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Background: In a pilot study, we showed that topical administration of a tetracycline could decrease oral bacteria levels for 6 hours in patients who underwent oral cancer surgery combined with tracheotomy and flap reconstruction. This multicenter, randomized control trial aimed to investigate the effectiveness of topical application of tetracycline ointment for prevention of surgical site infection (SSI) associated with major oral cancer surgery., Methods: One hundred seventeen patients who underwent oral cancer resection combined with neck dissection, flap reconstruction, and tracheotomy were divided randomly into an intervention group (n = 56) and a control group (n = 61). The intervention consisted of topical administration of tetracycline ointment on the dorsum of the tongue every 6 hours for 48 hours postoperatively. Factors relating to the occurrence of SSI in both groups were subjected to logistic regression analysis., Results: SSI occurred in 11 patients (19.6%) in the intervention group and 22 patients (36.1%) in the control group. Multivariate analysis showed that a longer operating time and not receiving topical tetracycline were independent risk factors for development of SSI., Conclusion: Administration of topical tetracycline for 48 hours postoperatively is an effective way of preventing SSI after oral cancer surgery.