Spontaneous osteoarthritis in dogs - clinical effects of single and multiple intra-articular injections of hyaluronic acid

Background: The treatments of osteoarthritis (OA) are commonly conservative and multimodal to relieve pain and improve movement. Intra-articular injection of hyaluronic acid (IAHA) has been studied as a treatment option for OA in dogs. IAHA helps restore the viscoelasticity of the synovial fluid and relieves the clinical symptoms of OA. However, the efficacy of IAHA in dogs is still a controversial subject. This study aims to confirm the IAHA effect in dogs with spontaneous OA and to compare the effectiveness depending on the number of injections. Materials, Methods & Results: Thirty dogs with spontaneous OA were assigned to a single injection group (n=17) and a 3-weekly injections group (n=13). Dogs weighing less than 10 kg were injected 1 mL of HA to the OA joint, and more than 10 kg dogs were injected 2 mL of HA. In the case of the 3-weekly injections group, the same amount was administered 3 times at 1-week intervals. After the injection, physical and orthopedic examinations were performed to check for complications. Radiographic OA score was evaluated before and 3 months after the injection to confirm and to evaluate the progression of OA. Clinical symptom evaluations were performed on pre-injection, 1-, 2-, and 3-months post-injection. They consisted of the clinical lameness score by veterinarians and Canine Brief Pain Inventory (CBPI) by owners. Results were compared with unpaired t-test, repeated-measures ANOVA with Tukey's or Sidak's multiple comparison test, or Wilcoxon test, with P < 0.05. Patients had a median age of 9 years (range 3 to 16 years) and a bodyweight of 4.8 kg (range 2 to 48 kg). No systemic side effects or major complications were detected during the trial period. IAHA produced temporary pain and discomfort in 6 cases. There was no change in the radiographic OA score before and 3 months after injections in both groups, and the difference between groups was not confirmed. In both groups, the clinical lameness score significantly decreased at 1, 2, 3 months after injection compared with pre-injection. The score was lower at 3 months after the injection than at 1 month. The clinical lameness score had no significant difference between the groups. Similarly, CBPI was all decreased in the single injection group and 3-weekly injections group compared to pre-injection, and the score at 3 months post-injection was lower than at 1 month. No significant differences between the groups were found in CBPI. Discussion: Most studies on the efficacy of IAHA in canine OA have been conducted using an experimental model, so studies on spontaneous canine OA are insufficient. This study confirmed that IAHA improves clinical symptoms such as pain relief and movement improvement in spontaneous OA dogs using CBPI and clinical lameness score. In order to confirm the optimal IAHA protocol, a single IAHA and 3-weekly IAHA were compared. The result shows that clinical symptoms improved in both single and 3-weekly injections groups, but no significant difference was confirmed during the 3-month study period. These findings may suggest that a single IAHA may have a similar effect to multiple IAHA, and repeated injections are unnecessary. In humans and canine OA models, it is reported that the effect of IAHA was maintained for 6 months. This study showed that the effect of IAHA was maintained for 3 months study period and that clinical symptoms improved at 3 months than at 1 month. In conclusion, these findings suggested that IAHA improves clinical symptoms in dogs with spontaneous OA, and a single IAHA showed a similar effect to 3 weekly IAHA.

Hyaluronic acid in tobacco-exposed rats. Inflammatory reaction, and duration of effect

Purpose:To evaluate the hyaluronic acid (HA) inflammatory reaction, fibroblasts, fibrosis and duration of effect in the dorsal region of tobacco-exposed rats.Methods:Ten Wistar rats were divided into two groups: tobacco-exposed-group (TEG;n=5) and air-control-group (CG;n=5). The TEG animals were tobacco-exposed twice a day, 30-minutes/session, during 60 days. After this period, all animals received 0.1 mL HA subcutaneous injection in the dorsal area. The volume of HA was measured immediately after HA injection and weekly using a hand-caliper in nine weeks. After this period, all the animals were euthanized, and a specimen of was collected to evaluate inflammatory cells, fibroblasts, and fibrosis by HE.Results:This study showed a higher inflammatory reaction in TEG than CG: inflammatory cell-count (CG: 1.07±0.9; TEG: 8.61±0.36, p<0.001); fibroblast count (CG: 2.92±0.17; TEG: 19.14±0.62, p<0.001), and fibrosis quantification (CG: 2.0; TEG: 3.75, p<0.001). The analysis of the HA volume in nine weeks in the dorsal region did not show a difference between groups (p=0.39).Conclusions:This study suggested that the HA injection in the TEG caused an increase in inflammatory cell count, fibroblast, and fibrosis quantification when compared to the CG. There was no difference in the duration of effect of HA between the groups.(AU)

Hyaluronic acid and synovial fluid in preventing adhesion formation after tenorrhaphy: An in vivo study on rabbit Achilles tendon / Ácido hialurônico e líquido sinovial na prevenção da formação de aderências após tenorrafia: Um estudo in vivo sobre o tendão de Aquiles de coelho

Peritendinous adhesions are a significant problem in tendon surgery. One of the most preferred methods for preventing adhesion formation is anti-adhesive applications. This study aimed to investigate the efficacy of natural synovial fluid as an alternative viscoelastic agent for preventing adhesion in the healing period compared to hyaluronic acid. Thirty adult New Zealand rabbits were used in the study to form three experimental models. In all groups, a rupture was induced in the Achilles tendons. Following tenorrhaphy, hyaluronic acid was applied in one group and synovial fluid in the other group for anti-adhesive purposes. In the control group, no viscoelastic application was made. Bandage was applied to the operated extremities for three weeks post-operatively. At the end of five weeks, euthanasia was performed on the animals using pentobarbital sodium. Surgical sites in all groups were macroscopically examined for healing status as well as the presence and extent of adhesions. Biomechanical tests and histopathological examinations were then performed on the tendons in all groups. Findings established once again the positive contribution of hyaluronic acid to preventing adhesion formation as well as to healing and tensile strength in tendon surgery. Although, we found that it is possible to use synovial fluid, which is a natural source of hyaluronic acid, as a viscoelastic material, it is not superior to commercial hyaluronic acid preparation.(AU)

