This study aimed to assess toothbrushing behavior and descriptively analyze the effect of age and gender. Two thousand and six people from the city of Sebha, Libya, aged 1 to 64 years (mean age 26.9 ± 11.6 years, 1,463 females and 543 males) constituted the study sample. Data was collected using a self-administered questionnaire. Overall, 85.3% were using a toothbrush and toothpaste as a cleaning aid, whereas 6.3% never brushed their teeth. Only 36.1% brushed twice daily. Age and gender were significantly associated with use of a toothbrush and frequency of toothbrushing. This data serves as a baseline to implement a "preventive self-care instruction program."