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

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.

Production of Cytotoxic Antibodies After Intra-Articular Injection of Allogeneic Synovial Membrane Mesenchymal Stem Cells With and Without LPS Administration.

Allogeneic mesenchymal stem cells (MSC) are widely used in clinical routine due to the shorter expansion time and reliability of its quality. However, some recipients can produce alloantibodies that recognize MSCs and activate the immune system, resulting in cell death. Although antibody production was already described after MSC injection, no previous studies described the immune response after intra-articular MSC injection in acute synovitis. This study aimed to evaluate the influence of inflammation on immune response after single and repeated intra-articular injections of synovial membrane MSC (SMMSC). Horses were divided in three groups: control group (AUTO) received autologous synovial membrane MSCs; whereas group two (ALLO) received allogeneic SMMSCs and group three (ALLO LPS) was submitted to acute experimental synovitis 8 h before SMMSCs injection. The procedure was repeated for all groups for 28 days. Physical and lameness evaluations and synovial fluid analysis were performed. Sera from all animals were obtained before and every 7 days after each injection up to 4 weeks, to perform microcytotoxicity assays incubating donor SMMSCs with recipients' sera. The first injection caused a mild and transient synovitis in all groups, becoming more evident and longer in ALLO and ALLO LPS groups after the second injection. Microcytotoxicity assays revealed significant antibody production as soon as 7 days after SMMSC injection in ALLO and ALLO LPS groups, and cytotoxicity scores of both groups showed no differences at any time point, being equally different from AUTO group. Although inflammation is capable of inducing MHC expression in MSCs, which enhances immune recognition, cytotoxicity scores were equally high in ALLO and ALLO LPS groups, making it difficult to determine the potentiation effect of inflammation on antibody production. Our findings suggest that inflammation does not display a pivotal role in immune recognition on first allogeneic MSC injection. In a translational way, since specific antibodies were produced against MSCs, patients that need more than one MSC injection may benefit from a first allogeneic injection followed by subsequent autologous injections.

Low Mg content on Ti-Nb-Sn alloy when in contact with eBMMSCs promotes improvement of its biological functions.

Magnesium is a metal used in the composition of titanium alloys and imparts porosity. Due to its osteoconductive, biocompatible and biodegradable characteristics, its application in the development of biomedical materials has become attractive. This study aimed to evaluate the influence of magnesium present in porous Ti-Nb-Sn alloys, which have a low elastic modulus in adhesive, osteogenic properties and the amount of reactive intracellular oxygen species released in mesenchymal stem cells derived from bone marrow equine bone (eBMMSCs). Mechanical properties of the alloy, such as hardness, compressive strength and elastic modulus, were analyzed, as well as surface morphological characteristics through scanning electron microscopy. The evaluation of magnesium ion release was performed by atomic force spectroscopy. The biological characteristics of the alloy, when in contact with the alloy surface and with the culture medium conditioned with the alloy, were studied by SEM and optical microscopy. Confirmation of osteogenic differentiation by alizarin red and detection of ROS using a Muse® Oxidative Stress Kit based on dihydroetide (DHE). The alloy showed an elastic modulus close to cortical bone values. The hardness was close to commercial Ti grade 2, and the compressive strength was greater than the value of cortical bone. The eBMMSCs adhered to the surface of the alloy during the experimental time. Osteogenic differentiation was observed with the treatment of eBMMMSCs with conditioned medium. The eBMMSCs treated with conditioned medium decreased ROS production, indicating a possible antioxidant defense potential of magnesium release.

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

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.

Physical and biological characterizations of TiNbSn/(Mg) system produced by powder metallurgy for use as prostheses material.

