Comparison of antibacterial effect of plantaricin 149 suspension and traditional root canal irrigation solutions in root canal infections in vitro.

Dental pulp and periapical diseases are common conditions in stomatology, caused by various pathogenic microorganisms. Antimicrobial peptides, as new antibiotics, offer promising applications in the irrigation and disinfection medicaments for root canals.One patient with chronic periapical periodontitis was selected to extract the clinical pathogenic bacteria. Porphyromonas gingivalis (Pg) (ATCC 33,277), Streptococcus mutans (Sm) (ATCC 25,175), and Prevotella intermedius (Pi) (ATCC 25,611) were used as test strains. The effects of plantaricin (Pln) 149 on the biofilm formation and growth in infected root canals were evaluated by RT-PCR, laser confocal scanning microscopy, and bacterial diversity analysis. In addition, the cytotoxicity of Pln 149 (100 µg/mL) to human dental pulp stem cells (hDPSCs) was assessed using an MTT assay. Pln 149 exhibited significant inhibitory effects on Pg Sm and Pi (P < 0.05), with significant differences in the biofilm images of the laser confocal scanning microscope (P < 0.05). There were no significant differences in hDPSCs viability or proliferation between the Pln 149 and control groups. Considering the excellent antimicrobial effects and low cytotoxicity, we suggest that Pln 149 might be a promising option for root canal irrigation solutions.

Adhesive Ability of Different Oral Pathogens to Various Dental Materials: An In Vitro Study.

Introduction: In dental treatments, the reason for secondary caries and the failure of root canal treatment is the microbial infection, which concerns most dentists. The challenge of how to reduce the number of bacteria at the filling materials and the number of residual bacteria in the root canal has become a research hotspot. In this study, the bacterial adhesion properties of several common dental materials were compared to provide a theoretical basis for the selection of antibacterial properties of dental materials. Methodology. Three commonly used dental restorative materials and five sealers in root canal treatment were selected. Each material block was immersed in the corresponding supragingival (Streptococcus mutans and Actinomyces viscosus) or subgingival (Porphyromonas gingivalis and Enterococcus faecalis) bacterial solution and cultured under anaerobic conditions at 37°C for 2, 4, 6, 8, 12, 16, 20, and 24 h. The adhesion of bacteria was observed, and the number of different bacteria adhering to various material model disks was calculated at different time intervals under a scanning electron microscope. The adherent CFU load of the materials was determined by colony counting. Results: Streptococcus mutans and Actinomyces viscosus exhibited the strongest adhesion ability to the resin material blocks. Porphyromonas gingivalis and Enterococcus faecalis exhibited the highest adhesion ability to the AH-Plus sealer block. Conclusions: In dental treatments, dental materials should be selected based on the chemical, physical, and biological properties of materials. In addition, it is necessary to develop new antibacterial dental materials.

Dental caries and associated factors in 3 to 5-year-old children in Zhejiang Province, China: an epidemiological survey.

BACKGROUND: Dental caries in preschool children is prevalent worldwide, but data regarding its magnitude and associated factors were not available for preschool children in Zhejiang Province, China. This study examines the dental caries situation and its associated factors in Zhejiang Province. METHODS: A total of 1591 children aged 3-5 years and their parents or caregivers were enrolled in this study. The condition of their teeth was assessed by three dental technicians qualified to WHO 2013 criteria. A structured questionnaire was completed by the children's parents or caregivers. A logistic regression analysis was used to analyze the risk factors that may be associated with dental caries occurring among preschool children. RESULTS: Caries prevalence (dmft> 0) of 3-5 year old children in Zhejiang Province was 70.4%. The mean decayed, missing and filled teeth (dmft) scores of the 3, 4 or 5 year old children surveyed were 2.96 ± 4.07, 4.42 ± 4.66, and 5.75 ± 5.19 respectively. The negative binomial regression model found that higher dental caries prevalence was found in children as age increased, with lower body mass index (BMI), with longer breastfeeding duration and with fewer hours of sleep. CONCLUSIONS: The dental caries prevalence and dmft score of 3-5-year-old children in Zhejiang Province was high, and it was associated with age, BMI, breastfeeding duration and hours slept.

