Effects of beta-aminopropionitrile fumarate and exercise on equine tendon healing: Gross and histological aspects

In the present study, the effects of intralesional injections of beta-aminopropionitrile fumarate (BAPN-F) was assessed in equine experimental tendinitis. BAPN-F is a lathyrogen which inhibits crosslinking of collagen, permitting more time for parallel reorientation of the repair tissue. Sixteen two-year-old Arabian horses without health problems were used in this experiment. The animals were divided into two groups: group one was left in box rest, and group two was submitted to controlled exercise during the experiment. Tendinitis was induced with collagenase in the superficial flexor tendon of both forelimbs. Twenty days after the induction of tendinitis, intralesional treatment with BAPN-F was performed and the contralateral limbs received saline. A biopsy was obtained and gross and histopathological analysis was performed on the 150th day of the experiment. The collagen fibrillar alignment pattern in the heating area was better in the BAPN-F group submitted to controlled exercise than in the other group, as observed by sonographic and histopathologic examination. The present results indicate that BAPN-F in combination with controlled loading improved sear remodeling and tendon wound collagen maturation.

Células mononucleares da medula óssea no tratamento da ruptura total dos tendões flexores e ligamento suspensor do boleto em equino

This paper describes the case of a horse treated in a Teaching Veterinary Hospital. The sutures of the flexor apparatus associated with cell therapy, the stem cell, provided a rapid response of repair compared to other cases treated in routine of the Hospital. The association of conventional treatments with the mononuclear fraction of bone marrow significantly contributed to the reduction of the hyperextension of the metatarsophalangeal joint. Ultrasound images showed rapid fill cell and decrease in inflammation.

Avaliação da resposta inflamatória local após aplicação de células tronco mesenquimais alogênicas em tendão flexor digital superficial de equinos

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Intralesional polysulphated glycosaminoglycan as treatment of equine collagenase induced tendinitis: clinical, ultrasonographic and histopathologic evaluation

Estudou-se o efeito de aplicações intratendíneas do polissulfato de glicosaminoglicanas (PSGAG) no tratamento de tendinite induzida pela colagenase. Dois grupos (GI e GII) de cinco eqüinos da raça Puro-Sangue Árabe, machos e fêmeas, com idades entre dois e seis anos, foram submetidos à tendinite do tendão flexor digital superficial do membro torácico esquerdo por aplicação intratendínea de 1,0ml de colagenase (2,5mg/ml). Decorridos sete dias da indução da lesão, os eqüinos do GI receberam cinco aplicações intralesionais de 1,0ml (125mg) de PSGAG, a intervalos de quatro dias, enquanto que os do GII receberam aplicações de solução fisiológica em igual volume e freqüência. Efetuaram-se avaliações clínicas e ultra-sonográficas, periodicamente, durante 150 dias. Todos os animais apresentaram claudicação e aumento local de sensibilidade, de temperatura e de volume 24 horas após a indução da lesão. Com exceção do aumento de volume, que permaneceu visível até o final do experimento, observou-se regressão de todos os sinais em todos os animais. A avaliação ultra-sonográfica evidenciou lesões de tamanho, forma e posição variados, de maior severidade entre o sétimo e 23º dia. Ao término do experimento, o grau de ecogenicidade encontrava-se entre 1 e 2, e o grau de paralelismo entre 0 e 2. A análise histopatológica evidenciou áreas cicatriciais com intensa fibroplasia e neovascularização, fibras colágenas pouco organizadas e endotendão hipercelular e espessado. Não se observou diferenças significativas entre os grupos quanto ao processo de reparação das lesões, concluindo-se que a aplicação intralesional de PSGAG não produziu efeito benéfico para tratar tendinite induzida por colagenase.

