title: Cyclic compressive mechanical stimulation induces sequential catabolic and anabolic gene changes in chondrocytes resulting in increased extracellular matrix accumulation.
abstract: Overcoming the limited ability of articular cartilage to self-repair may be possible through tissue engineering. However, bioengineered cartilage formed using current methods does not match the physical properties of native cartilage. In previous studies we demonstrated that mechanical stimulation improved cartilage tissue formation. This study examines the mechanisms by which this occurs. Application of uniaxial, cyclic compression (1 kPa, 1 Hz, 30 min) significantly increased matrix metalloprotease (MMP)-3 and MMP-13 gene expression at 2 h compared to unstimulated cells. These returned to constitutive levels by 6 h. Increased MMP-13 protein levels, both pro- and active forms, were detected at 6 h and these decreased by 24 h. This was associated with tissue degradation as more proteoglycans and collagen had been released into the culture media at 6 h when compared to the unstimulated cells. This catabolic change was followed by a significant increase in type II collagen and aggrecan gene expression at 12 h post-stimulation and increased synthesis and accumulation of these matrix molecules at 24 h. Mechanical stimulation activated the MAP kinase pathway as there was increased phosphorylation of ERK1/2 and JNK as well as increased AP-1 binding. Mechanical stimulation in the presence of the JNK inhibitor, SP600125, blocked AP-1 binding preventing the increased gene expression of MMP-3 and -13 at 2 h and type II collagen and aggrecan at 12 h as well as the increased matrix synthesis and accumulation. Given the sequence of changes, cyclic compressive loading appears to initiate a remodelling effect involving MAPK and AP-1 signalling resulting in improved in vitro formation of cartilage.
authors.affiliation: CIHR BioEngineering of Skeletal Tissues Team, Mount Sinai Hospital, Canada.
authors.forename: J N A
authors.initials: JN
authors.lastname: De Croos
authors.forename: S S
authors.initials: SS
authors.lastname: Dhaliwal
authors.forename: M D
authors.initials: MD
authors.lastname: Grynpas
authors.forename: R M
authors.initials: RM
authors.lastname: Pilliar
authors.forename: R A
authors.initials: RA
authors.lastname: Kandel
mesh_terms: D000818:Animals; D001696:Biomechanical Phenomena; D002356:Cartilage / Q000737:chemistry; D002417:Cattle; D019902:Chondrocytes / Q000378:metabolism*; D003094:Collagen / Q000737:chemistry; D003470:Culture Media; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005109:Extracellular Matrix / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic; D053509:Matrix Metalloproteinase 13 / Q000096:biosynthesis; D019278:Matrix Metalloproteinase 3 / Q000096:biosynthesis; D011312:Pressure; D013997:Time Factors; D023822:Tissue Engineering
chemical_list: D003470:Culture Media; D004791:Enzyme Inhibitors; D003094:Collagen; D053509:Matrix Metalloproteinase 13; D019278:Matrix Metalloproteinase 3