As a preliminary study, 1, 10, 100, and 1,000 U/disc of bovine testis-derived hyaluronidase (bovine t-HAase) were injected into normal canine intervertebral discs, and its chemonucleolytic effect and the establishment of the technical technique were examined. No changes in intervertebral discs were observed, and the inhibition of HAase activity was suggested. As the cause, we focused on the X-ray contrast agent used at the time of intradiscal administration. An enzyme reaction experiment between a hyaluronic acid (HA) substrate and bovine t-HAase with a contrast agent was conducted in vitro, and HAase activity was examined. When the concentrations of Omnipark (Iohexol), a contrast agent, were 0, 9.4, 20.3, 40.5, and 50%, the residual rates of HAase activity were 100, 87.1, 64.4, 43.7. and 50.4%, respectively. Further, when all seven contrast agents (four types) were tested, HAase activity inhibition was observed, tending to correlate with the iodine content. In conclusion, when a contrast agent and an enzyme are used in combination in X-ray fluoroscopy treatment, measurement of the enzymatic activity of the mixed solution is necessary in advance to prevent its inhibition.
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