Only a few studies have investigated use of the Dental Anxiety Scale in dental fear-induced neuroendocrine changes. The present study examined 19 female patients, each at two timepoints across an educational and a treatment session within a periodontitis therapy. Subjective measures included a visual analogue scale, the STAI State scale, and the Dental Anxiety Scale. Salivary cortisol was measured in parallel across all four timepoints. Although patients were significantly more aroused and anxious prior to the treatment session, salivary cortisol remained unchanged. However, patients with high Dental Anxiety were significantly more aroused and anxious and showed significantly higher salivary cortisol during the educational session than those with low scores on Dental Anxiety. In conclusion, the Dental Anxiety Scale differentiated mean neuroendocrine change between patients scoring low and high for Dental Anxiety and its use as an accurate tool to identify patients with high dental anxiety should be further studied predictively.