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)

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)

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)

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)

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)

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)

Ressonância magnética na avaliação de enxerto osteocondral autógeno na cartilagem articular de coelhos / Magnetic resonance in the evaluation of autogenous osteochondral graft in articular cartilage of rabbits

Este trabalho teve o objetivo de determinar as características e a aplicabilidade do exame de ressonância magnética na avaliação de enxerto ostecondral autógeno, em formato íntegro ou macerado, associado ou não ao fator de crescimento semelhante à insulina tipo 1 (IGF-1), utilizado no reparo de lesões induzidas na cartilagem articular de coelhos. Foram utilizados 9 coelhos da linhagem Nova Zelândia, em que as 18 articulações fêmoro-tíbio-patelares foram submetidas à enxertia osteocondral autógena no sulco troclear femoral. Estas foram divididas em quatro grupos, denominados como enxerto osteocondral íntegro + IGF-1 (n=5), enxerto osteocondral íntegro + solução fisiológica (n=4), enxerto osteocondral macerado + IGF-1 (n=5) e enxerto osteocondral macerado + solução fisiológica (n=4). Os animais foram eutanasiados em 12 semanas após a cirurgia e as articulações foram submetidas ao exame de ressonância magnética utilizando um aparelho scanner de 1,5 Tesla de alto campo magnético. Além disso, amostras dos locais de enxertia foram submetidas aos exames anatomopatológicos. O exame de ressonância magnética mostrou-se eficaz como um método não invasivo para avaliação do tecido de reparação em enxertos osteocondrais na cartilagem articular do fêmur de coelhos, fornecendo dados complementares aos exames macroscópicos e histológicos. Por meio destas imagens e dos exames anatomopatológicos, foram observados resultados satisfatórios em relação ao processo de reparação dos enxertos osteocondrais autógenos na cartilagem de coelhos, independentemente de seu formato ou da adição de IGF-1.(AU)

This study aimed to determine the characteristics and applicability of magnetic resonance imaging (MRI) in the evaluation of autogenous osteochondral graft in intact or macerated format, with or without insulin-like growth factor type 1 (IGF-1) used in repair of cartilage lesions induced in rabbits. Nine New Zealand rabbits were used, in which 18 stifle joints underwent grafting procedure in the femoral trochlear groove. These were divided into four groups, referred as intact osteochondral graft + IGF-1 (n=5), intact osteochondral graft + saline solution (n=4), macerated osteochondral graft + IGF-1 (n=5) and macerated osteochondral graft + saline solution (n=4). Animals were euthanized 12 weeks after surgery and the joints were subjected to MRI using a high magnetic field scanner of 1.5 Tesla. In addition, samples of grafting sites were subjected to anatomopathological examination. The MRI was effective as a noninvasive method to evaluate the repair tissue in osteochondral grafts in articular cartilage of the femur of rabbits by providing complementary data to macroscopic and histological examinations. Through these images and anatomopathological examinations satisfactory results were observed in relation to the repair process of autogenous osteochondral grafts in cartilage of rabbits, regardless of its format or the addition of IGF-1.(AU)

Cartogenina associada à microperfuração do osso subcondral na reparação da cartilagem articular em equinos / Kartogenin associated to micro of the subchondral bone in the joint cartilage repair in horses

A reconstituição da cartilagem articular danificada por doenças de desenvolvimento, trauma e osteoartrite tem sido um grande desafio na veterinária. O objetivo do presente estudo foi avaliar o uso da cartogenina, uma molécula capaz de induzir diferenciação de células mesenquimais em condrócitos, associado ou não à técnica de microfratura na reparação de defeitos condrais induzidos na tróclea femoral de equinos. Foram utilizados seis equinos pesando em média (±DP) 342±1,58kg, com idade de 7,2±1,30 anos e escore corporal de 7,1±0,75, os quais foram submetidos à videoartroscopia para indução da lesão condral de 1cm2 na tróclea lateral do fêmur e à realização da técnica de microperfuração do osso subcondral de ambos os joelhos. Foram realizadas quatro aplicações semanais com 20µM de cartogenina intra-articular em um dos joelhos (grupo tratado) e solução de ringer com lactato na articulação contralateral (grupo controle). Os animais foram submetidos a avaliações física, radiográfica, ultrassonográfica, por um período de 60 dias. Não houve qualquer diferença estatística entre as articulações tratadas e as controle. A terapia com cartogenina, segundo protocolo utilizado, não produziu melhora clínica em lesões osteocondrais induzidas e tratadas com microperfurações na tróclea lateral do fêmur em equinos.(AU)

