Glucosyltransferase-modulated Streptococcus mutans adhesion to different surfaces involved in biofilm formation by atomic force microscopy.

Biofilm on dental restorative materials is an important determinant in the etiology of secondary caries development. Formation of biofilm involves adhesion of bacteria onto substrate, bacterial cell, and biofilm surfaces. Glucosyltransferase B and C (GtfB and GtfC) are essential factors for regulation of Streptococcus mutans biofilm formation, but the mechanisms involving different kinds of bacterial adhesion still lack detailed description. In this study, nanoscale adhesion force measurement was performed using atomic force microscopy. Bacteria-coated cantilevers were used to probe S. mutans adhesion to substrates, bacterial cells, and early biofilms. Two representative dental materials, glass ionomer cement (GIC) and composite resin, served as substrates. It was found that deletion of gtfB and gtfC genes both reduced adhesion forces of S. mutans toward substrate and bacterial cell surfaces (P < 0.05). Notably, reduction of the gtfB gene remarkably decreased bacterial adhesion to biofilm surfaces (P < 0.05), while gtfC showed no obvious effect during this stage. Biofilms cultured on GIG further decreased cell-biofilm adhesion, compared with those on resin (P < 0.05). Confocal fluorescence images and scanning electron microscopy images showed that deletion of gtfB lead to reduced microcolony formation and less production of exopolysaccharides (EPSs) in the biofilm, and after bacterial culturing on GIC, the EPS content was further decreased. Our findings suggest that EPSs mainly mediate bacterial adhesion to early biofilm surface. Deletion of gtfB and coculture with GIC could significantly reduce the cell-biofilm adhesion, which is probably through decreasing of EPS production. gtfB exerts a critical role in the bacterial adhesion for the whole process of biofilm development, while gtfC possibly works only in the early stages.

Pegylated carbon nitride nanosheets for enhanced reactive oxygen species generation and photodynamic therapy under hypoxic conditions.

The application of photodynamic therapy (PDT) is of ever-increasing importance in the treatment of malignant tumors; however, there are several major constraints that make it impossible to achieve optimal therapeutic effects. Our objective is to develop a novel photosensitizing drug for skin cancer. In the experiment, we fabricated four-arm-poly ethylene glycol modified amino-rich graphite phase carbon nitride nanosheets (AGCN-PEG), which have good stability in physiological solution and show selective accumulation in tumor cells. Under hypoxic conditions, the AGCN-PEG induced PDT can effectively inhibit growth on A431 human epidermoid carcinoma cells in vivo and in vitro. What's more, after being combined with TMPyP4, the therapeutic effect of AGCN-PEG was greatly improved.

Research progress of electrically-enhanced membrane bioreactor (EMBR) in pollutants removal and membrane fouling alleviation.

Membrane bioreactor (MBR), as a biological unit for wastewater treatment, has been proven to have the advantages of simple structure and high pollutant removal rate. However, membrane fouling limits its wide application, and it is crucial to adopt effective membrane fouling control methods. As a new type of membrane fouling control technology, electrically-enhanced MBR (EMBR) has attracted more interest recently. It uses the driving force of electric field to make pollutants flocculate or move away from the membrane surface to achieve the purpose of inhibiting membrane fouling. This paper expounds the configuration of EMBR in recent years, including the location of membrane components, the way of electric field application and the selection of electrode and membrane materials, and provides the latest development information in various aspects. The enhanced effect of electric field on the removal of comprehensive and refractory pollutants is outlined in detail. And from the perspective of sludge properties (EPS, SMP, sludge particle size, zeta potential and microbial activity), the influence of electric field on sludge characteristics and the relationship between the changes of sludge properties in EMBR and membrane fouling are discussed. Moreover, the electrochemical mechanisms of electric field alleviating membrane fouling are elucidated from electrophoresis, electrostatic repulsion, electroflocculation, electroosmosis, and electrochemical oxidation, and the regeneration and stability of EMBR are assessed. The existing challenges and future research directions are also proposed. This review could provide theoretical guidance and further studies for subsequent topic, and promoting the wide engineering applications of EMBR.