Asderências peritendinares são um problema significativo na cirurgia tendínea. Um dos métodos mais empregados para evitar a formação de adesão são as aplicações anti-adesivas. Este estudo tem como objetivo investigar a eficácia do líquido sinovial natural como um agente viscoelástico alternativo para prevenir a adesão no período de cicatrização comparado ao ácido hialurônico. Trinta coelhos adultos da Nova Zelândia foram utilizados no estudo para formar três modelos experimentais. Em todos os grupos, uma ruptura foi induzida nos tendões de Aquiles. Após tenorrafia, o ácido hialurônico foi aplicado em um grupo e o líquido sinovial no outro grupo para fins anti-adesivos. No grupo controle, não foi feita qualquer aplicação viscoelástica. A bandagem foi aplicada às extremidades operadas por três semanas pós-operatório. No final de cinco semanas, a eutanásia foi realizada nos animais utilizando pentobarbital sódico. Os locais cirúrgicos em todos os grupos foram examinados macroscopicamente quanto ao estado de cura, bem como a presença e extensão das aderências. Testes biomecânicos e exames histopatológicos foram realizados nos tendões em todos os grupos. Os achados estabeleceram mais uma vez a contribuição positiva do ácido hialurônico para prevenir a formação de aderências, bem como para a cicatrização e resistência à tração na cirurgia do tendão. Embora se tenha constatado que é possível utilizar fluido sinovial, que é uma fonte natural de ácido hialurónico, como material viscoelástico, não é superior à preparação de ácido hialurónico comercial.(AU)

Evaluation of the effectiveness of sodium hyaluronate, sesame oil, honey, and silver nanoparticles in preventing postoperative surgical adhesion formation. An experimental study

Purpose: To evaluate the effectiveness of sodium hyaluronate, sesame oil, honey, and silver nanoparticles in preventing of postoperative surgical adhesion formation. Methods: Forty male Wistar rats were randomly assigned into five groups with eight rats in each group including control, hyaluronate, sesame, honey and silver groups. After two weeks the animals underwent laparotomy and were evaluated by two different blinded surgeons for severity of adhesions based on the two different classification scoring systems including Nair classification and cumulative adhesion scoring scale. Results: The scores of severity of adhesions in the hyaluronate and sesame groups were significantly lower than the control group based on the Nair classification (both P-values = 0.02), however based on the cumulative adhesion scoring scale just the score of severity of adhesions in the hyaluronate group was significantly lower than the control group (P-value = 0.02). In the hyaluronate group the severity of adhesions was decreased by 48% based on the cumulative adhesion scoring scale. Conclusions: Sodium hyaluronate and sesame oil may have a significant effect in preventing postoperative surgical adhesion formation.(AU)

Bullous keratopathy in a yellow-headed caracara (Milvago chimachima) treated with a modified third eyelid flap / Ceratopatia bolhosa em caracará (Milvago chimachima) tratado com flap de terceira pálpebra modificado

A Yellow-headed Caracara (Milvago chimachima) was submitted to the Ophthalmology Service of the Federal University of Bahia with a corneal abnormality. During ophthalmic evaluation the right cornea was stained positively with fluorescein; a blurred bullous lesion, with irregular surface, compatible with the diagnosis of bullous keratopathy was found. This is a rare condition in a bird which was treated successfully with a modified third eyelid flap associated with antibiotic and hyaluronic acid eye drops. The adopted therapeutic proved to be simple to implement and viable for repair of the bullous keratopathy in the Yellow-headed caracara.(AU)

Um gavião carrapateiro (Milvago chimachima) foi encaminhado ao Serviço de Oftalmologia Veterinária da Universidade Federal da Bahia com alteração em córnea. Durante a avaliação oftálmica a prova da fluoresceína foi positiva no olho direito e foi observado lesão bolhosa de aspecto irregular, compatível com o diagnóstico de ceratopatia bolhosa. Esta é uma alteração rara em aves e no presente caso foi tratada através de "flap" modificado de terceira pálpebra associado a utilização de colírio antibiótico e ácido hialurônico. A terapêutica adotada foi de fácil implementação e viável no reparo da ceratopatia bolhosa em gavião carrapateiro.(AU)

Bullous keratopathy in a yellow-headed caracara (Milvago chimachima) treated with a modified third eyelid flap / Ceratopatia bolhosa em caracará (Milvago chimachima) tratado com flap de terceira pálpebra modificado

A Yellow-headed Caracara (Milvago chimachima) was submitted to the Ophthalmology Service of the Federal University of Bahia with a corneal abnormality. During ophthalmic evaluation the right cornea was stained positively with fluorescein; a blurred bullous lesion, with irregular surface, compatible with the diagnosis of bullous keratopathy was found. This is a rare condition in a bird which was treated successfully with a modified third eyelid flap associated with antibiotic and hyaluronic acid eye drops. The adopted therapeutic proved to be simple to implement and viable for repair of the bullous keratopathy in the Yellow-headed caracara.(AU)

Um gavião carrapateiro (Milvago chimachima) foi encaminhado ao Serviço de Oftalmologia Veterinária da Universidade Federal da Bahia com alteração em córnea. Durante a avaliação oftálmica a prova da fluoresceína foi positiva no olho direito e foi observado lesão bolhosa de aspecto irregular, compatível com o diagnóstico de ceratopatia bolhosa. Esta é uma alteração rara em aves e no presente caso foi tratada através de flap modificado de terceira pálpebra associado a utilização de colírio antibiótico e ácido hialurônico. A terapêutica adotada foi de fácil implementação e viável no reparo da ceratopatia bolhosa em gavião carrapateiro.(AU)

Cashew apple juice as microbial cultivation medium for non-immunogenic hyaluronic acid production

In this work, natural cashew apple juice was used as cultivation medium as an alternative to substitute brain heart infusion medium. The effect of aeration and juice supplementation with yeast extract on the production of hyaluronic acid in batch fermentation was also investigated. Similar levels of cell mass were obtained in inoculum using cashew apple juice supplemented with yeast extract or the conventional brain heart infusion medium. Fermentation in Erlenmeyer flasks produced low biomass and hyaluronic acid concentrations. The hyaluronic acid concentration and viscosity increased from 0.15 g/L and 3.87 cP (no aeration or medium supplementation) to 1.76 g/L and 107 cP, when aeration (2 vvm) and 60 g/L of yeast extract were used. The results suggest the production of low-molecular weight hyaluronic acid oligomers instead of the high molecular weight polymer.

