Salivary levels of five microorganisms of root caries in nursing home elderly: a preliminary investigation.

BACKGROUND: Streptococcus, Bifidobacteria, Lactobacillus and Actinomyces are acidogenic aciduria that may be associated with root caries (RC). The aim of the study was to analyze Streptococcus mutans (S. mutans), Streptococcus sobrinus (S. sobrinus), Bifidobacterium spp., Lactobacillus spp. and Actinomyces naeslundii (A. naeslundii) in the saliva of nursing home elderly, to assess the correlation between bacterial composition and RC for five putative catiogenic organisms. METHODS: In this study, we collected 43 saliva samples and divided into two groups: the root caries group (RCG, n = 21) and the caries-free group (CFG, n = 22). Bacterial DNA was extracted from the saliva samples. The presence and abundance of the five microorganisms were detected by Quantitative real-time PCR (qPCR). Spearman correlation test was performed to evaluate the relationship between the numbers of root decayed filled surfaces (RDFS) and root caries index (RCI) and salivary levels of the bacteria. RESULTS: The salivary levels of S. mutans, S. sobrinus, Bifidobacterium spp. and Lactobacillus spp. were significantly higher in RCG than in CFG (p < 0.05). RDFS and RCI (RDFS/RCI) were positively associated with salivary levels of S. mutans, S. sobrinus and Bifidobacterium spp. (r = 0.658/0.635, r = 0.465/0.420 and r = 0.407/0.406, respectively). No significant differences in presence and amounts of A. naeslundii was observed between the two groups (p > 0.05). CONCLUSION: S. mutans, S. sobrinus and Bifidobacterium spp. in saliva appear to be associated with RC in the elderly. Taken together, the findings indicate that specific salivary bacteria may be involved in the progression of RC.

The Combinations Chitosan-Pam3CSK4 and Chitosan-Monophosphoryl Lipid A: Promising Immune-Enhancing Adjuvants for Anticaries Vaccine PAc.

We previously demonstrated that recombinant protein PAc could be administered as an anticaries vaccine. However, the relatively weak immunogenicity of PAc limits its application. In the present study, we investigated the effect of two adjuvant combinations of chitosan plus Pam3CSK4 (chitosan-Pam3CSK4) and of chitosan plus monophosphoryl lipid A (chitosan-MPL) in the immune responses to the PAc protein in vivo and in vitro PAc-chitosan-Pam3CSK4 or PAc-chitosan-MPL promoted significantly higher PAc-specific antibody titers in serum and saliva, inhibited Streptococcus mutans colonization onto the tooth surfaces, and endowed better protection effect with significantly less caries activities than PAc alone. Chitosan-Pam3CSK4 and chitosan-MPL showed no statistically significant differences. In conclusion, our study demonstrated that the chitosan-Pam3CSK4 and chitosan-MPL combinations are promising for anticaries vaccine development.

Identification of FimH derivatives as adjuvant vaccinated with PAc that enhance protection against Streptcoccus mutans colonization.

FimH is the adhesin of type I fimbriae expressed on Escherichia coli that can mediate specific adherence to host cells. High binding mutations in FimH are related to the adaptive evolution of bacteria. However, additional roles that these allelic variations may play remain elusive. To investigate novel biological functions of the mutations in FimH, we introduced four different variants of FimH by incorporating single amino acid substitutions at specific sites, namely A25P, G73R, A106, and T158P, respectively. In this study, adjuvant potential of FimH variants was evaluated by investigating their ability to trigger innate immune response to DC2.4 and adaptive immunity to improve immunological characteristics. The data revealed that purified A106 and T158P up-regulated the expression of co-stimulatory molecules critically involved in DC2.4 activation by interaction with TLR4, whereas A25P and G73R did not induce the phenotypic maturation of DC2.4. Besides, the culture of DC2.4 with A106 and T158P enhanced the release of cytokines and protein phagocytosis. When formulated with PAc, T158P elicited more robust PAc-specific IgG and IgA antibody responses compared to PBS, PAc and PAc+K12 groups and inhibited bacteria colonization. Collectively, the results confirmed that the T158P mutation located around the inter-domain interface of the protein induced a specific enhancement effect on adjuvant characteristics.

