação do proprietário foram história médica, exame ultrassonográfico e artroscópico. O protocolo de avaliação

The correlation between score-based protocol for equine joint assessment and subsequent arthroscopic intervention outcomes / Protocolo de avaliação baseado em escore das afecções articulares em equinos e correlação com o resultado do tratamento por artroscopia

This study aimed to grade joint abnormalities in horses submitted to therapeutic arthroscopy using score-based protocols for equine joint assessment, correlated with arthroscopic treatment outcomes and owner satisfaction. In this prospective study, we evaluated 126 joints of athletic horses referred for arthroscopy. The joints were scored according to findings of medical history and physical, radiographic, ultrasonographic and arthroscopic examination. Lameness, positive response to flexion test and decreased maximum joint flexion angle were detected in more than 50% of joints. Soft tissue swelling, sclerosis, subchondral bone osteolysis and single osteochondral fragments were the most common radiographic findings. Ultrasonographic examination revealed changes in synovial fluid volume and appearance, and subchondral bone irregularities. Increased vascularity of the synovial villi, chondral fibrillation, chondral fissures and superficial cartilage erosions were the most significant arthroscopic findings. The approaches that demonstrated greater sensitivity and correlation with treatment outcome and owner satisfaction were medical history, ultrasonographic and arthroscopic examination. The scoring protocol employed to grade joint abnormalities enabled the creation of a joint score system for the objective assessment, thus determining the most frequent findings and establishing an

O objetivo desse estudo foi avaliar as anormalidades articulares observadas em equinos submetidos à artroscopia terapêutica, utilizando protocolos baseados em pontuações e correlacionando os resultados do tratamento cirúrgico com a satisfação do proprietário. Neste estudo prospectivo, foram avaliadas 126 articulações de cavalos atletas, encaminhados para artroscopia. As articulações foram pontuadas de acordo com os achados da anamnese e exame físico, exame radiográfico, exame ultrassonográfico e avaliação artroscópica. A claudicação, resposta positiva ao teste de flexão e diminuição do ângulo máximo de flexão articular foram detectados em mais de 50% das articulações. Aumento de volume dos tecidos moles, esclerose subcondral, osteólise subcondral e fragmentos osteocondrais únicos foram os achados radiográficos mais comuns. O exame ultrassonográfico revelou, com frequência, as alterações no volume e na aparência do líquido sinovial, além de irregularidades osteocondrais. O aumento da vascularização das vilosidades sinoviais, fibrilação condral, fissuras condrais e erosões superficiais da cartilagem foram os achados artroscópicos mais significativos. As abordagens diagnósticas que demonstraram maior sensibilidade e melhor correlação entre o resultado do tratamento e satisfação do proprietário foram história médica, exame ultrassonográfico e artroscópico. O protocolo de avaliação

Direct inhibitory effect of caffeine on viability, synthesis activity and gene expression in cultures of chondrocytes extracted from the articular cartilage of rats / Efeito inibidor direto da cafeína sobre a viabilidade, atividade de síntese e expressão gênica em culturas de condrócitos da cartilagem articular de ratos

The aim of this study was to evaluate the effect of concentrations of caffeine on the viability, synthesis activity and gene expression in cultures of chondrocytes. Extracted articular cartilage from the femurs and tibias of 15 Wistar rats at three days old to isolate chondrocytes. Chondrocytes were cultured in chondrogenic medium (control) or supplemented with caffeine (0.5, 1.0, 2.0mM). Cell viability, alkaline phosphatase activity and collagen synthesis were assessed using colorimetric assays at 7, 14, 21 days. The chondrocyte cultures of all groups grown under coverslips were stained with hematoxylin-eosin to determine the percentage of cells/field and with PAS, safranin O, alcian blue to determine the percentage of matrix chondrogenic/field at 21 days. The expressions of gene transcripts for aggrecan, collagen-II, Sox-9, Runx-2 and alkaline phosphatase were also evaluated by RT-PCR at 21 days. The means were compared using Student-Newman-Keuls. Caffeine significantly reduced the conversion of MTT to formazan, percentage of cells/field, collagen synthesis, alkaline phosphatase activity, synthesis of PAS+, safranin O+ and alcian blue+ chondrogenic matrix, and the expression of aggrecan, Sox-9 and II collagen. It is concluded that caffeine at concentrations of 0.5, 1.0, 2.0mM has a direct inhibitory effect on chondrogenesis in cultures of chondrocytes from rats.(AU)

