New Insights into the Microbial Profiles of Infected Root Canals in Traumatized Teeth

Manoharan, Lokeshwaran; Brundin, Malin; Rakhimova, Olena; Paz, Luis Chávez de; Vestman, Nelly Romani

doi:10.3390/jcm9123877

Cited by 14 publications

(21 citation statements)

References 45 publications

(53 reference statements)

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“…Our characterization and analysis of pre‐treatment microbiological samples, using NGS data, showed that a homogeneous and reproducible microbial community was obtained between the experimental groups (alpha diversity). Although our experimental biofilm model may not completely reproduce the flora of the root canal system—which can vary substantially between individuals, infection stages and types (primary and secondary/persistent) (Manoharan et al, 2020; Sassone et al, 2008; Siqueira & Rôças, 2009)—the model showed to be reproducible and composed of bacterial genera implicated in endodontic infections.…”

Section: Discussionmentioning

confidence: 93%

In vitro efficacy of a non‐instrumentation technique to remove intracanal multispecies biofilm

et al. 2022

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Aim:The aim of this study was to assess the efficacy of a non-instrumentation technique to disinfect root canals infected by a human dental plaque-derived multispecies biofilm.Methodology: Twenty-two mandibular incisors were accessed, autoclaved and inoculated with dental plaque. The Center for Disease Control biofilm reactor was used to promote contamination of the root canal space. In the conventional technique (control), the specimens were instrumented until size 35/04 and irrigated with 6% NaOCl. In the non-instrumentation technique, a glide path was established using K-files size 10-20 and specimens were immediately cleaned with the GentleWave System. Samples were obtained for culture and 16S rRNA gene sequencing.Differences in abundances of genera were evaluated using Kruskal-Wallis test, and differences in alpha diversity were compared using anova. Alpha and beta diversity indices were calculated using mothur. The Shannon and Chao1 indices were used to measure alpha diversity. The Bray-Curtis dissimilarity was used to measure beta diversity. Differences in community composition were evaluated using analysis of similarity with Bonferroni correction for multiple comparisons. Results:The total numbers of reads in biological samples ranged from 126 to 45 286.Significantly fewer reads were obtained from samples following cleaning by either method (p < .0001), and significantly fewer reads were obtained in post-cleaning samples following conventional versus non-instrumentation cleaning regiment (p = .002). Communities in pre-treatment samples were similar in both groups; however, significantly greater relative abundances of Streptococcus, Veillonella and Campylobacter were observed following cleaning using non-instrumentation technique (Kruskal-Wallis p = .009, .033, and .001, respectively). Whilst no significant differences were observed in Shannon alpha diversity, the Chao1 index was significantly lower in post-cleaning samples.Conclusions: Significant shifts in composition were observed following cleaning by using both regimens, but the impact of this change was greater following a conventional cleaning technique.

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Section: Discussionmentioning

confidence: 93%

In vitro efficacy of a non‐instrumentation technique to remove intracanal multispecies biofilm

et al. 2022

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“…2016/520-31). Sample collection, processing, and characterization of isolates was performed as previously described ( Manoharan et al., 2020 ). Briefly, samples were collected from teeth isolated with a rubber dam and using strict aseptic techniques.…”

Section: Methodsmentioning

confidence: 99%

“…Fusobacterium nucleatum and S. exigua were the most prevalent isolated species from young infected and traumatized teeth ( Manoharan et al., 2020 ), while E. faecalis and A. gerensceriae were chosen for their role in root canal treatment failure ( Siqueira and Rocas, 2008 ). Lactobacillus gasseri , which displayed probiotic traits, had been previously isolated from the oral cavity of healthy infants ( Vestman et al., 2013 ).…”

Section: Methodsmentioning

confidence: 99%

Cytokine Secretion, Viability, and Real-Time Proliferation of Apical-Papilla Stem Cells Upon Exposure to Oral Bacteria

Rakhimova

Schmidt

Landström

et al. 2021

Front. Cell. Infect. Microbiol.

