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  • Author or Editor: S. A. Kincaid x
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Abstract

Objective

To evaluate the ability of hyaluronic acid (HA), with and without transforming growth factor β (TGF-β), to stabilize the catabolic processes associated with atrophy of articular cartilage.

Animals

20 adult, skeletally normal, hound-type dogs.

Procedure

Dogs (20 to 30 kg) were randomly assigned to 1 of 5 groups. One group served as untreated controls. Bivalve casts were placed on the left hind limbs of the remaining 16 dogs to limit weightbearing and motion of the limb for 92 days. One group served as the cast control. Beginning on day 56, 3 groups received aseptic intra-articular injections in the left stifles of either 5 mg of HA or 5 mg of HA containing either 20 or 50 μg of TGF-β. Intra-articular injections were repeated at 4-day intervals until the end of the study. On day 92, stifles were harvested at necropsy. Medial femoral condyles were histologically processed, and the articular cartilage was stained for the presence of proteoglycans, stromelysin, tumor necrosis factor (TNF) α, and TNF receptors (p55 and p75).

Results

Decreased metachromasia was evident in the cartilage matrix of all cast groups, with the smallest decrease in the HA-treated group. Stromelysin was immunolocalized in articular cartilage of the cast (left) limbs of cast control and both HA/TGF-β-treated groups. TNF-α was localized in articular cartilage of all cast (left) and right limbs, except those of the HA-treated group. Receptors for TNF were observed in both limbs of untreated control and cast control groups and cast limbs of HA/TGF-β-treated groups. The receptors were not localized in the right limbs of the HA with or without TGF-β-treated groups. TGF-β did not decrease stromelysin or TNF-α or receptors at the doses used.

Conclusions

HA may mediate a chondrostabilizing influence on articular cartilage by down-regulating TNF-α. Importantly, HA appeared to exert its inhibitory influence on TNF-α, as well as stromelysin and TNF receptors, on a systemic basis.

Clinical Relevance

Results provide insight into the mode of action of HA as a therapeutic agent for arthritis and its stabilizing influence on cartilage metabolism. (Am J Vet Res 1996;57:1488-1496)

Free access
in American Journal of Veterinary Research

Abstract

Objectives

To use lipopolysaccharide (LPS) to create synovitis in the midcarpal joint of ponies, and to assess the morphologic, histochemical, and immunohistochemical effects of synovitis on articular cartilage of the third carpal bone.

Animals

2- to 3-year-old ponies, 6 control (group 1) and 6 treated (group 2).

Procedure

Synovitis was induced in 1 midcarpal joint of group-2 ponies by intra-articular injections of LPS (0.02 μg/kg of body weight), morphine (0.1 mg/kg), and saline solution (group 2a) and morphine and saline solution alone in the contralateral midcarpal joint (group 2b). Articular cartilage sections and attached synovial membrane from the third carpal bones were examined by immunohistochemical distribution of interleukin 1β, tumor necrosis factor (TNF)-α, TNF receptors (P55, P75) and 3-B-3(–) epitopes, and by localization of proteoglycans (metachromatic staining). Proteoglycan extracts were assessed by metachromatic staining or western blotting and immunohistochemical staining, using anti-3-B-3 antibodies.

Results

Enhanced immunoreactivity for the cytokines and receptors was found in inflamed synovial membrane and noncalcified cartilage (group 2a more than 2b). Metachromasia of the noncalcified cartilage was greater in group-1 than in group-2a and group-2b specimens. In group 2a, chondrocyte hypertrophy and enhanced immunoreactivity for 3-B-3(–) epitope in areas of increased cytokine immunoreactivity suggested possible phenotypic change of the chondrocytes in response to synovitis. Immunohistochemical analysis by western blotting of proteoglycan extracts indicated strong 3-B-3(–) epitope immunolocalization in group-2a, weaker staining in group-2b, and barely detectable stain in group-1 specimens, which correlated with in situ immunolocalization.

Conclusions

Intra-articular administration of LPS may be used to induce a synovial environment conducive to increased immunoreactivity of interleukin 1β, TNF-α, and its receptors in equine synovial membrane and articular cartilage. These cytokines may be involved in the early phenotypic change of chondrocytes that is believed to occur in osteoarthritis and is characterized in this study by enhanced 3-B-3(–) epitope immunoreactivity and chondrocyte hypertrophy. (Am J Vet Res 1996;57:1080–1093)

Free access
in American Journal of Veterinary Research

Abstract

Objective

To ascertain the effects of locally injected immunostimulant and tripeptide-copper complex (TCC) on improving healing of pad wounds.

Design

Wounds in pads of large dogs were injected with either medication or physiologic saline solution (controls). Healing was evaluated.

Animals

12 mature English Pointers.

Procedure

Full-thickness 6 × 8-mm wounds in metatarsal and third and fourth digital pads were injected with immunostimulant or TCC at 0, 3, and 6 days after wounding. Wounds on control dogs were injected with physiologic saline solution. Using planimetric measurements at 0, 3, 6, 14, and 21 days, rates of healing were evaluated. Biopsy of the digital pad wounds at 3, 6, and 14 days was used to evaluate collagen content by hydroxyproline analysis. Biopsy specimens were also evaluated for type-I and type-III collagen, using Sirius red differential staining.

Results

Effect on healing rate and hydroxyproline content was best during the first week for immunostimulant. Immunostimulant- and TCC-injected wounds had more type-I collagen than did controls at 6 days; TCC-injected wounds had the most type-I collagen. At 14 days, the amount of type-I collagen in TCC-injected wounds was significantly greater than that in other wounds.

Conclusions

Tested medications had positive effects on healing of pad wounds.

Clinical Relevance

Intralesional injection of medications helps ensure their presence for enhancement of wound healing. The benefit could be lost with topical use in a bandage if the bandage is lost or becomes wet.(Am J Vet Res 1996;57:394-399)

Free access
in American Journal of Veterinary Research

Abstract

Objective—To study the musculoskeletal development of Great Dane puppies fed various dietary concentrations of calcium (Ca) and phosphorus (P) in fixed ratio by use of dual energy x-ray absorptiometry (DEXA), determination of serum insulin-like growth factor I and parathyroid hormone concentrations, radiography, and blood chemistry analysis results.

Animals—32 purebred Great Dane puppies from 4 litters.

Procedure—At weaning, puppies were assigned randomly to 1 of 3 diets. Blood was collected for biochemical analyses and hormone assays, and radiography and DEXA were performed through 18 months of age. Changes in body weight, bone mineral content, fat tissue weight, lean mass, result of serum biochemical analyses, hormonal concentrations, and radius lengths were analyzed through 18 months of age.

Results—Bone mineral content of puppies correlated positively with Ca and P content of the diets fed. Significant differences between groups in bone mineral content, lean mass, and body fat were apparent early. The disparity among groups increased until 6 months of age and then declined until body composition was no longer different at 12 months of age. Accretion rates for skeletal mineral content, fat, and lean tissue differed from each other and by diet group.

Conclusions and Clinical Relevance—Ca and P concentrations in the diet of young Great Dane puppies are rapidly reflected in the bone mineral content of the puppies until 5 to 6 months of age, after which hormonal regulation adjusts absorption and excretion of these minerals. Appropriate Ca and P concentrations in diets are important in young puppies < 6 months of age. (Am J Vet Res 2002;63:1036–1047)

Full access
in American Journal of Veterinary Research