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History

An approximately 12-hour-old Thoroughbred-Hanoverian crossbreed colt was evaluated by the University of Missouri Equine Ambulatory Service during a routine new foal examination. The dam of the foal was a maiden mare. The foal reportedly needed some assistance with rising initially but had nursed well since birth and eventually was able to rise on its own. The umbilicus had been dipped in chlorhexidine solution, and the foal had been observed to pass only a very small amount of meconium. The following morning, at approximately 24 hours after birth, the foal was found in lateral recumbency and unwilling to rise. The

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in Journal of the American Veterinary Medical Association
in Journal of the American Veterinary Medical Association

Abstract

Objective—To determine the effects of interleukin (IL)-1 and tumor necrosis factor (TNF)-α on canine chondrocytes cultured in an agarose-based 3-dimensional (3-D) system.

Sample Population—Humeral head articular cartilage chondrocytes obtained from 6 adult dogs.

Procedure—Chondrocytes were cultured in a 3-D system for ≤ 12 days in serum-free medium with IL-1α, IL-1β, or TNF-α at concentrations of 20, 50, or 100 ng/mL. After 1, 3, 6, and 12 days, glycosaminoglycan (GAG) concentrations in 3-D constructs; nitric oxide and prostaglandin E2 (PGE2) concentrations in media samples; and relative expressions of selected genes, including metalloproteinase (MMP)-13 and tissue inhibitor of metalloproteinase (TIMP)-1 and TIMP-2, were evaluated. Control specimens were comprised of chondrocytes cultured without proinflammatory cytokines.

Results—In control 3-D constructs, GAG content was significantly higher than for all other constructs. Compared with control values, relative expressions of MMP-13, TIMP-1, and TIMP-2 genes in the IL-1β (50 ng/mL) group were significantly higher at day 1; at all evaluations, media concentrations of nitric oxide were significantly higher in all TNF-α–treated cultures; and concentrations of PGE2 in media samples were significantly higher in the IL-1β (50 ng/mL) and IL-1β (100 ng/mL) groups at days 1 and 3, in the IL-1β (100 ng/mL) group at day 6, and in all TNF-α groups at days 1, 3, and 6.

Conclusions and Clinical Relevance—Results suggested that TNF-α more readily induces production of nitric oxide and PGE2 by canine chondrocytes, compared with IL-1β. In vitro, IL-1α appeared to have a minimal effect on canine chondrocytes. (Am J Vet Res 2005;66:1187–1196)

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in American Journal of Veterinary Research
History

A 13-year-old castrated male domestic shorthair cat was evaluated by a referring veterinarian because of progressive hair loss, anorexia, diarrhea, and profound weakness that developed over a period of a few weeks. The cat was referred to the Veterinary Medical Teaching Hospital at the University of Missouri for further assessment.

Clinical and Gross Findings

On initial evaluation, the cat's mentation was obtunded and mild dehydration was evident. Severe alopecia was present on the ventral aspects of the abdomen and neck, axillae, medial aspects of the forelimbs and hind limbs, periocular region, and nasal planum ( Figure 1 ). The

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in Journal of the American Veterinary Medical Association

Abstract

Objective—To assess the cellular, biochemical, and histologic effects of bipolar radiofrequency-generated heat on canine articular cartilage.

Sample Population—Articular cartilage explants (n = 72) from 6 canine cadavers and cultured articular chondrocytes from 5 canine cadavers.

Procedure—Cartilage explants were randomly assigned to receive no treatment or treatment with focal (3 seconds) or diffuse bipolar radiofrequency. Following treatment, methylene blue permeability assay was performed (n = 12) and remaining samples (60) were cultured. Immediately and 5, 10, and 20 days after treatment, cultured explants were assessed for glycosaminoglycan (GAG) and collagen contents, type II collagen and matrix metalloproteinase (MMP)-13 immunoreactivity, and modified Mankin histologic scores. Liquid culture media were collected every 4 days and GAG content measured. Additionally, cultured chondrocytes were exposed for 3 seconds to media preheated to 37°, 45°, or 55°C. Cell viability was determined via 2 different assays immediately and 24 hours after treatment.

Results—Radiofrequency-treated cartilage had reduced permeability and considerable histologic damage, compared with control samples; most treated samples had reduced collagen II staining and increased MMP-13 immunostaining. Compared with other treatments, less GAGs were released from cartilage after diffuse radiofrequency treatment throughout the study period. Cell viability was significantly different between controls and cells treated at 55°C immediately and 24 hours after heat treatment.

Conclusions and Clinical Relevance—In this study, bipolar radiofrequency treatment had detrimental effects on normal articular cartilage cells and extracellular matrix with probable long-term clinical consequences. The usefulness of radiofrequency for treatment of osteoarthritic articular cartilage requires further investigation. ( Am J Vet Res 2004;65:604–609)

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in American Journal of Veterinary Research

Abstract

Objective—To elucidate tissue inhibitor of metalloproteinase (TIMP)-mediated effects on chondrocytes.

Sample Population—Articular cartilage from humeral heads of 6 dogs.

Procedure—Chondrocytes from harvested specimens were cultured in 3-dimensional (3-D) agarose at 106 cells/mL. We prepared 3-D constructs exposed to only tumor necrosis factor (TNF)-α (50 ng/mL). Recombinant human TIMP-1 (255nM), -2 (285nM), or -3 (250nM) was added to liquid media bathing 3-D constructs cultured with TNF-α. Chondrocytes cultured without TIMP or TNF-α served as control samples. Samples of liquid media were collected on days 6, 9, 15, and 21 of culture for evaluation of glycosaminoglycan (GAG) and nitric oxide concentrations. The 3-D constructs were collected on days 9, 15, and 21 for evaluation of GAG, hydroxyproline (HP), and DNA contents.

