Matrix replenishment by intervertebral disc cells after chemonucleolysis in vitro with chondroitinase ABC and chymopapain.
BACKGROUND
One of the advantages of chemonucleolysis for the treatment of a herniated intervertebral disc is the potential for the disc to self-repair. It has been suggested that the enzymes used for chemonucleolysis differentially affect the potential of the disc cells to promote repair.


OBJECTIVE
To test the ability of nucleus pulposus and anulus fibrosus cells to repair the extracellular matrix degraded in vitro by either chondroitinase ABC or chymopapain.


METHODS
An alginate cell culture system was used to monitor the progress of matrix repair after chemonucleolysis in vitro.


METHODS
Rabbit nucleus pulposus or anulus fibrosus cells precultured for 10 days in alginate gel were briefly exposed to low concentrations of chondroitinase ABC or chymopapain and then returned to normal culture conditions for up to 4 weeks. At each time point, the contents of DNA and matrix macromolecules and proteoglycan synthesis were measured.


RESULTS
The DNA content of enzyme-treated alginate beads during the following 4 weeks of culture was higher in the chondroitinase ABC group than in the chymopapain group (NP, p<.01, and AF, p<.05). The content of proteoglycan in beads containing nucleus pulposus and anulus fibrosus cells in the chondroitinase ABC group was higher than that in the chymopapain group (NP and AF, p<.001). The rate of proteoglycan synthesis and the content of collagen did not, however, differ between those two groups.


CONCLUSIONS
Intervertebral disc cells exposed to chondroitinase ABC reestablish a matrix richer in proteoglycan than cells exposed to chymopapain. This may be because of differences in the substrate spectrum of each enzyme. Although these results cannot be translated directly to the in vivo situation, they suggest the possibility that cells in discs subjected to chondroitinase ABC-induced chemonucleolysis retain a greater ability to replenish their extracellular matrix with proteoglycans than cells in discs exposed to chymopapain.
Chiba
Kazuhiro
K
Department of Orthopedic Surgery, Rush Medical College at Rush University Medical Center, Chicago, IL 60612, USA.
Masuda
Koichi
K
Andersson
Gunnar B J
GB
Momohara
Shigeki
S
Thonar
Eugene J
EJ
D000464:Alginates; D000818:Animals; D002478:Cells, Cultured; D019765:Chondroitin ABC Lyase / Q000494:pharmacology*; D002917:Chymopapain / Q000494:pharmacology*; D003094:Collagen / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D005109:Extracellular Matrix / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology; D020723:Glucuronic Acid; D006603:Hexuronic Acids; D066298:In Vitro Techniques; D007403:Intervertebral Disc / Q000166:cytology*; D007404:Intervertebral Disc Chemolysis; D008863:Microspheres; D011509:Proteoglycans / Q000096:biosynthesis / Q000378:metabolism; D011817:Rabbits; D012038:Regeneration; D013431:Sulfates / Q000378:metabolism
D000464:Alginates; D006603:Hexuronic Acids; D011509:Proteoglycans; D013431:Sulfates; D020723:Glucuronic Acid; D003094:Collagen; D004247:DNA; D002917:Chymopapain; D019765:Chondroitin ABC Lyase