Nucleatum Bacteria Research Articles

Investigating the effect of Fusobacterium nucleatum on the aggressive behavior of cancer-associated fibroblasts in colorectal cancer

Fusobacterium nucleatum, (F. nucleatum) as a known factor in inducing oncogenic, invasive, and inflammatory responses, can lead to an increase in the incidence and progression of colorectal cancer (CRC). Cancer-associated fibroblasts (CAF) are also one of the key components of the tumor microenvironment (TME), which lead to resistance to treatment, metastasis, and disease recurrence with their markers, secretions, and functions. This study aimed to investigate the effect of F. nucleatum on the invasive phenotype and function of fibroblast cells isolated from normal and cancerous colorectal tissue. F. nucleatum bacteria were isolated from deep periodontal pockets and confirmed by various tests. CAF cells from tumor tissue and normal fibroblasts (NF) from a distance of 10 cm of tumor tissue were isolated from 5 patients by the explant method and were exposed to secretions and ghosts of F. nucleatum. The expression level of two markers, fibroblast activation protein (FAP), and α-smooth muscle actin (α-SMA), and the amount of production of two cytokines TGF-β and IL-6 from fibroblast cells were measured by flow cytometry and ELISA test, respectively before and after exposure to different bacterial components. The expression of the FAP marker was significantly higher in CAF cells compared to NF cells (P < 0.05). Also, the expression of IL-6 in CAF cells was higher than that of NF cells. In investigating the effect of bacterial components on the function of fibroblastic cells, after comparing the amount of IL-6 produced between the normal tissue of each patient and his tumoral tissue under 4 treated conditions, it was found that the amount of IL-6 production from the CAF cells of patients in the control group, treated with heat-killed ghosts and treated with paraformaldehyde-fixed ghosts had a significant increase compared to NF cells (P < 0.05). Due to the significant increase in FAP marker expression in fibroblast cells of tumor tissue compared to normal tissue, it seems that FAP can be used as a very good therapeutic marker, especially in patients with high levels of CAF cells. Various components of F. nucleatum could affect fibroblast cells differentially and at least part of the effect of this bacterium in the TME is mediated by CAF cells.

Liver Abscesses Due to Fusobacterium nucleatum: A Case Series

Abstract Fusobacteriae are anaerobic gram-negative bacilli, classically involved in the development of septic thrombophlebitis of the internal jugular vein in Lemierre syndrome. The goal of this study is to describe the various presentations, treatment course, and outcomes of patients with Fusobacterium liver abscess(es) and review available literature on this topic. We retrospectively reviewed our microbiologic database for positive cultures of Fusobacteriae species in a tertiary care hospital from July 2018 through November 2022 and identified 5 cases of liver abscess that involved Fusobacterium nucleatum species. Patients presented with a syndrome of fever and chills, abdominal pain, and nausea and vomiting. Laboratory investigations revealed leukocytosis and transaminitis, and imaging revealed liver abscess(es). The identification of the F. nucleatum bacteria involved invasive aspiration of the abscess in all but one case, where it was identified on blood cultures. One patient had a florid picture of sepsis and acute respiratory distress syndrome. All patients responded well with intravenous antibiotics and were discharged home to complete a prolonged course until radiographic resolution. In 3 patients, history and imaging were suggestive of possible sources that may have hematogenously spread to the liver, with history of recent appendectomy in 1 patient, possible appendicitis on computed tomography of the abdomen in another patient, and recent dental manipulation in the third patient. In conclusion, Fusobacteriae should be included in the differential diagnosis of liver abscess, especially in the setting of recent appendicitis or dental disease. Effective treatment includes surgical drainage when feasible and prolonged course of antibiotics with goal of radiologic resolution.

High-fat diet and oral infection induced type 2 diabetes and obesity development under different genetic backgrounds.