Cashew apple juice as microbial cultivation medium for non-immunogenic hyaluronic acid production

In this work, natural cashew apple juice was used as cultivation medium as an alternative to substitute brain heart infusion medium. The effect of aeration and juice supplementation with yeast extract on the production of hyaluronic acid in batch fermentation was also investigated. Similar levels of cell mass were obtained in inoculum using cashew apple juice supplemented with yeast extract or the conventional brain heart infusion medium. Fermentation in Erlenmeyer flasks produced low biomass and hyaluronic acid concentrations. The hyaluronic acid concentration and viscosity increased from 0.15 g/L and 3.87 cP (no aeration or medium supplementation) to 1.76 g/L and 107 cP, when aeration (2 vvm) and 60 g/L of yeast extract were used. The results suggest the production of low-molecular weight hyaluronic acid oligomers instead of the high molecular weight polymer.

Practical aspects for implementing in vitro embryo production and cloning programs in sheep and goats

In vitro embryo production has the potential to produce more offspring from genetically valuable animals than standard MO ET as it is capable of avoiding most of the causes of failure in MOET (poor response to superovulation, poor fertilization and premature luteolysis). It also allows repeating collection in the donor animals more often and more times in their reproductive life. However, consistent results are not easy to obtain when conducting large scale programs, mainly due to variability associated with the in vitro fertilization results. Cloning by somatic cell nuclear transfer has been used by a few groups to successfully produce genetic copies of individuals of high genetic merit, but remains to be a very inefficient reproduction technology even in the hand s of those that have been successful in producing multiple live clones. This review provides a collection of tips and points to consider when planning in vitro embryo production and cloning programs in sheep and goats.ien

Practical aspects for implementing in vitro embryo production and cloning programs in sheep and goats

In vitro embryo production has the potential to produce more offspring from genetically valuable animals than standard MO ET as it is capable of avoiding most of the causes of failure in MOET (poor response to superovulation, poor fertilization and premature luteolysis). It also allows repeating collection in the donor animals more often and more times in their reproductive life. However, consistent results are not easy to obtain when conducting large scale programs, mainly due to variability associated with the in vitro fertilization results. Cloning by somatic cell nuclear transfer has been used by a few groups to successfully produce genetic copies of individuals of high genetic merit, but remains to be a very inefficient reproduction technology even in the hand s of those that have been successful in producing multiple live clones. This review provides a collection of tips and points to consider when planning in vitro embryo production and cloning programs in sheep and goats.(AU)ien

Optimization of flask culture medium and conditions for hyaluronic acid production by a streptococcus equisimilis mutant nc2168

A mutant designated NC2168, which was selected from wild-type Streptococcus equisimilis CVCC55116by ultraviolet ray combined with60Co-γ ray treatment and does not produce streptolysin, was employed to produce hyaluronic acid (HA). In order to increase the output of HA in a flask, the culture medium and conditions for NC2168 were optimized in this study. The influence of culture medium ingredients including carbon sources, nitrogen sources and metal ions on HA production was evaluated using factional factorial design. The mathematical model, which represented the effect of each medium component and their interaction on the yield of HA, was established by the quadratic rotary combination design and response surface method. The model estimated that, a maximal yield of HA could be obtained when the concentrations of yeast extract, peptone, glucose, and MgSO4 were set at 3 g/100 mL, 2 g/100 mL, 0.5 g/100 mL and 0.15 g/100 mL, respectively. Compared with the values obtained by other runs in the experimental design, the optimized medium resulted in a remarkable increase in the output of HA and the maximum of the predicted HA production was 174.76 mg/L. The model developed was accurate and reliable for predicting the production of HA by NC2168.Cultivation conditions were optimized by an orthogonal experimental design and the optimal conditions were as follows: temperature 33ºC, pH 7.8, agitation speed 200 rpm, medium volume 20 mL.(AU)

Comparação terapêutica entre o ácido hialurônico e o plasma rico em plaquetas em potros brasileiro de hipismo com osteocondrose dissecante em crista intermedia da tíbia submetidos a vídeo artroscopia.

Pereira, M.F. Comparação terapêutica entre o ácido hialurônico e o plasma rico em plaquetas em potros brasileiro de hipismo com osteocondrose dissecante em crista intermédia de tíbia submetidos a vídeo artroscopia. 2017. 86f. Dissertação de Mestrado. Faculdade de Zootecnia e Engenharia de Alimentos, Universidade de São Paulo, Pirassununga, 2017. Enfermidade muito observada nas articulações dos equinos, o processo degenerativo articular crônico na articulação tibio-társica pode ser de origem primária, ou secundária à osteocondrose dissecante. A confirmação diagnóstica é feita por avaliação radiográfica. Importantes aliados diagnósticos são a avaliação do líquido sinovial, a avaliação ultrassonografia e a cirurgia de video-artroscopia, sendo esta última o tratamento para a enfermidade. Por sua evolução crônica e às vezes assintomática, a osteocondrose dissecante causa alterações no ambiente articular, como a perda de matriz cartilagínea, o que limita o potencial atlético do animal, acarretando em queda na performace e perdas econômicas. É comum o uso terapêutico intra-articular no periodo pós operatório recente de substâncias autólogas como o plasma rico em plaquetas (PRP), bem como o uso ácido hialurônico (HA), como adjuvante para o tratamento cirúrgico. O presente trabalho objetiva comparar e avaliar estes tratamentos. Foram utilizados 18 equinos, com idade variando entre 2 a 6 anos de idade, da raça Brasileiro de Hipismo, com peso corpóreo entre 400 e 650 kg, oriundos do mesmo criatório, com osteocondrose dissecante, na crista intermédia do osso tíbia. Estes animais no décimo dia pós operatório são dividos em três grupos experimentais: grupo AH, grupo PRP e grupo RCL como controle. Dados clínicos, ultrassonográficos e laboratoriais foram coletados em tempos definidos. Os resultados demonstraram que os animais do grupo controle grupo RCL, que receberam solução de ringer com lactato intraarticular, apresentaram respostas clínicas e laboratoriais que indicaram menor indução do processo inflamatório ao ambiente articular quando comparado aos grupos PRP e AH. Os animais que receberam PRP apresentaram maior efusão sinovial e resposta positiva ao teste de flexão quando comparados aos animais que receberam AH; ao passo que a administração intra-articular de AH induziu maiores alteração citológica do líquido sinovial quando comparadas aos animais que receberam PRP.

