The propagation of induced tendon lesions in the equine superficial digital flexor tendon: An ex vivo study

Reasons for performing study: Clinical tendon lesions usually enlarge during the first days to weeks after sustaining the injury due to enzymatic and biomechanical influences. Limiting this enlargement would positively influence prognosis related to lesion size. Objectives: To investigate the effect of cyclic loading on the propagation of enzymatically and physically induced tendon lesions and to assess the effect of immobilisation thereon in an ex vivo model. Methods: Equine cadaver limbs with either physically or collagenase‐induced SDFT lesions were placed in a pneumatic loading device. Groups consisted of unloaded limbs, cyclically loaded limbs and cyclically loaded limbs with fibreglass cast immobilisation. Lesion size was evaluated ultrasonographically and macroscopically and load at failure was determined for groups where lesions enlarged. Results: Physically induced lesions did not propagate after cyclical loading and were not further investigated. Collagenase‐induced lesions propagated significantly more after cyclical loading compared to unloaded limbs. This propagation could effectively be reduced by immobilisation through a lower limb cast. Immobilisation did not influence load at failure. Conclusions: The combination of enzymatic and mechanical stimulation caused the lesions to propagate most. Casting appeared effectively to reduce the enlargement of the original tendon lesions in this ex vivo model, while it did not affect ultimate tendon strength. Potential relevance: As prognosis of tendon injuries is related to lesion size, the outcome of this study warrants further investigations in vivo.     

The role of rituximab in the treatment of canine lymphoma: an ex vivo evaluation

Targeting the CD20 receptor that is common to many B-cell Non-Hodgkin's Lymphoma subtypes in people, rituximab is a chimeric monoclonal antibody which has significantly improved disease-free survival rates compared with the use of cytotoxic agents alone. This study evaluated ex vivo canine B cell binding and depletion by rituximab with flow cytometric technique as possible proof of concept for treatment of canine lymphoma. Despite immunohistochemistry supporting CD20 expression, rituximab did not bind or deplete canine B cells and it is unlikely that it will be added to the armamentarium of treatment options for canine lymphoma.     

Biomechanical characteristics of an implant used to secure semitendinosus-gracilis tendon grafts in a canine model of extra-articular anterior cruciate ligament reconstruction

Objective— To compare initial femoral fixation properties of a new implant, Graftgrab (GG), with 2 established methods of extra-articular femoral graft fixation, spiked washers (SW) and bone staples (ST).
Study Design— Experimental in vitro cohort study.
Methods— Canine semitendinosus–gracilis tendon grafts were passed through bone tunnels and fixed to the lateral surface of femoral condyles with spiked washers, bone staples, or new implant prototypes. The fixations were tested to failure with a single-cycle load at a rate of 50 mm/min. Failure and yield strength, stiffness, energy, and elongation were determined from load–displacement curves and failure modes were recorded.
Results— The graft failed midsubstance in 4 SW, 4 ST, and 1 GG fixations. In 3 SW, 3 ST, and 1 GG specimens, the graft slipped from the fixation. The graft ruptured at the clip (3) and the intra-articular (2) surface of the bone tunnel in the remaining GG specimens. There were no significant differences between fixation groups in femoral tunnel length, femoral width, or the mechanical properties evaluated.
Conclusions— The initial in vitro mechanical properties of the new fixation implant are comparable with those of spiked washers and bone staples.
Clinical Relevance— The initial mechanical performance of the new implant tested in this study was similar to those of comparable, established implants. The new implant is novel and may be useful for human anterior and veterinary cranial cruciate ligament graft reconstruction fixation.     