Type 2 diabetes (T2D) is an adult-onset and obese form of diabetes caused by an interplay between genetic, epigenetic, and environmental components. Here, we have assessed a cohort of 11 genetically different collaborative cross (CC) mouse lines comprised of both sexes for T2D and obesity developments in response to oral infection and high-fat diet (HFD) challenges. Mice were fed with either the HFD or the standard chow diet (control group) for 12 weeks starting at the age of 8 weeks. At week 5 of the experiment, half of the mice of each diet group were infected with Porphyromonas gingivalis and Fusobacterium nucleatum bacteria strains. Throughout the 12-week experimental period, body weight (BW) was recorded biweekly, and intraperitoneal glucose tolerance tests were performed at weeks 6 and 12 of the experiment to evaluate the glucose tolerance status of mice. Statistical analysis has shown the significance of phenotypic variations between the CC lines, which have different genetic backgrounds and sex effects in different experimental groups. The heritability of the studied phenotypes was estimated and ranged between 0.45 and 0.85. We applied machine learning methods to make an early call for T2D and its prognosis. The results showed that classification with random forest could reach the highest accuracy classification(ACC=0.91) when all the attributes were used. Using sex, diet, infection status, initial BW, and area under the curve (AUC) at week 6, we could classify the final phenotypes/outcomes at the end stage of the experiment (at 12 weeks).

Integrating Transcriptomics and Metabolomics to Explore the Novel Pathway of Fusobacterium nucleatum Invading Colon Cancer Cells

Colorectal cancer (CRC) is a malignancy with a very high incidence and mortality rate worldwide. Fusobacterium nucleatum bacteria and their metabolites play a role in inducing and promoting CRC; however, no studies on the exchange of information between Fusobacterium nucleatum extracellular vesicles (Fnevs) and CRC cells have been reported. Our research shows that Fusobacterium nucleatum ATCC25586 secretes extracellular vesicles carrying active substances from parental bacteria which are endocytosed by colon cancer cells. Moreover, Fnevs promote the proliferation, migration, and invasion of CRC cells and inhibit apoptosis; they also improve the ability of CRC cells to resist oxidative stress and SOD enzyme activity. The genes differentially expressed after transcriptome sequencing are mostly involved in the positive regulation of tumor cell proliferation. After detecting differential metabolites using liquid chromatography-tandem mass spectrometry, Fnevs were found to promote cell proliferation by regulating amino acid biosynthesis in CRC cells and metabolic pathways such as central carbon metabolism, protein digestion, and uptake in cancer. In summary, this study not only found new evidence of the synergistic effect of pathogenic bacteria and colon cancer tumor cells, but also provides a new direction for the early diagnosis and targeted treatment of colon cancer.

Antibacterial Effects of Various Concentrations of Natural Ingredients Snail Mucus (Achatina fulica) Against Inhibition Zones of Fusobacterium nucleatum Causes Periodontitis In Vitro

Periodontitis is a microorganism that invades the subgingival area, thereby triggering an inflammatory response of the periodontal tissue. Fusobacterium nucleatum is one of the most abundant gram-negative bacilli colonizing the subgingival plaque and closely associated with periodontal disease. The aim of study was to calculate the inhibition of snail mucus against Fusobacterium nucleatum bacteria with concentrations of 12.5%, 25%, 50%, and 100%. This study used a laboratory experimental test method and was conducted at the Oral Biology Laboratory, Faculty of Dentistry, Airlangga University. The steps were performed are: plant germ stock with sterile osse on BHI broth media then incubate for 48 hours, observe the turbidity of the germs then standardize with mc farland standard 0.5, plant germs on Hilton Muller agar media with the spreading technique, treat the test sample on sterile paper disk as much as 0, 01 ml with a sterile micropipette, then paste it on the surface of the media so that it is then incubated for 48 hours. Then, observe and measure the diameter of the clear zone. The results show that the average inhibition in the control group is 25.65 mm, and the snail mucus group is 12.5%, 0 mm, 25%, 12.40 mm, 50%, 16.70 mm and 100% 19.60 mm. There is a significant difference in the diameter of the inhibitory power between the treatment groups. The conclusion of the antibacterial effect of natural ingredients of snail mucus from the 25% treatment showed strong criteria, and the greatest inhibitory power was the concentration of 100%.

Antibacterial and Antioxidant Effects of Magnesium Alloy on Titanium Dental Implants.

