Introduction: Oral health knowledge ensures that the person has all the requisite information to find out which oral diseases are and how they occur and recognize the security measures that must be taken. This knowledge can lead to changes in attitude, which leads to changes in the person’s everyday life. This study aims to evaluate oral health-related knowledge and determine its associated factors among Universiti Putra Malaysia staff. Methods: A cross-sectional study involving 207 UPM staff. The study included the UPM Malaysian staff (permanent, temporary, contract), and excluded foreigners and staff on leave. Stratified random sampling was used to select a representative sample with five selected faculties. Oral health knowledge was assessed with a validated and pretested questionnaire. Results: The total respondents were 162 from 207 subjects, giving a response rate of 78.3%. In this study, more than half of the participants have satisfactory knowledge (56.2%). The results show a significant association between oral health knowledge and each of: level of education, monthly income, faculty, type of staff, and smoking. It is observed that postgraduate staff are more knowledgeable towards oral health with 63.2% compared to graduate level and below with 39.6% only. In addition, the staff with higher incomes are more likely to have satisfactory knowledge compared to the working staff with lower incomes with percentage up to 20%. Regarding the source of information association, none of the factors is found to have a significant association. Conclusion: More than half of the participants have satisfactory knowledge on oral health. Despite this, a significant proportion of staff have unsatisfactory knowledge, which suggests initiating an awareness program for the staff members to increase their knowledge of oral health.