Evaluation of wraps covering the distal aspect of pelvic limbs for prevention of bacterial strike-through in an ex vivo canine model

Objective— To determine differences in bacterial strike-through for materials commonly used to cover the distal aspect of the pelvic limb during operative site preparation.
Study Design— Randomized block design; ex vivo model.
Animals— Canine cadaveric pelvic limbs (n=40).
Methods— Pelvic limbs (n=40) were randomly assigned to 4 treatment groups: Group 1=Vetrap^™+sterile Coban^™; Group 2=latex glove+Vetrap^™+sterile Coban^™; Group 3=latex glove+Vetrap^™+sterile Coban^™+sterile latex glove+sterile Coban^™; and Group 4=latex glove+Vetrap^™+sterile disposable drape+sterile Coban^™. Limbs were contaminated with a standardized bacterial solution and routinely prepared using the assigned distal leg wrap. Bandages were fluid challenged with a saline (0.9% NaCl) solution-soaked laparotomy sponge for 30 seconds. The wrap surface was sampled for microbial culture before surgical preparation, immediately after, and 60 minutes after applying a sterile leg wrap.
Results— Bacterial growth occurred in all Group 1 cultures, 90% of Group 2 cultures, and none of the Group 3 and 4 cultures, 60 minutes after applying the sterile wrap.
Conclusion— A distal leg wrap of Vetrap^™+sterile Coban^™ is not effective in preventing bacterial strike-through.
Clinical Relevance— If similar results occur in the live animal, then a sterile impermeable barrier must be incorporated into the distal leg wrap to prevent bacterial strike-through.     

Short-term cast immobilisation is effective in reducing lesion propagation in a surgical model of equine superficial digital flexor tendon injury

Reasons for performing the study: Larger superficial digital flexor tendon (SDFT) injuries have a poorer prognosis than smaller lesions. During the inflammatory phase enlargement of the initial lesion is frequently noted, with biomechanical loading being recently proposed to play an important role. Objectives: To evaluate the effect of lower limb cast immobilisation on tendon lesion propagation in an equine model of surgically induced SDFT injury. Methods: Core lesions were surgically induced in both front SDFTs of 6 young mature horses. At the end of surgery, one leg was randomly placed in a lower limb cast and the other leg (control) was bandaged for 10 days. Computerised ultrasonographic tissue characterisation performed at Days 10, 15, 21, 28, 35 and 42 allowed measurement of lesion length (cm) and width (expressed as a percentage of whole tendon cross‐section). On Day 42 horses were subjected to euthanasia and both SDFTs were sectioned every centimetre to assess the lesion length macroscopically. Statistics were performed to compare cast vs. control legs with significance set at P<0.05. Results: When all time points were combined, lesion length was 19% shorter (P<0.0001) and lesion width 57% smaller (P = 0.0002) in the cast legs (6.13 ± 0.12 cm; 6.90 ± 0.64%) than in the control legs (7.30 ± 0.21 cm; 10.85 ± 1.22%). On Day 42 the lesion length on macroscopic evaluation was 19% shorter (P = 0.04) in the cast (7.00 ± 0.36 cm) than in the control legs (8.33 ± 0.33 cm). Conclusions: Cast immobilisation for 10 days effectively reduced lesion propagation (length and width) compared to bandaging in an in vivo model of artificially‐induced tendon lesions. Potential relevance: A short period of cast immobilisation during the early phase of tendon healing may be an easy and cost‐effective way to reduce the initial enlargement of lesion size and hence to improve prognosis.     

Biomechanical evaluation of a novel nucleus pulposus prosthesis in canine cadaveric spines

Partial disc replacement is a new surgical technique aimed at restoring functionality to degenerated intervertebral discs (IVDs). The aim of the present study was to assess biomechanically the behaviour of a novel nucleus pulposus prosthesis (NPP) in situ and its ability to restore functionality to the canine IVD after nuclectomy alone or after combined dorsal laminectomy and nuclectomy. Nine canine T13-L5 specimens (L2L3 group) and 10 L5-Cd1 specimens (LS group) were tested biomechanically in the native state, after nuclectomy (L2L3 group) or after combined dorsal laminectomy and nuclectomy (LS group), and after insertion of the NPP. Range of motion (ROM), neutral zone (NZ), and neutral zone stiffness (NZS) were determined in flexion/extension, lateral bending, and axial rotation. Nuclectomy alone and combined dorsal laminectomy and nuclectomy caused significant instability in all motion directions. Implantation of the NPP resulted in significant restoration of the parameters (ROM, NZ, and NZS) towards the native state; however, fragmentation/herniation of the NPP occurred in 47% of the cases. In conclusion, the NPP has the ability to improve functionality of the nuclectomized canine IVD. The high rate of NPP failure requires modifications directed at the integrity of the NPP and its confinement to the nuclear cavity.     

