Effect of chondroitinase ABC on matrix metalloproteinases and inflammatory mediators produced by intervertebral disc of rabbit in vitro

Makoto Sakuma, Norihisa Fujii, Toyomi Takahashi, Jirou Hoshino, Satoshi Miyauchi, Hisashi Iwata
Spine 2002 March 15, 27 (6): 576-80

STUDY DESIGN: Lumbar intervertebral discs in rabbit were cultured in the presence of chondroitinase ABC. The matrix metalloproteinases (MMPs) and inflammatory mediators produced in culture media were then analyzed.

OBJECTIVES: To investigate the effect of chondroitinase ABC on MMPs and inflammatory mediators produced by intervertebral disc of rabbit in vitro.

SUMMARY OF BACKGROUND DATA: The chemonucleolytic effect of chondroitinase ABC is caused by the decrease in the chondroitin sulfate, hyaluronan, and protein content of the nucleus pulposus in rabbit. The reason for the decreases in protein content remains unclear.

METHODS: Anulus fibrosus and nucleus pulposus were cultured for 72 hours with or without chondroitinase ABC stimulated or not stimulated by interleukin-1 after preculture for 4 days. Subsequently, the MMPs (gelatinases MMP-2, MMP-9, and collagenase) and inflammatory mediators (prostaglandin E2 and nitric oxide) produced in the culture media were analyzed.

RESULTS: In the anulus fibrosus chondroitinase ABC and interleukin-1 synergistically increased the collagenase activity, which was at a significantly higher level than the increment solely due to interleukin-1. In contrast, chondroitinase ABC counteracted the increase in nitric oxide production by interleukin-1. In the nucleus pulposus the collagenase and nitric oxide productions were not particularly affected by chondroitinase ABC and/or interleukin-1. In zymographic analysis MMP-2 was detected, but MMP-9 was only slightly detected in both tissues. There were no significant differences in both tissues for MMP-2 and prostaglandin E2 following incubation with or without chondroitinase ABC, whether stimulated by interleukin-1 or not.

CONCLUSIONS: The collagenase activity in the anulus fibrosus was increased by chondroitinase ABC with interleukin-1. This finding may support the hypothesis that some proteolytic activities are involved in the chemonucleolytic process by chondroitinase ABC treatment.

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