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Scutellaria baicalensis (SB) is a traditional Chinese medicine for treating infectious and inflammatory diseases. Our recent study shows potent antibacterial effects of nanoparticleencapsulated chlorhexidine (Nano-CHX). Herein, we explored the synergistic effects of the nanoparticle-encapsulated SB (Nano-SB) and Nano-CHX on oral bacterial biofilms. Loading efficiency of Nano-SB was determined by thermogravimetric analysis, and its releasing profile was assessed by high-performance liquid chromatographyusing baicalin (a flavonoid compound of SB) as the marker. The mucosal diffusion assay on Nano-SB was undertaken in a porcine model. The antibacterial effects of the mixed nanoparticles (Nano-MIX) of Nano-SB and Nano-CHX at 9:1 (w/w) ratio were analyzed in both planktonic and biofilm modes of representative oral bacteria. The Nano-MIX was effective on the mono-species biofilms of Streptococcus (S.) mutans, S. sobrinus, Fusobacterium (F.) nucleatum, and Aggregatibacter (A.) actinomycetemcomitans (MIC 50 μg/mL) at 24 h, and exhibited an enhanced effect against the multi-species biofilms such as S. mutans, F. nucleatum, A. actinomycetemcomitans, and Porphyromonas (P.) gingivalis (MIC 12.5 μg/mL) at 24 h that was supported by the findings of both scanning electron microscopy (SEM) and confocal scanning laser microscopy (CLSM). This study shows enhanced synergistic antibacterial effects of the Nano-MIX on common oral bacterial biofilms, which could be potentially developed as a novel antimicrobial agent for clinical oral/periodontal care. [ABSTRACT FROM AUTHOR]

To study the immune-inflammatory response and signaling mechanism of macrophages to purified Enterococcus faecalis (E. faecalis) lipoteichoic acid (LTA), intact LTA was obtained from an E. faecalis clinical strain P25RC using the butanol method and hydrophobic interaction chromatography purification. The fractions containing LTA were determined using phosphate detection. Contaminations with lipopolysaccharide and proteins were excluded using the Limulus amoebocyte lysate assay and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, respectively. LTA was analyzed using nuclear magnetic resonance. Prior to LTA stimulation assays, THP-1 monocytes were pretreated with phorbol 12-myristate 13-acetate to differentiate into macrophages. Macrophages were treated with LTA in concentration gradients and cells without LTA treatment as the control. Gene expression of TLR2, CD14 and MyD88 were evaluated by quantitative polymerase chain reaction. Tumor necrosis factor-α (TNF-α) and interleukin (IL)-10 were quantified using ELISA. The activated and total nuclear factor-αB (NF-αB) p65 and three mitogen-activated protein kinases (p38, ERK1/2 and JNK) were assessed using western blot analysis. E. faecalis LTA induced the gene expression of TLR2 and MyD88 whilst it downregulated CD14, suggesting a TLR2-dependent and CD14-independent immune-inflammatory activity. LTA stimulated the expression of pro-inflammatory cytokine TNF-α (P<0.05), but not the anti-inflammatory cytokine IL-10. In conclusion, E. faecalis LTA stimulated the expression of TNF-α in macrophages possibly through the NF-αB and p38 pathways. [ABSTRACT FROM AUTHOR]

The microbial diversity of biofilms formed on the surfaces of amalgam, glass-ionomer cement, and resin composite was analyzed bydenaturing gradient gel electrophoresis (DGGE). The V2-V3 region of salivary microbial 16S rDNA gene sequences of planktonic andbiofilm bacteria, after 1 day and 1 week of incubation, was amplified by polymerase chain reaction (PCR) and analyzed by DGGE.The amounts of strongly adherent phylotypes after 1 day and 1 week on the three dental restorative materials were more than thoseon hydroxyapatite. Streptococcus salivarius was detected in both loosely adherent and strong adherent groups of all 1-day samples.At 1 week, the amounts of loosely adherent and strongly adherent phylotypes present on the three restorative materials ranked inthis ascending order: glass-ionomer cement < resin composite < amalgam. Results of DGGE analysis suggested that glass-ionomercement was the best material of choice in terms of suppressing bacterial phylotypes in biofilms. [ABSTRACT FROM AUTHOR]

