Fusobacterium nucleatum (F. nucleatum) is an early pathogenic colonizer in periodontitis, but the host response to infection with this pathogen remains unclear. In this study, we built an F. nucleatum infectious model with human periodontal ligament stem cells (PDLSCs) and showed that F. nucleatum could inhibit proliferation, and facilitate apoptosis, ferroptosis, and inflammatory cytokine production in a dose-dependent manner. The F. nucleatum adhesin FadA acted as a proinflammatory virulence factor and increased the expression of interleukin(IL)-1β, IL-6 and IL-8. Further study showed that FadA could bind with PEBP1 to activate the Raf1-MAPK and IKK-NF-κB signaling pathways. Time-course RNA-sequencing analyses showed the cascade of gene activation process in PDLSCs with increasing durations of F. nucleatum infection. NFκB1 and NFκB2 upregulated after 3 h of F. nucleatum-infection, and the inflammatory-related genes in the NF-κB signaling pathway were serially elevated with time. Using computational drug repositioning analysis, we predicted and validated that two potential drugs (piperlongumine and fisetin) could attenuate the negative effects of F. nucleatum-infection. Collectively, this study unveils the potential pathogenic mechanisms of F. nucleatum and the host inflammatory response at the early stage of F. nucleatum infection.
Humans ; Fusobacterium nucleatum/metabolism* ; NF-kappa B/metabolism* ; Periodontal Ligament/metabolism* ; Signal Transduction ; Fusobacterium Infections/pathology* ; Stem Cells/metabolism*

Fusobacterium nucleatum is an opportunistic pathogenic bacterium that can be enriched in colorectal cancer tissues, affecting multiple stages of colorectal cancer development. The two-component system plays an important role in the regulation and expression of genes related to pathogenic resistance and pathogenicity. In this paper, we focused on the CarRS two-component system of F. nucleatum, and the histidine kinase protein CarS was recombinantly expressed and characterized. Several online software such as SMART, CCTOP and AlphaFold2 were used to predict the secondary and tertiary structure of the CarS protein. The results showed that CarS is a membrane protein with two transmembrane helices and contains 9 α-helices and 12 β-folds. CarS protein is composed of two domains, one is the N-terminal transmembrane domain (amino acids 1-170), the other is the C-terminal intracellular domain. The latter is composed of a signal receiving domain (histidine kinases, adenylyl cyclases, methyl-accepting proteins, prokaryotic signaling proteins, HAMP), a phosphate receptor domain (histidine kinase domain, HisKA), and a histidine kinase catalytic domain (histidine kinase-like ATPase catalytic domain, HATPase_c). Since the full-length CarS protein could not be expressed in host cells, a fusion expression vector pET-28a(+)-MBP-TEV-CarS_cyto was constructed based on the characteristics of secondary and tertiary structures, and overexpressed in Escherichia coli BL21-Codonplus(DE3)RIL. CarS_cyto-MBP protein was purified by affinity chromatography, ion-exchange chromatography, and gel filtration chromatography with a final concentration of 20 mg/ml. CarS_cyto-MBP protein showed both protein kinase and phosphotransferase activities, and the MBP tag had no effect on the function of CarS_cyto protein. The above results provide a basis for in-depth analysis of the biological function of the CarRS two-component system in F. nucleatum.
Humans ; Histidine Kinase/metabolism* ; Fusobacterium nucleatum/metabolism* ; Automobiles ; Protein Kinases/genetics* ; Escherichia coli/metabolism* ; Colorectal Neoplasms

OBJECTIVETo detect the presence and distribution of luxS gene in quorum sensing signal system in the periodontal pathogens.

METHODSThe total DNA of Porphyromonas gingivalis (Pg), Fusobacterium nucleatum (Fn), Actinobacillus acitinomycetimcomtans (Aa) were extracted. The presence of luxS was detected by polymerase chain reaction (PCR). The products of PCR were detected by electrophoresis, sequenced and identified by a Blast search of the GenBank database.