A poly(dimethylsiloxane)-based solid-phase microchip platform for dual detection of Pseudorabies virus gD and gE antibodies.

Pseudorabies caused by pseudorabies virus (PRV) infection is still a major disease affecting the pig industry; its eradication depends on effective vaccination and antibody (Ab) detection. For a more rapid and accurate PRV detection method that is suitable for clinical application, here, we established a poly(dimethylsiloxane)-based (efficient removal of non-specific binding) solid-phase protein chip platform (blocking ELISA) for dual detection of PRV gD and gE Abs. The purified gD and gE proteins expressed in baculovirus were coated into the highly hydrophobic nanomembrane by an automatic spotter, and the gray values measured by a scanner were used for the S/N (sample/negative) value calculation (gD and gE Abs standard, positive: S/N value ≤0.6; negative: S/N value >0.7; suspicious: 0.6 < S/N ≤ 0.7). The method showed an equal sensitivity in the gD Ab test of immunized pig serum samples compared to the neutralization test and higher sensitivity in the gE Ab test compared to the commercial gE Ab detection kit. In the clinical evaluation, we found an agreement of 100% (122/122) in the gD Ab detection compared to the neutralization test and an agreement of 97.5% (119/122) in the gE Ab detection compared to the commercial PRV gE Ab detection kit. In summary, the protein chip platform for dual detection of PRV gD and gE Abs showed high sensitivity and specificity, which is suitable for PRV immune efficacy evaluation and epidemic monitoring.

A novel theranostic nanoplatform for imaging-guided chemo-photothermal therapy in oral squamous cell carcinoma.

Oral squamous cell carcinoma (OSCC) is highly malignant and invasive, and current treatments are limited due to serious side effects and unsatisfactory outcomes. Here, we reported the terbium ion-doped hydroxyapatite (HATb) nanoparticle as a luminescent probe to encapsulate both the near-infrared (NIR) photothermal agent polydopamine (PDA) and anticancer doxorubicin (DOX) for imaging-guided chemo-photothermal therapy. The morphology, crystal structure, fluorescence, and composition of HATb-PDA-DOX were characterized. HATb-PDA showed a high DOX loading capacity. A theranostic nanoplatform showed pH/NIR responsive release properties and better antitumor outcomes in OSCC cells than monomodal chemotherapy or photothermal therapy, while keeping side effects at a minimum. Also, the luminescence signal was confirmed to be tracked and the increase of the red/green (R/G) ratio caused by the DOX release could be used to monitor the DOX release content. Furthermore, HATb-PDA-DOX plus NIR treatment synergistically promoted in vitro cell death through the overproduction of reactive oxygen species (ROS), cell cycle arrest, and increased cell apoptosis. Overall, this work presents an innovative strategy in designing a multifunctional nano-system for imaging-guided cancer treatment.

AS1411 aptamer modified carbon dots via polyethylenimine-assisted strategy for efficient targeted cancer cell imaging.

