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

NCT05581706 | Completed DMC

• 1 other identifier: Regenerative Endodontics
• Study Type: interventional
• Target: 23
• Locations: 1 country
• Sites: 1

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.

Conditions

Regenerative Endodontics

Keywords

regenerative endodontics; intracanal medicaments; MMP-8

Outcome Measures

Primary Outcomes (3)

• Clinical healing according to presence of clinical symptoms

  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.

  12 months

• Periapical healing according to change in amount of root hard tissues

  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.

  12 months

• MMP-8 levels from periapical tissue fluid samples

  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.

  14 day

Study Arms (4)

Regenerative endodontic treatment with calcium hydroxide

ACTIVE COMPARATOR

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.

Combination Product: Place the freshly prepared calcium hydroxide in to the root canals

Regenerative endodontic treatment with double antibiotic paste

ACTIVE COMPARATOR

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.

Combination Product: Place the freshly prepared double antibiotic paste in to the root canals

Radiographic evaluation

ACTIVE COMPARATOR

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.

Diagnostic Test: Taking standart periapical radiography with film holder

Periapical tissue exudate sample collection and Immunofluorometric assay MMP-8

ACTIVE COMPARATOR

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

Other: Sample collection for immunofluorometric assay

Interventions

Place the freshly prepared calcium hydroxide in to the root canals COMBINATION_PRODUCT

Calcium hydroxide (Merck, Darmstadt, Germany): prepared by mixing with sterile distilled water and then placed in the coronal 1/3 of the root canals.

Also known as: Ca(OH)2 medicament

Regenerative endodontic treatment with calcium hydroxide

Place the freshly prepared double antibiotic paste in to the root canals COMBINATION_PRODUCT

Double antibiotic paste (DAP): metronidazole (Flagyl, Sanofi-Aventis, Turkey) and ciprofloxacin (Cipro, Biofarma, Turkey) powdered antibiotics were stored and sealed in airtight containers. The same amount of each drug powder (1:1) was mixed and the mixed samples were 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.

Also known as: DAP medicament

Regenerative endodontic treatment with double antibiotic paste

Taking standart periapical radiography with film holder DIAGNOSTIC_TEST

Post-operative periapical radiographs were obtained with Digora Optime SPP system (Soredex Corp., Tuusula, Finland) using intraoral film holders to keep the plates parallel to the long axis of the teeth. A size #2 SPP was used for all exposures. SPPs were exposed with a Gendex Oralix DC dental x-ray unit (Gendex Dental Systems, Milan, Italy) operating at 60 kVp, 7 mA, 0.25 sec. and the plates were scanned immediately after exposure. The clinical and radiographical follow up was performed on 12th months. Image-J program (version 1.47, National Institutes of Health, Bethesda, MD) with TurboReg plug-in (Biomedical Imaging Group, Swiss Federal Institute of Technology, Lausanne, Switzerland) was used to determine the increase in root length, root width, and radiographic root area at 12th month follow up.

Also known as: Determination of rooth lenght, rooth width, radiographic root area

Radiographic evaluation

Sample collection for immunofluorometric assay OTHER

Periapical exudate samples were collected at the beginning of the RET and at 14th day. MMP-8 levels were measured by immunofluorometric assay (IFMA). Briefly, two monoclonal MMP-8-specific antibodies, 8708 and 8706 (Oy Medix Biochemica Ab, Espoo, Finland) were used as catching antibody and tracer antibody, respectively. Samples were diluted in enzyme buffer (50 mM Tris-HCl, pH 7.5; 0.2 mM NaCl, 1 mM CaCl2). Twenty microlitres of samples and 80 μl of assay buffer (20 mM Tris-HCl, pH 7.5, 0.5 M NaCl, 5 mM CaCl2, 0.5% BSA, 0.05% sodium azide and 20 mg/l diethylenetriaminepentaacetic acid with 2 μg/ml normal mouse serum) were pipetted into the wells. The tracer antibody was labelled using europium chelate. After adding the enhancement solution, fluorescence was measured using a 1234 Delfia Research Fluorometer (Wallac, Turku, Finland).

Also known as: IFMA

Periapical tissue exudate sample collection and Immunofluorometric assay MMP-8

Eligibility Criteria

• Age: 6 Years - 9 Years
• Sex: all
• Healthy Volunteers: Yes
• Age Groups: Child (0-17)

You may qualify if:

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

You may not qualify if:

• 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

Sponsors & Collaborators

• Ege University: lead

Study Sites (1)

Ege University Faculty of Dentistry Pediatric Dentistry

Izmir, Turkey (Türkiye)

Location

Related Publications (8)

• Torabinejad M, Alexander A, Vahdati SA, Grandhi A, Baylink D, Shabahang S. Effect of Residual Dental Pulp Tissue on Regeneration of Dentin-pulp Complex: An In Vivo Investigation. J Endod. 2018 Dec;44(12):1796-1801. doi: 10.1016/j.joen.2018.09.005.