Articular cartilage reconstruction is still a challenge in Veterinary Medicine. The aim was to evaluate the therapeutic effects of kartogenin, a small molecule that promotes chondrocyte differentiation, in the repair of induced chondral defects pretreated with subchondral drilling. Six horses with a mean (± SD) weight of 342 ± 1.58Kg, aging 7.2 ± 1.30 years, and with a mean 7.1 ± 0.75 body score condition were used. In both stifles, a 1cm2 chondral defect was induced in the lateral femoral trochlea followed by treatment with subchondral drilling. Four intra-articular injections with kartogenin (20mM) were performed weekly in one stifle (treated group). The same procedure using saline solution was performed in the contralateral joint (control group). Clinical, radiographic, ultrasound evaluations were performed for a period of 60 days. No statistical differences were detected between groups in any of the studied variables. Although kartogenin has shown to improve articular cartilage repair in laboratory animal models, the same was not observed in this equine model. In conclusion, the kartogenin therapy, according to the used protocol, did not promote any clinical benefit in equine femoral trochlear defects pretreated with subcondral drilling.(AU)

Cartogenina associada à microperfuração do osso subcondral na reparação da cartilagem articular em equinos / Kartogenin associated to micro of the subchondral bone in the joint cartilage repair in horses

A reconstituição da cartilagem articular danificada por doenças de desenvolvimento, trauma e osteoartrite tem sido um grande desafio na veterinária. O objetivo do presente estudo foi avaliar o uso da cartogenina, uma molécula capaz de induzir diferenciação de células mesenquimais em condrócitos, associado ou não à técnica de microfratura na reparação de defeitos condrais induzidos na tróclea femoral de equinos. Foram utilizados seis equinos pesando em média (±DP) 342±1,58kg, com idade de 7,2±1,30 anos e escore corporal de 7,1±0,75, os quais foram submetidos à videoartroscopia para indução da lesão condral de 1cm2 na tróclea lateral do fêmur e à realização da técnica de microperfuração do osso subcondral de ambos os joelhos. Foram realizadas quatro aplicações semanais com 20µM de cartogenina intra-articular em um dos joelhos (grupo tratado) e solução de ringer com lactato na articulação contralateral (grupo controle). Os animais foram submetidos a avaliações física, radiográfica, ultrassonográfica, por um período de 60 dias. Não houve qualquer diferença estatística entre as articulações tratadas e as controle. A terapia com cartogenina, segundo protocolo utilizado, não produziu melhora clínica em lesões osteocondrais induzidas e tratadas com microperfurações na tróclea lateral do fêmur em equinos.(AU)

Articular cartilage reconstruction is still a challenge in Veterinary Medicine. The aim was to evaluate the therapeutic effects of kartogenin, a small molecule that promotes chondrocyte differentiation, in the repair of induced chondral defects pretreated with subchondral drilling. Six horses with a mean (± SD) weight of 342 ± 1.58Kg, aging 7.2 ± 1.30 years, and with a mean 7.1 ± 0.75 body score condition were used. In both stifles, a 1cm2 chondral defect was induced in the lateral femoral trochlea followed by treatment with subchondral drilling. Four intra-articular injections with kartogenin (20mM) were performed weekly in one stifle (treated group). The same procedure using saline solution was performed in the contralateral joint (control group). Clinical, radiographic, ultrasound evaluations were performed for a period of 60 days. No statistical differences were detected between groups in any of the studied variables. Although kartogenin has shown to improve articular cartilage repair in laboratory animal models, the same was not observed in this equine model. In conclusion, the kartogenin therapy, according to the used protocol, did not promote any clinical benefit in equine femoral trochlear defects pretreated with subcondral drilling.(AU)