Candida is the most common fungal pathogen of humans worldwide and has become a major clinical problem because of the growing number of immunocompromised patients, who are susceptible to infection. Moreover, the number of available antifungals is limited, and antifungal-resistant Candida strains are emerging. New and effective antifungals are therefore urgently needed. Here, we discovered a small molecule with activity against Candida spp. both in vitro and in vivo. We screened a library of 50,240 small molecules for inhibitors of yeast-to-hypha transition, a major virulence attribute of Candida albicans. This screening identified 20 active compounds. Further examination of the in vitro antifungal and anti-biofilm properties of these compounds, using a range of Candida spp., led to the discovery of SM21, a highly potent antifungal molecule (minimum inhibitory concentration (MIC) 0.2 – 1.6 µg/ml). In vitro, SM21 was toxic to fungi but not to various human cell lines or bacterial species and was active against Candida isolates that are resistant to existing antifungal agents. Moreover, SM21 was relatively more effective against biofilms of Candida spp. than the current antifungal agents. In vivo, SM21 prevented the death of mice in a systemic candidiasis model and was also more effective than the common antifungal nystatin at reducing the extent of tongue lesions in a mouse model of oral candidiasis. Propidium iodide uptake assay showed that SM21 affected the integrity of the cell membrane. Taken together, our results indicate that SM21 has the potential to be developed as a novel antifungal agent for clinical use. [ABSTRACT FROM AUTHOR]

Purpose: Early pregnancy loss (EPL) is one of the most common complications encountered in clinical practice. As most of EPLs occur relatively early on during pregnancy, they are often misunderstood as an expected menstrual cycle. Thus, it is essential to investigate the diagnostic biomarkers for monitoring pregnancy loss for continuous non-invasive monitoring of EPL., Experimental Design: Unstimulated saliva was collected from 10 subjects with EPL and a matched cohort of healthy pregnant women as controls. Samples were analyzed using iTRAQ analysis, and ELISA was performed to validate results., Results: Enrichment analysis of the 38 differentially abundant proteins identified that regulation of nucleobase, nucleoside, nucleotide, and nucleic acid metabolism was significantly affected in EPL. The nucleosome assembly pathway was significantly underrepresented in EPL and was associated with depletion of histone proteins (H2B, H3, and H4). These results were validated with ELISA experiments. A depletion of histones can impair nucleosome assembly and cause the nuclear machinery to fail., Conclusion: Regulation of nucleosome is critical for the maintenance of genome stability and epigenetic information, lack of which may lead to pregnancy loss. Thus, assessing and monitoring salivary histone levels in patients with threatened miscarriage can be a quick and easy method of obtaining periodic diagnostic information that can speed up treatment decisions., Clinical Relevance: There is considerable uncertainty regarding the prognosis of threatened pregnancy, making it stressful for expecting mothers and healthcare professionals. Most EPLs are often misunderstood or ignored as an expected menstrual cycle. Thus it is essential to develop screenings and rapid detection devices using a medium that can be non-invasive and self-performed for continuous monitoring. Using saliva, we have identified that the nucleosome assembly gets affected in EPL with depletion of histone proteins (H2B, H3, and H4). With further verification, these findings can help saliva be utilized as a medium to determine which patients will/will not progress to miscarriage and at what point of their pregnancy. Assessing and monitoring EPL using salivary diagnostics can be a quick and easy method of obtaining periodic diagnostic information that can speed up treatment decisions. Hence, these findings need to be investigated further to improve the prediction of outcomes in women with threatened pregnancy., (© 2021 Wiley-VCH GmbH.)

Objective: To investigate the immunoinflammatory response in the crosstalk of human oral keratinocytes (HOKs) with selected periodontal commensals and pathogens., Methods: Four representative viable oral bacteria, including periodontal commensals (Streptococcus mutans, Sm; and Actinomyces israelii, Ai) and pathogens (Aggregatibacter actinomycetemcomitans, Aa; and Porphyromonas gingivalis, Pg), were selected. A viable bacteria-HOKs interactive model was tested under various conditions of oxygen, antibiotics, duration and multiplicity of infection (MOI). The expression of IL-6 and IL-8 in HOKs was assessed by real-time qPCR and ELISA., Results: An MOI of 1 was determined to be the appropriate ratio of bacteria and HOKs with substantial amounts of viable bacterial cells and HOKs in an antibiotic-free medium under aerobic conditions for 2 h. Sm and Pg significantly upregulated the expression of IL-6 and IL-8 (P < 0.05), while Ai and Aa could not induce significant levels of these cytokines with reference to the control., Conclusion: Within the limitations of this study, the current findings suggest that periodontal commensals and pathogens may differentially modulate immunoinflammatory response in human oral keratinocytes.