O objetivo deste estudo foi avaliar o efeito direto de concentrações de cafeína sobre a viabilidade, atividade de síntese e expressão gênica em culturas de condrócitos de ratos. As cartilagens dos fêmures e tíbias de 15 ratos Wistar com três dias foram extraídas para isolamento de condrócitos. Os condrócitos foram cultivados em meio condrogênico (controle) ou em meio acrescido de diferentes concentrações de cafeína (0,5, 1,0, 2,0mM). Foram avaliadas a viabilidade celular, a atividade da fosfatase alcalina e a síntese de colágeno por ensaios colorimétricos aos sete, 14 e 21 dias. Condrócitos cultivados sob lamínulas foram corados pela hematoxilina e eosina, para se determinar a porcentagem de células/campo, e pelo PAS, safranina O, alcian Blue, para se determinar a porcentagem de matriz condrogênica/campo aos 21 dias. Foi avaliada a expressão de transcriptos gênicos para Sox-9, Runx-2, agrecano, colágeno-II e fosfatase alcalina por qRT-PCR, aos 21 dias. As médias foram comparadas pelo Student-Newman-Keuls. A cafeína reduziu significativamente o MTT em cristais de formazan, a porcentagem de células/campo, a síntese de colágeno, a atividade da fosfatase alcalina e a síntese de matriz condrogênica PAS+, safranina O+, alcian blue+ e expressão de Sox-9 e colágeno-II. Conclui-se que a cafeína, nas concentrações de 0,5, 1,0, 2,0mM, apresenta efeito inibidor direto sobre a condrogênese em culturas de condrócitos de ratos.(AU)

Direct inhibitory effect of caffeine on viability, synthesis activity and gene expression in cultures of chondrocytes extracted from the articular cartilage of rats / Efeito inibidor direto da cafeína sobre a viabilidade, atividade de síntese e expressão gênica em culturas de condrócitos da cartilagem articular de ratos

The aim of this study was to evaluate the effect of concentrations of caffeine on the viability, synthesis activity and gene expression in cultures of chondrocytes. Extracted articular cartilage from the femurs and tibias of 15 Wistar rats at three days old to isolate chondrocytes. Chondrocytes were cultured in chondrogenic medium (control) or supplemented with caffeine (0.5, 1.0, 2.0mM). Cell viability, alkaline phosphatase activity and collagen synthesis were assessed using colorimetric assays at 7, 14, 21 days. The chondrocyte cultures of all groups grown under coverslips were stained with hematoxylin-eosin to determine the percentage of cells/field and with PAS, safranin O, alcian blue to determine the percentage of matrix chondrogenic/field at 21 days. The expressions of gene transcripts for aggrecan, collagen-II, Sox-9, Runx-2 and alkaline phosphatase were also evaluated by RT-PCR at 21 days. The means were compared using Student-Newman-Keuls. Caffeine significantly reduced the conversion of MTT to formazan, percentage of cells/field, collagen synthesis, alkaline phosphatase activity, synthesis of PAS+, safranin O+ and alcian blue+ chondrogenic matrix, and the expression of aggrecan, Sox-9 and II collagen. It is concluded that caffeine at concentrations of 0.5, 1.0, 2.0mM has a direct inhibitory effect on chondrogenesis in cultures of chondrocytes from rats.(AU)

O objetivo deste estudo foi avaliar o efeito direto de concentrações de cafeína sobre a viabilidade, atividade de síntese e expressão gênica em culturas de condrócitos de ratos. As cartilagens dos fêmures e tíbias de 15 ratos Wistar com três dias foram extraídas para isolamento de condrócitos. Os condrócitos foram cultivados em meio condrogênico (controle) ou em meio acrescido de diferentes concentrações de cafeína (0,5, 1,0, 2,0mM). Foram avaliadas a viabilidade celular, a atividade da fosfatase alcalina e a síntese de colágeno por ensaios colorimétricos aos sete, 14 e 21 dias. Condrócitos cultivados sob lamínulas foram corados pela hematoxilina e eosina, para se determinar a porcentagem de células/campo, e pelo PAS, safranina O, alcian Blue, para se determinar a porcentagem de matriz condrogênica/campo aos 21 dias. Foi avaliada a expressão de transcriptos gênicos para Sox-9, Runx-2, agrecano, colágeno-II e fosfatase alcalina por qRT-PCR, aos 21 dias. As médias foram comparadas pelo Student-Newman-Keuls. A cafeína reduziu significativamente o MTT em cristais de formazan, a porcentagem de células/campo, a síntese de colágeno, a atividade da fosfatase alcalina e a síntese de matriz condrogênica PAS+, safranina O+, alcian blue+ e expressão de Sox-9 e colágeno-II. Conclui-se que a cafeína, nas concentrações de 0,5, 1,0, 2,0mM, apresenta efeito inibidor direto sobre a condrogênese em culturas de condrócitos de ratos.(AU)

