Effects of human recombinant interleukin-1beta on canine articular chondrocytes in three-dimensional culture.
OBJECTIVE
To determine the effects of interleukin (IL)-1beta on matrix synthesis and degradation by chondrocytes cultured in a 3-dimensional (3-D) gel medium.


METHODS
Chondrocytes from 7 dogs.


METHODS
Articular chondrocytes were harvested and cultured in 3-D gel medium alone or with 10 or 20 ng IL-1beta/ml that was added beginning on day 0, 3, 6, or 9. On days 3, 6, 12, and 20 of 3-D culture, samples of the liquid medium were evaluated for glycosaminoglycan (GAG), prostaglandin E2 (PGE2), and matrix metalloprotease (MMP)-3 content. The 3-D plug in each well was evaluated for histologic characteristics of viability, cell morphology, and proteoglycan staining, immunohistochemically stained for collagen type II, and spectrophotometrically analyzed for GAG content.


RESULTS
Significant differences for all variables were detected between controls and each IL-1beta group, among groups with different IL-1beta concentrations, and among groups with IL-1beta added at various time points. Chondrocytes exposed to IL-1beta had loss of GAG, increased PGE2 and MMP-3 concentrations, and lack of collagen type-II synthesis. These IL-1beta effects appeared to be time and concentration dependent.


CONCLUSIONS
Addition of IL-1beta to chondrocytes in 3-D gel medium results in time- and concentration-dependent effects on matrix synthesis and degradation and provides an appropriate in vitro model for many of the pathophysiologic events associated with osteoarthritis.
Cook
J L
JL
Comparative Orthopaedic Laboratory, College of Veterinary Medicine, University of Missouri, Columbia 65211, USA.
Anderson
C C
CC
Kreeger
J M
JM
Tomlinson
J L
JL
D000818:Animals; D002358:Cartilage, Articular / Q000166:cytology / Q000187:drug effects*; D018929:Cell Culture Techniques / Q000379:methods; D019902:Chondrocytes / Q000187:drug effects*; D003094:Collagen / Q000032:analysis; D015232:Dinoprostone / Q000032:analysis; D004285:Dogs / Q000502:physiology*; D006025:Glycosaminoglycans / Q000032:analysis; D006801:Humans; D007124:Immunoenzyme Techniques / Q000662:veterinary; D007150:Immunohistochemistry; D007375:Interleukin-1 / Q000494:pharmacology*; D019278:Matrix Metalloproteinase 3 / Q000032:analysis; D008751:Methylene Blue / Q000031:analogs & derivatives / Q000737:chemistry; D011994:Recombinant Proteins / Q000494:pharmacology; D018709:Statistics, Nonparametric
D006025:Glycosaminoglycans; D007375:Interleukin-1; D011994:Recombinant Proteins; D003094:Collagen; D019278:Matrix Metalloproteinase 3; D015232:Dinoprostone; D008751:Methylene Blue; C016401:1,9-dimethylmethylene blue