Dental Caries Status and Caries Risk Factors in Students Ages 12-14 Years in Zhejiang, China.

BACKGROUND An accurate and valid caries prevention policy is absent in Zhejiang because of insufficient data. Therefore, the aim of this study was to investigate oral health status and related risk factors in 12- to 14-year-old students in Zhejiang, China. MATERIAL AND METHODS Using multi-stage, stratified, random sampling, we recruited a total of 4860 students aged 12 to 14 years old from 6 regions in Zhejiang in this cross-sectional study. Dental caries was measured using the Decayed, Missing and Filled Teeth (DMFT) index and the Significant Caries Index (SiC). Information concerning family background and relevant behaviors was collected in a structured questionnaire. Logistic regression analysis was used to study risk factors related to dental caries. RESULTS The overall prevalence of dental caries was 44% and the mean DMFT and SiC scores were 1.14 and 3.11, respectively. Female students had a higher level of dental caries than male students (P<0.01). The annual increase in caries prevalence was 3% with increasing age, and the DMFT score was 0.15. The results of logistic regression analysis showed that female sex, older age, snacks consumption once or more per day, fair or poor self-assessment of dental health, toothache experience, and dental visits were the most significant risk factors for dental caries, with odds ratios ranging from 1.24 to 2.25 (P<0.01). CONCLUSIONS The prevalence of dental caries in 12- to 14-year-old students in Zhejiang was low, with a tendency to increase compared with previous oral surveys. Female sex, older age, increased sugar intake, poor oral health self-assessment, and bad dental experience were the most important factors increasing dental caries risks.

Effect of Probiotic Lactobacilli on the Growth of Streptococcus Mutans and Multispecies Biofilms Isolated from Children with Active Caries.

BACKGROUND The purpose of this investigation was to evaluate the effect of probiotic lactobacilli on Streptococcus mutans (MS) and multispecies biofilms isolated from children with severe caries. MATERIAL AND METHODS Twenty children with active caries (DMFS ≥6) were selected as the experimental group and Streptococcus mutans (MS) were isolated from their saliva. After identification the MS strains were mixed with lactobacilli at 37°C, following which viable MS colonies were counted. At the same time dental plaques from the children were mixed with lactobacilli in vitro to form biofilms, and the population of nine common strains in the biofilms was enumerated after 24 hours of growth. RESULTS Lactobacillus casei Shirota, L. casei LC01, L. plantarum ST-III and L. paracasei LPC37 all had strong inhibitory effects on the majority the MS isolated from children with active caries, with the inhibition rate reaching approximately 70-90% (p<0.05). L. casei Shirota, L. casei LC01, L. plantarum ST-III, L. paracasei LPC37 also significantly reduced the numbers of MS, Streptococcus spp., S. sanguinis and total bacteria in mixed biofilms compared with the control group (p<0.05). CONCLUSIONS The four strains of lactobacilli were able to inhibit the growth of Streptococcus mutans and had effects on the composition of bacterial biofilms in vitro. Ingestion of probiotics may be a promising method of caries prevention.

Pyrosequencing analysis of oral microbiota shifting in various caries states in childhood.

Dental caries is one of the most prevalent childhood diseases worldwide, but little is known about the dynamic characteristics of oral microbiota in the development of dental caries. To investigate the shifting bacterial profiles in different caries states, 60 children (3-7-year-old) were enrolled in this study, including 30 caries-free subjects and 30 caries-active subjects. Supragingival plaques were collected from caries-active subjects on intact enamel, white spot lesions and carious dentin lesions. Plaques from caries-free subjects were used as a control. All samples were analyzed by 454 pyrosequencing based on 16S rRNA gene V1-V3 hypervariable regions. A total of 572,773 pyrosequencing reads passed the quality control and 25,444 unique phylotypes were identified, which represented 18 phyla and 145 genera. Reduced bacterial diversity in the cavitated dentin was observed as compared with the other groups. Thirteen genera (including Capnocytophaga, Fusobacterium, Porphyromonas, Abiotrophia, Comamonas, Tannerella, Eikenella, Paludibacter, Treponema, Actinobaculum, Stenotrophomonas, Aestuariimicrobium, and Peptococcus) were found to be associated with dental health, and the bacterial profiles differed considerably depending on caries status. Eight genera (including Cryptobacterium, Lactobacillus, Megasphaera, Olsenella, Scardovia, Shuttleworthia, Cryptobacterium, and Streptococcus) were increased significantly in cavitated dentin lesions, and Actinomyces and Corynebacterium were present at significant high levels in white spot lesions (P < 0.05), while Flavobacterium, Neisseria, Bergeyella, and Derxia were enriched in the intact surfaces of caries individuals (P < 0.05). Our results showed that oral bacteria were specific at different stages of caries progression, which contributes to informing the prevention and treatment of childhood dental caries.