Avaliações macroscópica e histológica do reparo da cartilagem articular equina tratada com microperfurações do osso subcondral associadas ou não à injeção intra-articular de cartogenina / Macroscopic and histological evaluations of equine joint cartilage repair treated with microperforation of the subchondral bone associated or not with intra-articular kartogenin

O objetivo deste estudo foi avaliar o reparo da cartilagem hialina equina, por meio de análises macroscópica (através de videoartroscopia) e histológica (através de fragmentos de biopsia), em defeitos condrais induzidos na tróclea lateral do fêmur tratados pela técnica de microperfurações subcondral associada ou não com administração intra-articular de cartogenina. Foram utilizados seis equinos pesando em média (±DP) 342±1,58 kg, com a idade aproximada de 7,2±1,30 anos e escore corporal de 7,1±0,75, que foram submetidos a videoartroscopia para indução da lesão condral de 1 cm2 na tróclea lateral do fêmur e realização da técnica de microperfuração do osso subcondral de ambos os joelhos. Foram realizadas quatro aplicações semanais com 20 μM de cartogenina intra-articulares em um dos joelhos (grupo tratado) e solução de ringer com lactato na articulação contralateral (grupo controle). Após o período de 60 dias, foram feitas as avaliações macroscópicas, através de videoartroscopias, e histológicas, através de biopsia. Não foram observadas diferenças significativas nos escores macroscópicos e histológicos para reparação condral entre animais dos grupos tratados e não tratados (P>0,05). De modo geral, a porcentagem média de cartilagem hialina no tecido de reparo (17,5%) foi condizente com a literatura internacional usando outros tipos de perfuração condral. Entretanto, não se observaram diferenças estatísticas entre grupos (P>0,05). A terapia com cartogenina, segundo protocolo utilizado, não produziu melhora do processo cicatricial em lesões condrais induzidas e tratadas com microperfurações na tróclea lateral do fêmur em equinos.(AU)

The aim of this study was to evaluate the joint cartilage repair by macroscopic (via arthroscopy) and histological (biopsy fragments) analyses in chondral defects induced into equine femoral trochlea treated by microperforation associated with or without intra-articular administration of kartogenin. Six horses weighing 342±1.58 kg (mean ± SD), aged approximately 7.2±1.30 years and with a body condition score of 7.1±0.75, were used. The horses underwent arthroscopy for induction of 1-cm2 chondral lesions in lateral femoral trochlea immediately treated by microperforation of the subchondral bone of both knees. Four weekly intra-articular injections of kartogenin (20μM) in one knee (treated group) and Ringer lactate solution in the contralateral joint (control group) were performed during the postoperative period. After 60 days, macroscopic evaluations were performed by video-arthroscopy, and biopsy samples of the repair tissue were taken for histopathological healing evaluation. No significant change was observed in macroscopic and histological scores for chondral healing between treated and untreated groups (P>0.05). The overall mean percentage of hyaline cartilage in both groups (17.5%) was consistent with other international studies using other types of chondral microperforation; however, no statistical differences were observed between groups (P>0.05). In conclusion, the therapy with kartogenin, according to the used protocol, did not produce any macroscopic and histological healing improvement in induced chondral lesions treated with microperforations in equine femoral trochlea.(AU)

Chondroitin sulfate and glucosamine in the cartilage and subchondral bone repair of dogs - Histological findings / Sulfato de condroitina e glucosamina na reparação da cartilagem e do osso subcondral de cães - Achados histológicos

