Objectives: To investigate if dental nurses perceive gender differences in the communication and interaction style used by their dentist in general practice. Methods: Data were collected from 64 Northern Irish dental nurses (70% response), and from 549 Dutch nurses (41% response). Respondents completed the Communication and Working Styles Questionnaire (CWSQ), a self-report questionnaire with 20 items on staff communication and working styles. Using principal component analysis, six distinguishable factors could be extracted:  gender-related (four items, Cronbach's α = 0.78),  friendly relation (four items, α = 0.66),  teamwork (four items, α = 0.44),  playfulness (two items, α = 0.69),  personal attention (three items, α = 0.60), and  business-like (two items, α = 0.32). Factors  and  were additionally analysed at item level given their low internal consistencies. Results: MANOVA indicated a main effect for dentist's gender [F(6,541) = 4.649, p < 0.001], and for country [F(6,541) = 37.601, p < 0.001] on the communication style scales. No interaction effect of gender with country was found. Nurses working with a male dentist had higher scores for the 'gender-related' communications styles, whereas those working with a female dentist had higher scores for 'friendly-relation' and 'personal attention' communication styles. Dutch nurses had higher scores on 'playfulness', whereas Northern Irish nurses had higher scores on 'friendly relation' and 'personal attention' communication styles. Some additional differences were found at item level. The age of the dental nurse explained some differences in communication and working styles [F(6,541) = 2.341, p = 0.031); younger dental nurses scored significantly higher for 'gender- related' communication and working style. Conclusion: Gender of the dentist does influence dental nurses' perceptions of the communication styles used in the primary dental care setting. Dental nurses working with female dentists report a style predominantly characterised by 'friendly-relation' and 'personal attention', whereas those working with a male dentist report 'gender-related' communication and working styles. Given the steadily growing percentage of females entering the dental profession, it is recommended to raise both dentists' and nurses' awareness on gender influences in professional communication by introducing these topics in their respective education.