Therapeutic comparison between hialuronic acid and platelet rich plasma in brasileiro de hipismo show jumpers young horses with osteochondrosis dissecans in tibial intermidiate ridge submitted to video arthroscopy surgery. Master Science Degree, 2017. 86 p. Faculdade Zootecnia e Engenharia de Alimentos, Universidade de São Paulo, Pirassununga, 2017. One of the most commom diseases in the equine joint, the degenerative joint disease in the tibio tarsal joint could be a primary disease, or secondary to osteochondrosis dissecans. The diagnosis is made by radiographic study. Allies in diagnosing this disease includes sinovial fluid evaluation, ultrasonographic evaluation and video arthroscopy surgery, which is the way to treat this disease. The osteochondrosis dissecans has a chronic evolution, and several times asymptomatic, which causes an alteration on articular environment that contributes to a loss in the cartilage matrix. Resulting in the limited development of the athletic potential of the animal, poor performance and economic losses. Is common to use therapeutical autholog substances intra articular as the platelet rich plasma, and hialuronic acid, in the early post surgical treatment as an adjuvant therapy to the surgical treatment. This study reaches to compare and evaluate those treatments. Were used 18 horses, with ages between 2 to 6 years old, Brasileiro de Hipismo breed, weighting between 400 and 650 kg, from the same breeding center, with osteochondrosis dissecans fragments from the intermediate tibial crest. Those patients, 10 days after the video arthroscopy surgery, were divided in three experimental groups: AH group, PRP group, and RCL group as control group. Laboratorial data, clinical signs and ultrassonographic exam were performed. The results proved that RCL group had show less induction of inflamatory process on articular enviroment, when we compare with PRP group and AH group. The PRP group had shown more synovial effusion, and more sensibility to the flexion test when we compare to AH group; but the AH group had shown more laboratorial alterations when we compare to the PRP group.

Engraftment of human adipose derived stem cells delivered in a hyaluronic acid preparation in mice / Implante de células tronco do tecido adiposo humano numa preparação de ácido hialurônico em camundongos

PURPOSE: To evaluate the implant of human adipose derived stem cells (ADSC) delivered in hyaluronic acid gel (HA), injected in the subcutaneous of athymic mice. METHODS: Control implants -HA plus culture media was injected in the subcutaneous of the left sub scapular area of 12 athymic mice. ADSC implants: HA plus ADSC suspended in culture media was injected in the subcutaneous, at the contra lateral area, of the same animals. With eight weeks, animals were sacrificed and the recovered implants were processed for extraction of genomic DNA, and histological study by hematoxilin-eosin staining and immunufluorescence using anti human vimentin and anti von Willebrand factor antibodies. RESULTS: Controls: Not visualized at the injection site. An amorphous substance was observed in hematoxilin-eosin stained sections. Human vimentin and anti von Willebrand factor were not detected. No human DNA was detected. ADSC implants - A plug was visible at the site of injection. Fusiform cells were observed in sections stained by hematoxilin- eosin and both human vimentin and anti von Willebrand factor were detected by immunofluorescence. The presence of human DNA was confirmed. CONCLUSION: The delivery of human adipose derived stem cells in preparations of hyaluronic acid assured cells engraftment at the site of injection.(AU)

OBJETIVO: Avaliar o implante de células tronco do tecido adiposo humano (CTTAH) em gel de ácido hialurônico (AH), injetados no tecido subcutâneo de camundongos atímicos. MÉTODOS: Implantes controle - HA com meio de cultura foram injetados no tecido subcutâneo da região infraescapular esquerda de 12 camundongos atímicos. Implantes de CTTAH: HA com CTTAH suspensas em meio de cultura foi injetado no subcutâneo da região contra lateral, dos mesmos animais. Com oito semanas, os animais foram sacrificados e os implantes recuperados foram processados para extração de DNA genômico, estudo histológico por coloração por hematoxilina eosina e imnuoflurescência utilizando anticorpos anti vimentina humana e anti fator de von Willebrand. RESULTADOS: Controles - implantes não visualizados no local da injeção. Uma substância amorfa foi observada nos cortes corados por hematoxilina eosina. Vimentina humana e fator anti von Willebrand não foram identificados. DNA humano não foi detectado. Implantes de CTTAH - Uma massa era visível no local da injeção. Células fusiformes foram observadas nos corte corados com hematoxilina eosina. Tanto vimentina humana quanto fator de von Willebrand foram identificados pela imunofluorescência. A presença de DNA humano foi confirmada. CONCLUSÃO: O implante de células tronco do tecido adiposo humano em veículo de ácido hialurônico gel assegurou a manutenção das células no local do implante.(AU)

Comparação dos efeitos da aplicação intra-articular de ácido hialurônico de diferentes pesos moleculares em modelo de sinovite aguda induzida por LPS em equinos