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The use of stem cells from the apical papilla (SCAPs) has been proposed as a means of promoting root maturation in permanent immature teeth, and plays a significant role in regenerative dental procedures. However, the role of SCAPs may be compromised by microenvironmental factors, such as hypoxic conditions and the presence of bacteria from infected dental root canals. We aim to investigate oral bacterial modulation of SCAP in terms of binding capacity using flow cytometry and imaging, real-time cell proliferation monitoring, and cytokine secretion (IL-6, IL-8, and TGF-β isoforms) under anaerobic conditions. SCAPs were exposed to key species in dental root canal infection, namely Actinomyces gerensceriae, Slackia exigua, Fusobacterium nucleatum, and Enterococcus faecalis, as well as two probiotic strains, Lactobacillus gasseri strain B6 and Lactobacillus reuteri (DSM 17938). We found that A. gerensceriae, S. exigua, F. nucleatum, and E. faecalis, but not the Lactobacillus probiotic strains bind to SCAPs on anaerobic conditions. Enterococcus faecalis and F. nucleatum exhibited the strongest binding capacity, resulting in significantly reduced SCAP proliferation. Notably, F. nucleatum, but not E. faecalis, induce production of the proinflammatory chemokine IL-8 and IL-10 from SCAPs. Production of TGF-β1 and TGF-β2 by SCAPs was dependent on species, cell line, and time, but secretion of TGF-β3 did not vary significantly over time. In conclusion, SCAP response is compromised when exposed to bacterial stimuli from infected dental root canals in anaerobic conditions. Thus, stem cell-mediated endodontic regenerative studies need to include microenvironmental conditions, such as the presence of microorganisms to promote further advantage in the field.

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“…Fusobacterium nucleatum is a high-frequency specie isolated from the oral cavity but usually absent or very rarely detected elsewhere in the human body [11]. F. nucleatum is closely related to the development of periodontitis [12] and endodontic infections [13] and is described as a possible oncogene related to colorectal cancer, pancreatic cancer, and oral cancers [14]. This Gram-negative bacterial specie is able to adhere to a variety of oral microbes and can produce a group of virulence factors and toxic metabolites, including lipopolysaccharides, porins, butyrate, and propionate ammonia [15].…”

Section: Of 23mentioning

confidence: 99%

“…The bacterial isolates were obtained as previously described [13]. Briefly, the contents of the root canal were absorbed into sterile paper points and transferred to fluid thioglycolate medium supplemented with agar.…”

Section: Bacterial Strains Supernatants and Growth Conditionsmentioning

confidence: 99%

Combined Transcriptomic and Protein Array Cytokine Profiling of Human Stem Cells from Dental Apical Papilla Modulated by Oral Bacteria

Zymovets

Razghonova²,

Rakhimova

et al. 2022

IJMS

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Stem cells from the apical papilla (SCAP) are a promising resource for use in regenerative endodontic treatment (RET) that may be adversely affected by oral bacteria, which in turn can exert an effect on the success of RET. Our work aims to study the cytokine profile of SCAP upon exposure to oral bacteria and their supernatants—Fusobacterium nucleatum and Enterococcus faecalis—as well as to establish their effect on the osteogenic and immunogenic potentials of SCAP. Further, we target the presence of key proteins of the Wnt/β-Catenin, TGF-β, and NF-κB signaling pathways, which play a crucial role in adult osteogenic differentiation of mesenchymal stem cells, using the Western blot (WB) technique. The membrane-based sandwich immunoassay and transcriptomic analysis showed that, under the influence of F. nucleatum (both bacteria and supernatant), the production of pro-inflammatory cytokines IL-6, IL-8, and MCP-1 occurred, which was also confirmed at the mRNA level. Conversely, E. faecalis reduced the secretion of the aforementioned cytokines at both mRNA and protein levels. WB analysis showed that SCAP co-cultivation with E. faecalis led to a decrease in the level of the key proteins of the Wnt/β-Catenin and NF-κB signaling pathways: β-Catenin (p = 0.0068 *), LRP-5 (p = 0.0059 **), and LRP-6 (p = 0.0329 *), as well as NF-kB (p = 0.0034 **) and TRAF6 (p = 0.0285 *). These results suggest that oral bacteria can up- and downregulate the immune and inflammatory responses of SCAP, as well as influence the osteogenic potential of SCAP, which may negatively regulate the success of RET.

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New Insights into the Microbial Profiles of Infected Root Canals in Traumatized Teeth

Cited by 14 publications

References 45 publications

In vitro efficacy of a non‐instrumentation technique to remove intracanal multispecies biofilm

In vitro efficacy of a non‐instrumentation technique to remove intracanal multispecies biofilm

Cytokine Secretion, Viability, and Real-Time Proliferation of Apical-Papilla Stem Cells Upon Exposure to Oral Bacteria

Combined Transcriptomic and Protein Array Cytokine Profiling of Human Stem Cells from Dental Apical Papilla Modulated by Oral Bacteria

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