Objectives In this study, a new type of dental implant by covering the surface of the titanium (Ti) implant with zinc-magnesium (Zn-Mg) alloy was designed, to study the antibacterial and antioxidant effects of Mg alloy on titanium (Ti) implants in oral implant restoration. Methods Human gingival fibroblasts (HGFs), S. sanguinis, and F. nucleatum bacteria were used to detect the bioactivity and antibacterial properties of Mg alloy-coated Ti implants. In addition, B6/J mice implanted with different materials were used to further detect their antibacterial and antioxidant properties. Results The results showed that Mg alloy could better promote the adhesion and proliferation and improve the alkaline phosphatase (ALP) activity of HGFs, which contributed to better improved stability of implant osseointegration. In addition, Mg alloy could better inhibit the proliferation of S. sanguinis, while no significant difference was found in the proliferation of F. nucleatum between the two implants. In the mouse model, the peripheral inflammatory reaction and oxidative stress of the Mg alloy implant were significantly lower than those of the Ti alloy implant. Conclusions Zn-Mg alloy-coated Ti implants could better inhibit the growth of Gram-positive bacteria in the oral cavity, inhibit oxidative stress, and facilitate the proliferation activity of HGFs and the potential of osteoblast differentiation, thus, better increasing the stability of implant osseointegration.

Fusobacterium nucleatum Extracellular Vesicles Modulate Gut Epithelial Cell Innate Immunity via FomA and TLR2.

Extracellular vesicles (EVs) derived from the gut microbiota are largely uncharacterized and their impacts on host intestinal physiology remain unresolved. Here, we isolated EVs from F. nucleatum for detailed characterization. Our analyses highlight the presence of the outer membrane protein porin FomA on EVs. Besides, we evaluated the impact of EVs on human intestinal epithelial cells (IECs) in a non-inflammatory context. Our results show no detrimental impact on the epithelial barrier. No internalization of EVs was observed. Moreover, we demonstrate that F. nucleatum EVs trigger innate immunity of IECs by promoting NF-κB activation via the dynamin-mediated endocytosis. The NF-κB activation was found to be TLR2-dependent yet, TLR4 was dispensable. Using competitive binding assays, we establish that FomA is involved in the NF-κB response. Taken together, our data indicate that EVs induce effects similar to those observed with whole F. nucleatum bacteria on IECs. In particular, our study highlights the role of TLR2 and FomA as major modulators of the gut epithelium immune responses to F. nucleatum.

The Effectiveness of Aloe vera Hydrogel Against Fusobacterium nucleatum

Bacteria can cause infectious diseases on oral health such as gingivitis and periodontitis. Scaling and rootplaning are effective against this disease, nevertheless several types of antimicrobial material can be added to increase the effectiveness of the initial treatment. Aloe vera is one of the natural antimicroba materials that is usually used. Hydrogel is a polymetric material that is stable in wet conditions, especially in the gingival sulcus. Sodium alginate is a natural polymer that is often used to produce hydrogels due to its biocompability. The aim of this study was to know the the effectiveness of Aloe vera hydrogel against Fusobacterium nucleatum by in vitro method. This research is experimental research with a total of 20 samples. The samples used were 5% and 10% sodium alginate based Aloe vera hydrogel which were placed in pure culture of F. nucleatum in MHA media for 24 hours. The inhibition zone diameter were measured using calipers. The results of the study using Shapiro-Wilk normality test showed that the data were not normally distributed. Then Kruskal Wallis test was performed which showed significant differences in each treatment group followed by Post Hoc Mann Whitney test to see a significant difference in the 5% group and 10%. Aloe vera hydrogel showed a significant inhibition against F. nucleatum bacteria at both 5% and 10% concentrations.Keywords: Alginate, Aloe vera, Antimicrobial material, Fusobacterium nucleatum, Hydrogel, Inhibitory zone

Antibacterial power effectiveness of calcium hydroxide and propolis mixture on Fusobacterium nucleatum bacteria

Background: Calcium hydroxide is a root canal dressing material that is widely used in dentistry because of its ability to regenerate hard tissue and eliminate bacteria. According to Ferreira et al. (2015), Fusobacterium nucleatum was found in 90% of teeth that had a root canal treatment done using calcium hydroxide as an intracanal medication. Due to this shortcoming of the antibacterial power of calcium hydroxide, additional research on alternative ingredients that can be combined with calcium hydroxide to improve its antibacterial power is necessary. Propolis is a natural material that has high antibacterial power and has long been used in dentistry. The addition of propolis to calcium hydroxide is expected to improve the antibacterial power of calcium hydroxide without eliminating its function in terms of regenerating hard tissue. Purpose: To find out how effective is the antibacterial power of a combination of calcium hydroxide and propolis against Fusobacterium nucleatum. Methods: The research was carried out using 4 treatment groups consisting of 6 samples for each group. Group 1 is given a combination of calcium hydroxide and propolis with a ratio of 1:1, group 2 with a ratio of 1:1.5, group 3 with a ratio of 1:2, and group 4 is a positive control of calcium hydroxide and sterile aquadest suspension. Each sample was put into a test tube containing BHIB and a suspension of Fusobacterium nucleatum, incubated at 37ºC for 24 hours, and vortexed for 1 minute. A total of 0.1 ml of bacterial inoculum was taken from each sample and then put into the MHA and grown for 24 hours. The number of Fusobacterium nucleatum colonies grown on MHA was calculated and expressed using the Colony Forming Unit (CFU). Results: There were fewer colonies of Fusobacterium nucleatum in the treatment group compared to the control group. Conclusion: The combination of calcium hydroxide and propolis has an effective antibacterial power against Fusobacterium nucleatum which the ratio of 1:2 is more effective than ratio of 1:1,5 and 1:1.

