The correlation between score-based protocol for equine joint assessment and subsequent arthroscopic intervention outcomes

This study aimed to grade joint abnormalities in horses submitted to therapeutic arthroscopy using score-based protocols for equine joint assessment, correlated with arthroscopic treatment outcomes and owner satisfaction. In this prospective study, we evaluated 126 joints of athletic horses referred for arthroscopy. The joints were scored according to findings of medical history and physical, radiographic, ultrasonographic and arthroscopic examination. Lameness, positive response to flexion test and decreased maximum joint flexion angle were detected in more than 50% of joints. Soft tissue swelling, sclerosis, subchondral bone osteolysis and single osteochondral fragments were the most common radiographic findings. Ultrasonographic examination revealed changes in synovial fluid volume and appearance, and subchondral bone irregularities. Increased vascularity of the synovial villi, chondral fibrillation, chondral fissures and superficial cartilage erosions were the most significant arthroscopic findings. The approaches that demonstrated greater sensitivity and correlation with treatment outcome and owner satisfaction were medical history, ultrasonographic and arthroscopic examination. The scoring protocol employed to grade joint abnormalities enabled the creation of a joint score system for the objective assessment, thus determining the most frequent findings and establishing an injury severity score for each joint.O objetivo desse estudo foi avaliar as anormalidades articulares observadas em equinos submetidos à artroscopia terapêutica, utilizando protocolos baseados em pontuações e correlacionando os resultados do tratamento cirúrgico com a satisfação do proprietário. Neste estudo prospectivo, foram avaliadas 126 articulações de cavalos atletas, encaminhados para artroscopia. As articulações foram pontuadas de acordo com os achados da anamnese e exame físico, exame radiográfico, exame ultrassonográfico e avaliação artroscópica. A claudicação, resposta positiva ao teste de flexão e diminuição do ângulo máximo de flexão articular foram detectados em mais de 50% das articulações. Aumento de volume dos tecidos moles, esclerose subcondral, osteólise subcondral e fragmentos osteocondrais únicos foram os achados radiográficos mais comuns. O exame ultrassonográfico revelou, com frequência, as alterações no volume e na aparência do líquido sinovial, além de irregularidades osteocondrais. O aumento da vascularização das vilosidades sinoviais, fibrilação condral, fissuras condrais e erosões superficiais da cartilagem foram os achados artroscópicos mais significativos. As abordagens diagnósticas que demonstraram maior sensibilidade e melhor correlação entre o resultado do tratamento e satisfação do proprietário foram história médica, exame ultrassonográfico e artroscópico. O protocolo de avaliação utilizado, baseado em um sistema de pontuação das anormalidades articulares observadas em cada exame, permitiu uma avaliação objetiva, ressaltando os achados mais frequentes e estabelecendo um escore de gravidade da lesão para cada articulação

Development of a rehabilitation protocol for inicial postoperative period of arthroscopy in horses

