Quantification of Bacterial DNA from Infected Human Root Canals Using qPCR and DAPI after Disinfection with Established and Novel Irrigation Protocols

Materials

Christian Hannig¹  Marie-Theres Weber¹  Sabine Basche¹  Yousef Alkhafaji¹  Anne Pioch¹  Torsten Sterzenbach¹  Evelyn Trips²  Martin Dannemann³  Alan Kilistoff⁴ 

[1] Clinic of Operative and Pediatric Dentistry, Medical Faculty Carl Gustav Carus, Technische Universität Dresden, Fetscherstraße 74, 01307 Dresden, Germany;Coordination Center for Clinical Studies Dresden, Medical Faculty Carl Gustav Carus, Technical University Dresden, Fetscherstraße 74, 01309 Dresden, Germany;Faculty of Automotive Engineering, Institute of Energy and Transport Engineering, Westsächsische Hochschule Zwickau, Scheffelstraße 39, 08012 Zwickau, Germany;Faculty of Medicine & Dentistry, University of Alberta, 11405 87th Ave NW, Edmonton, AB T6G 1C9, Canada;

关键词: irrigation protocol;    root canal system;    qPCR–DAPI;    silver diamine fluoride;    endodontics;   

DOI  :  10.3390/ma15051911

来源: DOAJ

【 摘 要 】

The removal of bacterial infections within the root canal system is still a challenge. Therefore, the cleansing effect of established and new irrigation-protocols (IP) containing silver diamine fluoride (SDF) 3.8% on the whole root canal system was analyzed using quantitative PCR (qPCR) and 4′,6-diamidino-phenylindole-(DAPI)-staining. Extracted human premolars were instrumented up to F2 (ProTaper Gold) under NaCl 0.9% irrigation and incubated with Enterococcus faecalis for 42 days. Subsequently, different ultrasonically agitated IP were applied to the roots: control (no irrigation), 1. NaOCl 3%, EDTA 20%, CHX 2%, 2. NaOCl 3%, EDTA 20%, 3. NaOCl 3%, EDTA 20%, SDF 3.8%, 4. SDF 3.8%, and 5. NaCl 0.9%. One half of the root was investigated fluorescent-microscopically with DAPI. The other half was grinded in a cryogenic mill and the bacterial DNA was quantified with qPCR. The qPCR results showed a statistically significant reduction of bacteria after the application of IP 1, 2, and 3 compared to the control group. While IP 4 lead to a bacterial reduction which was not significant, IP 5 showed no reduction. These data corresponded with DAPI staining. With qPCR a new molecular-biological method for the investigation of the complete root canal system was implemented. The novel IP 3 had an equally good cleansing effect as the already established IP.

【 授权许可】

Unknown