Helicobacter pylori is an infectious agent that colonizes the gastric mucosa of half of the population worldwide. This bacterium has been recognized as belonging to group 1 carcinogen by the World Health Organization for the role in development of gastritis, peptic ulcers, and cancer. Due to the increase in resistance to antibiotics used in the anti-H. pylori therapy, the development of an effective vaccine is an alternative of great interest, which remains a challenge. Therefore, a rational, strategic, and efficient vaccine design against H. pylori is necessary where the use of the most current bioinformatics tools could help achieve it. In this study, immunoinformatics approach was used to design a novel multiepitope oral vaccine against H. pylori. Our multiepitope vaccine is composed of cholera toxin subunit B (CTB) that is used as a mucosal adjuvant to enhance vaccine immunogenicity for oral immunization. CTB fused to 11 epitopes predicted of pathogenic (UreB170–189, VacA459–478, CagA1103–1122, GGT106–126, NapA30–44, and OipA211–230) and colonization (HpaA33–52, FlaA487–506, FecA437–456, BabA129–149, and SabA540–559) proteins from H. pylori. CKS9 peptide (CKSTHPLSC) targets epithelial microfold cells to enhance vaccine uptake from the gut barrier. All sequences were joined to each other by proper linkers. The vaccine was modeled and validated to achieve a high-quality three-dimensional structure. The vaccine design was evaluated as nonallergenic, antigenic, soluble, and with an appropriate molecular weight and isoelectric point. Our results suggest that our newly designed vaccine could serve as a promising anti-H. pylori vaccine candidate.

Helicobacter pylori is an infectious agent commonly associated with gastrointestinal diseases. The use of probiotics to treat this infection has been documented, however, their potential antimicrobial metabolites have not yet been investigated. In the present study, the effect of reuterin produced by Lactobacillus reuteri on H. pylori growth and virulence gene expression was evaluated. It was observed that reuterin caused significant (P < 0.05) H. pylori growth inhibition at concentrations from 0.08 to 20.48 mM, with minimal inhibitory concentrations (MICs) of 20.48 mM for H. pylori ATCC700824 and 10.24 mM for H. pylori ATCC43504. In a reuterin bacterial killing assay, it was observed that half of the MIC value for H. pylori (ATCC700824) significantly (P < 0.01) reduced colony numbers from 5.65 ± 0.35 to 3.78 ± 0.35 Log CFU/mL after 12 h of treatment and then increased them to 5.25 ± 0.23 Log CFU/mL at 24 h; at its MIC value (20.48 mM), reuterin abrogated (P < 0.01) H. pylori (ATCC700824) growth after 20 h of culture. In addition, reuterin significantly (P < 0.01) reduced H. pylori (ATCC 43504) colony numbers from 5.65 ± 0.35 to 4.1 ± 0.12 Log10 CFU/mL from 12 to 24 h of treatment and abrogated its growth at its MIC value (10.24 mM), after 20 h of treatment. Reuterin did not alter normal human gastric Hs738.St/Int cell viability at the concentrations tested for H. pylori strains. Furthermore, 10 μM reuterin was shown to significantly (P < 0.01) reduce mRNA relative expression levels of H. pylori virulence genes vacA and flaA at 3 h post-treatment, whose effect was higher at 6 h post-treatment, as measured by RT-qPCR. The observed direct antimicrobial effect and the downregulation of expression of virulence genes on H. pylori by reuterin may contribute to the understanding of the mechanisms of action of probiotics against H. pylori.

The variability in Helicobacter pylori vacA and cagA genes has been related to the progression of the gastrointestinal disease; also the presence of H. pylori in the oral cavity has been associated with periodontal disease in adults, but, in children without dyspeptic symptoms, little is known about this. We evaluated the prevalence of H. pylori and the presence of vacA/cagA genotypes in the oral cavity of Mexican children without dyspeptic symptoms. The gingival status was measured, and dental plaque samples (n = 100) were taken. 38% of children were positive for H. pylori 16S rRNA gene by qPCR. A significant association between H. pylori oral infection and gingival status was observed (P < 0.001). In 34.6% (9/26) of mild gingivitis cases, s1m2 genotype was found, while s1m1 was typed in 50% (3/6) of moderate gingivitis. The cagA prevalence among H. pylori-positive children was 80.8% (21/26), 83.3% (5/6), and 16.7% (1/6) of cases of mild gingivitis, moderate gingivitis, and nongingivitis, respectively (P < 0.001). The s1m1/cagA+ combinational genotype was the most detected in children with gingivitis. Our results suggest that the prevalence of H. pylori and detection of vacA/cagA genotypes-associated gastrointestinal disease in the oral cavity could be related to the progression of gingivitis in asymptomatic children.

Background/purpose Helicobacter pylori ( H. pylori ) infection is the most common in the world and is associated with various gastrointestinal pathologies, including chronic gastritis, peptic ulcers, and gastric cancer. The prevalence is associated with socioeconomic conditions, with this infection being more common in developing countries than in developed countries. The presence and permanence of H. pylori in the oral cavity has been reported, but its role is controversial. The aim of this study was to determine the prevalence of H. pylori in dental plaque of patients with periodontitis. Materials and methods A cross-sectional study was carried out and Periodontal Screening and Recording (PSR) index was determined. 38 dental plaque samples were taken and total DNA was extracted and qPCR was performed. Results 60.5% of the samples (n = 23) were positive for the presence of H. pylori by the amplification of the 16S rRNA and vacA genes. In addition, cagA gene was detected in 21.7% (n = 5) of H. pylori -positive. A significant relationship between periodontal status and H. pylori oral infection was found ( P ≤ 0.05); patients with initial and moderate periodontitis were the most affected with 39.1% and 30.4%, respectively. Conclusion Our results suggest that the prevalence of H. pylori in the oral cavity could be related to the progression of periodontal disease. Therefore, oral hygiene and treatment for the elimination of oral H. pylori could stop the progression of periodontal disease.

