4.7. Interventions

On the first day, all qualified subjects were assigned to brush their teeth 5 h before gargling with normal saline for 1 min. The oral rinse samples were collected and kept at 4 °C. The baselines of GI and PI scores of individual subjects were evaluated by a dentist using a periodontal probe and 6% erythrosine solution staining, respectively. After the assessment, all subjects were randomly assigned to one of the following five groups: group 1: placebo mouthwash, group 2: Kaffir lime (KL) leaf extract mouthwash, group 3: KL with Moringa (MO) leaf extract mouthwash, group 4: KL with Neem (NE) leaf extract mouthwash and group 5: 0.12% Chlorhexidine (CHX) mouthwash. The participants were asked to gargle with the mouthwash for 14 days in the amount of 10–15 mL twice daily for 30 s after brushing their teeth. Subsequent rinsing with water, drinking and eating were not allowed within 15 min. Use of commercial mouthwash during the study was prohibited.

To examine the change of gingival condition in terms of inflammation and plaque buildup, the gingival index (GI) and plaque index (PI) scoring system of Löe-Silness was used [5]. Gingival inflammation was examined based on the GI system, where inflammation of the gum is graded into four scales from 0 (normal) to 3 (severe gingival inflammation). To evaluate the formation of plaque, a 6% erythrosine solution was used, with scores from 0 (no debris or stain present) to 3 (soft debris covering more than two-thirds of the exposed tooth surface).