MicroRNA-195-5p Regulates Osteogenic Differentiation of Periodontal Ligament Cells Under Mechanical Loading.

Osteogenic differentiation and bone formation are tightly regulated by several factors, including microRNAs (miRNAs). However, miRNA expression patterns and function during mechanical loading-induced osteogenic differentiation of human periodontal ligament cells (PDLCs) remain unclear. Here, we investigated the differential expression of miRNA-195-5p in the periodontal tissues of mice under orthodontic mechanical loading and in primary human PDLCs exposed to a simulated tension strain. The miR-195-5p was observed to be down-regulated and negatively correlated with osteogenic differentiation. Overexpression of miR-195-5p significantly inhibited PDLC differentiation under cyclic tension strain (CTS), whereas the functional inhibition of miR-195-5p yielded an opposite effect. Further experiments confirmed that WNT family member 3A (WNT3A), fibroblast growth factor 2 (FGF2), and bone morphogenetic protein receptor-1A (BMPR1A), proteins important for osteogenic activity and stability, were direct targets of miR-195-5p. Mechanical loading increased the WNT3A, FGF2, and BMPR1A protein levels, while miR-195-5p inhibited WNT3A, FGF2, and BMPR1A protein expression. WNT, FGF, and BMP signaling were involved in osteogenic differentiation of PDLCs under CTS. Further study confirmed that reintroduction of WNT3A and BMPR1A can rescue the inhibition of miR-195-5p on osteogenic differentiation of PDLCs. Our findings are the first to demonstrate that miR-195-5p is a mechanosensitive gene that plays an important role in mechanical loading-induced osteogenic differentiation and bone formation.

Frequency of remnants of sealants left behind in pits and fissures of occlusal surfaces after 2 and 3 years.

OBJECTIVES: The null-hypothesis tested was that there was no difference in the frequency of remnants of high-viscosity glass-ionomer sealants left behind in pits and fissures of occlusal surfaces of first permanent molars and that of resin composite and glass-carbomer sealants. MATERIALS AND METHODS: Based on the results of a sealant trial, a sample of sealed teeth from which the material had apparently completely disappeared from at least one of the three sections into which the occlusal surface was divided, assessed through visible clinical examination, was also assessed from images of colour photographs and Scanning Electron Microscopy (SEM) as the reference image. The sample size consisted of 112 and 120 teeth from 59 and 98 children at evaluation years 2 and 3 respectively. Two examiners performed the assessments. Fisher's Exact Test was applied to test for the differences between the dependent variable and the sealant groups. RESULTS: The remnants of sealant material left in the deeper parts of pits and fissures were assessed from colour photograph and SEM images in five sections at year 2 and in eight sections at year 3. The assessment found no sealant group effect. CONCLUSION: The frequency of remnants of glass-ionomer sealant in pits and fissures of occlusal surfaces in first permanent molars is not higher than the frequency of glass-carbomer and resin sealants after 2 and 3 years. CLINICAL RELEVANCE: Contrary to the current assumption, there appears to be no significant difference in the frequency of remnants left behind in pits and fissures between glass-ionomer and resin sealants.

Retention Rate of Four Different Sealant Materials after Four Years.

