title: Articular cartilage glycosaminoglycans inhibit the adhesion of endothelial cells.
abstract: Articular cartilage undergoes severe loss of proteoglycan and its constituent glycosaminoglycans (GAGs) in osteoarthritis. We hypothesize that the low GAG content of osteoarthritic cartilage renders the tissue susceptible to pathological vascularization. This was investigated using an in vitro angiogenesis model assessing endothelial cell adhesion to GAG-depleted cartilage explants. Bovine cartilage explants were treated with hyaluronidase to deplete GAG content and then seeded with fluorescently tagged human endothelial cells (HMEC-1). HMEC-1 adherence was assessed after 4 hr and 7 days. The effect of hyaluronidase treatment on GAG content, chondrocyte viability, and biochemical composition of the extracellular matrix was also determined. Hyaluronidase treatment reduced the GAG content of cartilage explants by 78 ± 3% compared with that of controls (p < 0.0001). GAG depletion was associated with significantly more HMEC-1 adherence on both the surface (superficial zone) and the underside (deep zone) of the explants (both p < 0.0001). The latter provided a more favorable environment for extended culture of HMEC-1 compared with the articulating surface. Hyaluronidase treatment altered the immunostaining for chondroitin sulfate epitopes, but not for lubricin. Our results support the hypothesis that articular cartilage GAGs are antiadhesive to endothelial cells and suggest that chondroitin sulfate and/or hyaluronan are responsible. The loss of these GAGs in osteoarthritis may allow osteochondral angiogenesis resulting in disease progression.
authors.affiliation: The Robert Jones and Agnes Hunt Orthopaedic Hospital, NHS Foundation Trust and The Institute for Science and Technology in Medicine, Keele University, Oswestry, Shropshire, UK. jennifer.bara@rjah.nhs.uk
authors.forename: Jennifer Jane
authors.initials: JJ
authors.lastname: Bara
authors.forename: William Eustace Basil
authors.initials: WE
authors.lastname: Johnson
authors.forename: Bruce
authors.initials: B
authors.lastname: Caterson
authors.forename: Sally
authors.initials: S
authors.lastname: Roberts
mesh_terms: D000818:Animals; D002358:Cartilage, Articular / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D002417:Cattle; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology; D002470:Cell Survival / Q000187:drug effects; D019902:Chondrocytes / Q000187:drug effects / Q000502:physiology; D002809:Chondroitin Sulfates / Q000378:metabolism; D018450:Disease Progression; D000939:Epitopes / Q000187:drug effects; D005260:Female; D006023:Glycoproteins / Q000378:metabolism; D006025:Glycosaminoglycans / Q000032:analysis / Q000502:physiology*; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D006821:Hyaluronoglucosaminidase / Q000494:pharmacology; D018919:Neovascularization, Physiologic / Q000187:drug effects / Q000502:physiology*; D009924:Organ Culture Techniques; D010003:Osteoarthritis / Q000378:metabolism* / Q000473:pathology; D000099158:Proteoglycan 4
chemical_list: D002809:Chondroitin Sulfates; D000939:Epitopes; D006023:Glycoproteins; D006025:Glycosaminoglycans; D006821:Hyaluronoglucosaminidase; D000099158:Proteoglycan 4