Results—GAG content in control samples increased significantly during the study, whereas GAG content in 3-D constructs cultured with TNF-α or TNF-α plus TIMP did not increase. On day 9, GAG release from 3-D constructs cultured with TNF-α was significantly higher than that in other constructs. The HP content in control samples increased during the study and was significantly higher than that in all other constructs on day 21. Concentrations of nitric oxide were significantly lower in control samples on day 6, compared with concentrations for all other constructs.

Conclusions and Clinical Relevance—Addition of TIMPs did not counteract suppression of GAG and HP accumulation in 3-D constructs exposed to TNF-α. Apparently, adverse effects on chondrocytes exposed to TNF-α cannot be prevented by addition of TIMP alone. (Am J Vet Res 2004;65:1611–1615)

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in American Journal of Veterinary Research
History

A 4.5-year-old castrated male domestic shorthair cat was evaluated at the University of Missouri Veterinary Medical Teaching Hospital emergency service because of acute ataxia that progressed to recumbency and decreased responsiveness. The cat was housed at a local car mechanic garage and was last observed to behave normally 9 hours prior to the evaluation. Previous medical history was unremarkable, and no medications were being administered to the cat.

Clinical and Clinicopathologic Findings

Upon examination, the cat was assessed as 7% dehydrated, laterally recumbent, and minimally responsive, with a rectal temperature of 35.6°C (96.1°F). Both third eyelids were prolapsed; no

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in Journal of the American Veterinary Medical Association

Abstract

Objective—To investigate the presence or absence of Toll-like receptor (TLR)-2 and TLR-4 in synovial tissues collected from stifle joints (SJs) of dogs with or without osteoarthritis.

Animals—21 purpose-bred research dogs, 3 client-owned dogs with SJ osteoarthritis, and 3 dogs without SJ osteoarthritis.

Procedures—Research dogs underwent arthroscopic surgery in 1 SJ to induce osteoarthritis via cranial cruciate ligament transection (CrCLt; n = 5), femoral condylar articular cartilage groove creation (6), or release of the caudal horn of the medial meniscus (5); 5 dogs underwent sham surgery. Synovial tissue specimens were obtained from both stifle joints of each dog 12 weeks after surgery, and TLR-2 and TLR-4 gene expression were determined via real-time reverse transcription PCR assays. Expression of TLR-4 protein was determined via an immunofluorescence technique in additional specimens obtained from osteoarthritic SJs of dogs with cranial cruciate ligament insufficiency and from dogs with nonosteoarthritic SJs.

Results—Synovial tissues from CrCLt-treated joints had significantly higher TLR-4 gene expression, compared with the contralateral control SJs or any other joint group. TLR-2 gene expression did not differ significantly among groups. Toll-like receptor-4 protein was detected in synovial tissues of osteoarthritic SJs but was rarely evident in nonosteoarthritic SJs.

Conclusions and Clinical Relevance—Increased TLR-4 gene expression in the synovial tissue of SJs with osteoarthritis secondary to CrCLt suggests that activation of innate immunity may play a role in the pathophysiology of SJ osteoarthritis in at least a subset of dogs.

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in American Journal of Veterinary Research

Abstract

Objective—To characterize chondrocytes from naturally occurring osteochondrosis (OC) lesions of the humeral head of dogs.

Sample Population—15 cartilage specimens from 13 client-owned dogs with humeral head OC and 10 specimens from the humeral head of healthy dogs (controls).

Procedure—Chondrocytes were isolated and cultured in a 3-dimensional system. On days 7, 10, 15, 20, and 25, glycosaminoglycan and hydroxyproline content and cytologic characteristics were evaluated. Expression of collagen types I, II, and X was assessed by use of immunohistochemistry.

Results—Chondrocytes from OC lesions were less viable, compared with control chondrocytes. Glycosaminoglycan content in the OC group was significantly less than in the control group on all days except day 20. Hydroxyproline content was also significantly less in the OC group on days 10, 20, and 25. Expression of collagen type II was significantly less in the OC group, compared with the control group on all days, whereas expression of collagen type I was significantly greater in the OC group on days 20 and 25. Expression of collagen type X was significantly less in the OC group on all days except day 25.

Conclusions and Clinical Relevance—Chondrocytes from naturally occurring OC lesions of the humeral head of dogs cultured in a 3-dimensional system were less viable and less capable of producing appropriate extracellular matrix molecules than chondrocytes from unaffected dogs. Alterations in the synthetic capabilities of chondrocytes from OC-affected cartilage may be a cause or an effect of the disease process. (Am J Vet Res 2002;63:186–193)

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in American Journal of Veterinary Research
History

A 1.5-year-old sexually intact male sugar glider (Petaurus breviceps) underwent an emergency evaluation at the Kansas State University Veterinary Teaching Hospital because of decreased activity, tenesmus, and possible seizure-like activity. The sugar glider became increasingly lethargic and developed emesis, voided red-brown urine, and died within hours after arrival at the hospital. The body was submitted to the Kansas State Veterinary Diagnostic Laboratory for necropsy.

Clinical and Gross Findings

No clinicopathologic tests were performed prior to the death. At necropsy, the sugar glider weighed 85 g (0.19 lb) and was in fair body condition with mild postmortem autolysis.

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in Journal of the American Veterinary Medical Association