PURPOSE: To test the hypotheses: 1) cumulative survival rates of fully and partially retained high-viscosity glass-ionomer (HVGIC) ART sealants with heat application and glass-carbomer sealants on occlusal and free-smooth surfaces are both higher than that of resin sealants; 2) cumulative survival rate of fully and partially retained high-viscosity glass-ionomer ART sealant with heat application on occlusal and free-smooth surfaces is higher than that of comparable ART sealants without heat application. MATERIALS AND METHODS: The block-randomised clinical trial covered 405 eight-year-old children. The HVGIC was Ketac Molar Easymix, the glass carbomer was GlassCarbomer and the resin sealant was Clinpro. Retention rates of sealants on occlusal and free-smooth surfaces using conventional and modified categorisation (fully and partially retained sealants vs those completely lost [at least one-third of surface re-exposed]) were the dependent variables. The Kaplan-Meier survival method was used. RESULTS: The cumulative survival rate of completely and partially retained resin sealants on occlusal (81.2%) and free-smooth (81%) surfaces after 4 years was statistically significantly higher, and that of glass-carbomer sealants (10.8% and 21.1%, respectively) was statistically significantly lower than those of the other sealant groups. There was no statistically significant difference in survival rates of completely and partially retained high-viscosity glass-ionomer ART sealants with (56% for both surfaces) or without heat application (56%) on occlusal and free-smooth surfaces (55.7% and 59.1%, respectively). CONCLUSIONS: Resin sealants had the highest and glass-carbomer sealants the lowest retention rate after 4 years. Application of heat to high-viscosity glass-ionomer ART sealants did not result in a significantly higher sealant retention rate. Use of the modified categorisation for determining sealant retention is advocated.

IL-1R/TLR2 through MyD88 Divergently Modulates Osteoclastogenesis through Regulation of Nuclear Factor of Activated T Cells c1 (NFATc1) and B Lymphocyte-induced Maturation Protein-1 (Blimp1).

Toll-like receptors (TLR) and the receptor for interleukin-1 (IL-1R) signaling play an important role in bacteria-mediated bone loss diseases including periodontitis, rheumatoid arthritis, and osteomyelitis. Recent studies have shown that TLR ligands inhibit the receptor activator of NF-κB ligand (RANKL)-induced osteoclast differentiation from un-committed osteoclast precursors, whereas IL-1 potentiates RANKL-induced osteoclast formation. However, IL-1R and TLR belong to the same IL-1R/TLR superfamily, and activate similar intracellular signaling pathways. Here, we investigate the molecular mechanisms underlying the distinct effects of IL-1 and Porphyromonas gingivalis lipopolysaccharide (LPS-PG) on RANKL-induced osteoclast formation. Our results show that LPS-PG and IL-1 differentially regulate RANKL-induced activation of osteoclast genes encoding Car2, Ctsk, MMP9, and TRAP, as well as expression of NFATc1, a master transcription factor of osteoclastogenesis. Regulation of osteoclast genes and NFATc1 by LPS-PG and IL-1 is dependent on MyD88, an important signaling adaptor for both TLR and IL-1R family members. Furthermore, LPS-PG and IL-1 differentially regulate RANKL-costimulatory receptor OSCAR (osteoclast-associated receptor) expression and Ca(2+) oscillations induced by RANKL. Moreover, LPS-PG completely abrogates RANKL-induced gene expression of B lymphocyte-induced maturation protein-1 (Blimp1), a global transcriptional repressor of anti-osteoclastogenic genes encoding Bcl6, IRF8, and MafB. However, IL-1 enhances RANKL-induced blimp1 gene expression but suppresses the gene expression of bcl6, irf8, and mafb. Our study reveals the involvement of multiple signaling molecules in the differential regulation of RANKL-induced osteoclastogenesis by TLR2 and IL-1 signaling. Understanding the signaling cross-talk among TLR, IL-1R, and RANK is critical for identifying therapeutic strategies to control bacteria-mediated bone loss.

Cost-effectiveness, in a randomized trial, of glass-ionomer-based and resin sealant materials after 4 yr.