Predicting likelihood of in vivo chemotherapy response in canine lymphoma using ex vivo drug sensitivity and immunophenotyping data in a machine learning model

We report a precision medicine platform that evaluates the probability of chemotherapy drug efficacy for canine lymphoma by combining ex vivo chemosensitivity and immunophenotyping assays with computational modelling. We isolated live cancer cells from fresh fine needle aspirates of affected lymph nodes and collected post‐treatment clinical responses in 261 canine lymphoma patients scheduled to receive at least 1 of 5 common chemotherapy agents (doxorubicin, vincristine, cyclophosphamide, lomustine and rabacfosadine). We used flow cytometry analysis for immunophenotyping and ex vivo chemosensitivity testing. For each drug, 70% of treated patients were randomly selected to train a random forest model to predict the probability of positive Veterinary Cooperative Oncology Group (VCOG) clinical response based on input variables including antigen expression profiles and treatment sensitivity readouts for each patient's cancer cells. The remaining 30% of patients were used to test model performance. Most models showed a test set ROC‐AUC > 0.65, and all models had overall ROC‐AUC > 0.95. Predicted response scores significantly distinguished (P < .001) positive responses from negative responses in B‐cell and T‐cell disease and newly diagnosed and relapsed patients. Patient groups with predicted response scores >50% showed a statistically significant reduction (log‐rank P < .05) in time to complete response when compared to the groups with scores <50%. The computational models developed in this study enabled the conversion of ex vivo cell‐based chemosensitivity assay results into a predicted probability of in vivo therapeutic efficacy, which may help improve treatment outcomes of individual canine lymphoma patients by providing predictive estimates of positive treatment response.     

Biomechanical evaluation of cervical intervertebral plug stabilization in an ovine model

Objective— (1) To determine the biomechanical stability of cervical spinal segment C5–C6 after ventral slot (VS) decompression and distraction–stabilization by a modified polymethylmethacrylate (PMMA) intervertebral plug technique (IVP) in an ovine model. (2) To determine if the distraction of the disk space is maintained after VS‐IVP, and the frequency of bony intervertebral union. Study Design— Experimental in vivo study in an ovine model with an in vitro control study. Animals— Adult (>2 years age), intact sheep (n=18). Methods— Ten sheep had VS/IVP surgery of C5–6 with computed tomography (CT) images obtained immediately before and after surgery, as well as 8 (n=10) and 24 (n=5) weeks postoperatively. Cervical spines harvested 8 weeks (8‐week group; n=5) and 24 weeks (24‐week group; n=5) after surgery had three‐dimensional (3D) flexibility tests in flexion–extension, axial rotation, and lateral bending. Image data was analyzed qualitatively and quantitatively in 3D. Eight ovine cervical spines served as controls and had biomechanical testing in intact condition (n=5) and after in vitro VS/IVP (n=5). Results— Significantly decreased range of motion (ROM) was noted in all loading modes when comparing in vitro operated spines and the 24‐week group to intact spines. The 8‐week group was not significantly different from the intact group, except in lateral bending. Quantitative CT analysis of treatment groups showed progressive disk space collapse and ventral implant migration. Intervertebral bony union did not occur. Conclusion— Biomechanical stability was obtained immediately after VS/IVP surgery, but ROM at 8 weeks was not different from intact spines. However, at 24 weeks, the operated level had regained stability similar to the immediate postoperative level. Distraction of the disk space was not maintained and bony union did not occur in this ovine model of VS/IVP. Clinical Relevance— In clinical situations requiring constant increased stability of cervical segments for an extended time, more rigid stabilization techniques are required.     

Luxation of the superficial digital flexor tendon in a cat

Superficial digital flexor luxation has been described in dogs, horses, and cattle. To the authors' knowledge, it has not been reported in cats. In the case of this report, monofilament nonabsorbable suture material was used to repair a laterally luxating superficial digital flexor tendon in a cat. The repair was similar to that which has been described in dogs. Whereas many etiologies of superficial tendon luxation have been proposed in dogs, trauma was believed to have contributed to the tendon luxation in this cat.     