Titanium scaffolds with non-toxic ß stabilizing elements (Nb and Sn), Ti-34Nb-6Sn (TNS), and with magnesium as spacer (TNS/M), were processed by powder metallurgy, and sintered at 800 °C. The X-ray diffraction (XRD) pattern showed that materials are biphasic alloys, presenting 45 to 42% (wt %) in hcp (α-phase) and the rest is bcc (ß-phase), and the presence of a slight peak relating to TiO2 in both materials. Pores of approximately 50 µm for TNS and 300 µm to TNS/M were observed in the micrographic analysis by scanning electron microscopy (SEM). The wettability was higher for TNS/M compared to TNS. The elastic modulus was higher for TNS compared to TNS/M. Stem cells derived from equine bone marrow (BMMSCs) were used for in vitro assays. The morphologic and adhesion evaluation after 72 h, carried out by direct contact assay with the materials showed that the BMMSCs were anchored and adhered to the porous scaffolds, in the way the cytoplasmic extension was observed. The cellular migration, using the "wound healing" method, was significant for the groups treated with conditioned medium with materials in 24 h. Osteogenic differentiation of BMMSCs, assessed by calcium deposition and staining with Alizarin Red, was greater in the conditioned medium with TNS/M in 10 days of culture. Since the biological effects was good and the elastic modulus decreased in the system with magnesium is a promising new content titanium alloy for biomedical application.

Accuracy of differences in blood and peritoneal glucose to differentiate between septic and non-septic peritonitis in horses.

Difference in blood and peritoneal glucose (DBPG) is used in clinical practice to support a diagnosis of septic peritonitis in horses. It is inexpensive, easy and rapid to perform. The aim of this study was to evaluate the accuracy of the DBPG to differentiate between septic and non-septic peritonitis in horses. Blood and peritoneal fluids were harvested from suspected animals. Plasma and peritoneal glucose levels, total nucleated cell count, direct microscopic and microbiological examinations of the peritoneal fluid were evaluated. Using DBPG levels, the animals were classified into two groups: difference ≥ 50 mg/dL (positive test) and difference < 50 mg/dL (negative test). Positive microbiological examination and/or presence of bacteria in direct microscopic examination was used as a gold standard to detect septic peritonitis. The accuracy parameters analysed were: sensitivity, specificity, and positive/negative predictive values, for which the results were respectively: 0.23, 0.91, 0.60 and 0.67. Due to poor accuracy, other cut-off margins and peritoneal glucose concentrations were evaluated. The test was considered most accurate when the DBPG was zero with sensitivity, specificity, and positive/negative predictive values of 0.85, 0.82, 0.73, 0.90 respectively. Peritoneal glucose concentrations alone were not a reliable feature to detect peritonitis. DBPG ≥50 mg/dL, widely used for the diagnosis of septic peritonitis, does not have a good accuracy and the DBPG = 0 has a better accuracy for detecting the disease.

Association of intraperitoneal ceftriaxone, intravenous gentamicin and oral metronidazole in the treatment of an abdominal abscess and peritonitis in a horse: case report / Associação da ceftriaxona intraperitoneal, gentamicina intravenosa e do metronidazol oral no tratamento de abscesso abdominal e peritonite em equino: relato de um caso

Medical management of abdominal abscesses in horses requires prolonged antibiotic therapy and presents varied success rates. A 6-year-old male horse with a history of colic and multiple abdominal punctures to relieve gas was attended. At admission, tachycardia, tachypnea, hyperthermia, mucosal congestion, dehydration, and rigid gait were observed. The association of physical examination, laboratory and ultrasonographic findings allowed the diagnoses of peritonitis and abdominal abscess. Supporting treatment plus broad spectrum antibiotic therapy was performed: daily intraperitoneal ceftriaxone (25 mg/kg, 7 days); daily intravenous gentamicin (6.6 mg/kg, 7 days); per os metronidazole three times a day (15 mg/kg 12 days), followed by the same dose twice a day (15 mg/kg 33 days), totaling 45 days of treatment. Plasma fibrinogen and ultrasonographic examination were the most effective tools to evaluate abscess evolution. There was normalization of the physical examination 24 h after beginning the treatment, consecutive regression of the nucleated cell count in the peritoneal fluid, and regression of plasma fibrinogen and size of the abscess. On the 10th treatment day, the animal was discharged from the hospital, maintaining oral therapy with metronidazole every 12 h (15 mg / kg). When the animal returned on the 30th day, an abscess size regression was observed. However, there was no resolution, and therapy with metronidazole was maintained. On the 45th day of treatment, a new hospital evaluation was performed, where the abscess resolved, and metronidazole was suspended. It is highlighted that the therapeutic association used in the treatment of abdominal infection and abscess resulted in a rapid clinical response.(AU)