OBJECTIVES: Carbon dots (CDs), as a fascinating class of fluorescent carbon nanomaterials, have been proven to be powerful tools in the field of bioimaging and biosensing due to their small size, suitable photostability and favourable biocompatibility. However, the cellular uptake of free CDs lacks selectivity and the same negative charges as cell membranes may cause inefficient cell internalization. In this study, an efficient detecting and targeting nanosystem was developed based on the DNA aptamer AS1411 modified CDs with polyethyleneimine (PEI) as connecting bridge. MATERIALS AND METHODS: Hydrothermally prepared CDs were assembled with positive-charged PEI, followed by conjugation with AS1411 through electrostatic interaction to form CDs-PEI-AS1411 nanocomplexes. The CDs, CDs-PEI and CDs-PEI-AS1411 were characterized by transmission electron microscopy (TEM), fourier transform infrared (FTIR) spectra, UV-vis spectra, zeta potential measurements and capillary electrophoresis characterizations. The cytotoxicity investigation of the CDs-PEI-AS1411 and CDs-PEI in both MCF-7 and L929 cells was carried out by the CCK-8 assay. The cellular uptake of the CDs-PEI-AS1411 was studied with confocal microscopy and flow cytometry. RESULTS: The as-prepared nanosystem possessed good photostability and no obvious cytotoxicity. On the basis of the confocal laser scanning microscope observation and the flow cytometry studies, the cellular uptake of CDs-PEI-AS1411 nanosystem in MCF-7 cells was significantly higher than that of L929 cells, which revealed the highly selective detection ability of nucleolin-positive cells. CONCLUSIONS: The results of this study indicated that the CDs-PEI-AS1411 nanosystem had a potential value in cancer cell targeted imaging.

Nanoscale adhesion forces of glucosyltransferase B and C genes regulated Streptococcal mutans probed by AFM.

Glucosyltransferases (Gtfs), represented by GtfB and GtfC, are important virulence factors of Streptococcus mutans and the major etiologic pathogens of tooth decay. However, the individual roles of gtfB and gtfC in the initial attachment of S. mutans are not known. We used atomic force microscopy to explore the contribution of gtfB and gtfC, as well as enamel-surface roughness, on the initial attachment of S. mutans. Adhesion forces of four S. mutans strains (wild-type, ΔgtfB, ΔgtfC, and ΔgtfBC), onto etched enamel surfaces, were determined. Force curves showed that, with increasing etching time from 0 to 10 s, the forces of all strains increased accordingly with acid-exposure time, the adhesion forces of wild-type strains were significantly greater than those of mutant strains (p < .05), and the forces of the three mutants were similar (p < .05). When the etching time was increased from 10 to 30 s, difference in force between 20 and 30 s was not observed, and adhesion forces among ΔgtfB, ΔgtfC, and wild-type strains were not significantly different when the etching time was >20 s (p > .05). These data suggest that the roughness and morphology of enamel surfaces may have a significant influence upon the initial attachment of bacteria, and that gtfB and gtfC are essential for the adhesion activity of bacteria. Furthermore, gtfB seems to be more important than gtfC for bacterial-biofilm formation, and gtfB inactivation is an effective strategy to inhibit the virulence of cariogenic biofilms.

Effects of water and microbial-based aging on the performance of three dental restorative materials.

The objective of this study was to evaluate the performance changes of three restorative materials before and after three different aging treatments: storage in distilled water, Streptococcus mutans (S. mutans) and oral salivary microbes suspensions for one month. Resin composite (RC), giomer and glass ionomer cement (GIC) were chosen for aging procedures. Surface morphology, roughness average (Ra), color changes and mechanical properties were all determined before and after aging respectively. Biomass and metabolism difference of early attached biofilm on the material surface were tested through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and lactic acid measurement. The results showed that after S. mutans or salivary microbes aging treatments, GIC group displayed significant morphology changes, with Ra value significantly higher than that before aging (p < .001). Color changes of giomer and GIC group after S. mutans aging were not clinically acceptable. All materials after two microbial-based aging treatments had higher flexural strength than that before aging (p < .05). Giomer after salivary microbes aging had higher elastic modulus than the initial values (p < .05). Additionally, early attached biofilm biomass and lactic acid production in GIC group after S. mutans or salivary microbes aging were higher than that before aging (p < .05). While one-month water aging showed less influences on material performance to some extent. In conclusion, to better simulate the harsh oral environment, in vitro microbial-based aging models showed more advantages in evaluating dental restorative materials' degradation over time.

Tea Polyphenol-Functionalized Graphene/Chitosan as an Experimental Platform with Improved Mechanical Behavior and Bioactivity.

