Journal of Dental Research

Nanoparticles are normally considered to be of a size no greater than 100 nm, and the exploitation of their unique attributes to combat infection has increased markedly over the past decade. The potential of nanoparticles to control the formation of biofilms within the oral cavity, as a function of their biocidal, anti-adhesive, and delivery capabilities, is now coming under close scrutiny. Possible uses as constituents of prosthetic device coatings, as topically applied agents, and within dental materials are being explored. The latest insights into the application of nanoparticles in the control of oral infections, including their use in photodynamic therapy, will be discussed in this review. In particular, the use of nanoparticulate silver, copper, zinc, silicon, and their oxides will be considered in relation to their effects on bacterial populations. The recent interest in the applications of nanoparticulate polymers and calcium phosphates will also be assessed. Particular attention will be paid to the toxicity issues surrounding the potential impact of nanoparticles on oral and other tissues.

Dental caries is a highly prevalent diet-related disease and is a major public health problem. A goal of modern dentistry is to manage non-cavitated caries lesions non-invasively through remineralization in an attempt to prevent disease progression and improve aesthetics, strength, and function. Remineralization is defined as the process whereby calcium and phosphate ions are supplied from a source external to the tooth to promote ion deposition into crystal voids in demineralized enamel, to produce net mineral gain. Recently, a range of novel calcium-phosphate-based remineralization delivery systems has been developed for clinical application. These delivery systems include crystalline, unstabilized amorphous, or stabilized amorphous formulations of calcium phosphate. These systems are reviewed, and the technology with the most scientific evidence to support its clinical use is the remineralizing system utilizing casein phosphopeptides to stabilize and deliver bioavailable calcium, phosphate, and fluoride ions. The recent clinical evidence for this technology is presented and the mechanism of action discussed. Biomimetic approaches to stabilization of bioavailable calcium, phosphate, and fluoride ions and the localization of these ions to non-cavitated caries lesions for controlled remineralization show promise for the non-invasive management of dental caries.

Dental caries is influenced by a complex interplay of genetic and environmental factors, including dietary habits. Previous reports have characterized the influence of genetic variation on taste preferences and dietary habits. We therefore hypothesized that genetic variation in taste pathway genes (TAS2R38, TAS1R2, GNAT3) may be associated with dental caries risk and/or protection. Families were recruited by the Center for Oral Health Research in Appalachia (COHRA) for collection of biological samples, demographic data, and clinical assessment of oral health, including caries scores. Multiple single-nucleotide polymorphism (SNP) assays for each gene were performed and analyzed by transmission disequilibrium test (TDT) analysis (FBAT software) for three dentition groups: primary, mixed, and permanent. Statistically significant associations were seen in TAS2R38 and TAS1R2 for caries risk and/or protection.

Previous linkage analysis of an Asian population proposed possible candidate genes for mandibular prognathism, such as Matrilin-1 (cartilage matrix protein). To investigate the association between the single-nucleotide polymorphisms (SNPs) in Matrilin-1 and mandibular prognathism, we investigated three sequence variants (-158 T>C, 7987 G>A, 8572 C>T) in 164 mandibular prognathism patients and 132 control individuals with a normal occlusion. The results showed that the 8572 TT genotypes in Matrilin-1 showed increased risk of mandibular prognathism (OR = 9.28, 95% Cl = 1.19~197.57, P < 0.05), whereas the 7987 AA genotype showed a protective effect for mandibular prognathism (OR = 0.16, 95% Cl = 0.05~0.47, P < 0.05). Genotyping results showed that the Matrilin-1 polymorphism haplotype TGC (ht4; 158T, 7987G, and 8572C alleles) had a pronounced risk effect for mandibular prognathism compared with controls (OR = 5.16, 95% Cl = 2.03~13.93, P < 0.01). The results suggest that polymorphisms in Matrilin-1 could be used as a marker for genetic susceptibility to mandibular prognathism.

This study evaluates the accuracy of periodontitis prevalence determined by the National Health and Nutrition Examination Survey (NHANES) partial-mouth periodontal examination protocols. True periodontitis prevalence was determined in a new convenience sample of 454 adults ≥ 35 years old, by a full-mouth “gold standard” periodontal examination. This actual prevalence was compared with prevalence resulting from analysis of the data according to the protocols of NHANES III and NHANES 2001-2004, respectively. Both NHANES protocols substantially underestimated the prevalence of periodontitis by 50% or more, depending on the periodontitis case definition used, and thus performed below threshold levels for moderate-to-high levels of validity for surveillance. Adding measurements from lingual or interproximal sites to the NHANES 2001-2004 protocol did not improve the accuracy sufficiently to reach acceptable sensitivity thresholds. These findings suggest that NHANES protocols produce high levels of misclassification of periodontitis cases and thus have low validity for surveillance and research.