O tratamento conservativo dos abscessos abdominais em equinos requer antibioticoterapia prolongada e apresenta variadas taxas de sucesso. Foi atendido um cavalo de seis anos de idade, com histórico de cólica e múltiplas punções abdominais por agulha para esvaziamento de gás. Na admissão, foram observados taquicardia, taquipnéia, hipertermia, congestão mucosa, desidratação e marcha rígida. A associação do exame físico, achados laboratoriais e ultrassonográficos permitiu o diagnóstico de peritonite e abscesso abdominal. Foi realizado tratamento suporte e antibioticoterapia de amplo espectro: ceftriaxona intraperitoneal diária (25 mg/kg, 7 dias); gentamicina intravenosa diária (6,6 mg/kg, 7 dias); metronidazol oral três vezes ao dia (15 mg/kg, 12 dias), seguido de mesma dose duas vezes ao dia, por mais 33 dias, totalizando 45 dias de tratamento. O fibrinogênio plasmático e o exame ultrassonográfico foram os recursos mais eficazes para a avaliação da evolução do abscesso. Após 24 horas do início do tratamento foi constatada a normalização do exame fisico, regressão progressiva da contagem de células nucleadas no líquido peritoneal, do fibrinogênio plasmático e do tamanho do abscesso. No 10° dia de tratamento o animal recebeu alta hospitalar, mantendo-se a terapia oral com metronidazol a cada 12 horas (15 mg/Kg). Em retorno, ao 30° dia, observou-se regressão do tamanho do abscesso, entretanto, não houve resolução, tendo sido mantida a terapia com metronidazol. No 45º dia de tratamento, realizou-se nova avaliação hospitalar, onde foi observada a resolução do abscesso e a admnistração do metronidazol foi suspensa. Destaca-se, que a associação terapêutica utilizada no tratamento de infecção abdominal e abscesso resultou em rápida resposta clínica.(AU)

Experimental skin wound treatment with Copaifera langsdorffii Desf Kuntze (Leguminosae) extract and oil-resin in horses / Tratamento experimental de feridas cutâneas com extrato e óleo resina de Copaifera langsdorffii Desf Kuntze (Leguminosae) em equinos

Copaifera langsdorffii is a Brazilian native plant that is used for wound healing or as an anti-inflammatory agent. Non-healing wounds are an important health problem, particularly in horses, because they can cause the animal invalidity or even lead to death. In addition, horses respond to skin wounds with chronic inflammatory response and intense wound granulation, thereby delaying the healing process. By this way, our aim was to evaluate the healing potential of Copaifera langsdorffii hydroalcoholic extract (HE) and oil-resin creams (OR) in horse skin wounds. Four wounds were performed bilaterally in the lumbar region of six horses with a 2 cm punch and treated daily with the respective treatments: saline solution, vehicle, 10% HE creams or 10% OR cream. Daily planimetry analyses were performed to measure the wound area and clinical parameters. In four different experimental periods (3, 7, 14 and 21 d), wound biopsies were removed and used for microscopic analyses. SS wounds presented a significant small area at day 3 and 7, OR wounds presented significant small area in comparison with HE at 14 d, and no significant difference was observed between treatments at 21 d. A better microscopic and clinical healing activity of HE and OR was identified in comparison with the controls. The OR group showed better healing quality, specifically after 7 d of treatment. Therefore, Copaifera langsdorffii formulations demonstrated their wound healing potential in horse skin lesions, exhibiting an improvement of the macro- and microscopic parameters.(AU)

Copaifera langsdorffii é uma planta nativa brasileira usada para cicatrização de feridas e como agente anti-inflamatório. As feridas que não cicatrizam são um importante problema de saúde em equinos, devido à possibilidade de afastamento da carreira atlética ou até mesmo levar à morte. Além disso, os cavalos respondem às feridas com resposta inflamatória crônica e intensa granulação, atrasando o processo cicatricial. Dessa forma, o presente trabalho avaliou o potencial do extrato hidroalcoólico (EH) e cremes de óleo resina (OR) de Copaifera langsdorffii em feridas cutâneas de equinos. Quatro feridas de 2 cm foram realizadas bilateralmente na região lombar de seis equinos e tratadas diariamente com: solução salina, veículo, creme EH 10% ou creme OR 10%. Diariamente foi efetuada a análise macroscópica das feridas e planimetria para mensurar a área da ferida. Em quatro períodos experimentais diferentes (3, 7, 14 e 21 dias), as biópsias foram realizadas e utilizadas para análises microscópicas. As análises microscópica e clínica indicaram melhor resposta cicatricial nas feridas tratadas com HE e OR em comparação com os controles. O grupo OR apresentou melhor qualidade de cicatrização, especificamente após sete dias de tratamento. Portanto, as formulações de Copaifera langsdorffiiutilizadas apresentaram potencial de cicatrização de feridas em lesões de pele de cavalo, exibindo uma melhora nos parâmetros macro e microscópicos.(AU)

Evaluation of alginate hydrogel encapsulated mesenchymal stem cell migration in horses.

