Mechanoregulation of human articular chondrocyte aggrecan and type II collagen expression by intermittent hydrostatic pressure in vitro
Article first published online: 1 JAN 2006
Copyright © 2003 Orthopaedic Research Society
Journal of Orthopaedic Research
Volume 21, Issue 1, pages 110–116, January 2003
How to Cite
Ikenoue, T., Trindade, M. C. D., Lee, M. S., Lin, E. Y., Schurman, D. J., Goodman, S. B. and Smith, R. L. (2003), Mechanoregulation of human articular chondrocyte aggrecan and type II collagen expression by intermittent hydrostatic pressure in vitro. J. Orthop. Res., 21: 110–116. doi: 10.1016/S0736-0266(02)00091-8
- Issue published online: 1 JAN 2006
- Article first published online: 1 JAN 2006
- Manuscript Accepted: 10 MAY 2002
- Manuscript Received: 28 DEC 2001
- R&D VA Merit Review. Grant Number: A2128-RC (RLS)
- VA Medical Merit Funding (RLS)
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 × 1) or 4 h per day for four days (4 × 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 4 × 1 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 4 × 4 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 4 × 4 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.
© 2002 Orthopaedic Research Society. Published by Elsevier Science Ltd. All rights reserved.