Evaluation of polymyxin B in an ex vivo model of endotoxemia in horses

OBJECTIVE: To evaluate effects of polymyxin B sulfate (PMB) on response of horses to endotoxin, using an ex vivo model. ANIMALS: 8 healthy horses. PROCEDURE: In a crossover design, 3 doses of PMB (100, 1,000, and 10,000 U/kg of body weight) and physiologic saline solution (control) were evaluated. Prior to and for 24 hours after administration of PMB, blood samples were collected into heparinized tubes for use in 2 assays. For the endotoxin-induced tumor necrosis factor (TNF) assay, blood samples were incubated (37 C for 4 h) with 1 ng of Escherichia coli or Salmonella Typhimurium endotoxin/ml of blood. Plasma was harvested and assayed. For the residual endotoxin activity assay, plasma was collected into sterile endotoxin-free borosilicate tubes, diluted 1:10 with pyrogen-free water, and incubated for 10 minutes at 70 C. Escherichia coli endotoxin (0.1 or 1 ng/ml of plasma) was added to the thawed samples prior to performing the limulus ameobocyte lysate assay. Serum creatinine concentrations were monitored for 1 week. RESULTS: Compared with baseline values, PMB caused a significant dose- and time-dependent decrease in endotoxin-induced TNF activity. Compared with baseline values, residual endotoxin activity was significantly reduced after administration of 10,000 U of PMB/kg. Compared with baseline values, 1,000 and 5,000 U of PMB/kg should inhibit 75% of endotoxin-induced TNF activity for 3 and 12 hours, respectively. Serum creatinine concentrations remained within the reference range. CONCLUSION AND CLINICAL RELEVANCE: Results of the study suggest that PMB is a safe, effective inhibitor of endotoxin-induced inflammation in healthy horses.     

Tenosynovitis of the digital flexor tendon sheath and annular ligament constriction syndrome caused by longitudinal tears in the deep digital flexor tendon: a clinical and surgical report of 17 cases in warmblood horses

REASONS FOR PERFORMING STUDY: Inflammation of the digital flexor tendon sheaths is a chronic and nebulous condition often unresponsive to medical and surgical treatment. OBJECTIVES: To evaluate the incidence of longitudinal tears (LT) as the underlying cause of chronic tenosynovitis and annular ligament constriction syndrome (ALCS) in warmblood horses. METHODS: The records of 25 horses with chronic tenosynovitis and ALCS in which tenoscopical inspection of the digital flexor tendon sheath (DFTS) was performed between 1999-2000 were reviewed. Of 25 horses, 17 were diagnosed with an LT in the deep digital flexor tendon (DDFT). All horses had a history of distension of the DFTS and/or signs of an ALCS. All cases presented typical signs of a chronic inflamed DFTS on ultrasonography and 11 horses showed ultrasonographic changes (echogenic material or an irregular outlining) at the lateral or medial border of the DDFT. The diagnosis of LTs of the DDFT was established in all cases by tenoscopy. Surgical treatment consisted of removal of the torn collagen fibrils using a mechanical resector and decompressing the fetlock canal by a transection of the palmar annular ligament (PAL) using a hook knife under tenoscopic control. RESULTS: Ten horses became sound and resumed their previous level of work, 3 horses remained lame, 4 horses returned to previous level of work but needed intrasynovial treatment of the DFTS and reduced competition frequency to remain sound. CONCLUSIONS: Horses presented with chronic inflamed DFTS and/or ALCS might suffer from LTs in the DDFT; however, the diagnosis cannot be established with absolute certainty using only ultrasonography. Longitudinal tears should be suspected if ultrasonographic changes are present lateral or medial to the border of the DDFT but tenoscopical examination of the tendon sheath is essential to establish an accurate diagnosis and an effective treatment. POTENTIAL RELEVANCE: The presence of these LTs might explain why some cases of chronic tenosynovitis of the digital flexor tendon sheath and/or ALCS do not respond on surgical transection of the PAL alone without tenoscopy.