Purpose: The purpose of this study was to determine the preventive effect of joint mobilization on biphasic pain response induced formalin test. Methods: Sprague-dawley rats(n=30) were ramdomly divided into the control group without intervention, sham control group with application of hand contact without mobilization, joint mobilization group with application of hand contact with mobilization. Joint mobilization of knee procedure involved an grade III extension mobilization basically with anterior-posterior gliding of the tibia on the femur. Formalin injection caused biphasic pain response which is lated for 60 minute. The first phase result from primary afferent sensory fiber, wheareas the second phase has been proposed to central sensitization in the central nervous system. Behavioral analysis was performed by digital camera after 5% formalin subcutaneous injection into the dorsal foot. Results: Pain response of joint mobilization group show significant lower than control gorup and sham control group. Conclusion: This result suggest that pre-application of joint mobilization may be effective intervention to prevent the formalin induced pain.