Aim: The aim of this study was to assess knowledge, experience and confidence level in the use of local anesthesia among dentists. Materials and Methods: Dentists practicing in the Republic of Croatia were included in this cross-sectional study. The questionnaire consisted of seven parts (demographic and professional characteristics, knowledge of local anesthetics, experience in the use of local anesthetics, anesthetics and instruments used by dentists, self confidence level in the different techniques of local anesthesia, frequency of complications, use of topical anesthetic). Results: 441 dentists participated in the research. The median of age of the respondents was 35 (minimum 25, maximum 65, IQR 15). Median of knowlegde was 6 (minimum 0, maximum 12 out of 14) and median of self confidence in different techniques of local anesthesia was 54, with an interquartile range of 41 to 63 (min 17, max). Respondents of the youngest age group (≤ 30 years) had a statistically significantly higher level of knowledge (P<0.001) compared to respondents of the oldest age group (≥50 years). Specialists in endodontics and oral surgery had statistically significantly (P<0.001) higher level of knowledge compared to specialists in prosthodontics, pediatric dentistry, oral medicine, family dentistry and periodontics. Conclusion: Doctors of dental medicine who participated in the research showed insufficient knowledge about the use of local anesthesia and self-confidence when using different techniques. Based on the obtained results, it can be concluded that additional education on this topic is needed.