Osteoarthritis is an incapacitating disease characterized by pain and a progressive decrease in joint mobility. The implantation of mesenchymal stem cells (MSCs) has shown promising results for its treatment. The challenge remains to keep the cells longer at the site of action, increasing their therapeutic potential. The aim of this study was to evaluate the effectiveness of the Qtracker® 655 nanocrystal marking on allogeneic synovial membrane (SM) MSCs, encapsulated in alginate hydrogel, evaluating the migration of these cells. The 10 radiocarpal joints were submitted to arthroscopic surgery (D0), divided into two groups. The chondral defect was treated according to the group: GA free-labelled MSCSM and GB labelled MSCSM microcapsules. Seven days after lesion induction and implantation of labelled cells, biopsies of the lesion site were performed in two animals, and fragments of SM and joint capsule also collected, which were frozen and later processed for fluorescence microscopy. The synovial fluid of the three animals was analyzed by flow cytometry three times - 3, 7 and 21 days after application. The cellular marking with the nanocrystals allowed the visualization of the cells in cartilage, synovial membrane, synovial fluid and articular capsule, but with a predilection for the synovial membrane and the lesion site was scarce. The labelled MSCSM in microcapsules were scarce in the synovial fluid and could be related to the small quantity of MSCs leaving the pores of the microcapsules, also favorable results, as the cells release paracrine effects acting for a long period until the cellular differentiation.

Glossopharyngeal and vagus neuropathy associated to stylopharyngeus muscle compression in horse guttural pouch ­ case report / Neuropatia do glossofaringeo e vago associada à compressão pelo musculo estilofaringeo em bolsa gutural de equino ­ relato de caso

Neuropathies of pharyngeal branches of glossopharyngeal and vagus are often associated with guttural pouches diseases; however, these branches of injury due to stylopharyngeus muscle compression are not reported. A case was reported of a quarter horse mare, 8 years old, 450 kg, presenting dyspnea and respiratory noise associated with weight loss. Clinical examination observed mixed dyspnea, tachycardia, dysphagia, sialorrhea, lung crackles and submandibular and parotid lymphadenopathy. Endoscopic exam showed right arytenoid chondritis, nasopharyngeal collapse, generalized larynx edema and dorsal displacement of the soft palate. Right guttural pouch evaluation showed swelling in the origin of stylopharyngeus muscle with consequent compression of the XII, X and IX cranial nerves. Tracheotomy, systemic treatment with corticosteroids, beta lactams and aminoglycosides antibiotics were performed. No resolution was observed and, after 16 days, the animal showed clinical worsening, developed pleuropneumonia, uveitis, severe sepsis, acute renal failure and was euthanized. The mixed neuropathy resulted in rapid clinical deterioration of the animal, due to the difficulty in swallowing and consequent associated respiratory processes. This report emphasizes the importance of evaluating stylopharyngeus muscle origin in cases of nasopharyngeal collapse associated with dysphagia in horses, given the possibility that structural changes in this muscle can result in laryngeal neuropathy.(AU)