Concentrated tin- and fluoride-containing mouthrinses are effective erosion inhibitors in enamel and dentin. To test whether this is also true for solutions with lower concentrations, we conducted a randomized double-blind three-cell crossover in situ study with extra-orally performed erosive impacts (citric acid, 6 x 5 min/day) and an intra-oral rinsing protocol (1 x 30 sec/day) in 24 volunteers. The mouthrinses were a placebo, a NaF (500 ppm F^-), and an amine fluoride (AmF)/NaF/SnCl₂ mouthrinse (500 ppm F-, 800 ppm Sn²⁺). Compared with the placebo, the NaF mouthrinse reduced substance loss by 19% in enamel and 23% in dentin (p ≤ 0.01 each); the AmF/NaF/SnCl₂ mouthrinse reduced this parameter by 67% in enamel and 47% in dentin (p ≤ 0.001 each). AmF/NaF/SnCl₂ was significantly more effective than NaF in both tissues (p ≤ 0.01). The mouthrinse containing Sn and F exhibited good efficacy, even under severe erosive conditions.

Municipal water fluoridation has notably reduced the incidence of dental caries and is widely considered a public health success. However, ingested fluoride is sequestered into bone, as well as teeth, and data on the long-term effect of exposure to these very low doses of fluoride remain inconclusive. Epidemiological studies suggest that effects of fluoride on bone are minimal. We hypothesized that the direct measurement of bone tissue from individuals residing in municipalities with and without fluoridated water would reveal a relationship between fluoride content and structural or mechanical properties of bone. However, consonant with the epidemiological data, only a weak relationship among fluoride exposure, accumulated fluoride, and the physical characteristics of bone was observed. Analysis of our data suggests that the variability in heterogenous urban populations may be too high for the effects, if any, of low-level fluoride administration on skeletal tissue to be discerned.

Frequent consumption of cariogenic foods and bacterial infection are risk factors for early childhood caries (ECC). This study hypothesized that a short diet survey focused on frequency of foods, categorized by putative cariogenicity, would differentiate severe ECC (S-ECC) from caries-free children. Children’s diets were obtained by survey and plaque bacteria detected by PCR from 72 S-ECC and 38 caries-free children. S-ECC children had higher scores for between-meal juice (p < 0.01), solid-retentive foods (p < 0.001), eating frequency (p < 0.005), and estimated food cariogenicity (p < 0.0001) than caries-free children. S-ECC children with lesion recurrence ate fewer putative caries-protective foods than children without new lesions. Streptococcus mutans (p < 0.005), Streptococcus sobrinus (p < 0.005), and Bifidobacteria (p < 0.0001) were associated with S-ECC, and S. mutans with S. sobrinus was associated with lesion recurrence (p < 0.05). S. mutans-positive children had higher food cariogenicity scores. Food frequency, putative cariogenicity, and S. mutans were associated with S-ECC individually and in combination.

Few studies have examined, comprehensively and prospectively, determinants of oral-health-related quality of life. The aim of this study was to examine the relationships between psychosocial factors and oral health status, health perceptions, and quality of life. Measures of symptom and functional status, health perceptions, quality of life, oral health beliefs, and psychological (sense of coherence, self-esteem, health locus of control) and social factors (parents’ income and education) were collected from 439 12- and 13-year-olds at baseline and six-month follow-up, together with a clinical examination at baseline. Structural equation modeling indicated that increased levels of caries and more symptoms predicted more functional limitations, and, cross-sectionally, greater functional impact was associated with worse health perceptions, which were linked to lower quality of life. Sense of coherence was the most important psychosocial predictor. These factors are important in understanding how oral health affects young people’s daily lives.

Self-reported cognitive symptoms are frequent in persons with amalgam-related complaints, but few studies have focused on their cognitive function. The aim was to examine a symptom profile and whether participants with amalgam-related complaints have cognitive deficits in comparison with control individuals. We drew 342 participants with amalgam-related complaints and 342 one-to-one matched control individuals from a longitudinal population-based study. For 81 of the participants with amalgam-related complaints and controls, data were available approximately five years before the onset of complaints, making a longitudinal analysis possible. All participants were assessed by a self-reported health questionnaire and a comprehensive cognitive test battery. The participants with amalgam-related complaints reported more symptoms, mainly musculoskeletal and neuropsychological, compared with control individuals (p < 0.001). The results revealed no significant difference between the amalgam and control group, either cross-sectionally or longitudinally, for any of the cognitive tests. These results suggest that cognitive decline is not associated with amalgam-related complaints.

