Mapping Stresses on the Tibial Plateau Cartilage in an Ovine Model Using In-Vivo Gait Kinematics.

Understanding stresses within the knee joint is central to understanding knee function, and the etiology and progression of degenerative joint diseases such as post-traumatic osteoarthritis. In this study, in vivo gait kinematics of four ovine subjects were recorded using a highly accurate Instrumented Spatial Linkage (ISL) as each subject walked on a standard treadmill. The subjects were then sacrificed, and the right hind limbs removed. Ten purpose-built Fibre Bragg Grating (FBG) sensors were positioned within each stifle joint and used to measured contact stresses on the articulating surface of the tibial plateau as the recorded gait was replicated using a 6-degrees-of-freedom parallel robotic system. This study provides the first accurate, direct measurement of stress in a joint during in vivo gait replication. It was hypothesized that the results would indicate a direct link between gait kinematics and measured stress values. Contrary to this expectation no direct link was found between individualistic differences in kinematics and differences in stress magnitudes. This finding highlights the complex multifactorial nature of stress magnitudes and distribution patterns across articular joints. The results also indicate that stress magnitudes within the knee joint are highly position dependent with magnitudes varying substantially between points only a few mm apart.

Correlation of damage score in PTOA with changes in stress on cartilage in an ovine model.

Objective: There is a high risk of developing osteoarthritis (OA) following traumatic injury to the knee. Severe ligament injuries can disrupt the integrity of the multicomponent knee at both biological and biomechanical levels. We hypothesize changes in cartilages stresses could lead to tissue damage and development of OA. Design: The in-vivo gait kinematics of the stifle (knee) joint of four adult female ovine subjects were recorded prior to and at ten-and-twenty weeks following partial ACL-MCL transection. The subjects were sacrificed and the experimental joint from each subject was mounted on a parallel robotic system programmed with the kinematic findings. Ten custom-built Fibre Bragg Grating optic sensors were arranged to measure contact stresses on the surface of the tibial plateau articular cartilage. These sensors provide the first accurate stress measurements in a joint during gait replication using the previously recorded in-vivo kinematics. The relationship between the results obtained and observed focal damage was assessed. Results: The locations on the tibial plateaus that experienced the greatest change in contact stresses corresponded with the locations of focal damage development. No direct link was detected between individual animal differences in kinematics and variations in stress magnitudes or the development of focal cartilage damage. Conclusions: The findings highlight the importance of mechanical stress determinants in the integrated set point for the knee (with individual variation), and how injury-related stress changes correlate with development of PTOA.

Relative Surface Velocity of the Tibiofemoral Joint and Its Relation to the Development of Osteoarthritis After Joint Injury.

The relative velocity of the tibiofemoral surfaces during gait before and after partial-ACL and full MCL transection (p-ACL/MCL Tx) was examined in an ovine model (N = 5) and the relation between the variation in the relative sliding velocity component and gross morphological damage was investigated. We defined the in vivo kinematics of the tibiofemoral joints by using an instrumented spatial linkage and then determining the relative velocity components on the reconstructed femoral condyle surfaces. One major finding was that the magnitude of the relative velocity components was relatively high during the initial stance period of the gait and oscillated with a decaying envelope. Interestingly, for most subjects, the highest value of relative sliding velocity occurred during the stance phase, and not swing. The magnitude of the relative velocity components was increased in 3/5 subjects during stance after an injury. For the lateral compartment, there was a significant correlation (p value = 0.005) between the joint gross morphological damage and the increase in the maximum relative sliding velocity during stance. For the medial compartment, there was a trend (p value < 0.1) between the joint gross morphological score and the increase in the maximum relative sliding velocity during stance, 20 weeks after injury. In conclusion, a connection between an increase in the relative surface velocity and gross morphological damage might be due to an increase in the normal stress and the plowing friction between the surfaces.

Alterations in Joint Angular Velocity Following Traumatic Knee Injury in Ovine Models.