Chondroitin and glucosamine sulfate nutraceuticals are commonly used in the management of degenerative articular disease in veterinary routine. However, there are controversies on the contribution of these substances to articular cartilage. The purpose of this study was to evaluate the efficiency of a chondroitin and glucosamine sulfate-based veterinary nutraceutical on the repair of an induced osteochondral defect in a dog femoral condyle, by macroscopic, histological and histomorphometric analyses. The nutraceutical was orally administered the day following injury induction, every 24 hours (treated group, TG, n=24), compared with animals that did not receive the product (control group, CG, n=24). Six animals per group were anaesthetized for sample collection at 15, 30, 60 and 90 days after surgery. At 15 days, defects were macroscopically filled with red-pinkish tissue. After 30 days, whitish color tissue was observed, both in TG and CG animals, with firmer consistency to touch at 60 and 90 postoperative days. Histological analysis demonstrated that, in both groups, there was initial blood clot formation, which was subsequently substituted by a fibrin net, with capillary proliferation from the adjacent bone marrow and infiltration of mesenchymal cells in clot periphery. As cellular differentiation developed, repair tissue presented a fibrocartilage aspect most of the time, and new subchondral bone formation occurred in the deepest area corresponding to the defect. Histomorphometry suggested that the nutraceutical did not favor the articular cartilage repair process. It was concluded that nutraceutical did not significantly influence chondrocytes proliferation or hyaline architecture restoration.(AU)

Os nutracêuticos compostos de sulfato de condroitina e glucosamina são comumente utilizados no manejo da doença articular degenerativa na rotina veterinária. Entretanto, existem controvérsias sobre a contribuição dessas substâncias à cartilagem articular. O objetivo deste trabalho foi avaliar a eficácia de um nutracêutico veterinário à base de sulfato de condroitina e glucosamina na reparação de defeitos osteocondrais induzidos no côndilo femoral de cães, através de análises macroscópica, histológica e histomorfométrica. O nutracêutico foi administrado no dia seguinte à indução da lesão, pela via oral, a cada 24 horas (grupo tratado - GT, 24 animais), sendo comparado a animais que não receberam o produto (grupo controle - GC, de igual número de animais). Aos 15, 30, 60 e 90 dias após a cirurgia, seis animais por grupo foram anestesiados para ser realizada a coleta das amostras. Aos 15 dias, os defeitos eram macroscopicamente preenchidos por tecido de coloração rósea a avermelhada. Já a partir dos 30 dias, observou-se preenchimento por tecido de coloração esbranquiçada, tanto nos animais do GT quanto nos do GC, com consistência mais firme ao toque digital aos 60 e 90 dias de pós-operatório. A análise histológica revelou que, em ambos os grupos, houve inicialmente formação de coágulo sanguíneo que, posteriormente, foi substituído por uma rede de fibrina, com proliferação de capilares a partir da medula óssea adjacente e infiltração de células mesenquimais na periferia do coágulo. À medida que se processou a diferenciação celular, o tecido de reparação se apresentou na maioria das vezes com aspecto de fibrocartilagem e, na região mais profunda da área correspondente ao defeito, ocorreu formação de osso novo subcondral. A histomorfometria sugeriu que o nutracêutico não favoreceu o processo de reparação da cartilagem articular. Concluiu-se que o nutracêutico não influenciou consideravelmente na proliferação de condrócitos nem na restauração da arquitetura hialina.(AU)

Study on nano-structured hydroxyapatite/zirconia stabilized yttria on healing of articular cartilage defect in rabbit

PURPOSE: Articular Cartilage has limited potential for self-repair and tissue engineering approaches attempt to repair articular cartilage by scaffolds. We hypothesized that the combined hydroxyapatite and zirconia stabilized yttria would enhance the quality of cartilage healing. METHODS: In ten New Zealand white rabbits bilateral full-thickness osteochondral defect, 4 mm in diameter and 3 mm depth, was created on the articular cartilage of the patellar groove of the distal femur. In group I the scaffold was implanted into the right stifle and the same defect was created in the left stifle without any transplant (group II). Specimens were harvested at 12 weeks after implantation, examined histologically for morphologic features, and stained immunohistochemically for type-II collagen. RESULTS: In group I the defect was filled with a white translucent cartilage tissue In contrast, the defects in the group II remained almost empty. In the group I, the defects were mostly filled with hyaline-like cartilage evidenced but defects in group II were filled with fibrous tissue with surface irregularities. Positive immunohistochemical staining of type-II collagen was observed in group I and it was absent in the control group. CONCLUSION: The hydroxyapatite/yttria stabilized zirconia scaffold would be an effective scaffold for cartilage tissue engineering.(AU)