Periodontal infections have been associated with a state of chronic inflammation. To ascertain whether severe periodontitis and its treatment are associated with oxidative stress, we recruited 145 cases (periodontitis) and 56 controls in a case-control study. A further pilot intervention study of 14 cases (periodontal therapy) was performed. Blood samples were taken at baseline (case-control) and 1, 3, 5, 7, and 30 days after treatment (intervention). Diacron-reactive oxygen metabolites (D-ROM), anti-oxidant potential, C-reactive protein (CRP), interleukin-6, and lipid profiles were determined with high-sensitivity assays in serum. Patients with severe periodontitis exhibited higher D-ROM levels (P < 0.001) and lower total anti-oxidant capacity (P < 0.001) compared with healthy control individuals. These findings were independent of age, gender, smoking habits, ethnicity, and standard lipids differences. D-ROM levels were positively correlated with CRP (R = 0.4, P < 0.001) and clinical periodontal parameters (R = 0.20, P < 0.05). Acute increases of D-ROM (P < 0.01) were observed following periodontal therapy. Analysis of these data suggests a positive association between severe periodontitis and oxidative stress.

The subgingival microbiome is largely uncultivated, and therefore, cultivation-based and targeted molecular approaches have limited value in examining the effect of smoking on this community. We tested the hypothesis that the subgingival biofilm is compositionally different in current and never-smokers by using an open-ended molecular approach for bacterial identification. Subgingival plaque from deep sites of current and never-smokers matched for disease was analyzed by 16S sequencing. Smokers demonstrated greater abundance of Parvimonas, Fusobacterium, Campylobacter, Bacteroides, and Treponema and lower levels of Veillonella, Neisseria, and Streptococcus. Several uncultivated Peptostreptococci, Parvimonas micra, Campy-lobacter gracilis, Treponema socranskii, Dialister pneumosintes, and Tannerella forsythia were elevated in this group, while Veillonella sp. oral clone B2, Neisseria sp. oral clone 2.24, Streptococcus sanguinis, and Capnocytophaga sp. clone AH015 were at lower levels. The microbial profile of smoking-associated periodontitis is distinct from that of non-smokers, with significant differences in the prevalence and abundance of disease-associated and health-compatible organisms.

Although a large body of scientific literature shows that background color and luminance affect color perception, previous measurements of tooth color difference thresholds have not taken the effects of viewing context into account. The present study tested the hypothesis that differences in skin/gingival color influence individuals’ judgments of tooth color differences. Perceptibility and acceptability thresholds were determined in 10 individuals using a signal detection paradigm. They evaluated 500 pseudo-random presentations of two facial portraits: an African-American and a Caucasian. These portraits varied trial-to-trial only in the direction (CIELAB +L*, +a*, or +b*) or magnitude of the color difference between a portrait’s two central incisors. The individuals were significantly less sensitive to tooth color differences in the +L* direction in the Caucasian portrait than for any other combination of color direction or portrait type. Furthermore, comparable perceptibility and acceptability thresholds were generally not statistically significant from each other.

Of the various conservative treatment modalities available for temporomandibular disorders, we believe that therapeutic exercise has a good prognosis, especially for anterior disc displacement without reduction. Since its effectiveness has not been extensively evaluated, we conducted a comparative study to verify the hypothesis that treatment efficacy would not differ for exercise and occlusal splints. Fifty-two individuals with anterior disc displacement without reduction were randomly assigned to a splint or a joint mobilization self-exercise treatment group. Four outcome variables were evaluated: (i) maximum mouth-opening range without and (ii) with pain, (iii) current maximum daily pain intensity, and (iv) limitation of daily functions. All outcome variables significantly improved after 8 weeks of treatment in both groups. In particular, the mouth opening range increased more in the exercise group than in the splint group. This result demonstrates that therapeutic exercise brings earlier recovery of jaw function compared with splints.

In ethanol wet bonding, water is replaced by ethanol to maintain dehydrated collagen matrices in an extended state to facilitate resin infiltration. Since short ethanol dehydration protocols may be ineffective, this study tested the null hypothesis that there are no differences in ethanol dehydration protocols for maintaining the surface roughness, fibril diameter, and interfibrillar spaces of acid-etched dentin. Polished human dentin surfaces were etched with phosphoric acid and water-rinsed. Tested protocols were: (1) water-rinse (control); (2) 100% ethanol-rinse (1-min); (3) 100% ethanol-rinse (5-min); and (4) progressive ethanol replacement (50-100%). Surface roughness, fibril diameter, and interfibrillar spaces were determined with atomic force microscopy and analyzed by one-way analysis of variance and the Student-Newman-Keuls test (α = 0.05). Dentin roughness and fibril diameter significantly decreased when 100% ethanol (1-5 min) was used for rinsing (p < 0.001). Absolute ethanol produced collapse and shrinkage of collagen fibrils. Ascending ethanol concentrations did not collapse the matrix and shrank the fibrils less than absolute ethanol-rinses.

