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Evaluation of a nonthermal plasma needle to eliminate ex vivo biofilms in root canals of extracted human teeth.

ABSTRACT: To evaluate the efficacy of a nonthermal plasma (NTP) at atmospheric pressure on ex vivo biofilm in root canals of extracted teeth.Intracanal contents from three teeth with root canal infections were collected, pooled and grown in thirty-five microCT-mapped root canals of extracted and instrumented human teeth. One group of teeth was treated with NTP, another with 6% NaOCl and one set was left untreated. The intracanal contents from twenty-seven teeth (nine teeth in each group) were plated on agar and colony forming units were determined. Parametric test of one-way analysis of variance (anova) was used to analyse statistical significance. The remaining teeth were cut open, stained with LIVE/DEAD(®) and examined with confocal laser scanning microscopy.The untreated root canals were covered with biofilm of varying thickness. Treatment with nonthermal plasma decreased the number of viable bacteria in biofilms by one order of magnitude, whilst the NaOCl control achieved a reduction of more than four magnitudes. Both the NTP and the NaOCl treatment results were significantly different from the negative control (P < 0.05).The nonthermal plasma displayed antimicrobial activity against endodontic biofilms in root canals, but was not as effective as the use of 6% NaOCl.

SUBMITTER: Schaudinn C

PROVIDER: S-EPMC3687033 | biostudies-literature | 2013 Oct

REPOSITORIES: biostudies-literature

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Evaluation of a nonthermal plasma needle to eliminate ex vivo biofilms in root canals of extracted human teeth.

Schaudinn C C Jaramillo D D Freire M O MO Sedghizadeh P P PP Nguyen A A Webster P P Costerton J W JW Jiang C C

International endodontic journal 20130311 10

<h4>Aim</h4>To evaluate the efficacy of a nonthermal plasma (NTP) at atmospheric pressure on ex vivo biofilm in root canals of extracted teeth.<h4>Methodology</h4>Intracanal contents from three teeth with root canal infections were collected, pooled and grown in thirty-five microCT-mapped root canals of extracted and instrumented human teeth. One group of teeth was treated with NTP, another with 6% NaOCl and one set was left untreated. The intracanal contents from twenty-seven teeth (nine teeth ...[more]

PMID: 23480318

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Project description:Ultraviolet C (UVC) light emitting diode (LED) can kill the endodontic pathogen Enterococcus faecalis and has the potential to kill other oral microorganisms associated with endodontic infections. This same bacteriocidal device shows great promise in the stimulation of periapical healing and pain reduction resulting from inflammation in root canals. Previously, we found that 255 nm UVC LED killed E. faecalis and induced the production of cellular biomarkers in HEPM cells and gingival fibroblasts (Morio et al., 2019). Here, we extend those findings and hypothesize that UVC LED at other wavelengths and power levels kill microorganisms associated with root canal infections. Units emitting UVC LED at 265 nm (12 mW), 265 nm (22.5 mW), and 280 nm (8 mW) wavelenths were assembled and the energy levels of their emissions were measured. The energy doses in millijoules (mJ) were calculated from the power readings of the meter (µW) × time of exposure (seconds). Ex vivo models of root canals were prepared in extracted, instrumented, single canal human premolars. Five cultures of microorganisms were treated with 265 nm (12 mW), 265 nm (22.5 mW), or 280 nm (8 mW) UVC LED on discs in laboratory assays and 4 cultures of microorganisms were treated with 265 nm (22.5 mW) UVC LED in root canals of extracted, instrumented teeth. After UVC LED treatment, all microorganisms were cultivated on microbiological media. Colony forming units (CFU) of viable microorganisms treated with UVC LED were counted and compared with those of viable microorganisms not treated with UVC LED as controls. Tukey's Honestly Significant Difference was used to determine statistical significances (0.05). Units emitting UVC LED at 265 nm (12 mW), 265 nm (22.5 mW), and 280 nm (8 mW) killed Candida albicans, Staphylococcus aureus, methicillin-resistant S. aureus (MRSA), E. faecalis, and Streptococcus sanguinis after 30-90 seconds of exposure in laboratory assays (p < 0.05). Microbial killing differed among treatment times, UVC LED wavelengths, power levels of each unit, and specific microorganism. The unit emitting UVC LED at 265 nm (22.5 mW) killed C. albicans, S. aureus, MRSA, and E. faecalis in 30 s in root canals of extracted, instrumented teeth (p < 0.05). This dataset can be reused to assess the ability of other wavelengths and power levels to kill microorganisms as well as improve procedures for treating endodontic infections and inflammation in root canals.

Project description:Microorganisms are predominantly organized in biofilms, where cells live in dense communities and are more resistant to external stresses than are their planktonic counterparts. With in vitro experiments, the susceptibility of Candida albicans biofilms to a nonthermal plasma treatment (plasma source, kINPen09) in terms of growth, survival, and cell viability was investigated. C. albicans strain SC5314 (ATCC MYA-2876) was plasma treated for different time periods (30 s, 60 s, 120 s, 180 s, 300 s). The results of the experiments, encompassing CFU, fluorescence Live/Dead, and 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide salt (XTT) assays, revealed a negative influence of the plasma treatment on the proliferation ability, vitality, and metabolism of C. albicans biofilms, respectively. Morphological analysis of plasma-treated biofilms using atomic force microscopy supported the indications for lethal plasma effects concomitant with membrane disruptions and the loss of intracellular fluid. Yielding controversial results compared to those of other publications, fluorescence and confocal laser scanning microscopic inspection of plasma-treated biofilms indicated that an inactivation of cells appeared mainly on the bottom of the biofilms. If this inactivation leads to a detachment of the biofilms from the overgrown surface, it might offer completely new approaches in the plasma treatment of biofilms. Because of plasma's biochemical-mechanical mode of action, resistance of microbial cells against plasma is unknown at this state of research.IMPORTANCE Microbial communities are an increasing problem in medicine but also in industry. Thus, an efficient and rapid removal of biofilms is becoming increasingly important. With the aid of the kINPen09, a radiofrequency plasma jet (RFPJ) instrument, decisive new findings on the effects of plasma on C. albicans biofilms were obtained. This work showed that the inactivation of biofilms takes place mainly on the bottom, which in turn offers new possibilities for the removal of biofilms by other strategies, e.g., mechanical treatment. This result demonstrated that nonthermal atmospheric pressure plasma is well suited for biofilm decontamination.

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Evaluation of a nonthermal plasma needle to eliminate ex vivo biofilms in root canals of extracted human teeth.

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Evaluation of a nonthermal plasma needle to eliminate ex vivo biofilms in root canals of extracted human teeth.

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