Little effort has been directed towards the consequence of an injury on joint angular velocity. We hypothesized that the magnitude of knee joint angular velocity would be decreased after injury. Four injury groups were investigated in an ovine model: (1) anterior cruciate ligament (ACL) and medial collateral ligament (MCL) transection (ACL/MCL Tx) (n = 5), (2) lateral meniscectomy (Mx) (n = 5), (3) partial ACL transection (p-ACL Tx) (n = 5), and (4) partial-ACL and MCL transection (p-ACL/MCL Tx) (n = 5). The magnitude of the angular velocities decreased in the subjects of all groups at multiple points of the gait cycle. The maximum angular velocities during stance and the maximum extension angular velocities during swing were decreased in 15/20 and 17/20 subjects, respectively. There were strong correlations between morphological osteoarthritis scores and the reduction in the maximum extension angular velocities during swing 40 weeks post-p-ACL Tx and 20 weeks post-ACL/MCL Tx. There was no correlation between the decrease of the angular velocity and morphological osteoarthritis scores in the Mx group and the p-ACL/MCL Tx group. The reduction in angular velocity may be a helpful addition as a surrogate measure of OA risk after ACL injury, and could have clinical significance after further investigation in humans.

Anti-Inflammatory Benefits of Antibiotics: Tylvalosin Induces Apoptosis of Porcine Neutrophils and Macrophages, Promotes Efferocytosis, and Inhibits Pro-Inflammatory CXCL-8, IL1α, and LTB4 Production, While Inducing the Release of Pro-Resolving Lipoxin A4 and Resolvin D1.

Excessive accumulation of neutrophils and their uncontrolled death by necrosis at the site of inflammation exacerbates inflammatory responses and leads to self-amplifying tissue injury and loss of organ function, as exemplified in a variety of respiratory diseases. In homeostasis, neutrophils are inactivated by apoptosis, and non phlogistically removed by neighboring macrophages in a process known as efferocytosis, which promotes the resolution of inflammation. The present study assessed the potential anti-inflammatory and pro-resolution benefits of tylvalosin, a recently developed broad-spectrum veterinary macrolide derived from tylosin. Recent findings indicate that tylvalosin may modulate inflammation by suppressing NF-κB activation. Neutrophils and monocyte-derived macrophages were isolated from fresh blood samples obtained from 12- to 22-week-old pigs. Leukocytes exposed to vehicle or to tylvalosin (0.1, 1.0, or 10 µg/mL; 0.096-9.6 µM) were assessed at various time points for apoptosis, necrosis, efferocytosis, and changes in the production of cytokines and lipid mediators. The findings indicate that tylvalosin increases porcine neutrophil and macrophage apoptosis in a concentration- and time-dependent manner, without altering levels of necrosis or reactive oxygen species production. Importantly, tylvalosin increased the release of pro-resolving Lipoxin A4 (LXA4) and Resolvin D1 (RvD 1 ) while inhibiting the production of pro-inflammatory Leukotriene B4 (LTB4) in Ca2+ ionophore-stimulated porcine neutrophils. Tylvalosin increased neutrophil phospholipase C activity, an enzyme involved in releasing arachidonic acid from membrane stores. Tylvalosin also inhibited pro-inflammatory chemokine (C-X-C motif) ligand 8 (CXCL-8, also known as Interleukin-8) and interleukin-1 alpha (IL-1α) protein secretion in bacterial lipopolysaccharide-stimulated macrophages. Together, these data illustrate that tylvalosin has potent immunomodulatory effects in porcine leukocytes in addition to its antimicrobial properties.

Longitudinal evaluation of diagnostics in experimentally infected young calves during subclinical and clinical paratuberculosis.

Five calves were inoculated orally at 2 weeks of age with a dose of 5 × 10(9) colony-forming units of Mycobacterium avium subspecies paratuberculosis (MAP) on 2 consecutive days. Two calves developed clinical Johne's disease at 12 and 16 months of age after being consistently positive for MAP on fecal culture and antibody enzyme-linked immunosorbent assay (ELISA), starting 2 to 3 weeks and 4 to 5 months after inoculation, respectively.

