Effects of combined exogenous dextranase and sodium fluoride on Streptococcus mutans 25175 monospecies biofilms.

PURPOSE: To investigate the effects of exogenous dextranase and sodium fluoride on a S. mutans monospecies biofilm. METHODS: S. mutans 25175 was grown in tryptone soya broth medium, and biofilm was formed on glass slides with 1.0% sucrose. Exogenous dextranase and sodium fluoride were added alone or together. The biofilm morphology was analyzed by confocal laser scanning microscopy. The effects of the drug on the adhesion and exopolysaccharide production by the biofilms were evaluated by scintillation counting and the anthrone method, respectively. RESULTS: In this study, we found that the structure of initial biofilm and mature biofilm were partly altered by dextranase and high concentrations of sodium fluoride separately. However, dextranase combined with a low concentration of sodium fluoride could clearly destroy the typical tree-like structure of the biofilm, and led to less bacterial adhesion than when the dextranase or fluoride were used alone (P < 0.05). The amounts of soluble and insoluble exopolysaccharide were significantly reduced by combining dextranase with a low concentration of sodium fluoride, much more than when they were used alone (P < 0.05). These data indicate that dextranase and a low concentration of sodium fluoride may have synergistic effects against S. mutans biofilm and suggest the application of a low concentration of sodium fluoride in anticaries treatment.

Efficient Removal of Dental Plaque Biofilm from Training Typodont Teeth via Water Flosser.

Plaque biofilms play critical roles in the development of dental caries. Mechanical plaque control methods are considered to be most effective for plaque removal, such as brushing teeth or using flosser. Recently, water flosser has been paid much attention. Here, we tested the ability of a water flosser to remove the adhered sucrose and the dental plaque biofilms formed by Streptococcus mutans, Streptococcus sanguinis, and Actinobacillus viscosus. We found that the residual sucrose concentration was 3.54 mg/mL in the control group, 1.75 mg/mL in the syringe group (simulating the ordinary mouthwash), and 0 mg/mL in water flosser group. In addition, the residual bacterial concentration was 3.6 × 108 CFU/mL in the control group, 1.6 × 107 CFU/mL in the syringe group, and only 5.5 × 105 CFU/mL in the water flosser group. In summary, water flosser is effective for cleaning the teeth, which may have significant potential in preventing dental caries and maintaining oral health.

Microstructure and mechanical properties of glass-infiltrated Al2O3/ZrO2 nanocomposites.

This work was to investigate the effect of zirconia nanoparticles content on microstructure and mechanical properties of glass-infiltrated alumina/zirconia composites (AZGs). A series of slip-cast zirconia-toughened alumina (ZTA) compacts containing 10, 20, 30 wt% nano-zirconia, respectively, were partially sintered at 1,250°C for 2 h, then infiltrated with lanthanum borosilicate glass of lower thermal expansion at 1,180°C for 4 h. A porosity ranging from 21 to 25% mainly with submicron pore size was demonstrated in the partially-sintered ZTAs. Homogeneous distribution and micro-crystallization of intergranular glass phase was showed in the AZGs. The mechanical strength and fracture toughness of AZGs increased with zirconia content, the maximum (633.5 ± 41.7, 6.7 ± 0.6 MPa m(0.5)) were obtained in 30 wt% zirconia group, which were significantly higher than those in 10 wt% zirconia group (P < 0.05). The improved mechanical performance of AZGs containing 30 wt% zirconia was attributed to their larger zirconia content as well as thinner intergranular glass film.

Functional nanomaterials and their potentials in antibacterial treatment of dental caries.

Dental caries is one of the most common bacteria-related oral diseases, and the cariogenic bacteria play critical roles in its pathogenesis. Antibacterial therapy is regarded as an important approach for treating dental caries. However, the low local drug concentration and the potential generation of drug resistance limit the traditional antibacterial efficacy. Nanomaterials, such as polymeric nanoparticles and liposomes, have been widely used as functional carriers for drug delivery. The antibiotic-loaded nanomaterials have shown great potentials in enhancing the efficacy of antibacterial therapy. In addition, some functionalized nanomaterials, such as metal nanoparticles and graphene-based nanomaterials, can be used as direct antibacterial agents for physical antibacterial therapy, photothermal therapy and photodynamic therapy. Herein, we aim to provide a comprehensive understanding of dental caries and the related pathogenic bacteria, and summarize the recent progress in nanomaterials-based antibacterial treatment. Also, the antibacterial mechanisms and the concerns of nanomaterials in practical use are discussed.