O presente estudo teve por objetivo avaliar protocolo de reabilitação para o período pós-operatório inicial de artroscopias visando diminuir a inflamação no local operado e aumentar a mobilidade articular. Foram utilizados 12 equinos (total de 20 articulações) encaminhados para artroscopia com diagnóstico de osteocondrite dissecante. Dessas, dez articulações receberam protocolo de reabilitação nos primeiros cinco dias do período pós-operatório. O protocolo consistiu em crioterapia, movimentação passiva da articulação e exercício controlado de baixa intensidade, além de uso sistêmico de anti-inflamatório. O outro grupo, também composto por dez articulações, recebeu apenas a terapia utilizada rotineiramente no HOVET-USP, consistido de repouso em baia e antiinflamatório. As articulações foram avaliadas quanto à circunferência em centímetros, ângulo de flexão, termografia, grau de claudicação. Amostras de líquido sinovial foram coletadas imediatamente antes do procedimento cirúrgico (D1), após 48h (D3) e após 96h (D5) para análise física, qualidade do coágulo de mucina, e quantificação de biomarcadores (IL-1, IL-6 e IL-10, PGE2 e SAA). As análises de exame de claudicação, circunferência articular, ângulo de flexão articular e termografia não apresentaram diferenças significativas entre os grupos, nem entre os diferentes dias do mesmo grupo. Na análise do líquido sinovial, a cor e o aspecto apresentaram piora do D1 para o D3, de amarelo claro para avermelhado e de límpido para turvo, respectivamente, nos dois grupos. No entanto, no grupo tratado houve melhora do D3 para o D5, tanto para cor (de avermelhado para maioria xantocrômica e amarela) como aspecto (de maioria turva para ligeiramente turva). No grupo controle os líquidos permaneceram sem alteração em cor e aspecto de D3 para D5, e nas comparações entre os grupos não houve diferença para D1, D3 e D5. A viscosidade do líquido sinovial no grupo controle diminuiu significativamente quando comparados D1, D3 e D5. Já no grupo tratado a diminuição da viscosidade só foi observada quando comparados D1 e D5. O coágulo de mucina apresentou piora de D1 para D3 no grupo controle, com elevação não significativa de D3 para D5, enquanto que para o grupo tratado não houve diferença significativa de D1 para D3 e de D3 para D5, quando comparados o D5 dos dois grupos, o tratado obteve melhor qualidade. As concentrações de interleucina nas amostras não forneceram dados suficientes para análise. Na análise das concentrações de PGE2 não houve diferença entre os grupos nos diferentes momentos, ocorrendo elevação de D3 para D5 em ambos os grupos, porém, no grupo tratado não há diferença entre D1 e D5. Já para SAA os grupos apresentaram comportamento similar de resposta, com elevação de D1 para D3 e queda de D3 para D5, porém menos acentuado no grupo tratado, o que levou a diferença entre os grupos em D3. Pode-se concluir, que o protocolo de reabilitação, apesar de não gerar diferença significativa para as avaliações de exame físico dos animais, proporcionou melhor qualidade de líquido sinovial quanto a cor, aspecto, viscosidade e precipitado de mucina, além de evidenciar menores elevações nas concentrações de marcadores inflamatórios no liquido sinovial durante o período estudado.The purpose of this study was to evaluate a rehabilitation protocol for the initial postoperative period of metatarsophalangeal, metacarpophalangeal and tarsocrural´s arthroscopies, which seeks to, minimize local inflammation, diminish swelling, promote better joint range of motion and pain relief during such period. Twelve horses participated in this study - amounting to 20 joints - with dissecans ostheochondritis diagnosis. The first group was formed by ten joints, which were treated under rehabilitation protocol for the first 5 days as from the surgery (Treated group). The rehabilitation protocol consisted of cryotherapy, passive range of motion, low intensity exercise and non-steroidal anti-inflammatory drug. The second group also formed of ten joints received the standard HOVET-USP therapy, which consists of rest and non-steroidal anti-inflammatory drug Both groups were treated with the same non-steroidal anti-inflammatory drugs. The joints were measured for circumference, maximal flexion angle, thermography, and lameness score on the day before the surgery (D0) and during the first four days after the surgery. Synovial fluid samples were collected immediately before surgery (D1), within 48 hours (D3), and within 96 hours from the surgery (D5). The analysis evaluated gross appearance (color and aspect), viscosity and mucin clot quality, as well as biomarkers (Il-1, Il-6, Il- 10, PGE2, and SAA) quantification. Lameness examination, joint circumference, flexion angle and thermography evaluation were not significantly different between groups. In synovial fluid analyses de color and aspect have worsen from D1 (clear light yellow) to D3 (turbid hemorrhagic) in both groups. On treated group color and aspect improved from D3 (turbid hemorrhagic) to D5 (xanthochromic and yellow slightly turbid). On treated group there was no difference between D3 and D5. When the groups were compared, none significant differences was seen. The fluid viscosity of control group had significant decrease from D1, to D3 and from D1 and D5. In treated group this viscosity decrease was only seen between D1 and D5. The mucin clot formation worsened when D1 e D3 of control group was compared and remains similar from D3 to D5. In treatment group there were no differences when compared D1 with D3 and D3 with D5. The comparison between groups of D5 has shown treated group improved clot. The interleukin couldn´t be measured on sufficient number of samples for the statistics method. There were no differences between groups on all moments. The PGE2 response was similar in both group with a rise on concentration from D3 to D5. In treated group D1 was similar to D5. This results suggests more evident inflammatory response in the control group. For the SAA the groups have shown similar responses, with an increase from D1 to D3 and decrease from D3 to D5. The response on treated group was less intense and demonstrates lower values in D3 when compared with D3 control group. It was concluded with this study that rehabilitation protocol improved synovial fluid analyses for, color, aspect, viscosity and mucin clot. It even had promoted lower concentrations of inflammatory biomarkers for the treated group during the period