Autologous implant of bone marrow mononuclear cells as treatment of induced equine tendinitis

Superficial digital flexor tendonitis is an important cause of lameness in horses and its incidence ranges from 13% to 30%, depending on the horse's activity. This injury can occur in yearlings and compromise its carriers by reinjury or even impossibility to return to athletic life. In spite of the long period required for tendon repair, the scar tissue presents lack of elasticity and stiffness. As current treatment strategies produce only marginal results, there has been great interest in research of therapies that influence the quality or the speed of tendon repair. Stem cell therapy has shown promising results in degenerative diseases and cases of deficient healing processes. This study aims to evaluate the influence of autologous mesenchymal bone marrow stem cells in tendon healing, comparing treated and non-treated tendons. Superficial digital flexor tendonitis lesions were induced by collagenase infiltration in both forelimbs of 6 horses, followed by autologous implant in one of the forelimbs of each animal. The horses were evaluated using clinical, ultrasonographic, histopathologic, and immunohistochemical parameters. Tendon biopsies were performed at Day 48. Results found in the treatment group, such as high inflammatory cells infiltration, extracellular matrix synthesis, reduced amount of necrosis areas, small increase in cellular proliferation (KI-67/MIB-1), and low immunoreactivity to transforming growth factor P I, suggested the acceleration of tendon repair in this group. Further studies should be conducted in order to verify the influence of this treatment on later phases of tendon repair. Overall, after analysis of the results, we can conclude that cellular therapy with the mononuclear fraction of bone marrow has accelerated tendon repair at 48 days after treatment.

Cell-based Therapies for Tendon and Ligament Injuries

Tendon and ligament injuries have proved difficult to treat effectively. Cell-based therapies offer the potential to harness the complex protein synthetic machinery of the cell to induce a regenerative response rather than fibrous scarring. This article reviews the current state of play with respect to the clinically used cell preparations for the treatment of tendon and ligaments overstrain injuries.

Equine tendonitis therapy using mesenchymal stem cells and platelet concentrates: A randomized controlled trial

Introduction. Tendon injury is a major cause of lameness and decreased performance in athletic equines. Various therapies for tendonitis have been described; however, none of these therapies results in complete tissue regeneration, and the injury recurrence rate is high even after long recovery periods involving rest and physiotherapy. Methods. A lesion was induced with collagenase gel in the superficial digital flexor tendon in the center portion of the metacarpal region of eight equines of mixed breed. After two weeks, the lesions of the animals in the treated and control groups were treated through the intralesional administration of mesenchymal stem cells derived from adipose tissue (adMSCs) suspended in platelet concentrate (PC) and with phosphate buffered saline (PBS), respectively. Serial ultrasound analyses were performed every two weeks. After 16 weeks of therapy, a biopsy was performed for histopathological, immunohistochemical and gene expression (type I collagen (COL1A1), type III collagen (COL3A1), tenascin-C (TNC), tenomodulin (TNMD), and scleraxis (SCX)) analyses. Results: Differences in the ultrasound and histopathological analyses were observed between the groups. Improved results were reported in the group treated with adMSCs suspended in PC. There was no difference in the gene expression levels observed after the different treatments. The main results observed from the histopathological evaluation of the treated group were as follows: a prevention of the progression of the lesion, a greater organization of collagen fibers, and a decreased inflammatory infiltrate. A lack of progression of the lesion area and its percentage was observed in the ultrasound image, and increased blood flow was measured by Power Doppler. Conclusions: The use of adMSCs combined with PC for the therapy of experimentally induced tendonitis prevented the progression of the tendon lesion, as observed in the ultrasound examination, and resulted in a greater organization and decreased inflammation, as observed in the histopathological evaluation. These data demonstrate the therapeutic potential of this therapy for the treatment of equine tendonitis. © 2013 Carvalho et al.; licensee BioMed Central Ltd.

Mesenchymal stem cells modulate release of matrix proteins from tendon surfaces in vitro: a potential beneficial therapeutic effect

Aim: Injury of tendons contained within a synovial environment, such as joint, bursa or tendon sheath, frequently fails to heal and releases matrix proteins into the synovial fluid, driving inflammation. This study investigated the effectiveness of cells to seal tendon surfaces and provoke matrix synthesis as a possible effective injectable therapy. Materials & methods: Equine flexor tendon explants were cultured overnight in suspensions of bone marrow and synovium-derived mesenchymal stems cells and, as controls, two sources of fibroblasts, derived from tendon and skin, which adhered to the explants. Release of the most abundant tendon extracellular matrix proteins into the media was assayed, along with specific matrix proteins synthesis by real-time PCR. Results: Release of extracellular matrix proteins was influenced by the coating cell type. Fibroblasts from skin and tendon appeared less capable of preventing the release of matrix proteins than mesenchymal stems cells. Conclusion: The source of cell is an important consideration for cell therapy.

