Gene expression in hip soft tissues in incipient canine hip dysplasia and osteoarthritis.

Canine hip dysplasia and developmental dysplasia of the human hip share demographic, phenotypic, and clinical features including the predisposition to develop osteoarthritis in affected joints. To support the results of genetic mapping studies for CHD and its concomitant osteoarthritis with functional information, we performed RNA-seq on hip capsule and teres ligament of affected and unaffected dogs. RNA seq showed that expressed genes segregated according age, capsule or ligament, and hip phenotype. Expression of HHIP, DACT2, and WIF1 was significantly higher in capsule from control hips than dysplastic hips indicating a disruption of the hedgehog signaling pathway. Expression of SPON 1, a key component of the WNT pathway, was increased significantly in both dysplastic capsule and ligament while FBN2 and EMILIN3 were significantly increased in dysplastic capsule. Of genes associated with human hip osteoarthritis, expression of ACAN, IGF1, CILP2, COL11A1, COL8A1, and HAPLN was increased significantly in dysplastic capsule. The significant increase in expression of PLA2F, TNFRSF, TMEM, and IGFBP in dysplastic capsule indicated an injury response. Gene set enrichment analysis revealed that genes involved in extracellular matrix structure, epithelial to mesenchymal transition, myogenesis, growth factor signaling, cancer and immune pathways were enriched in dysplastic capsule. For teres ligament from dysplastic joints, genes in retinoic signaling pathways and those encoding extracellular matrix molecules, but not proteoglycans, were enriched. Hip tissues respond to abnormal mechanics early in dysplastic hip development and these pathways present targets for intervention in the early synovitis and capsulitis secondary to canine and human hip dysplasia. © 2018 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 37:313-324, 2019.

Direct and indirect markers of cartilage metabolism in synovial fluid obtained from dogs with hip dysplasia and correlation with clinical and radiographic variables.

OBJECTIVE: To compare activities of interleukin (IL)-1beta, IL-6, tumor necrosis factor (TNF)-alpha, and matrix metalloproteinase (MMP)-3 and contents of sulfated glycosaminoglycan (S-GAG) in joint fluid obtained from dogs with hip dysplasia (HD) and clinically normal dogs, evaluate correlations among these markers in joint fluid obtained from dogs with HD, and evaluate correlations between each marker and clinical and radiographic variables. Animals-26 dogs with HD (clinical group) and 43 clinically normal Beagles (control group). PROCEDURE: Joint fluid was aseptically collected from the hip joints of all dogs. For each dog in the clinical group, age, duration of lameness, radiographic osteoarthritis (OA) score, and Norberg angle in each affected joint were recorded. Activities of IL-1beta, IL-6, TNF-alpha, and MMP-3 and S-GAG contents were measured. Values were compared between groups by use of Mann-Whitney U tests, and the Spearman rank correlation test was used to evaluate correlations among markers and between each marker and clinical or radiographic variables. RESULTS: Values of all markers were significantly higher for the clinical group, compared with values for the control group. There was a moderate positive correlation between lameness duration and IL-6 activity and a strong negative correlation between the Norberg angle and IL-1beta activity. CONCLUSIONS AND CLINICAL RELEVANCE: Analysis of our results indicated that there was a significant increase in markers of OA in dogs with HD. Activities of IL-1beta and IL-6 in joint fluid of dogs with HD may be influenced by the severity of laxity in the hip joint and lameness duration, respectively.

The role of hyaluronic acid in joint stability--a hypothesis for hip dysplasia and allied disorders.

The concentration of hyaluronic acid (HA) and proteins in synovial fluids of hip and shoulder joints of a variety of canine breeds has been investigated. In the Australian Kelpie, a working dog with a low incidence of hip dysplasia, shoulder synovial fluid viscosity and HA concentration were higher than in similar joints of Alsatians in which hip dysplasia is relatively common. Moreover, the HA levels and viscosity in shoulder fluids of animals with clinically defined hip dysplasia were substantially lower than in all other breeds studied. On the basis of these findings, we propose that hip dysplasia and other joint abnormalities may arise as a consequence of a deficiency in the levels of HA in synovial fluids.

The type of collagen made by the articular cartilage in joints of dogs with degenerative joint disease.

The collagen in the degenerated articular cartilage from dysplastic canine hip joints was characterized as to type by examining cartilage directly and also after labeling cartilage slices with 14C-proline in vitro. Collagen analysis was by sodium dodecylsulfate-polyacrylamide gel electrophoresis of 14C-labeled collagen, and of 14C-peptides which had been cleaved by cyanogen bromide. Amino acid analysis was done on hydrolysates of whole cartilage and of purified collagens to quantify the ratio of hydroxyproline to hydroxylysine. Data revealed that type II collagen was the radiolabeled product synthesized in all samples of degenerated cartilage and was the only collagen detected in biochemical tests in the cartilage of joints with degenerative joint disease as well as in disease free hip joints. Results suggested that chondrocytes in degenerated cartilage of canine joints did not switch their collagen phenotype, but continued to synthesize type II collagen. Cartilage degeneration in joints of dogs with hip dysplasia proceeded without a change in collagen phenotype.

Metabolism of estradiol in greyhounds and German shepherd dogs. An investigation with special reference to hip dysplasia.

Metabolism of estradiol was investigated in 5 dogs, 3 female Greyhounds with radiographically perfect hip joints and 2 female German Shepherd dogs with hip dysplasia (one pregnant and the other non-pregnant). One of the Greyhounds was studied both when pregnant and non-pregnant. The non-pregnant dogs were injected with C14-labelled estradiol-17beta i.v. and 5 mg estradiol-17beta benzoate i.m. The pregnant dogs were given only radiolabelled estradiol-17beta. Twenty-four-hour-specimens of urine were collected from the dogs for 6--8 days. Determination of urinary estrone, estradiol-17beta, and estriol was made. It was found that most of the injected estradiol was excreted unmetabolized in all dogs. A significant amount of the injected estradiol was converted to estrone and a small amount to estriol. There was no significant difference in the excretion patterns of estrone, estradiol, and estriol between the Greyhounds with perfect hip joints and the German Shepherds with hip dysplasia, regardless whether the dogs were pregnant or not. The conclusion was drawn that the capacity of dogs with hip dysplasia to metabolize estradiol and to eliminate estradiol and metabolites is not impaired.