RESULTSElectrophoresis, sequencing and Blast searching indicated that the PCR products of Pg were highly consistent with the luxS gene in GenBank. The sequencing result of Fn was also identified with the target gene. The PCR product of Aa was the same as reference through electrophoresis.

CONCLUSIONSPg, Fn, Aa contain luxS gene. Further studies may be required to investigate the functions of luxS in the periodontal pathogens.

Aggregatibacter actinomycetemcomitans ; genetics ; metabolism ; Bacterial Proteins ; genetics ; isolation & purification ; metabolism ; Carbon-Sulfur Lyases ; genetics ; isolation & purification ; metabolism ; Fusobacterium nucleatum ; genetics ; metabolism ; Gene Expression Regulation, Bacterial ; Porphyromonas gingivalis ; genetics ; metabolism ; Quorum Sensing ; genetics ; Signal Transduction

OBJECTIVETo examine the expression of tumor necrosis factor in placenta of pregnant rats with chronic periodontitis.

METHODSTwenty Wistar female rats were randomly divided into two groups, control (n = 8) and experimental group (n = 12). The periodontitis model was established in the experimental group. The females and males in the two groups got together four weeks later. Nineteen days after pregnancy all rats were executed and placenta collected. The delivery time and neonatal birth weight were recorded and the pathological changes of periodontal tissue observed. Tumor necrosis factor (TNF) expression was examined in placenta by real-time quantitative polymerase chain reaction analysis.

RESULTSThe animal model of chronic periodontitis was successfully established. Experimental group delivered 30 offspring and the control group 56 offspring. The average number of pups born alive per litter in experimental group (4.1 ± 2.2) was significantly lower than that in control group (9.2 ± 2.2), P < 0.05. The birth weight of pups in experimental group [(5.01 ± 0.43) g] was significantly lower than that in the control group [(5.79 ± 0.53) g], P < 0.05. The relative quantitative expression of TNF was (1.807 ± 0.265) in experimental group the and (1.003 ± 0.021) in the control group (P = 0.001).

CONCLUSIONSChronic periodontitis may be related to preterm low birth weight.

Aggregatibacter actinomycetemcomitans ; Animals ; Animals, Newborn ; Birth Weight ; Chronic Periodontitis ; metabolism ; microbiology ; Disease Models, Animal ; Female ; Fusobacterium nucleatum ; Placenta ; metabolism ; Porphyromonas gingivalis ; Pregnancy ; Pregnancy Complications, Infectious ; metabolism ; microbiology ; Prevotella intermedia ; Random Allocation ; Rats ; Rats, Wistar ; Tumor Necrosis Factor-alpha ; metabolism

Human oral bacteria live in multispecies communities in the biofilm called dental plaque. This review focuses on the interactions of seven species and the ability of each species individually and together with other species to grow on saliva as the sole source of nutrient. Community formation in biofilms in flow cells is monitored using species-specific fluorophore-conjugated immunoglobulin G, and images are captured by confocal microscopy. Early colonizing veillonellae emerge from this review of interspecies interactions in saliva as a critical genus that guides the development of multispecies communities. Highly selective interspecies recognition is evident as initial colonizers pair with early and middle colonizers to form multispecies communities that grow on saliva.
Actinomyces ; growth & development ; Aggregatibacter actinomycetemcomitans ; growth & development ; Animals ; Biofilms ; growth & development ; Dental Enamel ; microbiology ; Dental Plaque ; metabolism ; microbiology ; Fluorescent Dyes ; metabolism ; Fusobacterium nucleatum ; growth & development ; Humans ; Microbial Consortia ; physiology ; Microbial Interactions ; physiology ; Microscopy, Confocal ; Polysaccharides, Bacterial ; chemistry ; Saliva ; metabolism ; microbiology ; Streptococcus oralis ; growth & development ; Veillonella ; growth & development