In Vitro Biological Performance of Alginate Hydrogel Capsules for Stem Cell Delivery

[EN] Encapsulation of biological components in hydrogels is a well described method for controlled drug delivery of proteins, tissue engineering and intestinal colonization with beneficial bacteria. Given the potential of tissue engineering in clinical practice, this study aimed to evaluate the feasibility of encapsulation of adipose tissue-derived mesenchymal stem cells (MSCs) of mules in sodium alginate. We evaluated capsule morphology and cell viability, immunophenotype and release after encapsulation. Circular and irregular pores were observed on the hydrogel surface, in which MSCs were present and alive. Capsules demonstrated good capacity of absorption of liquid and cell viability was consistently high through the time points, indicating proper nutrient diffusion. Flow cytometry showed stability of stem cell surface markers, whereas immunohistochemistry revealed the expression of CD44 and absence of MHC-II through 7 days of culture. Stem cell encapsulation in sodium alginate hydrogel is a feasible technique that does not compromise cell viability and preserves their undifferentiated status, becoming a relevant option to further studies of tridimensional culture systems and in vivo bioactive agents delivery.The Sao Paulo Research Foundation (FAPESP grant numbers 2017/ 14460-4, 2017/01369-9 and 2017/12815-0) funded the project execution and provided MSc scholarship to the author GR; The Coordination for the Improvement of Higher Education Personnel (CAPES funding code 001) funded the graduate programof the Sao Paulo State University. The National Council for Scientific and Technological Development (CNPq grant number 142322/2016-4 GD) provided the PhD scholarship to the author JS.De Souza, JB.; Rosa, GDS.; Correa Rossi, M.; Stievani, FDC.; Pfeifer, JPH.; Krieck, AMT.; Bovolato, ALDC.... (2021). In Vitro Biological Performance of Alginate Hydrogel Capsules for Stem Cell Delivery. Frontiers in Bioengineering and Biotechnology. 9:1-10. https://doi.org/10.3389/fbioe.2021.674581110

Presentation1_Biocompatibility of hydrogel derived from equine tendon extracellular matrix in horses subcutaneous tissue.PPTX

Tendinopathies account for a substantial proportion of musculoskeletal injuries. To improve treatment outcomes for partial and total tendon ruptures, new therapies are under investigation. These include the application of mesenchymal stem cells (MSCs) and biocompatible scaffolds derived from the Extracellular Matrix (ECM). Synthetic polymer hydrogels have not demonstrated results as promising as those achieved with ECM hydrogels sourced from the original tissue. This study aimed to evaluate the biocompatibility of a hydrogel formulated from equine tendon ECM. Six horses were administered three subcutaneous doses of the hydrogel, with a saline solution serving as a control. Biopsies were conducted on days 7, 14, and 56 post-application to gauge the hydrogel’s impact. Throughout the experiment, the horse’s physical condition remained stable. Thermographic analyses revealed a temperature increase in the treated groups compared to the control group within the initial 12 h. The von Frey test, used to measure the mechanical nociceptive threshold, also showed significant differences between the treated group and the control group at 6 h, 21 days, and 28 days. Histopathological analyses identified an inflammatory response on day 7, which was absent on days 14 and 56. Transmission electron microscopy indicated a decrease in inflammatory cellularity, while immunohistochemistry staining suggested an increased presence of inflammatory factors on day 14. In summary, the hydrogel is easily injectable, triggers a temporary local inflammatory response, and integrates into the adjacent tissue from day 14 onwards.</p