Regenerative Endodontics Using Different Intracanal Medicaments on Radiographic Outcomes and Periapical MMP-8 Levels

Not Applicable

Completed

• Conditions: Regenerative Endodontics
• Interventions: Combination Product: Place the freshly prepared calcium hydroxide in to the root canals; Other: Sample collection for immunofluorometric assay; Combination Product: Place the freshly prepared double antibiotic paste in to the root canals; Diagnostic Test: Taking standart periapical radiography with film holder

• Registration Number: NCT05581706
• Lead Sponsor: Ege University

Brief Summary

Two-session regenerative endodontic treatment was applied to 20 immature mandibular first molars with symptomatic irreversible pulpitis and symptomatic apical periodontitis. At the end of the first session, calcium hydroxide \[Ca(OH)2\] was applied to 10 randomly selected teeth and double antibiotic paste (DAP) intracanal medicaments were applied to the other 10 teeth. The effects of intracanal medicaments on periapical MMP-8 levels were determined by immunofluorometric assay (IFMA) in periapical tissue fluid samples taken from the distal root canal in the first and second sessions.

Detailed Description

The aim of the study was to investigate the effects of two different intracanal medicaments used during regenerative endodontic treatment (RET) on periapical MMP-8 in relation to the radiological outcomes of RET. After local anesthesia and rubber dam isolation were provided, disinfection procedures were applied gently in accordance with the current RET protocol of American Association of Endodontics (AAE) on the molar teeth. Periapical tissue fluid samples were taken with paper points placed to protrude 2 mm from the distal root canal tip. Calcium hydroxide \[Ca(OH)2\] was placed in 10 randomly selected teeth and double antibiotic paste (DAP) intracanal medicaments were placed in the other 10 teeth. Fourteen days later in the second session, the final sampling procedure was performed as in the first session. Then, the treatment was completed according to the current RET protocol of AAE and permanent restorations were made. The success of the treatment and its effect on root development were evaluated by comparing the standard radiographs taken at the end of the treatment and 12 months follow up. MMP-8 levels were measured by immunofluorometric assay (IFMA) in the periapical tissue fluid samples. First and second session MMP-8 levels were compared according to the groups of medicaments \[Ca(OH)2 and DAP groups\] placed in the root canals.

Study Timeline

• Study Start: 2017-09-01
• Primary Completion: 2020-02-01
• Study Completion: 2021-03-01
• Study First Submitted: 2022-10-12
• Study First Submitted QC: 2022-10-12
• Study First Posted: 2022-10-14
• Status Verified: 2023-12
• Last Update Submitted: 2023-12-26
• Last Update Posted: 2023-12-27

Recruitment & Eligibility

• Status: COMPLETED
• Sex: All
• Target Recruitment: 23

Inclusion Criteria

• Healthy children with irreversible pulpitis with symptomatic apical periodontitis (SIP-SAP), indication for RET and informed consent from the patients.

Exclusion Criteria

• Children with extra oral swelling or luxation with a deep periodontal pocket and whom application of RET was contraindicated.
• The teeth were indicated for vital pulp therapy (assesed during treatment)
• Children who have used antibiotics in the past 3 months and are uncooperative during treatment
• Children who have used any painkillers during the treatment

Study & Design

• Study Type: INTERVENTIONAL
• Study Design: PARALLEL

Arm && Interventions

Group	Intervention	Description
Regenerative endodontic treatment with calcium hydroxide	Place the freshly prepared calcium hydroxide in to the root canals	Regenerative endodontic treatment (RET) performed with procedures of American Association of Endodontics (AAE) in two sessions. At the end of the first session, periapical tissue fluid samples (baseline samples) were collected. Calcium hydroxide medicament was prepared by mixing with sterile distilled water and then placed in the coronal 1/3 of the root canals. In the second session (14 days after first treatment), the medicaments in the root canals were carefully removed with 5 ml sterile distilled water irrigation. Periapical tissue fluid samples were obtained from the distal canal using the same protocol as described previously (final samples). Then RET was finished according to the treatment protocol of AAE.
Periapical tissue exudate sample collection and Immunofluorometric assay MMP-8	Sample collection for immunofluorometric assay	At the end of the first session, periapical tissue fluid samples (baseline samples) were collected by introducing 3 sterile #45 paper points into the root canal until 2 mm passing through the root apex from the distal canal. After waiting for 1 min, the paper points were withdrawn, the tip was cut from 4 mm and were transferred to sterile Eppendorf tubes. At the beginnign of the second session periapical tissue fluid samples were obtained from the distal canal using the same protocol as described in the first session (final samples). Baseline and final MMP-8 concentrations were determined by a time-resolved immunofluorometric assay (IFMA). The monoclonal MMP-8 specific antibodies 8708 and 8706 (Oy Medix Biochemica Ab, Espoo, Finland) were used as a catching antibody and a tracer antibody, respectively. The tracer antibody was labeled using europium-chelate
Regenerative endodontic treatment with double antibiotic paste	Place the freshly prepared double antibiotic paste in to the root canals	Regenerative endodontic treatment (RET) performed with procedures of American Association of Endodontics (AAE) in two sessions. At the end of the first session, periapical tissue fluid samples (baseline samples) were collected. Double antibiotic paste (DAP) was prepared by mixing same amount of metronidazole and ciprofloxacin powdered antibiotics (1:1) and combined with sterile distilled water to form an ointment. DAP was introduced in roots canals using a lentulo to fill the entire root canal space. In the second session (14 days after first treatment) after removal of DAP in the root canals, periapical tissue fluid samples were obtained the same protocol as described previously (final samples). Then RET was finished according to the treatment protocol of AAE.
Radiographic evaluation	Taking standart periapical radiography with film holder	Pre-operative, post-operative and final recall (12th months follow up) standart digital radiographs were taken from patients with film holder. The radiographs saved and transferred to Image J software (version 1.47, National Institutes of Health, Bethesda, MD). The standardized radiographs were further aligned using the TurboReg plugin (Biomedical Imaging Group, Swiss Federal Institute of Technology, Lausanne, Switzerland) within the Image J toolkit to minimize any distortions caused by variability in the angulation. All images were calibrated according to size #2 SPP (vertical dimension 31 mm, horizontal dimension 41 mm) using the ''set scale'' option in Image J. Thereafter, the root lengths, root width and radiographic root area were measured on both preoperative and final recall images to evaluate treatment outcomes.

Primary Outcome Measures

Name	Time	Method
Clinical healing according to presence of clinical symptoms	12 months	All patients were called for follow up visits. The clinical healing was determined by examining clinical symptoms including presence of pain, tenderness to percussion and palpation, presence of sinus tract or swelling, and mobility. All teeth were asymptomatic at follow up appointments. If one of these symptoms occurred, it could considered as failure.
Periapical healing according to change in amount of root hard tissues	12 months	All periapical radiolucency detected due to apical periodontitis were healed. Significant increases in radiographic root length, root width and RRA was observed at 12th month.
MMP-8 levels from periapical tissue fluid samples	14 day	There was a statistically significant increase in MMP-8 levels on day 14 compared to baseline in both Ca(OH)2 and DAP groups. The increase in MMP-8 levels was higher in the DAP group compared to Ca(OH)2 group, however no statistically significant difference was observed between 2 groups regarding the final (14th day) MMP-8 levels.

Trial Locations

Locations (1)

Ege University Faculty of Dentistry Pediatric Dentistry; 🇹🇷 İzmir, Turkey