Can Triamcinolone acetonide, platelet-rich plasma, and pentosan polysulfate sodium induce oxidative stress in cultured equine chondrocytes? / Acetonido de triancinolona, plasma rico em plaquetas e pentosano polissulfato sódico podem ocasionar estresse oxidativo em cultivo de condrócitos de equinos?

Progressive deterioration and loss of articular cartilage are the final degenerative events common to osteoarthritis (OA). Reactive oxygen species (ROS) play an important role in this chondrocyte catabolic activity, leading to cell death and matrix components breakdown. Intra-articular corticosteroid injections such as triamcinolone acetonide have been used to control pain and inflammation associated with OA. New treatments for OA, platelet-rich plasma and pentosan polysulphate sodium have also been used and further investigations are necessary to determine their safety in joint cells. In this in vitro study, the use of these three substances (triamcinolone acetonide, platelet-rich plasma, and pentosan polysulphate sodium) in healthy chondrocytes did not alter the antioxidant status when compared to control groups, indicating that they could be considered safe in healthy conditions.(AU)

A deterioração progressiva e perda da cartilagem articular são os eventos finais da osteoartrite (OA). Espécies reativas de oxigênio (ROS) têm papel importante na atividade catabólica de condrócitos, levando a morte celular e quebra dos componentes da matriz. Injeções intra-articulares de corticosteroides, como com o acetonido de triancinolona, são usadas para controle da dor e inflamação associadas à OA. Novos tratamentos para a OA, como o plasma rico em plaquetas e o pentosano polissulfato sódico, também tem sido utilizados e necessitam de maiores investigações para determinar sua segurança para as células articulares de equinos. Neste estudo in vitro, o uso destas três substâncias (acetonido de triancinolona, pentosan polissulfato de sódio de plasma rico em plaquetas) em condrócitos saudáveis de equinos não alterou o status antioxidante quando comparado aos grupos controle, indicando que puderam ser considerados seguros em condições saudáveis.(AU)

Comparison of the effects of triamcinolone acetonide or platelet-rich plasma on expression of extracellular matrix-related genes in equine healthy chondrocytes in vitro / Comparação dos efeitos do acetonido de triancinolona ou plasma rico em plaquetas na expressão de genes relacionados à matriz extracelular de condrócitos in vitro de equinos adultos

In healthy cartilage, chondrocytes maintain an expression of collagens and proteoglycans and are sensitive to growth factors and cytokines that either enhance or reduce type II collagen synthesis. In osteoarthritis, pro-inflammatory cytokines, such as IL-6, induce overexpression of metalloproteinases (MMP) and decreasing synthesis of aggrecan. Use of chondroprotectors agents, such as Platelet-Rich Plasma (PRP) and triamcinolone (TA) are alternatives to reduce the progression of joint damage. In this study, we used chondrocytes extracted from metacarpophalangeal joints of healthy horses as the experimental model. Cells were treated in vitro with PRP or TA. No differences were observed between these treatments in comparison to the control group when the expressions of MMP9, MMP13, IL-6 and ACAN genes were evaluated (P<0.05). With these results, we can suggest that the treatments were not deleterious to equine cultured chondrocyte, once they did not stimulate MMPs and IL-6 synthesis or caused changes in ACAN.(AU)

Na cartilagem saudável, os condrócitos mantêm a expressão de colágenos e proteoglicanos, sendo sensíveis a fatores de crescimento e citocinas que aumentam ou reduzem a síntese de colágeno tipo II. Na osteoartrite, citocinas pró-inflamatórias, como a IL-6, estimulam a expressão de metaloproteinases (MMP) e reduzem a síntese de agrecano. O uso de condroprotetores, como o Plasma Rico em Plaquetas (PRP) e triancinolona (TA) é uma alternativa para se reduzir a progressão do dano articular. Neste estudo foram usados condrócitos extraídos das articulações metacarpofalangeanas de equinos saudáveis. As células foram tratadas in vitro com TA ou PRP. Não foram observadas diferenças entre os tratamentos comparando-se com o grupo controle quanto à expressão genética de MMP-9, MMP-13, IL-6 e ACAN (p<0,05). Assim, pode-se sugerir que os tratamentos não foram deletérios ao cultivo de condrócitos, uma vez que não estimularam a síntese de MMP e IL-6 e nem causaram alterações no ACAN.(AU)

