SCN9A Gene Expression and Inflammatory Cytokines

Jamia Millia Islamia

Status: Enrolling

Conditions: Pulpitis - Irreversible

Treatments: Procedure: Inferior Alveolar Nerve Block

Study type: Observational

Funder types: Other

Identifiers

NCT07532200

SCN9A IRRC- 173

Details and patient eligibility

About

Voltage-gated sodium channels, especially Nav1.7 encoded by the SCN9A gene, are key regulators of nociceptive transmission. Upregulation of SCN9A has been associated with increased neuronal excitability and heightened pain perception. In parallel, inflammatory cytokines such as interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), and interleukin-1 beta (IL-1β) are known to sensitize peripheral nociceptors and reduce the efficacy of local anesthetics by modifying tissue environment and ion channel activity. However, the combined influence of SCN9A expression and inflammatory cytokines on anesthetic success in SIP has not been fully elucidated.

This prospective case-control study aims to evaluate the association between SCN9A gene expression and inflammatory cytokine levels with the clinical success of IANB in patients with SIP affecting mandibular molars. Approximately 90-100 patients will be recruited and categorized into two groups based on anesthetic outcome: successful anesthesia and failed anesthesia. All patients will receive a standardized IANB using 2% lidocaine with 1:100,000 epinephrine. Anesthetic success will be determined based on the absence of pain during access cavity preparation and instrumentation.

Following access and pulp extirpation, pulpal tissue samples will be collected. SCN9A gene expression will be assessed using quantitative real-time polymerase chain reaction (RT-qPCR), with relative expression calculated using the 2^-ΔΔCt method. Inflammatory cytokine levels (IL-6, TNF-α, IL-1β) will be quantified using enzyme-linked immunosorbent assay (ELISA).

The primary outcome will be the difference in SCN9A expression between failed and successful anesthesia groups. Secondary outcomes will include comparison of cytokine levels and evaluation of correlations between SCN9A expression and inflammatory markers. Statistical analysis will include group comparisons, correlation analysis, logistic regression, and receiver operating characteristic (ROC) curve analysis to assess the predictive value of these biomarkers.

Full description

The present study is designed as a prospective case-control investigation to assess the association between SCN9A gene expression and levels of key inflammatory cytokines with the clinical success of IANB. A total of approximately 100 patients diagnosed with SIP in mandibular molars will be recruited and categorized into two groups based on anesthetic outcome: successful anesthesia and failed anesthesia. Standardized IANB will be administered using 2% lidocaine with 1:100,000 epinephrine, and anesthetic success will be determined based on absence of pain during access cavity preparation and instrumentation.

Following access cavity preparation and pulp extirpation, biological samples will be collected. Pulpal tissue samples will be used for RNA extraction and subsequent quantitative real-time polymerase chain reaction (RT-qPCR) analysis to assess SCN9A gene expression. Relative expression levels will be calculated using the 2^-ΔΔCt method with appropriate housekeeping genes. In parallel, inflammatory cytokine levels (IL-6, TNF-α, IL-1β) will be quantified using enzyme-linked immunosorbent assay (ELISA) from pulpal tissue homogenates or gingival crevicular fluid, depending on feasibility.

The primary outcome of the study will be the difference in SCN9A expression between failed and successful anesthesia groups. Secondary outcomes will include comparison of cytokine levels between groups and evaluation of correlations between SCN9A expression and inflammatory markers. Data will be analyzed using appropriate statistical tests, including independent t-tests or non-parametric equivalents, correlation analysis, and logistic regression modeling. Additionally, receiver operating characteristic (ROC) curve analysis may be performed to assess the predictive value of these biomarkers for anesthetic failure.

This study aims to provide mechanistic insights into anesthetic failure in SIP by integrating molecular and inflammatory pathways. The findings may contribute to the development of predictive biomarkers and targeted therapeutic strategies to improve anesthetic success in endodontic practice.

Enrollment: 100 estimated patients

Sex: All

Ages: 18 to 50 years old

Volunteers: Accepts Healthy Volunteers

Inclusion criteria

• Patients aged 18-60 years
• Systemically healthy individuals (ASA I or II)
• Presence of a mandibular first or second molar diagnosed with symptomatic irreversible pulpitis
• Vital tooth confirmed by positive response to pulp sensibility tests (cold test/EPT)
• Moderate to severe preoperative pain (Heft-Parker VAS)
• Patients requiring endodontic treatment under inferior alveolar nerve block
• Ability and willingness to provide informed consent

Exclusion criteria

• Patients who have taken analgesics, anti-inflammatory drugs, or antibiotics within the last 48 hours
• Presence of systemic diseases affecting pain perception or inflammation (e.g., diabetes, neuropathic disorders)
• Pregnant or lactating women
• Teeth with periapical abscess, swelling, or sinus tract
• Non-vital teeth or teeth with previous endodontic treatment
• Patients with known allergy to local anesthetic agents
• Patients with limited mouth opening or anatomical conditions affecting IANB administration
• Inability to understand pain assessment scales or comply with study protocol

Trial design

100 participants in 2 patient groups

IANB Success (Adequate Anesthesia) Group

Description:

This cohort will include patients with symptomatic irreversible pulpitis in mandibular molars who achieve successful pulpal anesthesia following administration of a standardized inferior alveolar nerve block (IANB) using 2% lidocaine with 1:100,000 epinephrine. Anesthetic success will be defined as the absence of pain (no or mild pain) during access cavity preparation and initial instrumentation without the need for any supplementary anesthetic techniques. Following confirmation of anesthesia, access cavity preparation and pulp extirpation will be performed, and pulpal tissue samples will be collected for analysis of SCN9A gene expression (RT-qPCR) and inflammatory cytokine levels (IL-6, TNF-α, IL-1β) using ELISA.

Treatment:

Procedure: Inferior Alveolar Nerve Block

IANB Failure (Inadequate Anesthesia) Group

Description:

This cohort will include patients with symptomatic irreversible pulpitis in mandibular molars who experience inadequate pulpal anesthesia following administration of a standardized inferior alveolar nerve block (IANB) using 2% lidocaine with 1:100,000 epinephrine. Anesthetic failure will be defined as the presence of moderate to severe pain during access cavity preparation or instrumentation, necessitating the use of supplementary anesthetic techniques (e.g., intraligamentary or intrapulpal injections). Pulpal tissue samples will be collected after access cavity preparation and pulp extirpation for analysis of SCN9A gene expression (RT-qPCR) and inflammatory cytokines (IL-6, TNF-α, IL-1β) using ELISA.

Treatment:

Procedure: Inferior Alveolar Nerve Block