BACKGROUND To increase the utilization of preventive dental care, it is essential to improve the knowledge and attitude of parents about such cares. The aim of this study was to evaluate the knowledge, attitude, and practice of school children's parents toward fissure sealant (FS) and professional fluoride therapy in Isfahan. METHODS In this cross-sectional study, school children's parents (n = 637) were selected based on proportional cluster sampling. A valid and reliable questionnaire was designed, including demographic section, questions about parents' experience and their knowledge and attitude about professional fluoride and FS therapy. Data were analyzed by ANOVA, regression, Chi-square, and correlation coefficient tests. RESULTS The means of total knowledge and knowledge about fluoride therapy and FS were 5.9 ± 4.1 out of 19, 3.3 ± 2.0 out of 9 and 2.6 ± 2.7 out of 10, respectively. The mean of attitude was 33.7 ± 5.8. The mean of knowledge toward FS therapy was significantly higher in academically educated parents (P = 0.023). The mean of total knowledge among those who received their knowledge by their dentist was also significantly higher than that of other resources such as mass media (P = 0.003). Total knowledge was positively correlated with attitude (P < 0.001, coefficient = 0.5). Of participants, 10.4% (54) and 23.2% (124) had the experience of FS and fluoride therapy, respectively. CONCLUSIONS Based on the low level of knowledge regarding professional preventive care in this study and the effectiveness of knowledge acquired through dentists and mass media consultations, it might be effective to require them to consider such training more seriously.