In this study, water-soluble, one-step highly reduced and functionalized graphene oxide was prepared via a facile, environment-friendly method by using tea polyphenol (TP), which acted as both reducing agent and stabilizer. The product obtained, that is, tea polyphenol-reduced graphene oxide (TPG), was used as a reinforcing building block for the modification of a mechanically weak chitosan (CS), TPG/CS. The morphology and physicochemical and mechanical properties of the composite were examined by various characterizations. The tensile strength and elastic modulus of CS were greatly improved by TPG, as compared to the findings for GO incorporation. Additionally, to our knowledge, this study is an in-depth analysis of the osteoblast functions of CS/TPG, including aspects such as cell cytotoxicity, proliferation, and expression of ossification genes, alkaline phosphatase (ALP), and Runt-related transcription factor (Runx2), which showed advantages in favorably modulating cellular activity. It was concluded that TPG/CS showed a higher elastic modulus, better hydrophilicity, and excellent biocompatibility than the pristine chitosan for promoting the proliferation and differentiation of osteoblasts, as well as for accelerating the expression of ALP and Runx2 (as shown by reverse transcription polymerase chain reaction (RT-PCR)). These results may provide new prospects for the use of TPG in the modification of biomaterials and for broadening the application of TPG in biological fields.

8DSS-promoted remineralization of demineralized dentin in vitro.

Dentin phosphoprotein (DPP) plays an extremely important role in the biomineralization of human tooth. The repetitive nucleotide sequence of aspartate-serine-serine (DSS) is the fundamental unit within DPP, and peptides containing 8 repeats of DSS (8DSS) have been shown to possess the ability to induce remineralization of the demineralized enamel. In this work, we coated the 8DSS peptide on the completely demineralized dentin to evaluate the effect of 8DSS peptide coating on dentin remineralization. Human dentin samples were demineralized with 37% phosphoric acid for 2 min, and then the 8DSS peptide (1 mg mL-1) was coated and the binding strength of the 8DSS peptide to demineralized dentin was examined using attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR). Then the coated dentin samples were immersed in artificial saliva for 3 weeks. After that, the remineralized dentin samples were characterized by scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS), X-ray diffraction (XRD) analysis and ATR-FTIR. The mechanical properties of the remineralized surfaces were determined by nano-indentation and atomic force microscopy (AFM). The results showed that the 8DSS peptide had good binding strength to demineralized dentin and could induce nano-crystals precipitation both on the surfaces and within the dentinal tubules. The mechanical properties of the 8DSS-coated samples were significantly improved. In contrast, there were hardly any newly generated minerals deposited on the samples without 8DSS peptide coating. In conclusion, the 8DSS peptide may be a promising biomaterial for restoring the demineralized human dentin.

Independent effect of polymeric nanoparticle zeta potential/surface charge, on their cytotoxicity and affinity to cells.

OBJECTIVES: Up to now, little research has been focussed on discovering how zeta potential independently affects polymeric nanoparticle (NP) cytotoxicity. METHODS: Polymeric nanoparticles of gradient zeta potential ranging from -30 mv to +40 mv were fabricated using the same poly-3-hydroxybutyrate-co-3-hydroxyhexanoate (PHBHHx) biopolymer. Interaction forces between nanoparticles and cells were measured by atomic force microscopy (AFM). Cytotoxicity of the nanoparticles to cells was investigated by using MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide) assay. RESULTS: Four kinds of nanoparticle with similar sizes and gradient zeta potentials, were fabricated. Those with positive surface charges were found to be more toxic than those with negative surface charges. Positively charged nanoparticles or nanoparticles with higher 'like' charges, offered higher interaction force with cells. CONCLUSION: This work proposes a novel approach for investigating interaction between NPs and cells, and discloses the importance of controlling zeta potential in developing NPs-based formulations in the future.