Relative increase of biglycan and decorin and altered chondroitin sulfate epitopes in the degenerating human intervertebral disc.
OBJECTIVE
Proteoglycans are major components of the extracellular matrix of the intervertebral disc. They are vital for the biomechanical properties of the tissue, and are subject to changes in disc degeneration. We aimed to further define these changes and their relationship to normal aging.


METHODS
Normal discs (age 13-53 years, n = 6) were analyzed from 5 different sites across the sagittal anterior-posterior direction. Degenerated anterior annulus fibrosus was collected from 7 patients aged 39-46 years. Extracted proteoglycans were separated using agarose and polyacrylamide gel electrophoresis and detected with toluidine blue staining and Western blotting.


RESULTS
The center of the disc showed the highest level of total proteoglycans, but lowest levels of decorin and biglycan. Western blots displayed reduced signal for both glycanated and nonglycanated biglycan and decorin after adolescence, while an increased signal of biglycan was observed in degenerated annuli. The 7D4(-) and 3B3(-) epitopes on native chondroitin sulfate chains were present in the large proteoglycans of intervertebral discs, but their signal intensity had no correlation to degeneration. Chondroitinase ABC digestion of the blots brought up 7D4(+) signal in the small proteoglycans of degenerated, but not in healthy tissue. Decrease or total loss of 2B6(+) epitope (indicating 4-sulfated stubs of chondroitin sulfate chains) were found in the large proteoglycans of all degenerated annuli.


CONCLUSIONS
Human intervertebral disc degeneration involves the accumulation of decorin and biglycan relative to other uronic acid containing proteoglycans, the disappearance of 4-sulfated core region in aggrecan-like large proteoglycans, and the emergence of a core structure in the chains of small proteoglycans reacting with the 7D4 antibody; these findings indicate a fundamental alteration in matrix properties that may contribute to the pathogenesis of the disease.
Inkinen
R I
RI
Department of Anatomy, University of Kuopio, Finland.
Lammi
M J
MJ
Lehmonen
S
S
Puustjärvi
K
K
Kääpä
E
E
Tammi
M I
MI
D000293:Adolescent; D000328:Adult; D058578:Biglycan; D002809:Chondroitin Sulfates / Q000378:metabolism*; D002810:Chondroitinases and Chondroitin Lyases / Q000378:metabolism; D058575:Decorin; D000939:Epitopes; D016326:Extracellular Matrix Proteins; D006801:Humans; D007403:Intervertebral Disc / Q000201:enzymology / Q000378:metabolism*; D008875:Middle Aged; D011509:Proteoglycans / Q000378:metabolism*
C546884:BGN protein, human; D058578:Biglycan; C546878:DCN protein, human; D058575:Decorin; D000939:Epitopes; D016326:Extracellular Matrix Proteins; D011509:Proteoglycans; D002809:Chondroitin Sulfates; D002810:Chondroitinases and Chondroitin Lyases