This study, conducted from a government program perspective, compared the incremental cost-effectiveness of oral health interventions, in particular their delivery to underserved populations in whom dental sealants constitute an important, high-yielding complement to toothbrushing in dental-caries prevention. The study data concern the relative cost-effectiveness of three sealant materials in four approaches to prevent cavitated dentine carious lesions in permanent molars in a community intervention trial among school-age children in Wuhan, China. The four approaches were high-viscosity glass-ionomer cement without heat application (HVGIC); high-viscosity glass-ionomer cement with heat application [light-emitting diode (LED) thermocured HVGIC]; glass-carbomer; and composite resin. The costs studied were: cost of sealing permanent molars; adverse event costs for restoring cavitated dentine carious lesions developing within 4 yr in study data; and projections of 1,000 sealants per group. Preventing one more cavitated dentine carious lesion cost US$105 for the study data when comparing HVGIC (n = 405) with composite resin (n = 396) and US$59 per 1,000 sealants in the projections; LED thermocured HVGIC compared with composite resin cost US$115 for one more cavitated lesion and US$52 per 1,000 sealants, respectively. Although more expensive than composite resin, LED thermocured HVGIC was identified as the most cost-effective among the sealant materials studied. Ease of application, minimal technical and infrastructure requirements, and cost-effectiveness make glass-ionomers a practicable option for governments making decisions under economic constraints.

Are Carious Lesions in Previously Sealed Occlusal Surfaces Detected as well on Colour Photographs as by Visual Clinical Examination?

PURPOSE: To compare the level of agreement between carious lesion assessments according to the visual clinical examination and the colour photograph methods. MATERIALS AND METHODS: Data on the presence of enamel/dentin carious lesions in previously sealed occlusal surfaces in first molars were obtained by two trained and calibrated examiners through visual clinical examination and from colour photographs 4 years after sealing. Kappa statistics were applied to calculate agreement between assessment methods. Data analysis was performed using sign, Bowker symmetry and McNemar's tests. RESULTS: The prevalence of dentin carious lesions was very low. The kappa coefficients for detecting enamel/dentin carious lesions using the two assessment methods were 0.65 (CI: 0.56-0.74) for examiner 1 and 0.70 (CI: 0.62-0.78) for examiner 2. Examiner 2 observed more enamel/dentin carious lesions on colour photographs than did examiner 1 (p = 0.008). Sensitivity analyses did not confirm this outcome. CONCLUSIONS: There was no difference in the detection of enamel/dentin carious lesions in previously sealed occlusal surfaces using colour photographs vs visual clinical examination. The colour photograph method is therefore equivalent to the visual clinical examination in detecting enamel/dentin carious lesions. More studies are required.

Oral squamous cancer cell exploits hnRNP A1 to regulate cell cycle and proliferation.

Oral squamous cell carcinoma (OSCC) is a common human malignant tumor with high mortality. So far, the molecular pathogenesis of OSCC remains largely unclear. Heterogeneous nuclear ribonucleoprotein (hnRNP) A1 is an important multi-function splicing factor and closely related to tumorigenesis. hnRNP A1 is overexpressed in various tumors, and promotes aerobic glycolysis and elongation of telomere, but the function of hnRNP A1 in cell cycle and proliferation remains unclear. We found that hnRNP A1 was overexpressed in OSCC tissues, and was required for the growth of OSCC cells. Moreover, hnRNP A1 was highly expressed in the G2/M cell cycle phase. Knockdown of hnRNP A1 induced G2/M arrest. DNA microarray assay result showed that hnRNP A1 regulated the expression of a number of target genes associated with G2/M phase. Moreover, hnRNP A1 controlled the alternative splicing of CDK2 exon 5. These findings suggested that hnRNP A1 plays key roles in the regulation of cell cycle progression and pathogenesis of OSCC.

Comparison of immunological and microbiological characteristics in children and the elderly with or without dental caries.

