Relationship Between Malocclusion Severity and Periodontitis Risk

After isolating the teeth with a cotton roll, supragingival plaque will be removed and a gentle airflow will be used to dry the teeth. GCF samples will be collected by inserting PerioPaper strips into the mesiobuccal gingival sulcus to a depth of approximately 1 mm for 30 seconds. A calibrated Periotron 8000 instrument will then be used to measure the total GCF volume. PerioPaper strips will then be stored at -80°C for future measurements.

Plaque Index (PI) This index is used to evaluate the amount of dental plaque accumulation on tooth surfaces. Each tooth's four surfaces (buccal, lingual, mesial, distal) are examined using a probe or visually. Each surface is scored from 0 to 3. The average score is calculated for each tooth.

The overall plaque index is obtained by averaging the scores from all examined teeth.

Score Description 0 No plaque.

• No visible plaque, but a slight film of plaque is detected when a probe is run along the gingival margin.
• Visible plaque along the gingival margin, forming a continuous band.
• Abundant plaque covering the gingival area and extending toward the middle of the tooth surface, including the interproximal spaces.

Papillary Bleeding Index (PBI) This index is used to assess bleeding of the interdental papilla-an early sign of periodontal disease-in order to evaluate the degree of gingival inflammation.

A periodontal probe is gently inserted into each interdental papilla (the triangular-shaped gum tissue between teeth). The probe is placed approximately 1-2 mm into the papilla and directed upward. Bleeding occurring within 10-30 seconds is observed and scored based on severity.

The scores from all papillae are totaled and divided by the number of examined sites to obtain the average PBI score.

Score Description 0 No bleeding

• Single-point, slight bleeding
• Thin line of blood
• Obvious and rapid bleeding across the entire papilla, slight spreading
• Severe, profuse bleeding that overflows the papilla, usually with evident inflammation

Bleeding on Probing (BOP) Index This index is used to assess the presence and severity of gingival inflammation by observing bleeding after gentle pressure is applied to the gingival sulcus using a periodontal probe. A thin periodontal probe is gently inserted into the gingival sulcus with light pressure (approximately 20-25 grams). Bleeding occurring within 10-30 seconds is observed. Each measurement site is recorded as either "bleeding present" or "bleeding absent." Presence of bleeding is an early and sensitive indicator of gingival inflammation.

Score Description 0 No bleeding

1 Bleeding present (mild or severe)

Probing Pocket Depth (PPD)

Using a Williams periodontal probe, the distance between the marginal gingival margin and the base of the periodontal pocket is measured in millimeters at six sites per tooth:

mesiobuccal, midbuccal, distobuccal, mesiolingual, midlingual, and distolingual.

All teeth are measured. The values obtained from each site are summed and then divided by the total number of sites measured.

This gives the average probing depth for the individual.

Measurement Interpretation 1-3 mm Healthy sulcus 4-5 mm Shallow to moderate pocket

≥6 mm Deep periodontal pocket (advanced disease)

Clinical Attachment Loss (CAL)

Using a Williams periodontal probe, the distance between the enamel-cementum junction and the base of the periodontal pocket is measured in millimeters at six sites per tooth:

mesiobuccal, midbuccal, distobuccal, mesiolingual, midlingual, and distolingual.

All teeth are measured. The values obtained from each site are summed and then divided by the total number of sites measured.

This gives the average probing depth for the individual.

CAL Value Interpretation 0-1 mm Healthy / minimal loss 2-3 mm Mild attachment loss 4-5 mm Moderate attachment loss

≥6 mm Severe attachment loss

Frozen paper strips will be thawed and diluted with 200 μL of phosphate-buffered saline (pH 7.4). The mixture will then be centrifuged at 13,000 g for 10 min at 4 °C. RANKL, Osteoprotegerin (OPG), Total Oxidative Status (TOS), and Total Antioxidant Capacity (TAC) kits will be analyzed by ELISA. Results will be expressed as pg/μL.