As neuropatias do glossofaríngeo e vago são frequentemente associadas com enfermidade das bolsas guturais. A ocorrência desta lesão secundária a compressão pelo músculo estilofaríngeo não é reportada. Relata-se o caso de uma égua quarto de milha, 8 anos de idade, 450 kg, apresentando dispneia, ruído respiratório e perda de peso. No exame clínico observou-se dispnéia mista, taquicardia, disfagia, sialorréia, crepitação pulmonar e linfadenopatia submandibular e parotídea. Ao exame endoscópico identificou-se condrite aritenóide direita, colapso nasofaríngeo, edema generalizado da laringe e deslocamento dorsal do palato mole. Na avaliação da bolsa gutural direita identificou-se aumento de volume na origem do músculo estilofaríngeo com consequente compressão dos nervos cranianos XII, X e IX. Foi realizada traqueotomia, tratamento sistêmico com corticosteróides e antibióticos. Nenhuma resolução foi observada, após 16 dias o animal apresentou piora clínica, pleuropneumonia, uveíte, sepse grave, insuficiência renal aguda e foi submetido à eutanásia. A neuropatia mista resultou em rápida deterioração clínica do animal, devido à dificuldade de deglutição e processos respiratórios associados. Salienta-se a importância de se avaliar a origem do músculo estilofaríngeo em casos de colapso nasofaríngeo associado a disfagia em cavalos, dada a possibilidade de que alterações estruturais nesse músculo possam resultar em neuropatia laríngea.(AU)

Correction to: Culture of mesenchymal stem cells derived from equine synovial membrane in alginate hydrogel microcapsules.

The original article [1] contained a minor error regarding the mean diameter of the alginate microcapsules described in relation to Fig. 4 in the Results section. The microcapsules had an actual mean diameter of 3000 µm instead of 1000 µm as mistakenly mentioned in the original article.

Allogeneic mesenchymal stem cell transplantation in healthy equine superficial digital flexor tendon: A study of the local inflammatory response.

The superficial digital flexor tendon (SDFT) is a structure frequently affected by injuries in high-performance athletic horses, and there are limited therapeutic options. Regenerative medicine has evolved significantly in treating different illnesses. However, understanding the cellular behaviour during mesenchymal stem cell (MSC) transplantation in healthy tissues is not fully known yet. To address the inflammatory response induced by allogeneic MSC transplantation, this study evaluated the local inflammatory response after the application of allogeneic adipose tissue-derived mesenchymal stem cells (AT-MSCs) in the equine tendon compared to an autologous transplant and the control group. Eighteen thoracic limbs (TL) in nine animals were divided into three groups and subjected to the application of AT-MSCs in the healthy tendon. In the allogeneic group (Gallog), the animals received an allogeneic AT-MSC application in the TL. The autologous group (Gauto) received an application of autologous cells in the TL, and in the control group (Gcont), phosphate-buffered saline (PBS) was applied. There were no significant differences among the evaluated groups in the physical, morphological, thermography, and ultrasonography analyses. A higher number of CD3-positive lymphocytes was observed in the Gauto group compared to the control (P < 0.05). Additionally, we did not observe different expressions of CD172 and microvascular density among the groups. The allogeneic transplantation of AT-MSCs did not result in an adverse or inflammatory reaction that compromised the use of these cells in this experiment. Their behaviour was similar to that of autologous transplantation.

Effect of phototherapy with light-emitting diodes (890 nm) on tendon repair: an experimental model in sheep.

The effect of phototherapy with 890-nm light-emitting diodes (LEDs) on the healing of experimentally induced tendinitis in sheep was evaluated in this study. Partial tenotomies measuring 0.2 cm wide × 0.5 cm long were performed on the second third of the superficial digital flexor tendons of 10 healthy sheep. The animals were divided into two groups: "treated" (TG), treated with LEDs at the aforementioned wavelength, and "control" (CG), a control group treated with a placebo. Kinesiotherapy, which consisted of 5-min walks on grassy ground, was performed on both groups. B-mode and power Doppler ultrasonographies (US) were performed to evaluate the tendon healing process during the first 14 days after surgery and on the 21st and 28th postoperative days. Biopsies were performed on day 28 for the histopathological assessment of neovascularisation and the pattern of the tendon fibres. The absence of lameness and a significant improvement (p < 0.05) in the sensitivity to pain during palpation were observed in the treated group. Furthermore, a significant reduction in oedema and an increased number of vessels (p < 0.05) were observed in this group with the B-mode and power Doppler US, respectively. No significant difference in the evolution of the lesion was found. There was a histological difference (p < 0.05) in neovascularisation in the treated group. Phototherapy with 890-nm light-emitting diodes decreases the inflammatory process.

Peritoneal reactivity evaluation in horses subjected to experimental small colon enterotomy and treated with subcutaneous heparin.