The purpose of this study was to determine the relationships among early childhood caries (ECC), root caries (RC), the quantity of Streptococcus mutans in saliva, and the concentrations of total and specific secretory IgA (sIgA). Saliva samples were collected from 70 children, 3-4 yr of age, with and without ECC, and from 43 adults, ≥60 yr of age, with and without RC. The decayed, missing, and filled teeth (dmft) and decayed, missing, and filled surfaces (dmfs) scores of each child, and the root decayed and filled teeth (RDFT) and root decayed and filled surfaces (RDFS) scores of each elderly subject, were determined. The S. mutans levels, total sIgA, and specific sIgA against two virulence antigens of S. mutans in saliva were analysed using quantitative real-time PCR (qPCR) and ELISAs. The quantity of S. mutans was significantly higher in caries-positive subjects within the two populations than in the caries-free subjects; and a positive correlation was found between the quantity of S. mutans and the dmft, dmfs, RDFT, and RDFS scores. In addition, the salivary total sIgA was significantly higher in children with severe early childhood caries (SECC) and in the elderly subjects with RC. Moreover, although the S. mutans level was significantly higher, the concentrations of specific sIgA against S. mutans antigens were significantly lower in samples from elderly subjects than in samples from children. These results support the concept that S. mutans is positively associated with ECC and RC. Furthermore, the levels of S. mutans-specific antibodies in saliva are too low to prevent infection with cariogenic bacteria and to inhibit development of ECC and RC.

BMP-2 induction of Dlx3 expression is mediated by p38/Smad5 signaling pathway in osteoblastic MC3T3-E1 cells.

Dlx3 is essential for osteoblast differentiation and bone formation, and its expression is regulated by bone morphogenetic protein-2 (BMP-2). However, the intimate mechanism of BMP-2 regulation of Dlx3 transcription in osteoblasts is still unknown. Considering the important roles of Smad5 and p38 in osteoblast differentiation, we hypothesized that Smad5 and p38 mediated BMP-2-induced Dlx3 transcription in osteoblasts. We found activation of Smad5 and p38 increased the expression of Dlx3, whereas knocking down Smad5 or inactivation of p38 inhibited BMP-2-induced Dlx3 expression. Both Smad5 and p38 were able to activate Dlx3 promoter activity and p38/Smad5 response elements were located from -698 to -368 in Dlx3 promoter. Two Smad5 binding sites (SBEI and SBEII, TGTCT box) were identified in this region by EMSA and ChIP assay. Deletions and mutagenesis study of the Dlx3 promoter region indicated that the TGTCT boxes are crucial for p38/Smad5-induced Dlx3 promoter activity. At last, we found a cross-talk between p38 and Smad5, and that activation of p38 is necessary for BMP-2-induced Smad5 phosphorylation and nuclear translocation. Overall, we provide a novel insight that BMP-2-induced Dlx3 expression is regulated by p38/Smad5 signaling pathway in osteoblasts.

Intranasal co-delivery of IL-6 gene enhances the immunogenicity of anti-caries DNA vaccine.

AIM: To investigate the effects of co-delivering IL-6 expressing plasmid pCI-IL-6 on the immunogenicity of the anti-caries DNA vaccine pCIA-P, which encodes the surface protein antigen PAc of Streptococcus mutans. METHODS: Plasmid pCI-IL-6 was constructed by inserting the murine IL-6 gene into the pCI vector. Expression of IL-6 in vitro was assessed using Western blot analysis. BALB/c mice were intranasally co-immunized with pCIA-P plus pCI-IL-6 on d 0 and 14. Anti-PAc IgG and secretory IgA (sIgA) were assessed by ELISA. Splenocytes from the mice were re-stimulated with the PAc protein, and IFN-γ and IL-4 production was measured using ELISA. Splenocyte proliferation was analyzed with flow cytometry. Rats were similarly immunized, and dental caries scores were determined using the Keyes method. RESULTS: Marked expression of IL-6 was found in COS-7 cells transfected with pCI-IL-6. In the pCI-IL-6 co-immunized mice, the specific IgG antibodies in serum and sIgA antibodies in saliva were significantly higher than those in the control mice at weeks 4 and 8. Moreover, the secretion of IFN-γ from splenocytes in response to re-stimulation with PAc protein was significantly higher in the pCI-IL-6 co-immunized mice than that in the control mice, whereas the secretion of IL-4 had no significant difference. The proliferation of splenocytes from the pCI-IL-6 co-immunized mice was significantly higher than that from the mice immunized with pCIA-P and pCI vector. In the rat caries model, the pCI-IL-6 co-immunization rats displayed lower caries scores than the control rats. CONCLUSION: Intranasal co-delivery of IL-6 gene significantly enhances the immunogenicity of the anti-caries DNA vaccine.

