Background: Vitamin D is crucial for mineralized tissue formation and immunological functions. The purpose of this study was to evaluate the association between vitamin D status and dental status in healthy children with vitamin D supplementation in infancy and at 6 years of age. Method: Eight-year-old children who had participated, in a vitamin D intervention project when they were 6 years old were invited to participate in a dental follow-up study. They had fair or darker skin complexion and represented two geographically distant parts of Sweden. 25-hydroxy vitamin D in serum had been measured at 6 years of age and after a 3-month intervention with 25, 10 or 2 (placebo) mu g of vitamin D-3 per day. Two years later, caries and enamel defects were scored, self-reported information on e.g., oral behavior, dietary habits and intake of vitamin D supplements was collected, and innate immunity peptide LL37 levels in saliva and cariogenic mutant streptococci in tooth biofilm were analyzed. The outcome variables were caries and tooth enamel defects. Results: Dental status was evaluated in 85 of the 206 children in the basic intervention study. Low vitamin D levels were found in 28% at baseline compared to 11% after the intervention, and 34% reported continued intake of vitamin D supplements. Logistic regression supported a weak inverse association between vitamin D status at 6 years of age and caries 2 years later (odds ratio 0.96; p = 0.024) with minor attenuation after an adjustment for potential confounders. Multivariate projection regression confirmed that insufficient vitamin D levels correlated with caries and higher vitamin D levels correlated with being caries-free. Vitamin D status at 6 years of age was unrelated to enamel defects but was positively associated with saliva LL37 levels. Conclusion: An association between vitamin D status and caries was supported, but it was not completely consistent. Vitamin D status at 6 years of age was unrelated to enamel defects but was positively associated with LL37 expression.