Involvement of TLR4 in the protective effect of intra-articular administration of curcumin on rat experimental osteoarthritis

Purpose: In view of the principal role of Toll-like receptor 4 (TLR4) in mediating sterile inflammatory response contributing to osteoarthritis (OA) pathogenesis, we used lipopolysaccharide (LPS), a known TLR4 activator, to clarify whether modulation of TLR4 contributed to the protective actions of intra-articular administration of curcumin in a classical rat OA model surgically induced by anterior cruciate ligament transection (ACLT). Methods: The rats underwent ACLT and received 50μl of curcumin at the concentration of 1 mg mL-1and 10 μg LPS by intra-articular injection once a week for 8 weeks. Morphological changes of the cartilage and synovial tissues were observed. Apoptotic chondrocytes were detected using TUNEL assay. The concentrations of IL-1β and TNF-ɑ in synovial fluid were determined using ELISA kits. The mRNA and protein expression levels of TLR4 and NF-κB p65 were detected by real-time PCR and Western blotting, respectively. Results: Intra-articular administration of curcumin significantly improved articular cartilage injury, suppressed synovial inflammation and down-regulated the overexpression of TLR4 and its downstream NF-κB caused by LPS-induced TLR4 activation in rat osteoarthritic knees. Conclusion: The data suggested that the inhibition of TLR4 signal might be an important mechanism underlying a protective effect of local curcumin administration on OA.(AU)

Histochemistry of equine damaged tendons, ligaments and articular cartilage

Background: The injury repair process in tendons and ligaments includes different phases such as inflammation, neovascularization, fibroblast proliferation and fibrosis. Collagen type and tissue characteristics of tendon and ligament repair are described such as type collagen differentiation and properties of the scars tissue. The degeneration of articular cartilage when, characterized by loss of the articular layers associated of the decreased of proteoglycans. The aim of this study is to describe by histochemistry techniques the characteristics of tissue scar, collagen type in the repair process of tendons and ligaments, as well as articular cartilage degeneration.Materials, Methods & Results: Tissue samples of equine tendons, ligaments and articular cartilage of the metacarpophalangeal joint region were evaluated by ultrasonography, macroscopically and prepared for routine histopathology (H&E staining). The inclusion criterion of the samples in this study was based on the presence of lesions characterized in H&E stain as fibroplasia, neovascularization, collagenolysis, chondroid metaplasia in tendons and ligaments and fibrillation and cartilaginous eburnation lesions in the articular cartilage samples. The Masson’s trichrome, Picrosirius red and Alcian blue staining techniques were also performed in addition to H&E. Pathologic findings in the tendons and ligaments included fibroplasia, collagenolysis, chondroid metaplasia and lymphohistioplasmacytic inflammation. Tendons and ligaments scars were composed of type III collagen but there was also some type I collagen. Fiber alignment of tendons and ligaments in the reorganization tissue was not flawless and the fiber appearance was characterized by a lack of the fiber crimp and parallelism. The fibroplasia was characterized by endotendinous tickening areas associated with the presence of loose connective tissue.[...]

Histochemistry of equine damaged tendons, ligaments and articular cartilage

Background: The injury repair process in tendons and ligaments includes different phases such as inflammation, neovascularization, fibroblast proliferation and fibrosis. Collagen type and tissue characteristics of tendon and ligament repair are described such as type collagen differentiation and properties of the scars tissue. The degeneration of articular cartilage when, characterized by loss of the articular layers associated of the decreased of proteoglycans. The aim of this study is to describe by histochemistry techniques the characteristics of tissue scar, collagen type in the repair process of tendons and ligaments, as well as articular cartilage degeneration.Materials, Methods & Results: Tissue samples of equine tendons, ligaments and articular cartilage of the metacarpophalangeal joint region were evaluated by ultrasonography, macroscopically and prepared for routine histopathology (H&E staining). The inclusion criterion of the samples in this study was based on the presence of lesions characterized in H&E stain as fibroplasia, neovascularization, collagenolysis, chondroid metaplasia in tendons and ligaments and fibrillation and cartilaginous eburnation lesions in the articular cartilage samples. The Massons trichrome, Picrosirius red and Alcian blue staining techniques were also performed in addition to H&E. Pathologic findings in the tendons and ligaments included fibroplasia, collagenolysis, chondroid metaplasia and lymphohistioplasmacytic inflammation. Tendons and ligaments scars were composed of type III collagen but there was also some type I collagen. Fiber alignment of tendons and ligaments in the reorganization tissue was not flawless and the fiber appearance was characterized by a lack of the fiber crimp and parallelism. The fibroplasia was characterized by endotendinous tickening areas associated with the presence of loose connective tissue.[...](AU)