Methods and preliminary findings of a cost-effectiveness study of glass-ionomer-based and composite resin sealant materials after 2 yr.

The cost-effectiveness of glass-carbomer, conventional high-viscosity glass-ionomer cement (HVGIC) [without or with heat (light-emitting diode (LED) thermocuring) application], and composite resin sealants were compared after 2 yr in function. Estimated net costs per sealant were obtained from data on personnel time (measured with activity sampling), transportation, materials, instruments and equipment, and restoration costs for replacing failed sealants from a community trial involving 7- to 9-yr-old Chinese children. Cost data were standardized to reflect the placement of 1,000 sealants per group. Outcomes were the differences in the number of dentine caries lesions that developed between groups. The average sealant application time ranged from 5.40 min (for composite resin) to 8.09 min (for LED thermocured HVGIC), and the average cost per sealant for 1,000 performed per group (simulation sample) ranged from $US3.73 (for composite resin) to $US7.50 (for glass-carbomer). The incremental cost-effectiveness of LED thermocured HVGIC to prevent one additional caries lesion per 1,000 sealants performed was $US1,106 compared with composite resin. Sensitivity analyses showed that differences in the cost of materials across groups had minimal impact on the overall cost. Cost and effectiveness data enhance policymakers' ability to address issues of availability, access, and compliance associated with poor oral-health outcomes, particularly when large numbers of children are excluded from care, in economies where oral health services are still developing.

Push-out bond strength of self-adhesive methacrylate resin-based sealers to root dentin.

This study examined the adhesive strength of two self-adhesive methacrylate resin-based sealers (MetaSEAL and RealSeal SE) to root dentin and compared them with RealSeal and AH Plus in properties. A total of 48 extracted human single-rooted teeth were used to prepare the 0.9-mm thick longitudinal tooth slice (each per tooth). Standardized simulated canal spaces of uniform dimensions were prepared in the middle of radicular dentin. After treated with 5.25% sodium hypochlorite (NaOCl) and 17% EDTA, tooth slices were allocated randomly to four groups (n=12) in terms of different sealers used: MetaSEAL, RealSeal SE, RealSeal, and AH plus groups. The simulated canal spaces were obturated with different sealers in each group. There were 10 slabs with 20 simulated canal spaces (n=20) used in each group for push-out testing. The failure modes and the ultrastructures of fractured sealer-dentin interfaces were examined. The remaining 2 slabs in each group underwent partial demineralization for observation of the ultrastructure of resin tags. The results showed that the push-out bond strength was 12.01±4.66 MPa in MetaSEAL group, significantly higher than that in the other three groups (P<0.05). Moreover, no statistically significant differences were noted in the push-out bond strength between RealSeal SE (5.43±3.68 MPa) and AH Plus (7.34±2.83 MPa) groups and between RealSeal SE and RealSeal (2.93±1.76 MPa) groups (P>0.05). Mixed failures were predominant in the fractured sealer-dentin interfaces in MetaSEAL and AH Plus groups, while adhesive failures were frequently seen in RealSeal SE and RealSeal groups. In conclusion, after complete removal of the smear layer, MetaSEAL showed superior bond ability to root dentin. The RealSeal SE is applicable in clinical practice, with its adhesive strength similar to that of AH Plus. The self-adhesive methacrylate resin-based sealer holds promise for use in endodontic treatment.