Activity of Ligustrum robustum (Roxb.) Blume extract against the biofilm formation and exopolysaccharide synthesis of Streptococcus mutans.

Ligustrum robustum (Roxb.) Blume is utilized as a traditional Chinese herbal tea with various health benefits and protective effects. Streptococcus mutans is an important cariogenic oral bacteria species. The present study aimed to assess the influence of Ligustrum robustum extract (LRE) on the biofilm formation of S. mutans and the mechanism of its action, as well as to identify its chemical components. For chemical identification, HPLC-MS and nuclear magnetic resonance were applied and four identified phytochemicals were reported (Ligurobustoside B, Ligurobustoside N, Ligurobustoside J, and Ligurobustoside C). The dose-dependent (0.5 to 2.0 µg/µL) antimicrobial toxicity of LRE against S. mutans biofilm formation and exopolysaccharide (EPS) synthesis was evaluated by confocal laser scanning microscopy (CLSM), Crystal violet stain, and CFU counting. The microstructure of S. mutans biofilm treated with LRE was investigated both on glass coverslips and ex vivo bovine dental enamel by scanning electron microscopy (SEM). Moreover, LRE downregulated the expression of S. mutans glucosyltransferase-encoding genes gtfB, gtfC, and gtfD, and the quorum sensing (QS) factors comD and comE, suggesting its toxic mechanism. In addition, the result of CCK-8 test on human oral cells revealed an acceptable biocompatibility of LRE. These findings indicated the possible application of this daily consumed herbal tea for caries prevention.

Drug-free and non-crosslinked chitosan scaffolds with efficient antibacterial activity against both Gram-negative and Gram-positive bacteria.

Treatment of oral pathogens is important for both oral and systemic health. The antimicrobial activity of chitosan (CS)-based scaffolds either loading antibiotics or compositing with other agents are well documented. However, the intrinsic antibacterial activity of CS scaffolds alone has never been reported. Herein, we fabricated the non-crosslinked CS scaffold and investigated its antibacterial activity against typical oral pathogens, Gram-negative Porphyromonas gingivalis and Gram-positive Streptococcus mutans. We found both pathogens were completely killed by 1 mg CS scaffolds at 6 h, due largely to the CS-induced time-dependent bacteria clustering. Interestingly, ß-glycerophosphate crosslinked scaffolds showed no antibacterial activity. In conclusion, the bactericidal activity of CS scaffolds alone is reported for the first time. Together with the biodegradability, physical stability, biocompatibility and great antibacterial activity, the non-crosslinked CS scaffolds may have great potentials not only in treating oral diseases but also in wound healing and tissue engineering.

Antibiofilm effect of drug-free and cationic poly(D,L-lactide-co-glycolide) nanoparticles via nano-bacteria interactions.

AIM: Recently, nano-bio interactions and their biomedical impacts have drawn much attention, but nano-bacteria interaction and its function are unknown. Herein, we aim to synthesize drug-free and cationic nanoparticles (CNPs) and investigate CNP-bacteria interaction and its antibiofilm effect. MATERIALS & METHODS: The bioactivity of CNPs against Streptococcus mutans was examined by colony-forming units counting and scanning electron microscopy. CNP-bacteria interaction force was measured by atomic force microscopy. RESULTS: CNPs (217.7 nm, 14.7 mv) showed a concentration-dependent activity against bacteria. Particularly, CNPs at 200 µg/ml completely inhibited planktonic bacterial growth and biofilm formation, and disrupted ∼70% mature biofilm. CNP-bacteria interaction force was up to 184 nN. CONCLUSION: CNPs have great potentials for convenient local use for prevention and treatment of bacteria-related oral diseases.

Organic composite-mediated surface coating of human acellular bone matrix with strontium.

