Matrix remodeling expression in anulus cells subjected to increased compressive load.
METHODS
Mechanobiology study of gene expression changes as a result of compressive overload of anular fibrochondrocytes.


OBJECTIVE
To test hypotheses regarding phenotype shift in genes coding for representative extracellular matrix (ECM) proteins and matrix modulators.


BACKGROUND
In degenerative disc disease, the transfer of compressive load through the disc shifts largely from the nucleus onto the anulus. In vivo models simulating this condition have shown derangement of the collagenous ultrastructure in the anulus. In vitro models of cultured anulus cells subjected to static compressive stress generally suggest a down-regulation of synthesis. This study evaluated the expression of specific isomers of genes responsible for mechanical viability and metabolism of the disc under cyclic compressive loads.


METHODS
Fibrochondrocytes were digested from the anuli of 3, 2-week-old pigs, embedded in 1.5% alginate gel, and hydrostatically compressed at 0.5 Hz for 3 hours to amplitudes of 10 and 30 atm. These levels represented nominal load transfer through the healthy disc and high load transfer through the degenerative disc. Ribonucleic acid was isolated, reverse transcribed, and evaluated by real-time polymerase chain reaction for expression of type I (C-I) and type II (C-II) collagen, aggrecan, the matrix metalloproteinase (MMP-1), and the transforming growth factor beta (TGFbeta-1). Results were expressed at percentages of uncompressed controls.


RESULTS
The lower pressure of 10 atm resulted in up-regulation of all ECM protein genes. C-I and C-II both averaged 141%, and aggrecan 121% of controls (P < 0.05). MMP-1 and TGFbeta-1 were essentially unchanged. With the pressure increased to 30 atm, C-II remained approximately at the level expressed under lower pressure, but C-I was reduced to 42% of controls (P < 0.05), indicating a phenotype shift. MMP-1 and TGFbeta-1 also were down-regulated to 71% and 54% of controls, respectively (P < 0.05).


CONCLUSIONS
The up-regulation of the ECM genes with nominal pressure highlights the mechanobiological importance of common activity in fibrocartilage homeostasis. Differential regulation of the 2 primary collagen types with high pressure indicates a capacity of the anulus to remodel according to pathomechanical conditions.
Wenger
Karl H
KH
Department of Rheumatology, University of Tennessee Health Science Center, Memphis, TN, USA. kwenger@mcg.edu
Woods
J Andrew
JA
Holecek
Arin
A
Eckstein
Eugene C
EC
Robertson
James T
JT
Hasty
Karen A
KA
D053674:Aggrecans; D000818:Animals; D000831:Animals, Newborn; D024042:Collagen Type I / Q000235:genetics / Q000378:metabolism; D024043:Collagen Type II / Q000235:genetics / Q000378:metabolism; D019245:Compressive Strength; D016326:Extracellular Matrix Proteins / Q000235:genetics / Q000378:metabolism*; D051445:Fibrocartilage / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D006874:Hydrostatic Pressure; D007403:Intervertebral Disc / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D037181:Lectins, C-Type / Q000235:genetics / Q000378:metabolism; D020781:Matrix Metalloproteinase 1 / Q000235:genetics / Q000378:metabolism; D023421:Models, Animal; D011509:Proteoglycans / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D013552:Swine; D016212:Transforming Growth Factor beta / Q000235:genetics / Q000378:metabolism; D053773:Transforming Growth Factor beta1; D015854:Up-Regulation; D016474:Weight-Bearing
D053674:Aggrecans; D024042:Collagen Type I; D024043:Collagen Type II; D016326:Extracellular Matrix Proteins; D037181:Lectins, C-Type; D011509:Proteoglycans; D012333:RNA, Messenger; D016212:Transforming Growth Factor beta; D053773:Transforming Growth Factor beta1; D020781:Matrix Metalloproteinase 1