Effects and mechanisms of polystyrene micro- and nano-plastics on the spread of antibiotic resistance genes from soil to lettuce.

Effects of microplastics (MPs) and nanoplastics (NPs) on the spread of antibiotic resistance genes (ARGs) in soil-plant systems are still unclear. To investigate the spread and mechanisms of ARGs from soil to lettuce, lettuce was exposed to soil spiked with two environmentally relevant concentrations of polystyrene MPs (100 µm) and NPs (100 nm). Results showed that microorganisms that carried ARGs in soil were increased after exposure to MPs/NPs, which led to an increase in ARGs in roots. NPs were absorbed by roots and can be transported to leaves. Analysis of transcriptomics, proteomics and metabolomics indicated that high concentration of NPs regulated the expression of related genes and proteins and improved the accumulation of flavonoids in the lettuce, therefore decreased the abundance of microorganisms that contained ARGs. Our work emphasizes the size and dose influences of MPs and NPs on the spread of ARGs from soil to plant.

Toxic effects and mechanisms of engineered nanoparticles and nanoplastics on lettuce (Lactuca sativa L.).

Engineered nanoparticles (ENPs) and nanoplastics (NPs) are typical nanoparticles in terrestrial environments. Till now, few studies have compared their toxicity and mechanism to plants. Here we investigated the effects of CuO, nZVI ENPs and polystyrene (PS) NPs on lettuce growth, metabolic functions, and microbial community structure. Results showed that low concentrations of nanoparticles decreased root biomass and promoted photosynthetic indicators, whereas increased reactive oxygen species (ROS) were detected in roots exposed to high concentrations of nanoparticles. High-dose CuO ENP exposure significantly raised the MDA content by 124.6 % compared to CK, causing the most severe membrane damage in the roots among the three types of nanoparticles. Although linoleic acid metabolism was down-regulated, the roots alleviated CuO stress by up-regulating galactose metabolism. Uptake of PS by roots similarly caused ROS production and activated the oxidative stress system by altering amino acid and vitamin metabolism. Faster microbial responses to nanoparticles were observed in the nZVI and PS networks. The root toxicity was indirectly mediated by ion release, NP uptake, or ROS generation, ultimately impacting root cell metabolism, rhizospheric microorganism and plant growth. These findings provide theoretical basis for assessing environmental impact of nanoparticles and their possible ecological risks.

Different effects and mechanisms of polystyrene micro- and nano-plastics on the uptake of heavy metals (Cu, Zn, Pb and Cd) by lettuce (Lactuca sativa L.).

Heavy metals are widely distributed in soil ecosystems, posing a potential threat to soil biota. Micro- and nano-plastics (MNPs) can impact the accumulation of heavy metals in plants through changing soil microbial community and cause injury to plants. In this work, two concentrations (100 and 1000 mg/kg) polystyrene microplastics (PS-MPs) and nanoplastics (PS-NPs) were adopted to explore the effects and mechanisms of MNPs on the uptake of Cu, Zn, Pb and Cd in lettuce (Lactuca sativa L.). MPs increased the uptake of heavy metals in lettuce by increasing the relative abundance of the key metal-activation bacteria in rhizospheric soil. At the end of experiment, the contents of Cu, Zn, Pb and Cd in NP treatments were significantly (p < 0.05) higher than that of MPs, particularly in 1000 mg/kg of NPs, with concentrations of 52.6, 174, 10.3, and 33.2 mg/kg, respectively. Biomarkers and gene expression reveled that 1000 mg/kg of NPs caused more severe injuries to lettuce plant at the end. Moreover, metabolomic analysis demonstrated that NPs disturbed the metabolism of ATP-binding cassette transporter (ABC transporter) and plant hormone signal transduction of lettuce root, causing increased uptake of heavy metals by lettuce. This work reveals that MPs may increase accumulation of heavy metals by altering the rhizosphere microorganisms, whereas NPs increase accumulation of heavy metals by causing more severe injuries to lettuce plant.

