Inappropriate use of antibiotics may lead to adverse side effects. This cross-sectional survey aimed to investigate the knowledge and attitude of female non-medical students regarding the medical and dental use of antibiotics. Four hundred validated self-administered questionnaires were distributed in Princess Norah Bint-Abdurrahman University, Riyadh, Saudi Arabia. The questionnaire included questions about accessibility, attitude toward usage, efficacy, side effects, resistance, and usage for dental issues. Knowledge was estimated for every respondent by counting the correct answers, which were considered as points. The scores were categorized as poor, moderate, and high. Of the respondents, 77.8% answered they get antibiotics according to a doctor's prescription; however, 31% stops taking antibiotics when they feel well. Only 38.8% of respondents knew that antibiotics may cause allergic reactions while 59.8% believed the human body can be resistant to antibiotics. The percentages of answers related to dental issues were: antibiotics relieve dental pain (68.8%), antibiotics can be harmful for children's teeth (27.3%), antibiotics are best avoided in pregnancy (56.7%) and no need for antibiotics after scaling (33.8%), root canal treatment (16%), or simple extraction (40.3%). Of respondents, 68% had poor scores about antibiotics efficacy, side effects, and resistance while 86.8% had poor scores related to dental problems. This study noticed a bad attitude related to antibiotics usage, with many misconceptions and poor knowledge. Moreover, the necessity of antibiotics for treatment of dental disease or after dental procedures was totally unclear for the respondents. Community campaigns are recommended every university semester to educate students about the indications, efficacy, and side effects of antibiotics.