Plasma rico em plaquetas no tratamento de lesões condrais articulares induzidas experimentalmente em equinos: avaliação clínica, macroscópica, histológica e histoquímica / Platelet-rich plasma in the treatment of articular chondral defects experimentally induced in horses: clinical, macroscopic, histopathological and histochemical evaluation

Estudou-se a eficácia do plasma rico em plaquetas (PRP) no tratamento de lesões condrais articulares, experimentalmente induzidas em equinos. Para isso, foi induzida uma lesão condral, na tróclea medial femoral dos dois membros pélvicos de quatro animais. Após 30 dias da indução, as oito articulações foram divididas em dois grupos. Os animais do grupo 1 receberam o tratamento intralesional e intra-articular com PRP, e os do grupo 2 foram tratados apenas com solução fisiológica. As avaliações clínicas, constituídas de exames de claudicação e análises do líquido sinovial, foram realizadas antes da indução da lesão - tempo zero -, quinzenalmente, até 120 dias e aos 150 dias. Avaliações macroscópicas, histológicas e histoquímicas foram realizadas no tempo zero e aos 150 dias. Os equinos do grupo 1 apresentaram melhora do grau de claudicação em relação aos do grupo 2. Os exames macroscópicos, histológicos e histoquímicos revelaram melhor tecido de reparação igualmente no grupo 1. Concluiu-se que a administração de PRP apresentou efeitos benéficos no tratamento de lesões condrais experimentais de equinos.

The use of platelet-rich plasma (PRP) in the treatment of articular cartilage defects induced experimentally in horses was studied. For this purpose, both patellofemoral joints of four animals were approached through arthroscopic surgery to perform a cartilage defect on the medial femoral trochlea. After 30 days of induction the eight joints were divided into two groups. Group 1 animals received intralesional and intra-articular treatment with PRP and Group 2 animals were treated only with saline solution. The clinical assessments, constituted by lameness signs and synovial fluid analysis, were performed before induction of injury (time zero) and every 15 days in 120 days, with last analysis on day 150. The macroscopic and morphologic analysis were performed at 0 and 150 days. During the experiment Group 1 animals showed improvement in lameness when compared to Group 2 animals. The macroscopic and morphological analysis showed a better tissue repair in the treated horses. Thus, the implantation of autologous PRP showed beneficial effects in the treatment of chondral lesions, experimentally induced in horses.

Plasma rico em plaquetas no tratamento de lesões condrais articulares induzidas experimentalmente em equinos: avaliação clínica, macroscópica, histológica e histoquímica / Platelet-rich plasma in the treatment of articular chondral defects experimentally induced in horses: clinical, macroscopic, histopathological and histochemical evaluation

Estudou-se a eficácia do plasma rico em plaquetas (PRP) no tratamento de lesões condrais articulares, experimentalmente induzidas em equinos. Para isso, foi induzida uma lesão condral, na tróclea medial femoral dos dois membros pélvicos de quatro animais. Após 30 dias da indução, as oito articulações foram divididas em dois grupos. Os animais do grupo 1 receberam o tratamento intralesional e intra-articular com PRP, e os do grupo 2 foram tratados apenas com solução fisiológica. As avaliações clínicas, constituídas de exames de claudicação e análises do líquido sinovial, foram realizadas antes da indução da lesão - tempo zero -, quinzenalmente, até 120 dias e aos 150 dias. Avaliações macroscópicas, histológicas e histoquímicas foram realizadas no tempo zero e aos 150 dias. Os equinos do grupo 1 apresentaram melhora do grau de claudicação em relação aos do grupo 2. Os exames macroscópicos, histológicos e histoquímicos revelaram melhor tecido de reparação igualmente no grupo 1. Concluiu-se que a administração de PRP apresentou efeitos benéficos no tratamento de lesões condrais experimentais de equinos.(AU)

The use of platelet-rich plasma (PRP) in the treatment of articular cartilage defects induced experimentally in horses was studied. For this purpose, both patellofemoral joints of four animals were approached through arthroscopic surgery to perform a cartilage defect on the medial femoral trochlea. After 30 days of induction the eight joints were divided into two groups. Group 1 animals received intralesional and intra-articular treatment with PRP and Group 2 animals were treated only with saline solution. The clinical assessments, constituted by lameness signs and synovial fluid analysis, were performed before induction of injury (time zero) and every 15 days in 120 days, with last analysis on day 150. The macroscopic and morphologic analysis were performed at 0 and 150 days. During the experiment Group 1 animals showed improvement in lameness when compared to Group 2 animals. The macroscopic and morphological analysis showed a better tissue repair in the treated horses. Thus, the implantation of autologous PRP showed beneficial effects in the treatment of chondral lesions, experimentally induced in horses.(AU)