Heparin is routinely administered in postoperative abdominal surgery aiming to prevent adhesions formation; however, there is no consensus indicating its effectiveness. This study evaluated the effect of heparin on peritoneal reactivity after abdominal surgery, through the association between peritoneal fluid features and ultrasonographic and laparoscopic examination. Ten adult horses were used: control group (CG) and treated group (TG). Both groups underwent laparotomy and small colon enterotomy. TG received subcutaneous heparin at 150 IU/kg every 12 hours for 5 days. The animals underwent ultrasonography and peritoneal fluid examination prior to enterotomy (M0) 12 hours (M1), 1 day (M2), 2 days (M3), 4 days (M4), 6 days (M5), 10 days (M6), and 14 days after enterotomy (M7) with laparoscopic examination being performed on the fifth postoperative day. Peritoneal inflammatory response was observed in both groups. The peritoneal fluid of TG animals showed higher echogenicity during heparin therapy. No inflammatory difference was observed between groups through peritoneal fluid features, except for the higher D-dimer concentration in CG. On laparoscopy, slightly diffuse peritoneal reactivity for both groups was observed, being higher for TG. Laparoscopy and ultrasonography association allowed detailed access to the abdominal cavity. Ultrasonography assessed the diffuse peritoneal inflammation, and laparoscopy allowed the detailed analysis of the segments. No gross beneficial reactions resulting from the use of heparin on peritoneal reactivity were observed; however, it was observed by D-dimer evaluation that the TG had less fibrin deposition, which is directly related to a lower rate of abdominal adhesions formation.

Peritoneal response to abdominal surgery: the role of equine abdominal adhesions and current prophylactic strategies.

Intra-abdominal adhesions constitute a significant clinical and surgical problem that can lead to complications such as pain and bowel occlusion or subocclusion. These adhesions are frustrating and potentially fatal, representing a major postoperative complication in abdominal surgery. It is estimated that 32% of horses undergoing laparotomy will present clinical symptoms due to adhesions, but the true prevalence is not known because a large proportion of animals with postoperative recurrent colics are medically treated or submitted to euthanasia without necropsy. Adhesions are highly cellular, vascularized, dynamic structures that are influenced by complex signaling mechanisms. Understanding their pathogenesis could assist in applying better therapeutic strategies and in developing more effective antiadhesion products. Currently, there are no definitive strategies that prevent adhesion formation, and it is difficult to interpret the results of existing studies due to nonstandardization of an induction model and evaluation of their severity. The best clinical results have been obtained from using minimally traumatic surgical techniques, anti-inflammatory agents, antimicrobials, anticoagulants, and mechanical separation of serosal surfaces by viscous intraperitoneal solutions or physical barriers. This paper aims to review adhesion formation pathogenesis, guide the understanding of major products and drugs used to inhibit adhesion formation, and address their effectiveness in the equine species.

Aspectos clínicos, ultra-sonográficos e venográficos da tromboflebite jugular experimental em equinos / Equine experimental thrombophlebitis: clinical, ultrasonographic and venographic evaluation

A tromboflebite jugular ocorre frequentemente em equinos, decorrendo geralmente de processos mórbidos associados à iatrogenia, podendo levar a perda de função, edema cefálico, diminuição do desempenho atlético e ainda causar o óbito. Esta enfermidade nos equinos apesar de frequente é pouco conhecida quanto à sua evolução e tratamentos. O objetivo deste trabalho foi avaliar a evolução da tromboflebite jugular experimental em equinos, quanto às alterações clínicas e estruturais envolvidas na enfermidade, observando-se os aspectos clínicos, ultra-sonográficos e venográficos no contexto do trombo e do vaso, quanto à possibilidade de recanalização do trombo produzido e da vascularização compensatória. A tromboflebite da veia jugular foi induzida, unilateralmente, em 05 equinos nos quais previamente à indução da tromboflebite e diariamente após foram observadas manifestações clínicas e realizados exames ultra-sonográficos. Venografias foram feitas nos momentos pré-indução, na indução e a cada seis dias após a indução da tromboflebite, verificando-se a recanalização do trombo oclusivo e a presença de vasos na drenagem sanguínea compensatória. Observou-se a ocorrência de edema moderado das regiões parotídea, massetérica e supra-orbitária e discreto edema submandibular que reduziram até o 6º dia, permanecendo apenas discreto aumento parotídeo. O ingurgitamento da jugular cranial a região da indução permaneceu durante todo o período de avaliação. A porção caudal à tromboflebite mostrou ingurgitamento frente ao garrote na entrada do tórax desde o primeiro dia após a indução. Os exames ultra-sonográficos mostraram formação de trombo oclusivo total durante todo o período de observação em 3 animais e o restabelecimento parcial do fluxo na jugular em 2 animais e a presença de vasos colaterais conduzindo o sangue da porção cranial para a porção caudal à obstrução. As venografias revelaram fluxo sanguíneo "linear" normal no momento pré-operatório, constatando nos momentos pós-operatórios a presença oclusiva do trombo, com o contraste preenchendo os vasos tributários compensatórios direcionados à porção caudal à oclusão da veia ou ainda estagnado cranialmente ao trombo. Conclui-se que a trombose oclusiva na tromboflebite jugular experimental e unilateral sofre recanalização e compensação vascular por vasos tributários de drenagem, com redução gradativa dos sinais decorrentes da estase sanguínea de retorno, especificamente as regiões cefálicas com edema. Estudos envolvendo a tromboflebite jugular nos equinos devem evoluir nos aspectos experimental e clínico.