Diversos experimentos em humanos e equinos discutem a eficácia do uso intra-articular do ácido hialurônico (AH). Pouco se sabe a respeito de seu potencial pró ou anti-inflamatório, bem como sobre a eficácia das moléculas de AH de diferentes pesos moleculares. O objetivo deste estudo foi avaliar os efeitos anti-inflamatórios, anabólicos e antioxidantes da aplicação intra-articular de AH em sinovite aguda induzida por lipopolissacarídeo (LPS) em equinos. Foram utilizadas 24 articulações metacarpofalangeanas hígidas de 12 equinos jovens, avaliadas por exame físico, radiográfico e ultrassonográfico. As articulações foram divididas aleatoriamente em três grupos de oito, sendo o grupo controle tratado com triancinolona (TA) (10 mg), o grupo AH de baixo peso molecular (AH-BPM), e o grupo AH de alto peso molecular (AH-APM). Inicialmente, foram aplicados 0,25 ng de LPS por via intra-articular (momento 0). Após uma hora (momento 1), coletou-se líquido sinovial (LS) para contagem celular e imediatamente após realizado o tratamento com TA, AH-BPM ou AH-APM. Posteriormente, LS foi coletado nos momentos 8, 24 e 48 horas para contagem celular, avaliação do burst oxidativo, quantificação de prostaglandina E2 (PGE2), condroitim sulfato (CS), AH e mensuração do peso molecular do AH. Exame físico dos animais, incluindo mensuração da circunferência articular e avaliação de claudicação por meio do equipamento Lameness Locator® foi realizado em todos os momentos. As frequências cardíaca e respiratória dos animais aumentaram em todos os grupos às 8 horas (P<0,05), sendo menor no grupo TA. Em relação à circunferência articular o grupo AH-BPM apresentou maior aumento às 24 e 48 horas em relação aos outros grupos (p<0,05). O grau de claudicação do grupo AH-APM foi maior que o do grupo TA às 8 e 24 horas, mas similar ao grupo AH-BPM. O grupo TA apresentou maior quantidade de células nucleadas no LS do que o grupo AH-BPM as 24 e 28 horas e do que o grupo AH-APM às 48 horas. A concentração de PGE2 foi menor no grupo TA em relação ao grupo AH-APM às 8 horas, e a ambos os grupos que receberam AH às 48 horas (p<0,05). O grupo AH-APM mostrou maior concentração de AH às 8 horas do que os outros grupos, contudo às 48 horas o grupo TA mostrou maior concentração de AH do que os outros grupos (p<0,05). Também às 48 horas o grupo TA foi o que apresentou a menor porcentagem de AH de alto peso molecular (p<0,05). A maior concentração de CS no LS às 24 e 48 horas ocorreu no grupo TA, também observou-se o grupo TA foi o único em que a concentração de CS aumentou as 8 horas em relação aos valores basais. O grupo AH-APM foi o que apresentou menor concentração de CS às 8 horas (p<0,05). O grupo tratado com AH-BPM apresentou maior intensidade de fluorescência nos momentos 24 e 48 horas (p<0,05). Apesar da triancinolona possuir efeito superior ao ácido hialurônico no controle da inflamação, ela possui efeito catabólico sobre a cartilagem articular, demonstrado pelo aumento da concentração de CS, além de quebra da molécula de AH no líquido sinovial. O AH-APM é mais indicado para terapia intra-articular em equinos uma vez que demonstrou menor efeito catabólico e oxidativo do que o AH-BPM

Several experiments have compared the efficacy of intra-articular usage of hyaluronic acid (HA) within horses. Little is known about the effectiveness of different subtypes of HA in terms of its pro- and anti-inflammatory potential. The goal of this study was to investigate the anti-inflammatory, anabolic and antioxidant effects of intra-articular application of HA in acute synovitis induced by lipopolysaccharide (LPS) in horses. 24 metacarpophalangeal joints of 12 young horses without musculoskeletal alterations were subjected to lameness examination (Lameness Locator®), radiography and ultrasound. These joints were divided into three groups: a positive control group treated with triamcinolone acetonide (TA) (10mg) and 2 experimental groups treated with either hyaluronic acid of low molecular weight (HA- LMW) or hyaluronic acid of high molecular weight (HA- HWM). Initially, all joints were injected with 0.25ng of LPS. After an hour, synovial fluid (SF) was collected for cell counting. TA, HA- HWM or HA- LMW treatments were then applied to the joints. After treatment applications, SF was collected at 3 different time points (8, 24 and 48 hours) for cell counts, oxidative burst evaluation, prostaglandin E2 (PGE2) quantification, chondroitin sulfate (CS), HA and molecular weight measurements. Physical examinations, circumference measurements and lameness evaluations were carried out at each time point. The heart and respiratory rate of the animals increased in all groups at 8 hours (p <0.05), that increase was lower in the TA group. Regarding the circumference of the joint the HA-LMW group showed a greater increase at 24 and 48 hours compared to the other groups (p <0.05). The lameness of the HA-HMW group was higher than the TA group at 8 and 24 hours, but similar to the HA-LMW group. The TA group had a higher nucleated cells count in the SF compared to the HA-LMW group at 24 hours and 28 hours, and higher than the HA-HMW group at 48 hours. The PGE2 concentration was lower in the TA group in relation to the HA-HMW group at 8 hours, and lower than both groups that received HA at 48 hours (p <0.05). The HA-HMW group showed higher HA concentration at 8 hours than the other groups at 48 hours. However, the TA group showed higher HA concentration than the other groups (p <0.05). Also, at 48 hours the TA group was the one with the lowest percentage of high molecular weight HA (p <0.05). The highest concentration of CS in SF at 24 and 48 hours occurred in the TA group, and it was also observed in the TA group was the only one in which the concentration of CS increased at eight hours compared to baseline. The HA-HMW group showed the lowest concentration of CS at 8 hours (p <0.05). The group treated with HA-LMW showed higher fluorescence intensity at 24 and 48 hours (p <0.05). Although triamcinolone have an superior effect of hyaluronic acid in controlling inflammation, it has catabolic effects on cartilage, shown by increased concentration of CS, and breakage of the HA molecule in synovial fluid. The HA-HMW is recommended for intra-articular therapy in equine joints since it showed less oxidative and catabolic effects.

EFICÁCIA CLÍNICA DA VISCOSUPLEMENTAÇÃO COMPARATIVAMENTE AO TRATAMENTO CONSERVATIVO TRADICIONAL EM CÃES COM DISPLASIA COXOFEMORAL

A viscossuplementação é um tratamento que consiste na administração intra-articular (IA) de derivados do ácido hialurônico, cuja eficácia clínica tem sido confirmada em seres humanos. Objetivou-se comparar a eficácia clínica da viscossuplementação em relação ao tratamento conservativo convencional (TCC) em cães com displasia coxofemoral (DCF). Quinze cães foram distribuídos aleatoriamente em dois grupos: AH (n=8) tratamento IA com ácido hialurônico (5 mg a 10 mg); Controle (n=7): tratamento IA com solução salina (0,5 mL a 1,0 mL), além do TCC por via oral com condroprotetor (750-1000 mg, BID, 90 dias) e carprofeno (2,2 mg/kg BID, 15 dias). Os sinais clínicos da DCF foram avaliados de forma encoberta, antes do tratamento (basal), 15, 30, 60 e 90 dias após a injeção IA, por sistema de escore pelo pesquisador e mediante questionários respondidos pelos proprietários dos cães, empregando-se o Breve Inventário de Dor Canina (BIDC) e o Indicador de Dor Crônica de Helsinque (IDCH). Intervenção analgésica foi permitida se o somatório dos escores do BIDC e/ou IDCH excedesse 50%. Os resultados foram analisados pelo teste t não-pareado, teste de Mann-Whitney e teste de Friedman (P < 0,05). Os escores avaliados pelo médico veterinário foram inferiores (P = 0,0005) aos 30, 60 e 90 dias em relação ao basal no grupo AH, e aos 15 dias no grupo Controle (P = 0,0003). Na comparação entre grupos, escores inferiores foram observados aos 30, 60 e 90 dias no grupo AH em relação ao grupo controle, pela avaliação do pesquisador e pelo BIDC. Na comparação ao longo tempo, os escores totais do BIDC foram inferiores aos 60 e 90 dias em relação ao basal nos grupos Controle e AH, respectivamente (P= 0,0145, AH e P = 0,0291, Controle). Na avaliação pelo IDCH escores inferiores foram observados aos 15 e 90 dias em relação ao basal nos grupos Controle e AH, respectivamente (P= 0,0054, AH e P = 0,041, Controle). Intervenção analgésica não foi necessária durante o período de avaliação. Ambos os tratamentos reduziram os sinais clínicos associados à DCF, porém resultados mais expressivos foram alcançados com a viscossuplementação.

