Effects of glucosamine and chondroitin sulfate on bovine cartilage explants under long-term culture conditions

Am J Vet Res. 2007 Jul;68(7):709-15. doi: 10.2460/ajvr.68.7.709.

Abstract

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.

Sample population: Articular cartilage explants harvested from carpal joints of 4 Holstein steers after slaughter.

Procedures: 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 and clinical relevance: Treatment with GLN and CS consistently downregulated mRNA expression for inflammatory mediators and matrix degrading enzymes while increasing TIMP-3 transcripts.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ADAM Proteins / biosynthesis
  • ADAM Proteins / genetics
  • ADAMTS4 Protein
  • Animals
  • Cartilage, Articular / drug effects*
  • Cartilage, Articular / metabolism
  • Cartilage, Articular / physiology
  • Cattle
  • Chondroitin Sulfates / pharmacology*
  • Collagen Type II / biosynthesis
  • Collagen Type II / genetics
  • Culture Media
  • Cyclooxygenase 2 / biosynthesis
  • Cyclooxygenase 2 / genetics
  • Dinoprostone / metabolism
  • Drug Therapy, Combination
  • Gene Expression Regulation / drug effects
  • Glucosamine / pharmacology*
  • Interleukin-1beta / pharmacology
  • Intramolecular Oxidoreductases / biosynthesis
  • Intramolecular Oxidoreductases / genetics
  • Male
  • Matrix Metalloproteinases / biosynthesis
  • Matrix Metalloproteinases / genetics
  • Nitric Oxide Synthase Type II / biosynthesis
  • Nitric Oxide Synthase Type II / genetics
  • Nitrites / metabolism
  • Procollagen N-Endopeptidase / biosynthesis
  • Procollagen N-Endopeptidase / genetics
  • Prostaglandin-E Synthases
  • RNA, Messenger / biosynthesis
  • RNA, Messenger / genetics
  • Reverse Transcriptase Polymerase Chain Reaction
  • Tissue Culture Techniques
  • Tissue Inhibitor of Metalloproteinase-3 / biosynthesis
  • Tissue Inhibitor of Metalloproteinase-3 / genetics

Substances

  • Collagen Type II
  • Culture Media
  • Interleukin-1beta
  • Nitrites
  • RNA, Messenger
  • Tissue Inhibitor of Metalloproteinase-3
  • Chondroitin Sulfates
  • Nitric Oxide Synthase Type II
  • Cyclooxygenase 2
  • ADAM Proteins
  • Matrix Metalloproteinases
  • Procollagen N-Endopeptidase
  • ADAMTS4 Protein
  • Intramolecular Oxidoreductases
  • Prostaglandin-E Synthases
  • Dinoprostone
  • Glucosamine