title: Chondrocytes harvested from osteochondritis dissecans cartilage are able to undergo limited in vitro chondrogenesis despite having perturbations of cell phenotype in vivo.
abstract: Our objective was to characterize the variation in gene expression for key genes associated with chondrogenic phenotype of osteochondrosis (OC)-affected and normal chondrocytes, and to identify whether OC chondrocytes can redifferentiate and regain a phenotype similar to normal chondrocytes if appropriate chondrogenic signals are given. Equine articular cartilage removed at surgery to treat clinically significant OC lesions was collected (n = 10), and the gene expression evaluated and compared to aged-matched normal samples (n = 10). Cartilage was harvested from normal (n = 4) and OC (n = 3) joints from horses at necropsy. Chondrogenic pellet cultures were established following monolayer proliferation. After 14 days in culture, the pellets were assessed by histochemical and pellet weight analysis, assay of glycosaminoglycan (GAG) content, and gene expression. Chondrocytes from OC cartilage expressed significantly more Coll-I, -II, -III, and -X than chondrocytes from normal cartilage (all p < 0.0001). Furthermore, OC chondrocytes expressed significantly more MMP-13, ADAMTS-4 (both p < 0.0001), and TIMP-1 (p < 0.001) and significantly less TIMP-2 and TIMP-3. Pellets created from OC chondrocytes contained significantly less GAG (p = 0.0069) and expressed significantly less Sox9 and significantly more superficial zone protein (SZP) (p = 0.0105) than pellets created from normal cartilage. The results suggest that chondrocytes from OC cartilage at the time of surgical treatment have perturbations in phenotype compared to cells from normal cartilage. Despite these differences, following monolayer expansion and pellet culture under chondrogenic conditions, chondrocytes derived from OC cartilage retain some ability to undergo chondrogenic differentiation and synthesize an appropriate cartilage-like matrix. However, this chondrogenic differentiation potential is inferior to that seen in aged-matched normal chondrocytes.
authors.affiliation: Musculoskeletal Research Group, The University of Liverpool Veterinary Teaching Hospital, Leahurst, Neston, Wirral, United Kingdom. e.garvican@knac.com
authors.forename: E R
authors.initials: ER
authors.lastname: Garvican
authors.forename: A
authors.initials: A
authors.lastname: Vaughan-Thomas
authors.forename: C
authors.initials: C
authors.lastname: Redmond
authors.forename: P D
authors.initials: PD
authors.lastname: Clegg
mesh_terms: D000818:Animals; D002356:Cartilage / Q000166:cytology / Q000502:physiology; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D019902:Chondrocytes / Q000473:pathology / Q000502:physiology*; D020219:Chondrogenesis / Q000502:physiology*; D003094:Collagen / Q000235:genetics; D016326:Extracellular Matrix Proteins / Q000235:genetics; D015870:Gene Expression / Q000502:physiology; D006734:Horse Diseases / Q000473:pathology / Q000503:physiopathology*; D006736:Horses; D066298:In Vitro Techniques; D020782:Matrix Metalloproteinases / Q000235:genetics; D010003:Osteoarthritis / Q000473:pathology / Q000503:physiopathology / Q000662:veterinary; D010008:Osteochondritis Dissecans / Q000473:pathology / Q000503:physiopathology* / Q000662:veterinary*; D010641:Phenotype; D015398:Signal Transduction / Q000502:physiology; D019714:Tissue Inhibitor of Metalloproteinases / Q000235:genetics
chemical_list: D016326:Extracellular Matrix Proteins; D019714:Tissue Inhibitor of Metalloproteinases; D003094:Collagen; D020782:Matrix Metalloproteinases