Mass-based trophic transfer of polystyrene nanoplastics in the lettuce-snail food chain.

To investigate the potential transfer of nanoplastics (NPs) from water to plants and subsequently to a higher trophic level, we established a food chain and evaluated the trophic transfer of polystyrene (PS) NPs based on mass concentrations by pyrolysis gas chromatography-mass spectrometry. Lettuce plants were cultivated in Hoagland solution with varying concentrations of PS-NPs (0.1, 1, 10, 100 and 1000 mg/L) for a period of 60 d and then a total of 7 g lettuce shoot was fed to snails for 27 d. Shoot biomass exposed at 1000 mg/L PS-NPs was reduced by 36.1 %. No significant change in root biomass was observed, however, root volume was reduced by 25.6 % at 100 mg/L. Moreover, PS-NPs were detected in both lettuce roots and shoots across all concentrations. Additionally, PS-NPs were transferred to snails and primarily found in feces (>75 %). Only 28 ng/g of PS-NPs were detected in the soft tissue of snails indirectly exposed at 1000 mg/L. Although PS-NPs were bio-diluted when transferred to species at higher trophic levels, they significantly inhibited the growth of snails, indicating that their potential risk to high trophic levels cannot be ignored. This study provides key information on trophic transfer and patterns of PS-NPs in food chains and helps to evaluate risk of NPs in terrestrial ecosystem.

Quantification of nanoplastics uptake and transport in lettuce by pyrolysis gas chromatography-mass spectrometry.

Nanoplastics (NPs) can enter the edible parts of crop and threaten human health, which attract widespread attention. However, the precise quantification of NPs in crop is still a tremendous challenge. Herein, a method with Tetramethylammonium hydroxide (TMAH) digestion, dichloromethane extraction combined with pyrolysis gas chromatography-mass spectrometry (Py-GC/MS) quantification was present to quantify polystyrene (PS) NPs uptake in lettuce (Lactuca sativa). 25% of TMAH was optimized as extraction solvent and 590 °C was selected as pyrolysis temperature. Recoveries of 73.4-96.9% were obtained for PS-NPs at spiking level of 4-100 µg/g in control samples (RSD < 8.6%). The method exhibited good intra-day and inter-day reproducibility, detection limits of 34-38 ng/g and linearity with 0.998-0.999. The reliability of Py-GC/MS method was verified by europium-chelated PS using inductively coupled plasma mass spectrometry (ICP-MS). To simulate different environmental conditions, hydroponic culture and soil incubated lettuce were exposed to different concentrations of NPs. Higher levels of PS-NPs were detected in roots and very few was transferred to the shoots. NPs in lettuce were confirmed by laser scanning confocal microscopy (LSCM). The developed method provides new opportunities for the quantification of NPs in crops.

Biodegradable microplastics impact the uptake of Cd in rice: The roles of niche breadth and assembly process.

Biodegradable microplastics (MPs) can impact the accumulation of cadmium (Cd) by plants, however, its mechanisms have not been fully understood. In this study, two biodegradable MPs, polypropylene carbonate (PPC) and polylactic acid (PLA), were used to examine their influences on the uptake of Cd in rice plants. Results showed that PPC significantly reduced the accumulation of Cd in rice root and aerial part, whereas PLA increased the Cd concentrations in rice root. The random forest analysis revealed that the bacterial biomarkers enriched by two MPs were different at genus level. Niche breadths were significantly reduced under Cd stress, and PPC alleviated this environmental pressure for entire bacterial community, whereas PLA reduced the niche breadth for whole community and abundant taxa, which was further verified by co-occurrence network and normalized stochasticity ratio model. The abundant taxa of group PPC were primarily governed by deterministic process while rare taxa were more driven by stochastic process. Structural equation model and Mantel analysis identified that the niche breadth imposed a strong selection on Cd accumulation after co-exposure. This study reveals the underlying mechanism of assembly process and niche breadth of rice rhizosphere microbiome on Cd accumulation by rice plants.