Acellular bone matrix (ACBM) provides an osteoconductive scaffold for bone repair, but its osteoinductivity is poor. Strontium (Sr) improves the osteoinductivity of bone implants. In this study, we developed an organic composite-mediated strontium coating strategy for ACBM scaffolds by using the ion chelating ability of carboxymethyl cellulose (CMC) and the surface adhesion ability of dopamine (DOPA). The organic coating composite, termed the CMC-DOPA-Sr composite, was synthesized under a mild condition, and its chemical structure and strontium ion chelating ability were then determined. After surface decoration, the physicochemical properties of the strontium-coated ACBM (ACBM-Sr) scaffolds were characterized, and their biocompatibility and osteoinductivity were determined in vitro and in vivo. The results showed that the CMC-DOPA-Sr composite facilitated strontium coating on the surface of ACBM scaffolds. The ACBM-Sr scaffolds possessed a sustained strontium ion release profile, exhibited good cytocompatibility, and enhanced the osteogenic differentiation of mesenchymal stem cells in vitro. Furthermore, the ACBM-Sr scaffolds showed good histocompatibility after subcutaneous implantation in nude mice. Taken together, this study provided a simple and mild strategy to realize strontium coating for ACBM scaffolds, which resulted in good biocompatibility and improved osteoinductivity.

Adenovirus encoding BMP-7 immobilized on titanium surface exhibits local delivery ability and regulates osteoblast differentiation in vitro.

OBJECTIVE: The local delivery of growth factors such as bone morphogenetic protein-7 (BMP-7) into the tissues around dental implants may improve their osseointegration. We have designed a new method of attaching BMP-7 to a titanium surface and assessed both the retention of the BMP-7 and its effect on osteoblast differentiation. DESIGN: Adenoviral vector expressing BMP-7 was attached to dental titanium discs by hexon-specific antibodies in a type I collagen-avidin gel. FITC-labelled secondary antibody was used to measure the continuing adherence of the coating after repeated rinsing. Osteoblasts were harvested and seeded on the titanium discs. Gene transduction efficiency and targeting ability were assessed after 24h. Surface morphology was observed by SEM. Cell proliferation and alkaline phosphatase (ALP) activities were measured. RESULTS: The anti-adenohexon antibody adhered strongly to the collagen-avidin gels. BMP-7 gene expression was localized precisely to cells growing on the gels bound by the hexon-specific antibody. Osteoblasts on the titanium containing Ad-BMP-7 had a higher ALP activity than those without Ad-BMP-7. CONCLUSIONS: This study describes a novel technique for the precise attachment of BMP-7 to titanium surfaces. The process may enhance the osseointegration of dental implants.

Mineral densities and elemental content in different layers of healthy human enamel with varying teeth age.

OBJECTIVES: This study aims to evaluate the mineral density, calcium (Ca), and phosphorus (P) weight percent of healthy human enamel of varying human teeth age and enamel layers. METHODS: Twenty human teeth were divided into "young" (18-24 years) and "old" (55 ≤ years) age groups. From each tooth, one enamel slab (approximately 8 mm × 2 mm × 1.5 mm) was obtained. All enamel slabs were embedded and scanned by micro-computed tomography. The scanned slices were reconstructed into a 3D image and regions of interest (ROIs) were defined. Then, the mineral density for each ROI was calculated. The Ca and P weight percent in the outer, middle, and inner enamel layers were quantified by scanning electron microscopy with energy dispersive spectroscopy. Finally, the data were analysed using one-way ANOVA and Student's t-test. RESULTS: In the two age groups, the mineral density, Ca and P weight percent decreased from outer to the inner enamel layer. The mineral density, Ca and P weight percent in the outer enamel layer in the old age group were significantly higher than those in the young age group (P<0.05); however, no age-dependent differences were observed for these properties in the middle and inner enamel layers (P>0.05). CONCLUSIONS: The mineral density, Ca and P weight percent decreased from the outer to the inner enamel layers in the two age groups. In the outer enamel layer, the differences in these properties between the two age groups were significantly different; however, no region-dependent differences were observed in the middle and inner enamel layers.

[Effects of different concentrations of ethylenediamine tetraacetic acid paste on removing root canal smear].

