The large proteoglycans of the human intervertebral disc. Changes in their biosynthesis and structure with age, topography, and pathology.
METHODS
The structure and extracellular assembly of the newly synthesized aggregating proteoglycans of the human intervertebral disc were examined using an explant culture system.


OBJECTIVE
The objective was to study the changes with aging, topography, and pathology, comparing newly synthesized with endogenous proteoglycans.


BACKGROUND
No detailed studies of the biosynthesis of human disc proteoglycans have been previously reported.


METHODS
A method of explant culture that minimizes swelling and matrix loss was used to maintain the tissue architecture. Slices of postmortem and pathologic disc tissues were incubated in medium containing polyethylene glycol at appropriate concentrations to balance the swelling pressure of the tissue. The disc slices were contained in small-pore dialysis tubing to prevent penetration of the polyethylene glycol into the tissue. The newly synthesized proteoglycans were radiolabeled with [35S]-sulphate. Proteoglycans were then extracted from the tissue slices and characterized with gel chromatographic and electrophoretic techniques.


RESULTS
It was found that a single, high molecular weight proteoglycan is the major 35S-labeled synthesis product of disc cells at all ages. However, biosynthetic changes do occur: the monomer made by fetal and newborn disc cells was larger than that of adults. Furthermore, adult disc cells made other minor large 35S-labeled products, the synthesis pattern of which varied between regions.


CONCLUSIONS
These results provide the first evidence that biosynthetic changes contribute to the age-related increase in the heterogeneity of the human disc proteoglycan population.
Johnstone
B
B
Skeletal Research Center, Case Western Reserve University, Cleveland, Ohio, USA.
Bayliss
M T
MT
D000293:Adolescent; D000328:Adult; D000367:Age Factors; D000368:Aged; D000369:Aged, 80 and over; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D005865:Gestational Age; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D007403:Intervertebral Disc / Q000737:chemistry / Q000378:metabolism*; D008875:Middle Aged; D011509:Proteoglycans / Q000096:biosynthesis* / Q000302:isolation & purification
D011509:Proteoglycans