  PMID: 30477665 BACKGROUND

• Sabeti M, Golchert K, Torabinejad M. Regeneration of Pulp-Dentin Complex in a Tooth with Symptomatic Irreversible Pulpitis and Open Apex Using Regenerative Endodontic Procedures. J Endod. 2021 Feb;47(2):247-252. doi: 10.1016/j.joen.2020.09.021. Epub 2020 Oct 10.

  PMID: 33045267 BACKGROUND

• Wahlgren J, Salo T, Teronen O, Luoto H, Sorsa T, Tjaderhane L. Matrix metalloproteinase-8 (MMP-8) in pulpal and periapical inflammation and periapical root-canal exudates. Int Endod J. 2002 Nov;35(11):897-904. doi: 10.1046/j.1365-2591.2002.00587.x.

  PMID: 12453017 BACKGROUND

• Kuula H, Salo T, Pirila E, Tuomainen AM, Jauhiainen M, Uitto VJ, Tjaderhane L, Pussinen PJ, Sorsa T. Local and systemic responses in matrix metalloproteinase 8-deficient mice during Porphyromonas gingivalis-induced periodontitis. Infect Immun. 2009 Feb;77(2):850-9. doi: 10.1128/IAI.00873-08. Epub 2008 Nov 24.

  PMID: 19029300 BACKGROUND

• Bose R, Nummikoski P, Hargreaves K. A retrospective evaluation of radiographic outcomes in immature teeth with necrotic root canal systems treated with regenerative endodontic procedures. J Endod. 2009 Oct;35(10):1343-9. doi: 10.1016/j.joen.2009.06.021. Epub 2009 Aug 15.

  PMID: 19801227 BACKGROUND

• Reynaud af Geijersstam A, Sorsa T, Stackelberg S, Tervahartiala T, Haapasalo M. Effect of E. faecalis on the release of serine proteases elastase and cathepsin G, and collagenase-2 (MMP-8) by human polymorphonuclear leukocytes (PMNs). Int Endod J. 2005 Sep;38(9):667-77. doi: 10.1111/j.1365-2591.2005.01011.x.

  PMID: 16104981 BACKGROUND

• Martinho FC, Nascimento GG, Leite FR, Gomes AP, Freitas LF, Camoes IC. Clinical influence of different intracanal medications on Th1-type and Th2-type cytokine responses in apical periodontitis. J Endod. 2015 Feb;41(2):169-75. doi: 10.1016/j.joen.2014.09.028. Epub 2014 Nov 11.

  PMID: 25453567 BACKGROUND

• Pekpinarli B, Kaval ME, Cogulu D, Ilhan B, Sorsa T, Tervahartiala T, Oncag O. The effect of calcium hydroxide and double antibiotic paste on radiographic outcomes and periapical MMP-8 levels in regenerative endodontic procedures: a randomized clinical trial. J Appl Oral Sci. 2024 Sep 20;32:e20240122. doi: 10.1590/1678-7757-2024-0122. eCollection 2024.

  PMID: 39319904 DERIVED

MeSH Terms

Interventions

Specimen Handling; Fluoroimmunoassay

Intervention Hierarchy (Ancestors)

Clinical Laboratory Techniques; Diagnostic Techniques and Procedures; Diagnosis; Investigative Techniques; Fluorescent Antibody Technique; Immunohistochemistry; Histocytochemistry; Cytological Techniques; Histological Techniques; Immunoassay; Immunologic Techniques

Study Officials

• Burç Pekpınarlı

  Ege University Faculty of Dentistry

  PRINCIPAL INVESTIGATOR

Study Design

• Study Type: interventional
• Phase: not applicable
• Allocation: RANDOMIZED
• Masking: SINGLE
• Who Masked: OUTCOMES ASSESSOR
• Purpose: TREATMENT
• Intervention Model: PARALLEL
• Sponsor Type: OTHER
• Responsible Party: PRINCIPAL INVESTIGATOR
• PI Title: Principal investigator, Asst. Dr

Study Record Dates

• First Submitted: October 12, 2022
• First Posted: October 14, 2022
• Study Start: September 1, 2017
• Primary Completion: February 1, 2020
• Study Completion: March 1, 2021
• Last Updated: December 27, 2023

Record last verified: 2023-12

Data Sharing

• IPD Sharing: Will not share

Locations

TU (1)