Enxerto osteocondral autógeno associado ao IGF-1 emlesão induzida na cartilagem articular de coelhos / Autogenous osteochondral graft associated with IGF-1 in induced articular cartilage lesion in rabbits

Background: Articular cartilage has a limited capacity for regeneration and of the various treatments proposed, none have reached appropriate therapeutic effectiveness. This study aimed to evaluate autogenous osteochondral grafts in intact or macerated format, in association with or without insulin-like growth factor type-1 (IGF-1) in the repair of osteochondral defects induced in the femoral trochlear groove of rabbits.Materials, Methods & Results: Seventeen healthy White New Zealand rabbits were selected for this study. The rabbits were female, six months old, and had an average body weight of 4.5 kg. All 34 stifle joints were subjected to autogenous osteochondral grafting in the femoral trochlear groove. The joints were divided into four groups designated as intact osteochondral graft with IGF-1 (INT + IGF), intact osteochondral graft with physiological solution (INT + FIS), macerated osteochondral graft with IGF-1 (MAC + IGF), and macerated osteochondral graft with physiological solution (MAC + FIS). Serial evaluations were performed by orthopedic and radiographic examination. After 6 and 12 weeks postoperatively, the grafted area was subjected to macroscopic, histological, and immunohistochemical analyses. Although no statistically significant differences were found between the groups in relation to clinical, macroscopic, histological, and immunohistochemical aspects, a tendency of IGF-1 to promote tissue repair was evident. In the radiographic evaluation, the articular surface and the recipient site in both groups with IGF-1 showed significantly more effective filling (P ≤ 0.05). Regardless of the group, collagen type 2 production, as assessed by immunohistochemistry, was found to be appropriate on the grafted articular surface.[...]

Enxerto osteocondral autógeno associado ao IGF-1 emlesão induzida na cartilagem articular de coelhos / Autogenous osteochondral graft associated with IGF-1 in induced articular cartilage lesion in rabbits

Background: Articular cartilage has a limited capacity for regeneration and of the various treatments proposed, none have reached appropriate therapeutic effectiveness. This study aimed to evaluate autogenous osteochondral grafts in intact or macerated format, in association with or without insulin-like growth factor type-1 (IGF-1) in the repair of osteochondral defects induced in the femoral trochlear groove of rabbits.Materials, Methods & Results: Seventeen healthy White New Zealand rabbits were selected for this study. The rabbits were female, six months old, and had an average body weight of 4.5 kg. All 34 stifle joints were subjected to autogenous osteochondral grafting in the femoral trochlear groove. The joints were divided into four groups designated as intact osteochondral graft with IGF-1 (INT + IGF), intact osteochondral graft with physiological solution (INT + FIS), macerated osteochondral graft with IGF-1 (MAC + IGF), and macerated osteochondral graft with physiological solution (MAC + FIS). Serial evaluations were performed by orthopedic and radiographic examination. After 6 and 12 weeks postoperatively, the grafted area was subjected to macroscopic, histological, and immunohistochemical analyses. Although no statistically significant differences were found between the groups in relation to clinical, macroscopic, histological, and immunohistochemical aspects, a tendency of IGF-1 to promote tissue repair was evident. In the radiographic evaluation, the articular surface and the recipient site in both groups with IGF-1 showed significantly more effective filling (P ≤ 0.05). Regardless of the group, collagen type 2 production, as assessed by immunohistochemistry, was found to be appropriate on the grafted articular surface.[...](AU)

Xenogeneic mesenchymal stem cells in the formation of hyaline cartilage in osteochondral goat failure