Viscosupplementation is one of the treatment modalities for osteoarthritis which consist in an intraarticular (IA) injection of hyaluronic composts. The aim of this randomized, double-blind study was to compare the clinical efficacy of viscosupplemantion with conventional conservative treatment (CCT) in dogs with hip dysplasia (HD). Fifteen dogs were distributed into two groups: HA (n=8): IA injection of hyaluronic acid (5-10 mg); Control (n=7): IA injection with saline solution (0.5-1.0 mL) in combination with a CCT using an oral nutraceutical (750-1000 mg, every 12 h, 90 days) and carprofen (2.2 mg/kg, every 12 h, 15 days). The clinical signs of HD were evaluated prior to treatment (baseline), 15, 30, 60, and 90 days after the IA injection by the veterinary using a score system and by the owners with a questionnaire about their dogs condition using the Canine Brief Pain Inventory (CBPI) and Helsinki Chronic Pain Index (HCPI). The data were analyzed using test unpaired-t, Mann-Whitney-U test and Friedman test, followed by post-hoc Dunns test (P < 0.05). The scores measured by the veterinary decreased from 30 to 90 days in relation to the baseline in the HA group (P = 0.0005), and at 15 days in the Control group (P = 0.0003). The scores evaluated by veterinary and by CBPI were significantly lower in the HA than in the Control from 30 to 90 days. Compared with baseline, both groups had lower total CBPI scores from 60 to 90 days after IA injection respectively (P= 0.0145, HA and P = 0.0291, Control). The HCPI scores were significantly lower in the Control and HA groups at 15 and 90 days after treatment, respectively (P= 0.0054, HA and P = 0.041, Control). Both treatments reduced the clinical signs of HD, however better results were reached with viscosupplementation.

EFICIÊNCIA DO PROTOCOLO SUPERESTIMULATÓRIO P-36 COM A UTILIZAÇÃO DE DUAS ADMINISTRAÇÕES DE FSH, EM ANIMAIS DA RAÇA ANGUS (Bos taurus)

MARTINS BONOTTO, Ramiro. Eficiência do protocolo superestimulatório P-36 com a utilização de duas administrações de FSH, em animais da raça angus (Bos taurus). 2015. 71f. Dissertação (Mestrado em Ciência Animal Área: Produção Animal) Universidade do Estado de Santa Catarina. Programa de Pós-Graduação em Ciência Animal, Lages, 2015. A transferência de embriões é uma biotecnologia amplamente utilizada em vacas Bos taurus. Este princípio envolve a realização de um protocolo de superestimulação para indução de ovulações múltiplas. O protocolo P-36 é um dos mais utilizados com este propósito, pois permite a inseminação artificial em tempo fixo (IATF) facilitando o manejo de doadoras e receptoras de embriões. A superestimulação é feita com oito doses decrescentes de pFSH a cada 12h, porém estudos mostram que a administração de pFSH com uma absorção lenta diminui o número de aplicações, mantendo a mesma produção de embriões. Assim o objetivo deste trabalho foi comparar a superovulação de vacas utilizando o protocolo P36 com redução no número de manejos. Foram utilizadas 24 vacas doadoras da raça Angus, distribuídas aleatoriamente em 2 grupos: Controle e MAP-5 (ácido hialurônico), em um delineamento cross over. Em dia aleatório do ciclo estral (D0), as doadoras receberam um dispositivo intravaginal contendo 1,0 g de progesterona (DIV) e benzoato de estradiol (2 mg, via IM). No grupo controle, os animais foram superestimulados com pFSH (via IM, dose total = 200 mg) duas vezes ao dia em doses decrescentes do D4 ao D7, enquanto que no grupo MAP-5 foram realizadas somente duas administrações no D4 (75% da dose) e D6 (25% da dose). As coletas de embriões foram realizadas no D15 do protocolo. A análise estatística foi realizada com o pacote estatístico SAS utilizando o procedimento Mixed com nível de significância a 5%, sendo consideradas as variáveis: número de folículos no dia da indução de ovulação, número de corpos lúteos no momento da coleta, número de estruturas totais coletadas, número de estruturas fertilizadas e número de embriões viáveis. O grupo controle demonstrou-se significativamente melhor que o grupo MAP-5 quanto ao número de folículos (10,08±0,61 vs. 8,25±0,61), corpos lúteos (7,25±0,59 vs. 3,25±0,59) o que resultou em uma taxa de ovulação de 71,90% para o grupo controle e 39,39% para o grupo MAP-5. Quanto ao número de estruturas totais (8,29±0,98 vs. 3,08±0,98), estruturas fertilizadas (6,50±0,90 vs. 2,50±0,90) e embriões viáveis (4,21±0,72 vs. 2,00±0,72) o grupo controle também demonstrou diferença significativa quando comparado ao MAP-5. O que demonstra que a redução dos manejos com a dose de 200 mg de FSH proposta como grupo MAP-5 não foi eficaz para doadoras da raça Angus.