Evaluation of mesenchymal stem cell migration after equine tendonitis therapy

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Use of power doppler for the assessment of vascularization in equine tendinitis treated with mesenchymal stem cells

Tendinous lesions are very common in athlete horses. The process of tendon healing is slow and the quality of the new tissue is often inferior to the original, leading in many cases to recurrence of the lesion. One of the main reasons for the limited healing capacity of tendons is its poor vascularization. At present, cell therapy is used in equine practice for the treatment of several disorders including tendinitis, desmitis and joint disease. However, there is little information regarding the mechanisms of action of these cells during tissue repair. It is known that Mesenchymal Stem Cells (MSCs) release several growth factors at the site of implantation, some of which promote angiogenesis. Comparison of blood flow using power Doppler ultrasonography was performed after the induction superficial digital flexor tendon tendinitis and implantation of adipose tissue-derived MSCs in order to analyze the effect of cell therapy on tendon neovascularization. For quantification of blood vessel histopathological examinations were conducted. Increased blood flow and number of vessels was observed in treated tendons up to 30 days after cell implantation, suggesting promotion of angiogenesis by the cell therapy.

Anatomofisiologia da bainha tendínea sinovial digital dos eqüinos

Os autores abordam a descrição morfológica da bainha tendínea sinovial digital dos eqüinos e sua função, bem como o processo de formação de seu líquido sinovial, indicando a importância do estudo de tal estrutura.

Repair of lamellar scleral lesions in dogs with preserved equine renal capsule: short report

OBJETIVO: Avaliar o uso da cápsula renal de eqüino preservada em glicerina 98% no reparo de lesões lamelares esclerais em cães. MÉTODOS: Foram utilizados 12 cães, machos e fêmeas, com peso médio de 12kg. Foram realizadas avaliações clínica e morfológica aos 1, 3, 7, 15, 30 e 60 dias de pós-operatório. Após anestesia geral e procedimentos padrões de preparo do campo operatório, foi realizada cantotomia temporal, seguida de incisão conjutival e escleral com área de 0,5x0,5 cm na posição de 1hora, próxima ao limbo. em seguida, um fragmento de mesma dimensão de cápsula renal de eqüino preservada em glicerina, previamente hidratado em solução salina, foi aplicado ao defeito escleral criado sendo fixado com pontos simples isolados com vicryl 7-0®. RESULTADOS: A avaliação clínica revelou blefaroespasmo/fotofobia até o sétimo dia de pós-operatório. Foi observado edema conjuntival até o quinto dia, acompanhado de secreção ocular mucóide, que persistiu até o décimo dia de pós-operatório. Não foram observados sinais clínicos de rejeição do enxerto em todos os animais, em todos os períodos avaliados. Os segmentos anterior e posterior do bulbo ocular não apresentaram sinais de inflamação. A análise morfológica revelou exsudação inflamatória aguda nos períodos precoces e intermediários da avaliação e inflamação crônica nos períodos tardios da observação. Houve incorporação do enxerto ao leito receptor. CONCLUSÃO: Os resultados sugerem que a cápsula renal de eqüino preservada pode ser mais uma alternativa de membrana biológica para o reparo de lesões esclerais lamelares em cães e no homem.