The presence of Helicobacter pylori in the oral cavity has been associated to the failure of antimicrobial therapy in patients with gastrointestinal infection and the development of oral diseases. However, it has been reported that the maintenance of good oral hygiene can improve the therapeutic success rates, where the use of mouthwashes with anti-Helicobacter activity would help to achieve it. The aim was to evaluate the antimicrobial activity of OxOral® mouthwash against H. pylori and its effect on biofilm formation. The minimum inhibitory concentration (MIC) of OxOral® (pH = 6.4–7.5, ORP = 650–900 mV) against H. pylori was calculated testing serial dilutions 0.117–15 ppm against 1 × 108 CFU/mL of H. pylori (ATCC® 700824™) by broth microdilution method using 96‐well plates. The H. pylori biofilm formation was determined by the optical density measurement at 600 nm from coverslips stained with 0.1% crystal violet. The gene expression of ureA, luxS, flaA, omp18, and lpxD were analyzed by RT‐qPCR. OxOral® cytotoxicity was evaluated in a human gingival fibroblast cell line by MTT assay. MIC was of 3.75 ppm, with 99.7 ± 7.7% bacterial growth inhibition. In the negative control, the biofilm formation was observed, whereas when bacteria were treated with OxOral® at 0.234, 0.469, and 0.938 ppm, an inhibition of 35.5 ± 0.9%, 89.1 ± 1.2%, and 99.9 ± 5.5% were obtained, respectively. The gene expression analysis showed that flaA, omp18, and lpxD genes were down‐regulated with OxOral® compared with control (p<0.05). Low cytotoxicity of 16.5 ± 7.6% was observed at the highest dose (15 ppm); no significant differences were observed from 15 to 0.469 ppm compared to the control of untreated cells (p>0.05). Our results reveal an important anti-Helicobacter activity of OxOral® and open the possibility of its therapeutic use new studies, which would increase the success rate of conventional therapies against H. pylori.

In this paper we report on the influence of polysaccharides’ molecular structure on the antibacterial activity and cytotoxicity of composites based on silver nanoparticles (AgNPs) immobilized into carboxymethyl-cellulose (CMC). These composites were green synthesized from the reduction of silver ions into aqueous solutions of the polysaccharide, using CMC with different degree of substitution (DS) and molecular weight (Mw). The composites were characterized by transmission electron microscopy (TEM), as well as infrared (ATR-FTIR), ultraviolet (UV-Vis), Raman, and X-ray photo-electron (XPS) spectroscopic techniques. The antibacterial activity was evaluated with minimum inhibitory concentration against Enterococcus faecalis. The cytotoxicity of composites was assessed against human gingival fibroblast. Experimental evidence suggests that particle size distribution and morphology of AgNPs change according to the quantity of silver precursor added to the reaction, as well as the DS and Mw of CMC used for composites preparation. This is related to the dispersion of silver precursor into aqueous solutions of the polysaccharide and the formation of Ag-O coordination bonds among AgNPs and COO− moieties of CMC. Moreover, these coordination bonds modify the ability of nanoparticles to produce and release Ag+ into aqueous dispersion, adjusting their antibacterial activity and the induction of cytotoxicity into the tested biological environments.

Cobalt–chromium (Co-Cr) alloys have been used in a wide variety of biomedical applications, including dental, cardiovascular, and orthopedic devices. In vitro studies have shown that the mineralization of cells involved in osteogenesis is regulated by boron. The development of a new cobalt-chromium-boron (Co-Cr-B) alloy improves the mechanical properties of the metal, such as wear resistance, and meets biocompatibility requirements. Therefore, the objective of this study was to evaluate the osteogenic differentiation and biocompatibility in in vitro assays. Human dental pulp mesenchymal cells (hDPSCs) were isolated from volunteers and then co-cultured with the Co-Cr plus boron alloy from 0.3% to 1% for 15 days, while the formation of calcium deposits was quantified by Alizarin red staining and the expression of genes was related to osteodifferentiation by RT-qPCR. Simultaneously, the cytotoxicity of our alloy was evaluated by MTT assay and the change in the gene expression of cytokines commonly associated with inflammatory processes. The results showed low cytotoxicity when cells were treated with the Co-Cr-B alloy, and no change in the gene expression of IL-1β, TNF-α, IL-6, and IL-8 was observed compared to the untreated control (p > 0.05). The osteoinduction results shown an increase in mineralization in hDPSCs treated with Co-Cr-B alloy with 1.0% B. In addition, a significant increase in mRNA levels for collagen type 1 in with 0.3% boron and alkaline phosphatase and Runx2 with 0.6% boron were observed. The addition of Boron to the ASTM F75 Co-Cr base alloy improves the biocompatible characteristics. No cytotoxicity and any change of the expression of the pro-inflammatory cytokines IL-1β, TNF-α, IL-6, and IL-8 in human peripheral blood mononuclear cells treated with the cobalt-chromium-boron alloy was observed in vitro assays. Furthermore, our alloy acts as an osteoinductive in osteogenic differentiation in vitro. Therefore, our results could set the standard for the development of in vivo trials and in the future, it could be considered as an alternative for regenerative therapy.

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