Objective: To examine the correlations among oral health knowledge, attitude, practices and oral disease among pregnant Chinese women in Singapore. Methods : A descriptive correlational study was conducted in pregnant Chinese women in Singapore. A questionnaire was used to collect data of oral health knowledge, attitude and practices. Plaque index scores were used to assess the oral health of subjects. Results : A total of 82 pregnant women participated in the study, out of whom 38% showed adequate oral health knowledge, nearly half of them achieved adequate and oral health attitude and practice scores while 34% had good Plaque index scores. The lower income group had higher experience of self-reported dental problems during pregnancy than those in the higher income group ( p = 0.03). There were significant positive correlations between scores of oral health practice, attitude and oral health knowledge levels. The plaque index scores negatively correlated with the oral health practice scores ( p = 0.02). Conclusions : Our findings provided evidence that oral health knowledge, attitude and practices among Chinese pregnant women were not optimal which implies the importance of promoting their oral health during pregnancy through the improvement of knowledge and attitudes. This would facilitate formulation and implementation of appropriate oral health promotion policies., Competing Interests: All the authors declare that they have no conflict of interest.

Scutellaria baicalensis (SB) is a traditional Chinese medicine for treating infectious and inflammatory diseases. Our recent study shows potent antibacterial effects of nanoparticle-encapsulated chlorhexidine (Nano-CHX). Herein, we explored the synergistic effects of the nanoparticle-encapsulated SB (Nano-SB) and Nano-CHX on oral bacterial biofilms. Loading efficiency of Nano-SB was determined by thermogravimetric analysis, and its releasing profile was assessed by high-performance liquid chromatographyusing baicalin (a flavonoid compound of SB) as the marker. The mucosal diffusion assay on Nano-SB was undertaken in a porcine model. The antibacterial effects of the mixed nanoparticles (Nano-MIX) of Nano-SB and Nano-CHX at 9:1 ( w / w ) ratio were analyzed in both planktonic and biofilm modes of representative oral bacteria. The Nano-MIX was effective on the mono-species biofilms of Streptococcus ( S. ) mutans , S. sobrinus , Fusobacterium ( F. ) nucleatum , and Aggregatibacter ( A. ) actinomycetemcomitans (MIC 50 μg/mL) at 24 h, and exhibited an enhanced effect against the multi-species biofilms such as S. mutans , F. nucleatum , A. actinomycetemcomitans , and Porphyromonas ( P. ) gingivalis (MIC 12.5 μg/mL) at 24 h that was supported by the findings of both scanning electron microscopy (SEM) and confocal scanning laser microscopy (CLSM). This study shows enhanced synergistic antibacterial effects of the Nano-MIX on common oral bacterial biofilms, which could be potentially developed as a novel antimicrobial agent for clinical oral/periodontal care.

Dental plaque is an archetypical biofilm composed of a complex microbial community. It is the aetiological agent for major dental diseases such as dental caries and periodontal disease. The clinical picture of these dental diseases is a net result of the cross-talk between the pathogenic dental plaque biofilm and the host tissue response. In the healthy state, both plaque biofilm and adjacent tissues maintain a delicate balance, establishing a harmonious relationship between the two. However, changes occur during the disease process that transform this 'healthy' dental plaque into a 'pathogenic' biofilm. Recent advances in molecular microbiology have improved the understanding of dental plaque biofilm and produced numerous clinical benefits. Therefore, it is imperative that clinicians keep abreast with these new developments in the field of dentistry. Better understanding of the molecular mechanisms behind dental diseases will facilitate the development of novel therapeutic strategies to establish a 'healthy dental plaque biofilm' by modulating both host and microbial factors. In this review, the present authors aim to summarise the current knowledge on dental plaque as a microbial biofilm and its properties in oral health and disease.