Aims: Recently, Vitamin D deficiency is pandemic now. The main reason of vitamin D deficiency is inadequate exposure to sunlight. Vitamin D level in nutrients is low and it can be ineffective to meet the daily requirements. Vitamin D is synthesized in the skin by ultraviolent radiation. In the present study, the authors aimed to investigate serum 25(OH) D levels of the medical staff working in different positions in the Faculty of Dentistry. Subjects and Methods: A total of 51 dentists, 28 dental assistants, 11 secretaries, and 10 nurses working in the Faculty of Dentistry, Erciyes University between November and December 2014 were included to the study. The serum 25(OH) D levels of the participants were evaluated. Results: Of the participants, 62 were females and 38 were males with a mean age of 30.17 5.77 (range: 20u49) years. The mean vitamin D levels were found to be 12.1 8.37 ng/mL (range: 2.1u38.3). A total of 51 participants had severe vitamin D insufficiency, while three participants had normal vitamin D levels with only one dentist. Conclusion: According to the results of present study, dental staff should be considered as a high-risk group for vitamin D deficiency. Vitamin D deficiency and insufficiency is a common problem among medical staffs as in several working groups such as white-collar workers and bankers. Results of present study suggest that insufficient exposure to the sunlight may lead to severe vitamin D deficiency in dental professionals. Vitamin D supplementation may be recommended to the dental staff.