title: P38 mitogen-activated protein kinase promotes dedifferentiation of primary articular chondrocytes in monolayer culture.
abstract: Articular cartilage is an avascular tissue with poor regenerative capacity following injury, a contributing factor to joint degenerative disease. Cell-based therapies for cartilage tissue regeneration have rapidly advanced; however, expansion of autologous chondrocytes in vitro using standard methods causes 'dedifferentiation' into fibroblastic cells. Mitogen-activated protein kinase (MAPK) signalling is crucial for chondrocyte metabolism and matrix production, and changes in MAPK signals can affect the phenotype of cultured cells. We investigated the effects of inhibition of MAPK signalling on chondrocyte dedifferentiation during monolayer culture. Blockade of extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK) signalling caused a significant increase in cartilage gene expression, however, also caused up-regulation of fibrotic gene expression. Inhibition of p38 MAPK (p38) caused a significant up-regulation of collagen type II while suppressing collagen type I expression. P38 inhibition also resulted in consistently more organized secretion of collagen type II protein deposits on cell culture surfaces. Follow-on pellet culture of treated cells revealed that MAPK inhibition reduced cell migration from the pellet. ERK and JNK inhibition caused more collagen type I accumulation in pellets versus controls while p38 inhibition strongly promoted collagen type II accumulation with no effect on collagen type I. Blockade of all three MAPKs caused increased GAG content in pellets. These results indicate a role for MAPK signalling in chondrocyte phenotype loss during monolayer culture, with a strong contribution from p38 signalling. Thus, blockade of p38 enhances chondrocyte phenotype in monolayer culture and may promote more efficient cartilage tissue regeneration for cell-based therapies.
authors.affiliation: Soft Tissue Biophysics Laboratory, Department of Chemical Engineering, McGill University, Montreal, QC H3A 2B2, Canada.
authors.forename: Derek H
authors.initials: DH
authors.lastname: Rosenzweig
authors.forename: Sing J
authors.initials: SJ
authors.lastname: Ou
authors.forename: Thomas M
authors.initials: TM
authors.lastname: Quinn
mesh_terms: D000818:Animals; D000873:Anthracenes / Q000494:pharmacology; D002358:Cartilage, Articular / Q000166:cytology*; D002417:Cattle; D054337:Cell Dedifferentiation; D049109:Cell Proliferation; D002470:Cell Survival; D019902:Chondrocytes / Q000502:physiology*; D020219:Chondrogenesis; D005109:Extracellular Matrix / Q000378:metabolism; D005419:Flavonoids / Q000494:pharmacology; D015870:Gene Expression; D006025:Glycosaminoglycans / Q000378:metabolism; D007093:Imidazoles / Q000494:pharmacology; D020935:MAP Kinase Signaling System; D020928:Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D010641:Phenotype; D061251:Primary Cell Culture; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011725:Pyridines / Q000494:pharmacology; D048051:p38 Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000502:physiology*
chemical_list: D000873:Anthracenes; D005419:Flavonoids; D006025:Glycosaminoglycans; D007093:Imidazoles; D047428:Protein Kinase Inhibitors; D011725:Pyridines; C432165:pyrazolanthrone; D020928:Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases; C093642:SB 203580; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one