Clinical, Pathological, and Molecular Characteristics of CpG Island Methylator Phenotype in Colorectal Cancer: A Systematic Review and Meta-analysis

BACKGROUND: CpG island methylator phenotype (CIMP) tumors, comprising 20% of colorectal cancers, are associated with female sex, age, right-sided location, and BRAF mutations. However, other factors potentially associated with CIMP have not been robustly examined. This meta-analysis provides a comprehensive assessment of the clinical, pathologic, and molecular characteristics that define CIMP tumors. METHODS: We conducted a comprehensive search of the literature from January 1999 through April 2018 and identified 122 articles, on which comprehensive data abstraction was performed on the clinical, pathologic, molecular, and mutational characteristics of CIMP subgroups, classified based on the extent of DNA methylation of tumor suppressor genes assessed using a variety of laboratory methods. Associations of CIMP with outcome parameters were estimated using pooled odds ratio or standardized mean differences using random-effects model. RESULTS: We confirmed prior associations including female sex, older age, right-sided tumor location, poor differentiation, and microsatellite instability. In addition to the recognized association with BRAF mutations, CIMP was also associated with PIK3CA mutations and lack of mutations in KRAS and TP53. Evidence of an activated immune response was seen with high rates of tumor-infiltrating lymphocytes (but not peritumoral lymphocytes), Crohn-like infiltrates, and infiltration with Fusobacterium nucleatum bacteria. Additionally, CIMP tumors were associated with advance T-stage and presence of perineural and lymphovascular invasion. CONCLUSION: The meta-analysis highlights key features distinguishing CIMP in colorectal cancer, including molecular characteristics of an active immune response. Improved understanding of this unique molecular subtype of colorectal cancer may provide insights into prevention and treatment.

Antibacterial effects of Pluchea indica Less leaf extract on E. faecalis and Fusobacterium nucleatum (in vitro)

Background: Enterococcus. faecalis (E. faecalis) and Fusobacterium nucleatum (F. nucleatum) are the most common bacteria found in infected tooth root canal. Most of these bacteria often cause failure in endodontic treatments. Pluchea indica Less leaf is a species of plants that has several chemical properties. It consists of flavonoids, tannins, polyphenols, and essensial oils which have been reported as antibacterial agents. Because of its benefits, the extract of Pluchea indica Less leaves may be potentially developed as one of root canal sterilization dressing. Purpose: This study aimed to determine antibacterial activity of Pluchea indica Less leaves extract against E. faecalis and F. nucleatum bacteria. Method: Dilution method was conducted first to show Minimum Inhibitory Concentration (MIC) of the extract against E. faecalis and F. nucleatum. The antibacterial activity test on Pluchea indica Less leaves extract was performed on E. faecalis and F. nucleatum bacteria using agar diffusion method. The Pluchea indica Less leaves extract used for antibacterial activity test was at a concentrations of 100%, 50%, 25%, 12.5%, and 6.25%. Thirty-five petridiscs were used and divided into five groups based on the extract concentration. Result: The results showed strong and moderate antibacterial effects of the Pluchea indica Less leaves extract on E. faecalis at the concentrations of 100% and 50%, while on F. nucleatum only at the concentration of 100% with moderate effect. Conclusion: Pluchea indica Less leaves extract has antibacterial activity against E. faecalis and F. nucleatum bacteria with strong-moderate effect.