OBJECTIVE: To evaluate the effect of different concentrations of ethylenediamine tetraacetic acid (EDTA) paste on the removing root canal smear layers and the degrees of erosion on the surface of the root canal walls at the different portions of canal. METHODS: Sixty human teeth with single root were instrumented using step-back technique, then were divided into six groups and treated with different concentrations EDTA paste and NaCLO solution. Group A: 0.9% saline; group B: 5.25% NaC10+5% EDTA; group C: 5.25% NaC10+10% EDTA; group D: 5.25% NaC10+15% EDTA; group E: 5.25% NaC10+17% EDTA; group F: 5.25% NaClO+20% EDTA. Then the teeth were split, the root canals with different treatments were examined with scanning electron microscope at the coronal, middle and apical thirds for smear layers removal and the degrees of erosion. RESULTS: The effect of EDTA paste on removing smear layers at the coronal, middle thirds of the root canal increased with the concentrations. 15% EDTA paste could remove the smear layers at the coronal and middle thirds of the root canal, but the effect on apical third was invalid, and no erosion could be found in the root canal wall. 17% and 20% EDTA paste produced the erosion to the root canal wall. CONCLUSION: The concentrations of the EDTA paste can influence the effect of the removing smear layers, and the concentration beyond 17% would produce the erosion to root canal wall.

Ectopic bone formation in adipose-derived stromal cell-seeded osteoinductive calcium phosphate scaffolds.

The phenomenon of osteoinduction by biomaterials has been proven and used in animals. However, whether the ability of a biomaterial to initiate bone formation in ectopic implantation sites improves the performance of such osteoinductive biomaterial as a scaffold for tissue-engineered (TE) bone remains unclear. In this study, we compared ectopic bone formation by combining autologous adipose-derived stromal cells (ADSCs) with an osteoinductive and a nonosteoinductive biphasic calcium phosphate (BCP) ceramic to create a tissue engineering construct in the muscle of dogs. Two groups of BCP scaffolds (BCP1 and BCP2) were prepared. In each group, ADSCs were seeded, and the scaffolds without seeded cells served as controls. All implants were implanted in the back muscle of 10 adult dogs for 8 weeks and 12 weeks. Microcomputed tomography (Micro-CT) analysis and histomorphometry were performed to evaluate and quantify ectopic bone formation. The results indicated that the osteoinductive BCP1 performed significantly better compared to the nonosteoinductive BCP2 in cell-based TE bone formation ectopically. The ADSCs had a significantly positive effect on the ectopic bone formation. In addition, the usefulness of Micro-CT for the efficient and nondestructive analysis of mineralized bone and calcium phosphate scaffold was confirmed.

[The relationship in growth between A.actinomycetemcomitans and cariogenic bacteria in vitro].

PURPOSE: To investigate the correlation in growth between A.actinomycetemcomitans and cariogenic bacteria of S.mutans, L.acidophilus A.naeslundii A.viscosus in vitro. METHODS: Using improvement agar diffusion method and dual-species (A.actinomycetemcomitans with each of cariogenic bacteria) incubation in BHI broth, we studied the relationship between A.actinomycetemcomitans and 4 kinds cariogenic bacteria in growth. The characteristic and percentage of A.actinomycetemcomitans in dual-species biofilms were detected with SEM and CFU(colony forming units).The data were analyzed for significance using an independent two-sample t test with SPSS 10.0 software package. RESULTS: The agar diffusion essay showed that A.actinomycetemcomitans had no effect on the growth of S.mutans, L.acidophilus A.naeslundii and A.viscosus. While these 4 kinds of cariogenic bacteria could inhibit the growth of A.actinomycetemcomitans. The ratio of A.actinomycetemcomitans in the dual-species suspension decreased gradually.In dual-species BHI broth, the proportion of A.actinomycetemcomitans cultured alone ahead of 12 hours decreased as well, which came to 0 at 24-hour in S.mutans and L.acidophilus groups. There was significant difference between different time groups(P<0.05).The single A.actinomycetemcomitans was failed to form three-dimensional biofilm and just a small quantity of cells accumulated and adhered. On the contrary, the 4 kinds of single cariogenic bacteria could form mature biofilms respectively by themselves at 48-hour,of which the appearance of biofilms had no apparent change to that of single cariogenic bacteria after mixing A.actinomycetemcomitans. CFU of dual-species biofilms indicated that the percentage of A.actinomycetemcomitans in biofilms decreased gradually by changing medium per 12 hours. There were significant difference among the three time spots(P<0.05). CONCLUSION: The study shows that S.mutans, L.acidophilus A.naeslundii and A.viscosus could inhibit the growth of A.actinomycetemcomitans in vitro.