Anti-caries DNA vaccine-induced secretory immunoglobulin A antibodies inhibit formation of Streptococcus mutans biofilms in vitro.

AIM: To investigate the effects of anti-caries DNA vaccine-induced salivary secretory immunoglobulin A (S-IgA) antibodies on Streptococcus mutans (S. mutans) adherence and biofilms formation in vitro. METHODS: Adult female Wistar rats were intranasally immunized with the anti-caries DNA vaccine pGJA-P/VAX. Their saliva samples were collected at different times after the immunization, and S-IgA antibody level in the saliva and its inhibition on S. mutans adherence were examined. The effects of S-IgA in the saliva with the strongest inhibitory effects were examined at 3 different stages, ie acquired pellicles, biofilm formation and production of mature biofilms. The number of viable bacteria and depth of the biofilm at 16 h in each stage were determined using counting colony forming units and using a confocal laser scanning microscopy (CLSM). The participation of S-IgA in acquired pellicles and its aggregation with S. mutans were also observed under CLSM. RESULTS: The S-IgA titer in saliva reached its peak and exhibited the strongest inhibition on S. mutans adhesion at 10 weeks after the immunization. The colonies and depth of the biofilm in the saliva-pretreated group were 41.79% and 41.02%, respectively, less than the control group. The colonies and depth of the biofilm in the co-culture group were 27.4% and 22.81% less than the control group. The assembly of S. mutans and S-IgA was observed under CLSM after co-cultivation. In the mature-stage biofilm, no differences were observed between the different groups. CONCLUSION: These results demonstrate that the anti-caries DNA vaccine induces the production of specific S-IgA antibodies that may prevent dental caries by inhibiting the initial adherence of S. mutans onto tooth surfaces, thereby reducing the accumulation of S. mutans on the acquired pellicles.

Co-delivery of ccl19 gene enhances anti-caries DNA vaccine pCIA-P immunogenicity in mice by increasing dendritic cell migration to secondary lymphoid tissues.

AIM: To investigate how co-delivery of the gene encoding C-C chemokine ligand-19 (CCL-19) affected the systemic immune responses to an anti-caries DNA vaccine pCIA-P in mice. METHODS: Plasmid encoding CCL19-GFP fusion protein (pCCL19/GFP) was constructed by inserting murine ccl19 gene into GFP-expressing vector pAcGFP1-N1. Chemotactic effect of the fusion protein on murine dendritic cells (DCs) was assessed in vitro and in vivo using transwell and flow cytometric analysis, respectively. BALB/c mice were administered anti-caries DNA vaccine pCIA-P plus pCCL19/GFP (each 100 µg, im) or pCIA-P alone. Serum level of anti-PAc IgG was assessed with ELISA. Splenocytes from the mice were stimulated with PAc protein for 48 h, and IFN-γ and IL-4 production was measured with ELISA. The presence of pCCL19/GFP in spleen and draining lymph nodes was assessed using PCR. The expression of pCCL19/GFP protein in these tissues was analyzed under microscope and with flow cytometry. RESULTS: The expression level of CCL19-GFP fusion protein was considerably increased 48 h after transfection of COS-7 cells with pCCL19/GFP plasmids. The fusion protein showed potent chemotactic activity on DCs in vitro. The level of serum PAc-specific IgG was significantly increased from 4 to 14 weeks in the mice vaccinated with pCIA-P plus pCCL19/GFP. Compared to mice vaccinated with pCIA-P alone, the splenocytes from mice vaccinated with pCIA-P plus pCCL19/GFP produced significantly higher level of IFN-γ, but IL-4 production had no significant change. Following intromuscular co-delivery, pCCL19/GFP plasmid and fusion protein were detected in the spleen and draining lymph nodes. Administration of CCL19 gene in mice markedly increased the number of mature DCs in secondary lymphoid tissues. CONCLUSION: CCL19 serves as an effective adjuvant for anti-caries DNA vaccine by inducing chemotactic migration of DCs to secondary lymphoid tissues.

