Evaluation of PTHrP, RANKL, and OPG Levels in Periodontal Health and Diseases

Evaluation of Gingival Crevicular Fluid Parathyroid Hormone-Related Protein (PTHrP), RANKL, and OPG Levels in Periodontal Health, Gingivitis, and Periodontitis.

This observational study aims to evaluate the levels of specific biological markers-Parathyroid Hormone-Related Protein (PTHrP), RANKL, and Osteoprotegerin (OPG)-in the gingival crevicular fluid of individuals with different gum health conditions. The study includes a total of 81 participants, divided equally into three groups: 27 with healthy gums, 27 with gingivitis, and 27 with periodontitis. During a routine dental visit, participants undergo a standard periodontal examination to measure clinical signs such as plaque levels, gum bleeding, and pocket depths. Additionally, small sterile paper strips are gently placed in the gum crevices for 30 seconds to collect fluid samples. These fluid samples are then analyzed in a laboratory to understand how these specific proteins relate to gum health and the severity of gum disease.

Periodontitis is a chronic inflammatory disease characterized by the destruction of tooth-supporting tissues and alveolar bone loss, primarily driven by an imbalance in bone metabolism. The regulation of bone turnover relies heavily on the balance between RANKL, which promotes osteoclast activation, and OPG, its natural inhibitor. Furthermore, PTHrP acts as a critical autocrine and paracrine regulator in periodontal tissue health and alveolar bone repair. The primary rationale of this research is to investigate the dynamic role of PTHrP in periodontal pathogenesis during both the inflammatory and tissue-destruction phases, evaluating its potential as a biological marker. Following a comprehensive clinical periodontal examination by a single calibrated clinician, specific sampling sites are isolated with cotton rolls and gently air-dried. Supragingival plaque is carefully removed prior to sampling. Gingival crevicular fluid (GCF) is then collected from these designated interproximal sites using sterile filter paper strips, which are inserted into the sulcus until mild resistance is felt and left in place for 30 seconds. Samples contaminated with blood or saliva are immediately excluded. The exact volume of the collected fluid is quantified using an electronic volume meter (Periotron 8010). All valid strips are subsequently transferred to microcentrifuge tubes and preserved at -80°C until laboratory processing. The biochemical quantification of PTHrP, RANKL, and OPG levels is performed via commercially available Enzyme-Linked Immunosorbent Assay (ELISA) kits in accordance with the manufacturer's protocols.

Inclusion Criteria:

• Individuals aged 18 years and older.
• Presence of at least 20 natural teeth, excluding third molars.
• For the Periodontally Healthy GrouP: PD ≤ 3 mm, BOP < 10%, and no interproximal CAL or RBL.
• For the Gingivitis Group: PD ≤ 3 mm, BOP ≥ 30%, and no interproximal CAL or RBL.
• For the Periodontitis Group: Diagnosed with Stage III periodontitis (interproximal CAL ≥ 5 mm, PD ≥ 6 mm, and RBL extending to the middle third of the root or more apically).

Exclusion Criteria:

• Presence of systemic diseases that may affect periodontal tissues (e.g., cardiovascular diseases, atherosclerosis, immune system diseases, rheumatoid arthritis, obesity, cancer, and diabetes).
• Use of antibiotics within the last 3 months.
• Use of any medication that may affect periodontal tissues.
• Presence of infectious diseases such as HIV, HBV, or tuberculosis.
• Pregnancy or lactation.
• History of periodontal treatment within the last 6 months.
• Smoking