Effects of glucosamine and chondroitin sulfate on bovine cartilage explants under long-term culture conditions.
OBJECTIVE
To determine effects of glucosamine (GLN) and chondroitin sulfate (CS) on expression of genes encoding putative mediators of osteoarthritis in bovine cartilage explants cultured for 2 weeks.


METHODS
Articular cartilage explants harvested from carpal joints of 4 Holstein steers after slaughter.


METHODS
Cartilage disks were treated as follows: fetal bovine serum only (control treatment), human recombinant interleukin (IL)-1beta (50 ng/mL; IL-1 treatment), GLN (5 microg/mL) with addition of CS (20 microg/mL; GLN-CS treatment), and human recombinant IL-1beta (50 ng/mL) with addition of GLN and CS (IL-1-GLN-CS treatment). Media were analyzed for nitric oxide and prostaglandin E(2) (PGE(2)) release. Explants were subjected to quantitative real-time PCR analysis; expressions of mRNA for inducible nitric oxide synthase, cyclooxygenase-2, microsomal prostaglandin E synthase 1, matrix metalloproteinase (MMP)-3 and -13, aggrecanase-1 and -2, tissue inhibitor of metalloproteinase (TIMP)-3, type II collagen, and aggrecan were assessed.


RESULTS
IL-1-GLN-CS and GLN-CS treatments decreased nitrite release, compared with IL-1 treatment; IL-1-GLN-CS treatment decreased IL-1-induced PGE(2) release. Expressions of inducible nitric oxide synthase, cyclooxygenase-2, and microsomal prostaglandin E synthase 1 mRNA were abrogated by GLN-CS and IL-1-GLN-CS treatments. Interleukin-1-induced mRNA expressions of proteolytic enzymes were diminished by IL-1-GLN-CS treatment. Compared with control treatment, GLN-CS treatment decreased MMP-3 and aggrecanase-2 mRNA expression. Transcripts of TIMP-3 were increased by IL-1-GLN-CS treatment, compared with IL-1 treatment. Genes encoding type II collagen and aggrecan on day 14 were upregulated by GLN-CS and IL-1-GLN-CS treatments, compared with control treatment.


CONCLUSIONS
Treatment with GLN and CS consistently downregulated mRNA expression for inflammatory mediators and matrix degrading enzymes while increasing TIMP-3 transcripts.
Chan
Pooi-See
PS
Department of Animal Science, Michigan State University, East Lansing, MI 48824, USA.
Caron
John P
JP
Orth
Michael W
MW
D051722:ADAM Proteins / Q000096:biosynthesis / Q000235:genetics; D000071121:ADAMTS4 Protein; D000818:Animals; D002358:Cartilage, Articular / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology; D002417:Cattle; D002809:Chondroitin Sulfates / Q000494:pharmacology*; D024043:Collagen Type II / Q000096:biosynthesis / Q000235:genetics; D003470:Culture Media; D051546:Cyclooxygenase 2 / Q000096:biosynthesis / Q000235:genetics; D015232:Dinoprostone / Q000378:metabolism; D004359:Drug Therapy, Combination; D005786:Gene Expression Regulation / Q000187:drug effects; D005944:Glucosamine / Q000494:pharmacology*; D053583:Interleukin-1beta / Q000494:pharmacology; D019746:Intramolecular Oxidoreductases / Q000096:biosynthesis / Q000235:genetics; D008297:Male; D020782:Matrix Metalloproteinases / Q000096:biosynthesis / Q000235:genetics; D052247:Nitric Oxide Synthase Type II / Q000096:biosynthesis / Q000235:genetics; D009573:Nitrites / Q000378:metabolism; D011348:Procollagen N-Endopeptidase / Q000096:biosynthesis / Q000235:genetics; D000071876:Prostaglandin-E Synthases; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D046509:Tissue Culture Techniques; D019717:Tissue Inhibitor of Metalloproteinase-3 / Q000096:biosynthesis / Q000235:genetics
D024043:Collagen Type II; D003470:Culture Media; D053583:Interleukin-1beta; D009573:Nitrites; D012333:RNA, Messenger; D019717:Tissue Inhibitor of Metalloproteinase-3; D002809:Chondroitin Sulfates; D052247:Nitric Oxide Synthase Type II; D051546:Cyclooxygenase 2; D051722:ADAM Proteins; D020782:Matrix Metalloproteinases; D011348:Procollagen N-Endopeptidase; D000071121:ADAMTS4 Protein; D019746:Intramolecular Oxidoreductases; D000071876:Prostaglandin-E Synthases; D015232:Dinoprostone; D005944:Glucosamine