Reparação de defeitos osteocondrais de cães com implante de cultura de condrócitos homólogos e membrana biossintética de celulose: avaliação clínica, ultrassonográfica e macroscópica / The repair of osteochondral defects in dogs with homologous articular chondrocytes and biosynthetic cellulose membrane: clinical, ultrasound and macroscopic evaluation

Avaliou-se o implante de condrócitos homólogos em lesões osteocondrais, utilizando a membrana biossintética à base de celulose (MBC) como revestimento. Dez cães adultos e clinicamente sadios foram submetidos à artrotomia das articulações fêmoro-tíbio-patelares. Defeitos de quatro milímetros de diâmetro por quatro milímetros de profundidade foram induzidos na tróclea femoral de ambos os membros. A MBC foi aplicada na base e superfície das lesões. Os defeitos do membro direito foram preenchidos com condrócitos homólogos cultivados e formaram o grupo tratado (GT); e os defeitos do membro esquerdo, sem implante celular, formaram o grupo controle (GC). Os animais foram avaliados clínica e ultrassonograficamente aos 30 e 60 dias. A evolução pós-operatória dos cães foi analisada com especial interesse nos processos de reparação da lesão, por meio de macroscopia. Não houve diferença clínica e ultrassonográfica entre os grupos. Entretanto, à macroscopia, ocorreu maior prevalência de formação de tecido cicatricial esbranquiçado no GT. O tecido neoformado apresentou melhor qualidade associado ao implante homólogo de condrócitos, mas não promoveu reparação por cartilagem hialina.(AU)

The aim of the study was to evaluate the repair of deep cartilaginous defects made in the femoral trochlear sulcus of dogs, using the cellulose biosynthetic membrane (CBM) as coating. Ten healthy adult dogs without locomotor disorders were used. All animals were submitted to arthrotomy of stifle joints and defects with four millimeters diameter x four millimeters deep were done in the femoral trochlear sulcus of both limbs. CBM was applied in the lesion's base and surface of all limbs. In the treated group (TG), the defects of the right limb were filled with cultivated homologous chondrocytes, and in control group (CG), the defects of the left limb were filled without cellular implant. The animals were evaluated by physical examination and ultrasound at 30 and 60 days. The postoperative follow up of the dogs was done by macroscopy with special interest in the healing process of the osteochondral defect. No clinical and ultrasonographic differences were observed in both groups. In the macroscopic evaluation higher prevalence of whitish scar tissue formation was noted in TG, but without statistical difference. The neoformed tissue showed slightly higher quality in TG, but without promoting repair by the hyaline cartilage.(AU)

Effect of dithiocarbamate thiram on Wistar rat growth plate and articular cartilage / Efeito do ditiocarbamato tirame sobre a placa de crescimento e cartilagem articular de ratos Wistar

Avaliou-se o efeito do tirame, ditiocarbamato largamente utilizado na agricultura como antifúngico e repelente de roedores, na ossificação endocondral de mamíferos, usando, como modelo, ratos Wistar. Não foram observadas lesões na cartilagem articular, nem nas placas de crescimento, o que pode ser atribuído à dose utilizada e à duração do ensaio. A diminuição da altura da placa de crescimento nos animais aos quais foi administrado o tirame parece traduzir o atraso verificado no crescimento em geral, e não um efeito específico na cartilagem, uma vez que as diferentes zonas da placa epifisária mantiveram as proporções dos animais do grupo-controle. Embora não tenham sido verificados, no presente trabalho, os efeitos registrados para outras espécies nos tecidos cartilaginosos, sugere-se a avaliação dos efeitos crónicos do tirame no crescimento e no desenvolvimento dos ossos longos em mamíferos.(AU)