Equine chronic proliferative synovitis (villonodular synovitis): clinical, radiographic and ultrasonographic aspects: relate of case

Doria, R.G.S.; Canola, P.A.; Freitas, S.H. & Canola, J.C. [Equine chronic proliferative synovitis (villonodular synovitis): clinical, radiographic and ultrasonographic aspects: Relate of case.] Sinovite proliferativa cronica (sinovite vilonodular) em equino: aspectos clinicos, radiograficos e ultra-sonograficos: Relato de caso. Revista Brasileira de Medicina Veterinciria 30(3):157-161, 2008. Departamento de Cirurgia, Faculdade de Medicina Veterinaria, Universidade de Cuiaba, UNIC, Av. Antartica 788, Casa 26, Residencial Villas Boas, Ribeirao da Ponte, Cuiaba, MT 78040-500. Brasil. E-mail: redoria@uol.com.brThe development of intracapsullar masses at the dorsal aspect of the metacarpofalangeal joint for a period of several months is commonly secondary to the chronic synovitis. Although it is known as villonodular synovitis in horses probably is better to refer it as chronic proliferative synovitis. The most common causes are the non-treated osteocondral fractures of the dorsal portion of the proximal phalanx. In addition, the development of villonodular masses follows the degenerative process in the joint. A case of a lame animal is reported at the present study. The correct diagnosis and the adequate therapeutic propositions were given based on the clinical examination, therapeutic local-anesthetic test and radioghaphic and ultrasonographic imaging exams. The development of a criterious identification of this disease must be based on clinical findings, radiographic and ultrasonographic exams which assume fundamental importance to the treatment and prognostic. The aim of this study is to describe the clinical, radiographic and ultrasonographic findings' allowing the identification and diagnosis of chronic proliferative synovitis at the thoracic metacarpofalangeal joint in the horse.

Características físico-químicas e citológicas do líquido sinovial da bainha tendínea digital de eqüinos

Foram estudadas as características físico-químicas e citológicas do líquido sinovial da bainha tendínea digital de nove eqüinos hígidos. Verificou-se que o líquido é viscoso, amarelo claro, límpido, livre de partículas e que não coagula à temperatura ambiente. Sua concentração média de ácido hialurônico foi 60,20mg/dl, a taxa de glicose, similar à plasmática e sua concentração protéica não ultrapassou 1,74g/dl, com relação média albumina:globulina de 0,94. O número médio de células nucleadas foi de 313 células/µl, com predominância de grandes células mononucleares e linfócitos. Houve correlação significativa (r = - 0,649, P<0,01) entre o aumento da concentração de ácido hialurônico e a diminuição percentual de linfócitos. As mensurações das características pesquisadas no líquido sinovial da bainha tendínea digital de eqüinos são de execução simples e passíveis de implantação na rotina de atendimentos clínico-cirúrgicos.

Serologic evidence of equine granulocytic anaplasmosis in horses from central West Brazil

A Erliquiose é uma doença zoonótica causada por bactérias gram-negativas e intracelulares obrigatórias. A Anaplasmose Granulocítica Equina - AGE (anteriormente denominada Erliquiose Granulocítica Equina, EGE) é uma enfermidade sazonal, normalmente auto-limitante em equinos. No Brasil, existem poucos relatos deste agente erliquial, bem como de seus vetores naturais. Atualmente, veterinários têm levantado a suspeita de casos de AGE em equinos com sinais clínicos sugestivos de erliquiose e não responsivos ao tratamento para a piroplasmose equina. O objetivo do presente estudo foi identificar equinos expostos a A. phagocytophilum por meio de técnicas sorológicas e moleculares. Vinte amostras de sangue e soro de equinos da região Centro-oeste do Brasil foram avaliados por meio do exame microscópico de capa leucocitária, ensaio imunoenzimático indireto (ELISA), reação de imunofluorescência indireta (RIFI) e reação em cadeia da polimerase (nested PCR). Adicionalmente, o diagnóstico sorológico de Theileria equi pela RIFI e ELISA foram realizados, assim como o diagnóstico molecular pelo nPCR. Treze (65%) amostras de soro foram positivas para A. phagocytophilum pelo teste de ELISA, entretanto nenhum equino foi positivo pelo exame microscópico da capa leucocitária ou nPCR. Anticorpos IgG anti-T. equi foram detectados em 18 (90%) e 17 (85%) equinos pela RIFI e ELISA, respectivamente e o agente foi detectado em 9 (45%) animais pelo nPCR. Estes dados sugerem importante informação para o entendimento da ocorrência da AGE e piroplasmose equina no Centro-oeste do Brasil.