Effects of carprofen and dexamethasone on canine chondrocytes in a three-dimensional culture model of osteoarthritis.
OBJECTIVE
To determine effects of carprofen and dexamethasone on chondrocytes in a culture model of osteoarthritis (OA).


METHODS
Chondrocytes isolated from articular cartilage of the humeral head of 5 adult dogs.


METHODS
Chondrocytes were harvested, cultured and subcultured in monolayer, and then cultured in a 3-dimensional (3-D) medium. Cells from each dog were distributed into 6 groups with differing content of liquid medium for each 3-D construct (agarose [AG], AG plus interleukin [IL]-1beta, AG plus carprofen [4 microg/mL], AG plus dexamethasone [1 mg/mL], AG plus IL-1beta [20 ng/mL] plus carprofen [4 microg/mL], and AG plus IL-1beta (20 ng/mL) plus dexamethasone (1 mg/mL). On days 3, 6, 12, and 20 of culture, samples from all groups were collected. Liquid media were assayed for glycosaminoglycan, prostaglandin (PG)E2, matrix metalloprotease (MMP)-3, and MMP-13 concentrations. All 3-D constructs were evaluated for viability, cell morphology, proteoglycan staining, and collagen type-II concentration. Total glycosaminoglycan content in each 3-D construct was quantitated by spectrophotometric assay.


RESULTS
Addition of IL-1beta caused a significant loss of cell viability and matrix production. Addition of carprofen or dexamethasone caused significant decreases in PGE2 in the liquid media, and each was minimally effective in protecting chondrocytes against negative effects of IL-1beta.


CONCLUSIONS
Human recombinant IL-1beta resulted in loss of cell viability, alterations in extracellular matrix components, and production of PG and MMP Carprofen and dexamethasone had little effect on cell and matrix variables but did decrease PGE2 concentrations and primarily affected the inflammatory pathway of osteoarthritis.
Dvorak
Laura D
LD
Department of Veterinary Medicine and Surgery, College of Veterinary Medicine, University of Missouri, Columbia 65211, USA.
Cook
James L
JL
Kreeger
John M
JM
Kuroki
Keiichi
K
Tomlinson
James L
JL
D000818:Animals; D002227:Carbazoles / Q000494:pharmacology* / Q000627:therapeutic use; D002358:Cartilage, Articular / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D002478:Cells, Cultured; D019902:Chondrocytes / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D017364:Collagenases / Q000378:metabolism; D003907:Dexamethasone / Q000494:pharmacology* / Q000627:therapeutic use; D015232:Dinoprostone / Q000378:metabolism; D004283:Dog Diseases / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology; D004285:Dogs; D006025:Glycosaminoglycans / Q000378:metabolism; D007150:Immunohistochemistry; D053509:Matrix Metalloproteinase 13; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D010003:Osteoarthritis / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology / Q000662:veterinary*; D013997:Time Factors
D002227:Carbazoles; D006025:Glycosaminoglycans; D003907:Dexamethasone; D017364:Collagenases; C507079:MMP13 protein, human; D053509:Matrix Metalloproteinase 13; D020778:Matrix Metalloproteinase 2; C007005:carprofen; D015232:Dinoprostone