MARTINS BONOTTO, Ramiro. Efficiency protocol superstimulatory P-36 with the use of two FSH administration in Angus cows (Bos taurus) . 2015. 71f. Thesis (MS in Animal Science - Area: Animal Production) University of the State of Santa Catarina. Graduate Program in Animal Science, Lages, 2015. Embryo transfer is a biotechnology widely used in Bos taurus cows. This principle involves making an over stimulation protocol for inducing multiple ovulations. The P-36 protocol is one of the most used for this purpose as it allows artificial insemination in fixed time (TAI) facilitating the management of donor and recipient embryo. The overstimulation is made with eight pFSH decreasing doses every 12 hours, but studies show that the administration pFSH with a slow absorption decreases the number of applications, maintaining output similar embryos. So the aim of this study was to compare the superovulation of cows using the P36 protocol with reduction in the number of handlings. They were used 24 donor cows of Angus, randomly assigned to two groups: Control and MAP-5 (hyaluronic acid), in a crossover design. At random stage of the estrous cycle (D0), donors received an intravaginal device containing 1.0 g progesterone (DIV) and estradiol benzoate (2 mg, IM). In the control group, the animals were over stimulated with pFSH (IM, total dose = 200 mg) twice daily in decreasing doses of D4 to D7, while the MAP-5 group were carried out only two administrations in the D4 (75% of dose) and D6 (25% of dose). The embryo collections were made in the protocol D15. Statistical analysis was performed with SAS statistical package using the Mixed procedure with 5% significance level, and the variables considered: number of follicles on the day of ovulation induction, the number of corpora lutea at collection time, total number of collected structures, structures and number of fertilized number of viable embryos. The control group showed significantly better than the MAP-5 group and the number of follicles (10.08 ± 0.61 vs. 8.25 ± 0.61), corpora lutea (7.25 ± 0.59 vs. 3.25 ± 0.59) which resulted in a 71.90% ovulation rate for the control group and 39.39% for the MAP-5 group. As for the total number of structures (8.29 ± 0.98 vs. 3.08 ± 0.98), fertilized structures (6.50 ± 0.90 vs. 2.50 ± 0.90) and viable embryos (4 21 ± 0.72 vs. 2.00 ± 0.72) the control group also showed a significant difference when compared to MAP-5. This shows that the reduction of handlings with 200 mg of FSH proposed with MAP-5 group was not effective donor Angus breed.

Estudo do efeito da injeção de PRP e concentrado de medula óssea sobre o reparo de defeitos condrais experimentalmente induzidos e tratados com microfraturas e ácido hialurônico

Defeitos de cartilagem e a mais comum doença articular, a osteoartrite, são caracterizadas pela destruição da cartilagem articular, e consequentemente na perda da função articular em humanos e animais. As estratégias atuais de tratamento, conservativas e cirúrgicas, são insuficientes: não resultam em restauração total da cartilagem hialina, e, portanto trazem um prognóstico reservado a longo prazo. O presente estudo tem por objetivo avaliar os efeitos do administração conjunta de concentrado de medula óssea, plasma rico em plaqueta sobre lesões condrais experimentalmente induzidas e tratadas com microfraturas e ácido hialuronico. Foram utilizadas as articulações metacarpofalangeana de 6 éguas, as quais foram divididas em 2 grupos aleatoriamente e cego. Foram feitos defeitos condrais totais através de artroscopia e, todos foram tratados com microfraturas e ácido hialurônico no transoperatório (M 0) sendo, esse repetido após 15 dias (M 15) e 30 dias (M 30). Grupo C (controle) e grupo T (tratado). O grupo T foi tratado com aspirado concentrado de células tronco de medula óssea adicionada ao plasma rico em plaquetas (PRP), os quais foram injetados na articulação no final da cirurgia (M0). O grupo T recebeu ainda 2 aplicações articulares adicionais de PRP em 15 dias (M 15) e 30 dias (M 30). As seguintes avaliações foram realizadas: exame clínico de claudicação, ultrassonografia, estudo radiográfico, avaliações de líquido sinovial (físico, bioquímico e citológico). As avaliações foram realizadas antes da cirurgia (M 0), com 3, 5 e 7 dias. Posteriormente a cada 15 dias (M 15, M 30, M 45 e M 60) e os 3 últimos momentos foram aos 90 (M90), 120 (M120) e 210 (M210) dias. Ao final do experimento os animais foram enviados para abate comercial. Foram verificadas diferenças estatísticas (p<0,05) entre o grupo tratado e controle para avaliação de proteínas no líquido sinovial corrigido por uréia em 3, 5 e 7 dias; para PGE2 no líquido sinovial em 3 e 5 dias onde para ambas as variáveis com maiores valores para o grupo tratado. Já a concentração de ácido hialurônico apresentou maiores valores (p<0,05) em 3, 45 e 90 dias no grupo controle. Para as demais variáveis não houve diferença estatística entre o grupo tratado e controle. Porém, notou-se medianas maiores para condroitin sulfato em 3 e 5 dias no grupo controle. Notavelmente, o grupo tratado apresentou melhor escore macroscópico na avaliação do tecido de reparo. Contudo, a administração intra-articular de concentrado de medula óssea e plasma rico em plaquetas sobre lesões condrais induzidas e tratadas com microfraturas e ácido hialurônico produziu uma reação articular transitória, principalmente nos primeiros 60 dias, e foi evidenciado pelo aumento de PGE2 e proteínas no líquido sinovial, bem como, claudicação, dor a flexão passiva, diminuição da mobilidade articular e aumento de volume articular. No entanto, o tratamento produziu um efeito condroprotetor e anabólico sobre tecido de reparo formado, uma vez que o grupo tratado apresentou menor concentração de ácido hialurônico 3, 45 e 90 dias e melhor escore macroscópico ICRS aos 210 dias.