Jugular thrombophlebitis is a common complication of disease processes associated with repeated venipuncture, injection of irritant solutions, and the use of indwelling catheters, especially with bacterial contamination. Bilateral thrombophlebitis may result in edema of the soft tissues of the head, reduction of athletic performance and even death of the animal. This disease, although common in horses, is not much known regarding its evolution and treatment. The aim of this study was to evaluate the clinical and structural changes of experimentally induced jugular thrombophlebitis in horses, through clinical examination, ultrasound and venography of the thrombus and the vessel, verifying the possibility of thrombus recanalization and compensatory produced blood flow. The jugular thrombophlebitis was induced unilaterally into 5 horses, monitored by clinical (general, regional and local) and ultrassonographycs exams. Venographs were made at pre-induction, induction and every 6 days after induction of thrombophlebitis, in order to observe recanalization of the occlusive thrombus and presence of blood vessels in the drainage allowance. Occurrence of moderate edema was observed in the parotid, masseter and supra orbital regions, and mild edema in the submandibular region. The jugular engorgement of the cranial region of induction persisted throughout the period of evaluation. The caudal portion to the thrombophlebitis showed engorgement with compression on the vein at the thorax entrance since the first day after induction. The ultrasound examinations showed total occlusive thrombus formation of 3 animals, partial recirculating flow in the jugular vein in 2 animals, and collateral blood vessels from the cranial obstruction to the caudal portion. The venography revealed normal linear blood flow in the preoperative and occlusive thrombus with contrast directed filling of the vessels to the compensatory portion caudal to the vein occlusion or cranial to the thrombus in the postoperative moments. After vein resection of the segment containing the thrombus, the cephalic edema was less intense than after the induction of the thrombophlebtits. The ultrassonography and venography post resection showed vascularity increase in this region. It was concluded that there is recanalization with endothelialization and vascular compensation made by pre-existing vessels necessary for drainage.

Avaliação histológica e morfométrica do retalho axial oris angularis e da terapia por ondas de choque aplicados a defeito palpebral experimental em cães / Histologic and morfometric evaluation of oris angularis axial skin flap and extracorporeal shockwave on experimental eyelids defects in dogs

Os retalhos de padrão axial têm como característica significativa vascularização intrínseca, considerada uma vantagemsobre outras técnicas. Considerando que complicações isquêmicas podem afetar os retalhos cutâneos, técnicas desalvamento são descritas, dentre estas, a terapia por ondas de choque extracorpóreas (TOCE), descrita como capazde modular a vascularização e cicatrização dos retalhos. O presente estudo avaliou histológica e morfometricamente21 amostras de pele; destas, 14 foram submetidas à confecção do retalho axial, sendo sete tratadas também pelaTOCE, obtidas da região distal do retalho axial oris angularis, utilizado para a reconstrução de defeitos palpebraisexperimentais extensos em cães. Foram avaliadas também sete amostras de pele normal da mesma região acimadescrita (grupo controle). Não foram evidenciadas diferenças histológicas significativas no infiltrado inflamatório eatrofia epidérmica microscopicamente. Na análise morfométrica, o número de vasos, a área vascular total e a área médiaforam semelhantes entre os grupos experimentais. O retalho oris angularis associado ou não à TOCE não apresentoucaracterísticas microscópicas de complicações inflamatórias e atróficas significativas. Sinais de integridade tecidual evascularização sanguínea adequados foram observados em ambos os grupos tratados, demonstrando efetividade doretalho oris angularis. A aplicação da TOCE no retalho oris angularis, em dose única de 2500 impulsos a 0,15 mJ/mm2no pós-operatório imediato, não promoveu efeitos colaterais deletérios