Evaluation of the antibacterial effects and mechanism of Plantaricin 149 from Lactobacillus plantarum NRIC 149 on the peri-implantitis pathogens.

Peri-implantitis is a common reversible disease after tooth implantation, caused by a variety of pathogenic microorganisms. Based on non-surgical or surgical treatment principles, supplementation by local or systemic drugs might enhance treatment efficacy. Porphyromonas gingivalis (Pg) (ATCC 33,277) and Prevotella intermedius (Pi) (ATCC 25,611) were used as test strains. The effects of Pln 149 on the biofilm formation and growth of four periodontal pathogens were evaluated by RT-PCR, fluorescence microscopy, and scanning electron microscopy. The antibacterial mechanism was tested by the patch-clamp technique. The cytotoxicity of Pln 149 (125 µg/ml) to bone marrow stromal cell (BMSC) was assessed using an MTT assay. Pln 149 exhibited significant inhibitory effects on Pg and Pi (P < 0.05), with significant differences in the biofilm images of fluorescence microscope and scanning electron microscope (P < 0.05). Pln 149 could change the sodium channel currents and exerted no cytotoxicity on bone marrow stromal cell. Pln 149 could inhibit the biofilm formation and growth of periodontal pathogens. Considering the absence of antimicrobial resistance and cytotoxicity, we suggest that the Pln 149 from Lactobacillus plantarum 149 might be a promising option for managing peri-implantitis.

Profiling of Oral Microbiota in Early Childhood Caries Using Single-Molecule Real-Time Sequencing.

Background: Alterations of oral microbiota are the main cause of the progression of caries. The goal of this study was to characterize the oral microbiota in childhood caries based on single-molecule real-time sequencing. Methods: A total of 21 preschoolers, aged 3-5 years old with severe early childhood caries, and 20 age-matched, caries-free children as controls were recruited. Saliva samples were collected, followed by DNA extraction, Pacbio sequencing, and phylogenetic analyses of the oral microbial communities. Results: Eight hundred and seventy six species derived from 13 known bacterial phyla and 110 genera were detected from 41 children using Pacbio sequencing. At the species level, 38 species, including Veillonella spp., Streptococcus spp., Prevotella spp., and Lactobacillus spp., showed higher abundance in the caries group compared to the caries-free group (p < 0.05). The core microbiota at the genus and species levels was more stable in the caries-free micro-ecological niche. At follow-up, oral examinations 6 months after sample collection, development of new dental caries was observed in 5 children (the transitional group) among the 21 caries free children. Compared with the caries-free children, in the transitional and caries groups, 6 species, which were more abundant in the caries-free group, exhibited a relatively low abundance in both the caries group and the transitional group (p < 0.05). We conclude that Abiotrophia spp., Neisseria spp., and Veillonella spp., might be associated with healthy oral microbial ecosystem. Prevotella spp., Lactobacillus spp., Dialister spp., and Filifactor spp. may be related to the pathogenesis and progression of dental caries.

Inflammatory cytokine levels in patients with periodontitis and/or coronary heart disease.

The purpose of this study was to investigate systemic and local levels of four classic inflammatory cytokines (IL-1ß, MCP-1, VEGF, PDGF) in patients with periodontitis and coronary heart disease (CHD). 109 volunteers were enrolled and the condition of their periodontal tissue and coronary artery were assessed. The patients were then divided into four distinct groups: periodontitis only, CHD only, periodontitis with CHD, and healthy controls. Gingival crevicular fluid (GCF) and venous blood were collected. The concentrations of cytokines were detected meanwhile by specific ELISA. The IL-1ß and MCP-1 concentrations in the serum and GCF of the three disease groups were significantly higher than those in the control group (P ≤ 0.05). Serum VEGF concentrations of the patients with existing disease was lower than that of the controls. VEGF levels in the GCF of all disease groups were significantly higher than that of the control group (P ≤ 0.05).