Cartilage defects and the most common joint disease, osteoarthritis, are characterized by destruction of articular cartilage, and consequently in loss of joint function in humans and animals. Current strategies of conservative and surgical treatment are insufficient: they dont result in complete restoration of hyaline cartilage, and bring a poor prognosis on the long term. This study aims to evaluate the effects of co-administration of bone marrow concentrate, platelet rich plasma on experimentally induced chondral lesions and treated with microfractures and hyaluronic acid. The metacarpophalangeal joints of 6 mares were used, which were divided into 2 groups at random and blind. Total chondral defects were made using arthroscopy, and all were treated with microfractures and hyaluronic acid during surgery (M 0) and the hyaluronic acid was repeated after 15 days (M 15) and 30 days (M 30). Group C (control) and T group (treated). Group T was treated with concentrated aspirated bone marrow stem cells added to the platelet rich plasma (PRP), which were injected into the joint at surgery (M0). The T group had another 2 additional joint PRP applications in 15 days (M 15) and 30 days (M 30). The following evaluations were performed: clinical examination of lameness, ultrasound, radiographic studies and synovial fluid analysis (physical, biochemical and cytological). The evaluations were performed before surgery (M 0), 3, 5 and 7 days. Then, every 15 days (M 15, M 30, M 45 and M 60) and the last 3 evaluation were at 90 (M90), 120 (M120) and 210 (M210) days. At the end of the experiment the animals were sent to commercial slaughter. Statistical differences were found (p <0.05) between the treated and control group for evaluation of protein in synovial fluid corrected by urea at 3, 5 and 7 days; for PGE2 in the synovial fluid in 3 to 5 days where both variables had higher values for the treated group. The hyaluronic acid concentration was higher (p <0.05) at 3, 45 and 90 days in the control group. For the other variables there were no statistical difference between the treated and control groups. However, greater medians were noticed for chondroitin sulfate in 3 to 5 days in the control group. Notably, the treated group showed better macroscopic score in the evaluation of the repair tissue. In conclusion, intra-articular administration of bone marrow concentrate and platelet-rich plasma on induced chondral lesions and treated with microfractures and hyaluronic acid produced a transient response joint, especially during the first 60 days, and it was evidenced by the increase in PGE2 and proteins of the synovial fluid, as well as lameness, pain passive flexion, decreased joint mobility and joint swelling. Besides that, the treatment produced an anabolic chondroprotective effect on repair tissue formed once the treated group showed lower concentration of hyaluronic acid 3, 45 and 90 days, and better ICRS macroscopic scoring at 210 days.

Comparative study between the effects of hyaluronic acid and acid galactan purified from eggs of the mollusk Pomacea sp in wound healing

PURPOSE: To compare the effect of hyaluronic acid (HA) and of AG on the healing of intestine wounds. METHODS: The semi-purified extract of the eggs of the mollusc was obtained by fractionation with ammonium sulfate and purification for ion-exchange chromatography. The obtained galactans were eluted in water (neutral galactan) and in 0.1 and 0.2M NaCl (acidic galactans). The in vivo study was performed with 45 "Wistar" rats, separated in three groups (n=15). Solutions containing HA 1%, GA 1% or saline solution 0,9%, was placed topically on the sutures of wounds in the small intestine of the rats. After 05, 10 and 21 days the animals were sacrificed and biopsy of the healing tissue was done. RESULTS: The hystologic grading was more significant for HA and AG groups when compared to the group C. AG stimulated the appearance of macrophages, giant cells and increase in the concentration of collagen in the area of the wound when compared to HA. CONCLUSION: The topical use of GA in intestinal wounds promoted the anticipation of events that are important in the wound healing.

OBJETIVO: Comparar o efeito de ácido hialurônico (AH) e do GA na cicatrização de feridas intestinais. MÉTODOS: O extrato semipurificado dos ovos do molusco foi obtido por fracionamento com sulfato de amônio e purificação por cromatografia de troca-iônica. Os galactanos obtidos foram eluídos em água (galactano neutro) e em 0.1 e 0.2M NaCl (galactanos acídicos). O estudo em vivo foi executado com 45 ratos Wistar, separados em três grupos (n=15). Soluções contendo AH 1%, GA 1% ou solução salina 0,9%, foram aplicadas topicamente nas suturas das feridas longitudinais localizadas no intestino delgado dos ratos. Depois de 05, 10 e 21 dias os animais foram sacrificados e biópsias dos tecidos cicatrizados foram realizadas. RESULTADOS: A graduação histológica foi mais significante para os grupos do AH e GA quando comparado ao grupo C. O GA estimulou o aparecimento de macrófagos, células gigantes e aumentou a concentração de colágeno na área da ferida quando comparado ao AH. CONCLUSÃO: O uso tópico de GA em feridas intestinais promoveu a antecipação de eventos que são importantes na cicatrização de feridas.

Tratamento tópico de queimaduras do dorso de ratos com ácido hialurônico

Previous studies have sugested that hyaluronic acid, a naturally occuring glycosaminoglycan in high doses and for a long time on extracelular matrix of healing wounds of fetus, is the responsible for the result of the fetal healing without scar and wound contraction. The present study investigated the topical effect of hyaluronic acid in burns of adult rats, until the complete epitelization of the lesions. Twenty Wistar rats weighting 225±15g were used. A 5 cm2 back burn was done in each rat under anesthesia. In the I group (n=10) the burns were treated with daily topical application of 1ml of 1% hyaluronic acid and in the II group (n=10) the treatment was done with 1ml of saline 0,9%, until the complete healing. The healing time was 29±1,33 days in the I group and 38±2,58 days in the II group . The histologic score was 27,0±2,78 in the I group and 18,1±3,66 in the II group. The differences were significant (p 0,05). The analysis of the aesthetic result revealed larger deformities in the scars of the II group (controls). In conclusion, the topic hyalurinic acid contributed to accelerate the healing rate, it improved the histologic evolution and turned better the aesthetic result in burns of adult rats.

Estudos prévios têm demonstrado que o ácido hialurônico, um glicosaminoglicano que ocorre em doses elevadas e por tempo prolongado na matriz extracelular dos tecidos em cicatrização do feto, é o responsável pelo resultado da cicatrização fetal sem marcas residuais e sem contração. O presente estudo investigou a ação tópica do ácido hialurônico em queimaduras de ratos adultos, até a epitelização completa das lesões. Foram usados 20 ratos Wistar com peso médio 225± 15g, nos quais foi provocada queimadura de 5 cm2 na pele do dorso, sob anestesia com éter sulfúrico. No grupo I (n=10) as queimaduras foram tratadas com aplicação tópica diária de 1ml de ácido hialurônico 1% e no grupo II (n=10) com 1ml de solução salina 0,9%, até a completa epitelização. O tempo de cicatrização foi de 29± 1,33 dias no grupo I e 38± 2,58 no grupo II. A média dos escores histológicos foi de 27,0± 2,78 no grupo I e de 18,1± 3,66 no grupo II. As diferenças mostraram-se significantes (p 0,05). A análise do resultado estético revelou maiores deformidades nas cicatrizes do grupo II (controle). Concluiu-se que o ácido hialurônico tópico contribuiu para acelerar o tempo de cicatrização, melhorou a evolução histológica e o resultado estético em queimaduras do dorso de ratos adultos.