English

Increasing resistance to several antibiotics has crucial importance. Many researchers are trying to discover alternative methods or agents to develop antimicrobial effectiveness. In this study, antimicrobial activities of four diverse synthesized perylene diimides were tested, because they are well known dyes and pigments for useful various applications and also antimicrobial activities. These perylene diimides were determined by using disc diffusion and microdilution methods against 21 microorganisms including three yeast cells. The antimicrobial activity of 1,7-dibromo-N,N&rsquo;-(L-alanine tert-butylester)-3,4:9,10-perylene diimide (2) have shown effect on Staphylococcus aureus, Streptococcus pyogenes, Bacillus cereus, Corynebacterium sp. and Fusobacterium nucleatum bacteria. In addition, the other perylene diimide, 1,7-dibromo-N,N&rsquo;-(L-glutamicacid tert-butylester) 3,4:9,10-perylene diimide (1) was only active against S. aureus and B. cereus. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of perylene diimides for these bacteria were determined, respectively. These results are novel about perylene diimides, and further improvements on these first attempts may produce a practically applicable antimicrobial activity. Key words: Perylene dimide, antimicrobial activity, antibacterial activity

Host Susceptibility to Periodontitis

Host susceptibility to periodontal infection is controlled by genetic factors. As a step toward identifying and cloning these factors, we generated an A/J x BALB/cJ F2 mouse resource population. A genome-wide search for Quantitative Trait Loci (QTL) associated with periodontitis was performed. We aimed to quantify the phenotypic response of the progenies to periodontitis by microCT analysis, to perform a genome-wide search for QTL associated with periodontitis, and, finally, to suggest candidate genes for periodontitis. We were able to produce 408 F2 mice. All mice were co-infected with Porphyromonas gingivalis and Fusobacterium nucleatum bacteria. Six weeks following infection, alveolar bone loss was quantified by computerized tomography (microCT) technology. We found normal distribution of the phenotype, with 2 highly significant QTL on chromosomes 5 and 3. A third significant QTL was found on chromosome 1. Candidate genes were suggested, such as Toll-like receptors (TLR) 1 and 6, chemokines, and bone-remodeling genes (enamelin, ameloblastin, and amelotin). This report shows that periodontitis in mice is a polygenic trait with highly significant mapped QTL.

Pulmonary botryomycosis on a lung cavity: a rare pulmonary infection mimicking cancer

Lung botryomycosis is a rare disease. We report what is to our knowledge the first case occurring on a lung cavity. In a 42-year-old man suffering asthenia and cough, a chest radiograph revealed a right upper lobe opacity. Computed tomography scan showed a necrotic mass which was also spiculated. Repeated research for Mycobacterium tuberculosis was negative. The patient underwent a lobectomy. Histological and bacteriological examinations made the diagnosis of botryomycosis, because the cavity presented numerous colonies of pyogenic Fusobacterium nucleatum bacteria. Botryomycosis is a difficult diagnosis that clinically mimics actinomycosis, tuberculosis or cancer. In most cases, surgery is necessary to assess diagnosis and treatment.

The Role of Aggregation in Fusobacterium nucleatum– induced Immune Cell Death

Introduction Fusobacterium nucleatum, an anaerobic oral bacterium, has been shown to be highly abundant in endodontic infections. Its role in these infections remains unclear. Previous studies have shown that F. nucleatum could aggregate immune cells. We have demonstrated that F. nucleatum can induce significant apoptosis in peripheral blood mononuclear cells (PBMCs). In this in vitro study, we sought to determine what role this aggregation phenomenon has on the induction of apoptosis in PBMCs. Methods F. nucleatum bacteria were treated as follows: formaldehyde-fixed, heat-treated, or sonicated before co-culturing with PBMCs. Cell aggregation and apoptosis of the PBMCs were assessed under light microscopy and analyzed by flow cytometry, respectively. PBMCs were then immobilized with a Matrigel matrix before treatment with F. nucleatum. Aggregation and apoptosis were assessed as before. Surface staining of activation marker CD69 was assessed by flow cytometry. The apoptosis and CD69 data underwent one-way analysis of variance, followed by post hoc Bonferroni test and χ 2 test, respectively, to determine statistical significance. Results Viable and formaldehyde-treated but not sonicated or heat-treated F. nucleatum bacteria were able to cause severe aggregation and apoptosis of the immune cells. Disruption of F. nucleatum mediated aggregation by immobilization of the cells with a Matrigel matrix resulted in a significant diminution of cell death but not cell activation when assessed by using surface expression of CD69 early activation antigen. Conclusions F. nucleatum’s ability to induce cell death in immune cells appears to be mediated through the immune cells being aggregated, which might have important implications for its pathogenesis.