Considering that the cutaneous flap can be affected by isquemic complications the extra corporeal shock wavetherapy (ESWT) was described as rescue techniques. The present study was developed to analyze histological andwith morfometry, twenty one skin samples treated or not with the shock wave therapy, obtained from flap’s distalborder, used in this study to repair eyelids’ experimental defects in dogs. The flap with or without ESWT did notshow any histological sign of inflammatory or atrophic alterations. Both group treated showed similar morphometricalcharacteristics. The ESWT with the protocol used in this study (2500 impulses at 0,15 mJ/mm2) did not demonstratesignificant clinical outcomes as a rescue technique when applied over the oris angularis flap, however results showed nosignals of collateral deleterious effects

Determinação da densidade mineral e da elasticidade óssea após a aplicação de ondas de choque extracorpóreas no terceiro metacarpiano de equinos atletas / Mineral density and bone elasticity determination after extracorporeal shock waves application in third metacarpus of athlete equines

Objetivou-se neste estudo avaliar os efeitos da aplicação de ondas de choque extracorpóreas (ESWT) no terceiro metacarpiano de equinos hígidos através da determinação da elasticidade óssea. Foram utilizados 20 equinos Puro Sangue Inglês, machos e fêmeas, com dois anos de idade, em início de treinamento e selecionados quanto ao estado de higidez. No início do experimento (D0), todos os animais foram submetidos à avaliação da elasticidade óssea do terceiro metacarpiano. Os animais foram divididos em dois grupos (Grupo Controle - GC e Grupo Experimental - GE). A aplicação das ESWT foi realizada no membro torácico direito dos animais do GE, na região coincidente à da avaliação da elasticidade óssea. Foi utilizado aparelho para terapia de ondas extracorpóreas com densidade de fluxo de energia de 0,15mJ/mm2 e 2000 pulsos com probe E6R20, com característica do foco da onda de choque de 20 mm. A ESWT foi repetida a cada 21 dias totalizando três sessões (D0, D21 e D42). A análise da determinação da elasticidade óssea foi realizada no D0, D21, D42 e D72. A média da velocidade ultrassonográfica diferiu entre os grupos no D21, D42 e D72, sendo que os animais do GE apresentaram menor densidade mineral óssea após as aplicações das ESWT. Houve igualmente, diferença na massa óssea entre os grupos no D21 e D42, parâmetro onde os animais do GE apresentaram diminuição significativa da massa óssea. O risco de fratura apresentou-se maior nos animais do GE no D21. Concluiuse que, a ESWT é capaz de promover alteração da densidade mineral óssea.

The porpoise of this study was to evaluate the effects of extracorporeal shock waves in third metacarpus bone from healthy horses by determination of bone elasticity. It were used 20 Thoroughbred horses, male and female, with two years old, on top of training and selected as the state healthy. At the beginning of the experiment (D0), all animals were submitted for evaluation of bone elasticity held in the third metacarpus bone. The animals were divided into two groups (Control Group - CG and Experimental Group - EG). The application of extracorporeal shock wave therapy (ESWT) was performed on the right forelimb of the animals in the experimental group in the same place evaluated for bone elasticity and was used apparatus for extracorporeal therapy of waves with 0.15 mJ/mm2 energy flux density and 2000 pulses with E6R20 probe, with focus feature of the shock wave of 20 mm. The ESWT were repeated every 21 days, a total of three sessions (D0, D21 and D42). The analysis of bone elasticity determination was realized at D21, D42 and D72. The average of speed ultrasound differed between groups at D21, D42 and D72, and the animals from EG had lower bone mineral density after applications of ESWT. There was also difference in the analysis of bone mass (Z-Score) between the groups at D21 and D42, which animals from EG showed a significant decrease in bone mass. The risk of fracture was higher in animals from experimental group at D21. It was concluded that ESWT is able to promote change in bone mineral density.