The important factors contributing to the effectiveness of fluoride varnish are the amount of fluoride ion release and the retention time of the varnish on the tooth surface. Commercial fluoride varnishes are susceptible to mechanical removal; therefore, patients are informed to avoid eating or performing oral hygiene for at least 12-24 h, which results in patient inconvenience. However, cyanoacrylate-based fluoride varnish would not have these disadvantages. This study compared the daily fluoride ion release, abrasion resistance to brushing, and toxicity to human gingival fibroblasts (hGFs) between a newly-developed cyanoacrylate-based fluoride varnish and conventional fluoride varnish (Duraphat varnish). The results demonstrated that the cyanoacrylate varnish had a significantly higher fluoride release for 9 days after application, higher abrasion resistance to brushing, and slightly less toxicity to hGFs compared with Duraphat varnish. This novel cyanoacrylate varnish could be an alternative fluoride varnish for preventing dental caries.