Background: Osteochondral knee failures are among the most common causes of disability among the elderly human population and animal athletes. The xenogeneic transplantation of mesenchymal stem cells is a questionable therapeutic alternative that, despite the low expression of Major Histocompatibility Complex type II by these cells, still has relevant uncertainties about the safety and clinical efficacy. The main objective of the present study was to investigate whether the xenogeneic transplantation of mesenchymal stem cells induces hyaline cartilage formation, without histopathological evidence of rejection, in osteochondral failures of goats.Materials, Methods & Results: Five female goats were used, submitted to three surgical osteocondral failures in the right knee, treated with xenogenic mesenchymal stem cells of dental pulp, xenogenic platelet-rich plasma and hemostatic sponge of hydrolyzed collagen, respectively. The lesions were evaluated after 60 days of treatment, aiming to identify the presence of hyaline cartilage or fibrocartilage and the subchondral bone pattern (regenerated or disorganized). Transplantation of xenogenic mesenchymal stem cells induced predominant formation of hyaline cartilage (P 0.05). Macroscopically, the lesions of the stem cell treated group showed formation of firm repair tissue, opaque staining, integrated with adjacent cartilage and with the failure filling almost completely. The groups treated with PRP and hemostatic sponge of hydrolyzed collagen presented, on average, partial filling of the lesion, with irregular shape and darkened coloration.[...]

Xenogeneic mesenchymal stem cells in the formation of hyaline cartilage in osteochondral goat failure

Background: Osteochondral knee failures are among the most common causes of disability among the elderly human population and animal athletes. The xenogeneic transplantation of mesenchymal stem cells is a questionable therapeutic alternative that, despite the low expression of Major Histocompatibility Complex type II by these cells, still has relevant uncertainties about the safety and clinical efficacy. The main objective of the present study was to investigate whether the xenogeneic transplantation of mesenchymal stem cells induces hyaline cartilage formation, without histopathological evidence of rejection, in osteochondral failures of goats.Materials, Methods & Results: Five female goats were used, submitted to three surgical osteocondral failures in the right knee, treated with xenogenic mesenchymal stem cells of dental pulp, xenogenic platelet-rich plasma and hemostatic sponge of hydrolyzed collagen, respectively. The lesions were evaluated after 60 days of treatment, aiming to identify the presence of hyaline cartilage or fibrocartilage and the subchondral bone pattern (regenerated or disorganized). Transplantation of xenogenic mesenchymal stem cells induced predominant formation of hyaline cartilage (P < 0.05), with no histopathological evidence of inflammation when compared to the other treatments. Therapies with xenogeneic platelet-rich plasma and hemostatic sponge of hydrolyzed collagen induced greater formation of fibrocartilaginous cartilage, with no significant difference between them (P > 0.05). Macroscopically, the lesions of the stem cell treated group showed formation of firm repair tissue, opaque staining, integrated with adjacent cartilage and with the failure filling almost completely. The groups treated with PRP and hemostatic sponge of hydrolyzed collagen presented, on average, partial filling of the lesion, with irregular shape and darkened coloration.[...](AU)

Patellar luxation and articular lesions in dogs: a retrospective: study research article / Luxação de patela e lesões articulares em cães: estudo retrospectivo

This study describes lesions that occur in the stifle joints of dogs with patellar luxation. These lesions are associated with the animal's age, body weight, and degree of luxation. The rate of redislocation was also evaluated. The patellar lesions found include articular cartilage erosion, subchondral bone exposure, a flattened or concave patellar surface, and enthesophytes. Extra-patellar lesions included synovitis, osteophytes, blunting of the trochlear groove, an absent trochlea, erosion of the condylar margins, capsule thickening, a long digital extensor tendon injury, cranial cruciate ligament rupture, and meniscal prolapse. Such lesions were frequently found in animals with Grade II or III luxation who were aged 24 months or more, and they were more severe in dogs weighing more than 15 kg. Patellar luxation causes changes that favor articular degeneration and should be treated surgically. Conservative treatment relieves pain, but does not address tissue alterations.(AU)

O estudo descreve as lesões articulares em cães com luxação de patela. Elas foram associadas com a idade do animal, massa corporal e grau de luxação. Foi avaliada também a porcentagem de casos com recidiva. As lesões patelares observadas foram erosão da cartilagem articular, exposição óssea subcondral, superfície patelar achatada ou côncava e entesófitos. As lesões extra patelares incluíram sinovite, osteófitos, ausência do sulco troclear, erosão das bordas condilares, espessamento da cápsula, lesão do tendão do músculo extensor digital, ruptura do ligamento cruzado cranial e prolapso de menisco. As lesões foram encontradas com maior frequência em animais com luxação de Grau II ou III e idade de 24 meses ou mais, sendo mais graves em cães com massa corporal superior a 15 kg. A luxação patelar ocasiona alterações que favorecem a degeneração articular e devem ser tratadas cirurgicamente. O tratamento conservativo alivia a dor, mas não corrige as alterações teciduais.(AU)