This randomized clinical trial aimed to compare residual oral bacterial DNA detection after ultrasonic irrigant activation and 980-nm diode laser irrigant activation during root canal treatment of necrotic single-rooted teeth. Sixteen patients requiring endodontic treatment were randomly assigned into two groups: ultrasonic activation or 980-nm diode laser activation. Root canals were prepared using standardized chemomechanical procedures and irrigated with 3% sodium hypochlorite activated by the assigned method. Microbial samples were collected before treatment and after irrigant activation. Bacterial DNA was identified using 16S rRNA polymerase chain reaction (PCR) sequencing. The primary outcome was the presence or absence of detectable oral bacterial DNA after treatment, while the secondary outcome was the taxonomic pattern of detected bacteria before and after activation. The study was designed to explore the comparative antibacterial effects of ultrasonic and diode laser activation in infected root canals.

Pulp necrosis is associated with microbial colonization and biofilm formation within the root canal system. Residual bacteria that persist after root canal treatment may contribute to persistent infection and unfavorable clinical outcomes. Although chemomechanical preparation and sodium hypochlorite irrigation are essential components of endodontic disinfection, anatomical complexities may limit complete bacterial elimination. Therefore, irrigant activation techniques have been introduced to improve irrigant penetration, biofilm disruption, and antimicrobial effectiveness.

Ultrasonic activation enhances irrigant movement through acoustic streaming and cavitation, whereas diode laser activation may produce additional photothermal antibacterial effects and deeper penetration into dentinal tubules. However, evidence comparing residual bacterial detection after these activation methods remains limited, particularly when evaluated using molecular identification techniques.

This exploratory randomized clinical trial compared ultrasonic irrigant activation and 980-nm diode laser irrigant activation in necrotic single-rooted teeth using 16S rRNA PCR sequencing. The study was conducted at the Dental and Oral Hospital of Hasanuddin University and the Hasanuddin University Medical Research Center, Indonesia. Sixteen necrotic single-rooted teeth were randomly allocated into two groups: ultrasonic activation (n = 8) and 980-nm diode laser activation (n = 8).

After rubber dam isolation and access preparation under aseptic conditions, initial microbial samples were collected before chemomechanical preparation. Root canal instrumentation was performed using ProGlider and ProTaper Gold rotary instruments up to F2. In both groups, canals were irrigated using 3% sodium hypochlorite. In the ultrasonic group, irrigant activation was performed using ultrasonic activation at 45 kHz for 20 seconds per cycle for three cycles. In the diode laser group, activation was performed using a 980-nm diode laser at 1.5 W for 20 seconds per cycle for three cycles. Post-treatment microbial samples were then collected.

Bacterial DNA was extracted and amplified using universal 16S rRNA primers, followed by sequencing and taxonomic identification. The primary outcome was the presence or absence of detectable oral bacterial DNA after treatment. Secondary outcomes included identification of bacterial taxa detected before and after treatment and comparison of bacterial profile reduction between activation methods.

The study was designed as an exploratory clinical-molecular investigation intended to generate preliminary evidence regarding the antibacterial effects of ultrasonic and diode laser irrigant activation in root canal disinfection.

Inclusion Criteria:

• Adults aged 18 years or older
• Patients requiring endodontic treatment for mature single-rooted teeth diagnosed with pulp necrosis
• Physically and mentally able to undergo treatment
• No use of analgesics or antibiotics within one week before treatment
• Teeth without clinical or radiographic evidence of apical periodontitis
• Diagnosis confirmed by clinical examination, pulp vitality testing, and periapical radiography

Exclusion Criteria:

• Teeth with mobility
• Periodontal pockets greater than 4 mm
• Root fracture or fracture during treatment
• Internal or external root resorption
• Canal calcification or canal obliteration
• Patients with systemic conditions requiring antibiotic prophylaxis for routine dental treatment
• Recent antimicrobial exposure that could influence microbial findings

The IPD will not be shared because the researchers have guaranteed the confidentiality of patient data

None (Open Label) Not applicable. No masking was applied in this study