Évaluation longitudinale du diagnostic lors de paratuberculose subclinique et clinique chez de jeunes veaux infectés expérimentalement. Cinq veaux ont été inoculés oralement à l'âge de 2 semaines avec une dose de 5 × 109 unités formatrices de colonies de Mycobacterium avium sous-espèce paratuberculosis (MAP) pendant 2 jours consécutifs. Deux veaux ont développé la maladie de Johne clinique à l'âge de 12 et de 16 mois après avoir obtenu des résultats constamment positifs pour MAP aux cultures de fèces et à l'ELISA, commençant entre 2 et 3 semaines et 4 et 5 mois après l'inoculation, respectivement.(Traduit par Isabelle Vallières).

Immunomodulatory effects of tulathromycin on apoptosis, efferocytosis, and proinflammatory leukotriene B4 production in leukocytes from Actinobacillus pleuropneumoniae-or zymosan-challenged pigs.

OBJECTIVE: To investigate the anti-inflammatory and immunomodulatory properties of tulathromycin in vitro and in experimental models of Actinobacillus pleuropneumoniae-induced pleuropneumonia and zymosan-induced pulmonary inflammation in pigs. ANIMALS: Blood samples from six 8- to 30-week-old healthy male pigs for the in vitro experiment and sixty-five 3-week-old specific pathogen-free pigs. PROCEDURES: Neutrophils and monocyte-derived macrophages were isolated from blood samples. Isolated cells were exposed to tulathromycin (0.02 to 2.0 mg/mL) for various durations and assessed for markers of apoptosis and efferocytosis. For in vivo experiments, pigs were inoculated intratracheally with A pleuropneumoniae, zymosan, or PBS solution (control group) with or without tulathromycin pretreatment (2.5 mg/kg, IM). Bronchoalveolar lavage fluid was collected 3 and 24 hours after inoculation and analyzed for proinflammatory mediators, leukocyte apoptosis, and efferocytosis. RESULTS: In vitro, tulathromycin induced time- and concentration-dependent apoptosis in neutrophils, which enhanced their subsequent clearance by macrophages. In the lungs of both A pleuropneumoniae- and zymosan-challenged pigs, tulathromycin promoted leukocyte apoptosis and efferocytosis and inhibited proinflammatory leukotriene B4 production, with a concurrent reduction in leukocyte necrosis relative to that of control pigs. Tulathromycin also attenuated the degree of lung damage and lesion progression in A pleuropneumoniae-inoculated pigs. CONCLUSIONS AND CLINICAL RELEVANCE: Tulathromycin had immunomodulatory effects in leukocytes in vitro and anti-inflammatory effects in pigs in experimental models of A pleuropneumoniae infection and nonmicrobial-induced pulmonary inflammation. These data suggested that in addition to its antimicrobial properties, tulathromycin may dampen severe proinflammatory responses and drive resolution of inflammation in pigs with microbial pulmonary infections.

Anti-inflammatory effects of retinoids and carotenoid derivatives on caspase-3-dependent apoptosis and efferocytosis of bovine neutrophils.

OBJECTIVE: To evaluate immunomodulatory properties of all-trans retinoic acid and a fully oxidized ß-carotene dietary product in calves with Mannheimia haemolytica-induced pneumonia. ANIMALS: Twenty-five 6- to 10-week-old male Holstein calves for experimental inoculations and three 8- to 30-week-old Angus heifers for blood donations. PROCEDURES: In vitro, neutrophils and monocyte-derived macrophages isolated from blood of healthy Angus heifers were treated with all-trans retinoic acid (1 µM) or fully oxidized ß-carotene (8.3 µg/mL) for various times and assessed for markers of cellular death, antimicrobial function, and production of proinflammatory leukotriene B4. Following 28 days of dietary supplementation with fully oxidized ß-carotene, Holstein calves were experimentally inoculated with M haemolytica. Bronchoalveolar lavage fluid was collected at 3 and 24 hours after challenge inoculation and analyzed for markers of apoptosis. RESULTS: In vitro, all-trans retinoic acid and fully oxidized ß-carotene induced cell-selective, caspase-3-dependent apoptosis in neutrophils, which subsequently enhanced efferocytosis in macrophages. Conversely, neither treatment altered phorbol 12-myristate 13-acetate-induced oxidative burst, phagocytosis of nonopsonized zymosan (complement or antibody independent), or M haemolytica-induced leukotriene B4 production in bovine neutrophils. In vivo, fully oxidized ß-carotene enhanced leukocyte apoptosis in bronchoalveolar lavage fluid as well as subsequent efferocytosis by macrophages without altering numbers of circulating leukocytes. CONCLUSIONS AND CLINICAL RELEVANCE: Neutrophil apoptosis and subsequent efferocytosis by macrophages are key mechanisms in the resolution of inflammation. Findings for the present study indicated that all-trans retinoic acid and fully oxidized ß-carotene could be novel nutraceutical strategies that may confer anti-inflammatory benefits for cattle with respiratory tract disease.