A new gcrR-deficient Streptococcus mutans mutant for replacement therapy of dental caries.

BACKGROUND: gcrR gene acts as a negative regulator related to sucrose-dependent adherence in S. mutans. It is constructive to test the potential capacity of mutans with gcrR gene deficient in bacteria replacement therapy. METHODS: In this study, we constructed the mutant by homologous recombination. The morphological characteristics of biofilms were analyzed by confocal laser scanning microscopy. S. mutans UA159 and the mutant MS-gcrR-def were inoculated, respectively, or together for competitive testing in vitro and in rat model. RESULTS: Adhesion assay showed that the adhesion ability of the mutant increased relative to the wild type, especially in the early stage. MS-gcrR-def out-competed S. mutans UA159 in vitro biofilm, and correspondingly coinfection displayed significantly fewer caries in vivo. The former possessed both a lower level of acid production and a stronger colonization potential than S. mutans UA159. CONCLUSION: These findings demonstrate that MS-gcrR-def appears to be a good candidate for replacement therapy.

Effect of different protocols for treating cavities in primary molars on the quality of life of children in Brazil--1 year follow-up.

The aim of this study was to test the hypothesis that the conventional restorative treatment (CRT) and the atraumatic restorative treatment (ART) protocols, in comparison with the ultra-conservative treatment (UCT) protocol, would increase the quality of life of children over a period of 1 year. Cavitated primary molars of 302 children 6-7 years of age were treated according to the CRT, ART and UCT protocols at the school compound. Children's parents completed the Brazilian version of the Early Childhood Oral Health Impact Scale (B-ECOHIS) at baseline and one year later. Paired t-test, Chi-square test and ANOVA were applied in analysing the data. Questionnaires from 277 and 160 children were collected at baseline and after 1 year, respectively. A statistically significant difference in B-ECOHIS scores over the 1-year period was found for domains 'child symptoms' (P = 0.03) and 'child psychology' (P = 0.02). Treatment protocols did not statistically significantly influence the changes in B-ECOHIS scores over the 1-year period (P = 0.78). It can be concluded that the UCT protocol was as good as the two restorative protocols. All treatment protocols were effective in reducing children's experience of pain, their sleeping problems and their irritability and/or frustration levels over the 1-year period.

What happens to cavitated primary teeth over time? A 3.5-year prospective cohort study in China.

OBJECTIVES: Data showed that among 5-year-old Chinese, 96.7% of cavitated primary teeth were left untreated. The study aimed to report on the course of cavitated primary teeth within the Chinese health-care system over a period of 3.5 years. METHODS: Selection of high caries risk children for inclusion in a sealant comparison study was based on the presence of cavitated dentine lesions in their primary teeth. At the 6-month sealant evaluation point many of these cavitated dentine lesions had not been treated. This necessitated monitoring these cavitated teeth 6-monthly for those exfoliated, restored, with a cavity left open, having caused toothache (symptom) and having (or having had) an abscess or fistulae (symptom). Care-seeking instruction was given at every evaluation point. anova and t-test were used in analysing the data. RESULTS: A total of 1012 cavitated primary teeth in 305 children (7.6 to 9.3 years old), were followed for 3.5 years. A total of 92.9% of cavitated primary teeth were left open, while 7.1% were restored at some stage during the observation period; 98.5% of restored teeth and 95.5% of cavitated teeth left open exfoliated and 93.9% of restored teeth and 81.5% of cavitated primary teeth left open exfoliated without any symptoms. Having (or having had) toothache was the symptom most frequently related to exfoliated restored teeth and to exfoliated cavitated teeth left open. Restored primary teeth survived statistically significantly longer than cavitated primary teeth left open: 1.99 ± 0.07 years and 1.68 ± 0.03 years, respectively. CONCLUSION: The large majority of cavitated primary teeth in this child population exfoliated without symptoms.