Mechanoregulation of human articular chondrocyte aggrecan and type II collagen expression by intermittent hydrostatic pressure in vitro.
This study addressed the hypothesis that duration and magnitude of applied intermittent hydrostatic pressure (IHP) are critical parameters in regulation of normal human articular chondrocyte aggrecan and type II collagen expression. Articular chondrocytes were isolated from knee cartilage and maintained as primary, high-density monolayer cultures. IHP was applied at magnitudes of 1, 5 and 10 MPa at 1 Hz for durations of either 4 h per day for one day (4 x 1) or 4 h per day for four days (4 x 4). Total cellular RNA was isolated and analyzed for aggrecan and type II collagen mRNA signal levels using specific primers and reverse transcription polymerase chain reaction (RT-PCR) nested with beta-actin primers as internal controls. With a 4x1 loading regimen, aggrecan mRNA signal levels increased 1.3- and 1.5-fold at 5 and 10 MPa, respectively, relative to beta-actin mRNA when compared to unloaded cultures. Changing the duration of loading to a 4x4 regimen increased aggrecan mRNA signal levels by 1.4-, 1.8- and 1.9-fold at loads of 1, 5 and 10 MPa, respectively. In contrast to the effects of IHP on aggrecan, type II collagen mRNA signal levels were only upregulated at loads of 5 and 10 MPa with the 4x4 loading regimen. Analysis of cell-associated protein by western blotting confirmed that IHP increased aggrecan and type II collagen in chondrocyte extracts. These data demonstrate that duration and magnitude of applied IHP differentially alter chondrocyte matrix protein expression. The results show that IHP provides an important stimulus for increasing cartilage matrix anabolism and may contribute to repair and regeneration of damaged or diseased cartilage.
Ikenoue
Takashi
T
Rehabilitation R&D Center, Veterans Affairs Palo Alto Health Care System, Palo Alto, CA 94304, USA.
Trindade
Michael C D
MC
Lee
Mel S
MS
Lin
Eric Y
EY
Schurman
David J
DJ
Goodman
Stuart B
SB
Smith
R Lane
RL
D053674:Aggrecans; D015153:Blotting, Western; D002358:Cartilage, Articular / Q000166:cytology; D019902:Chondrocytes / Q000737:chemistry / Q000502:physiology*; D024043:Collagen Type II / Q000032:analysis / Q000235:genetics*; D016326:Extracellular Matrix Proteins; D005786:Gene Expression Regulation / Q000502:physiology; D006801:Humans; D006874:Hydrostatic Pressure; D066298:In Vitro Techniques; D037181:Lectins, C-Type; D011509:Proteoglycans / Q000032:analysis / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis
D053674:Aggrecans; D024043:Collagen Type II; D016326:Extracellular Matrix Proteins; D037181:Lectins, C-Type; D011509:Proteoglycans; D012333:RNA, Messenger