Transcutaneous intraluminal impedance measurement for minimally invasive monitoring of gastric motility: validation in acute canine models.

Transcutaneous intraluminal impedance measurement (TIIM) is a new method to cutaneously measure gastric contractions by assessing the attenuation dynamics of a small oscillating voltage emitted by a battery-powered ingestible capsule retained in the stomach. In the present study, we investigated whether TIIM can reliably assess gastric motility in acute canine models. Methods. Eight mongrel dogs were randomly divided into 2 groups: half received an active TIIM pill and half received an identically sized sham capsule. After 24-hour fasting and transoral administration of the pill (active or sham), two force transducers (FT) were sutured onto the antral serosa at laparotomy. After closure, three standard cutaneous electrodes were placed on the abdomen, registering the transluminally emitted voltage. Thirty-minute baseline recordings were followed by pharmacological induction of gastric contractions using neostigmine IV and another 30-minute recording. Normalized one-minute baseline and post-neostigmine gastric motility indices (GMIs) were calculated and Pearson correlation coefficients (PCCs) between cutaneous and FT GMIs were obtained. Statistically significant GMI PCCs were seen in both baseline and post-neostigmine states. There were no significant GMI PCCs in the sham capsule test. Further chronic animal studies of this novel long-term gastric motility measurement technique are needed before testing it on humans.

Suture marker lesion detection in the colon by self-stabilizing and unmodified capsule endoscopes: pilot study in acute canine models.

BACKGROUND: Capsule endoscopy is a noninvasive method for examining the small intestine. Recently, this method has been used to visualize the colon. However, the capsule often tumbles in the wider colon lumen, resulting in potentially missed pathology. In addition, the capsule does not have the ability to distend collapsed segments of the organ. Self-stabilizing capsule endoscopy is a new method of visualizing the colon without tumbling and with the ability to passively distend colon walls. OBJECTIVE: To quantitatively compare the detection rate of intraluminal suture marker lesions for colonoscopy by using a custom-modified, self-stabilizing capsule endoscope (SCE); an unmodified capsule endoscope (CE) of the same brand; and a standard colonoscope. DESIGN: Four mongrel dogs underwent laparotomy and the implantation of 5 to 8 suture markers to approximate colon lesions. Each dog had both capsule endoscopy and self-stabilizing capsule endoscopy, administered consecutively in random order. In each case, the capsule was inserted endoscopically into the proximal lumen of the colon followed by pharmacologically induced colon peristalsis to propel it distally through the colon. Blinded standard colonoscopy was performed by an experienced gastroenterologist after the capsule endoscopies. SETTING: Experimental study in a live canine model. SUBJECTS: Four dogs. INTERVENTION: Laparotomy, capsule endoscopy, colonoscopy. MAIN OUTCOME MEASUREMENTS: Comparison of the marker detection rate of the SCE to that of the unmodified MiroCam CE and a colonoscope. RESULTS: The average percentages of the marker detection rate for unmodified capsule endoscopy, self-stabilizing capsule endoscopy, and colonoscopy, respectively, were 31.1%, 86%, and 100% (P < .01), with both self-stabilizing capsule endoscopy and colonoscopy performing significantly better than the unmodified capsule endoscopy. LIMITATIONS: Acute canine model, suture markings poorly representative of epithelial polyps, limited number of animals. CONCLUSION: The proposed self-stabilizing capsule endoscope delivered a significant improvement in detection rates of colon suture markings when compared with the unmodified capsule endoscope.