Little information is available about whether the presence of residual caries beneath an occlusal restoration affects fracture strength of the tooth. This in vitro study tested the hypothesis that restored teeth after incomplete excavation have lower fracture strengths than restored teeth after complete excavation. Fourteen pairs of molars were randomly assigned to an experimental (incomplete excavation) or to a control group (complete excavation) and loaded vertically (after cyclic loading). Failure load and fracture mode were recorded. Restored teeth in the incomplete excavation group resulted in reduced fracture strength (p < 0.001) of the tooth-restoration complex (1276 N ± 626 N) compared with the control group (2768 N ± 710 N). Teeth in the complete excavation group all fractured vertically, while in the experimental group, cracks in the restoration were observed, characterized as ‘ice-cracks’. The fracture strength of teeth restored over incomplete caries excavation was significantly reduced, possibly resulting in long-term clinical failure.

To define the optimized material properties of caries-infiltrating resins (infiltrants), we evaluated the effects of infiltrant composition and penetration coefficient (PC) on the inhibition of the progression of natural approximal caries lesions in vitro. Non-cavitated natural enamel lesions were etched (15% HCl gel) and infiltrated for 5 min with experimental resins varying in PC (cm/sec): PC63, PC185, PC204, and PC391. As a positive control (PosC), lesions were covered with flowable composite resin, whereas untreated lesions served as negative controls (NegC). Specimens were exposed to a demineralizing solution (pH 4.95) for 400 days. Progression of mineral loss was analyzed by transverse wavelength-independent microradiography. Percentage progressions of NegC [median (25th/75th percentile), 24 (10/56)], PC63 [9 (3/39)], and PC185 [12 (9/24)] were significantly higher compared with those of PosC [1 (-8/8)], PC204 [-2 (-4/2)], and PC391 [0 (-5/9)] (p < 0.05). Thus, infiltrants with PC > 200 cm/sec are capable of inhibiting lesion progression.

To understand the mechanism of 10-methacryloyloxydecyl dihydrogen phosphate (MDP) hydrolysis, we investigated the degradation of 2-methacryloyloxyethyl dihydrogen phosphate (MEP), because the MEP molecule has the methacryloxy and phosphate ester portions of MDP but, unlike the latter, is water-soluble. The MEP-N-methcryloyl glycine (NMGly), MDP-NMGly, and MDP-2-hydroxyethyl methacrylate (HEMA) primers were designed, stored for different periods, and then analyzed. Our null hypotheses were that (1) the mechanism of MDP hydrolysis differs from that of MEP and (2) the type of hydrophilic monomer—NMGly or HEMA—has no effect on the MDP hydrolysis rate. Similar to the production of methacrylic acid (MA) and 2-hydroxyethyl dihydrogen phosphate (HEP) during MEP hydrolysis, MDP produced MA and 10-hydroxydecyl dihydrogen phosphate (HDP) during hydrolysis. However, the rate of MDP hydrolysis depended on the type of hydrophilic monomer: Compared with HEMA, NMGly significantly increased the rate of MDP hydrolysis.

The continuous growth of rodent incisors requires the presence of stem cells capable of generating ameloblasts and odontoblasts. While epithelial stem cells giving rise to ameloblasts have been well-characterized, cells giving rise to the odontoblasts in incisors have not been fully characterized. The goal of this study was to gain insight into the potential population in dental pulps of unerupted and erupted incisors that give rise to odontoblasts. We show that pulps from unerupted incisors contain a significant mesenchymal-stem-cell (MSC)-like population (cells expressing CD90+/CD45-, CD117+/CD45-, Sca-1+/CD45-) and few CD45+ cells. Our in vitro studies showed that these cells displayed extensive osteo-dentinogenic potential, but were unable to differentiate into chondrocytes and adipocytes. Dental pulps from erupted incisors displayed increased percentages of CD45+ and decreased percentages of cells expressing markers of an MSC-like population. Despite these differences, pulps from erupted incisors also displayed extensive osteo-dentinogenic potential and inability to differentiate into chondrocytes and adipocytes. These results provide evidence that continuous generation of odontoblasts and dentin on the labial and lingual sides of unerupted and erupted incisors is supported by a progenitor population and not multipotent MSCs in the dental pulp.

Although fundamentally similar to other bones, the jaws demonstrate discrete responses to developmental, mechanical, and homeostatic regulatory signals. Here, we hypothesized that rat mandible vs. long-bone marrow-derived cells possess different osteogenic potential. We established a protocol for rat mandible and long-bone marrow stromal cell (BMSC) isolation and culture. Mandible BMSC cultures formed more colonies, suggesting an increased CFU-F population. Both mandible and long-bone BMSCs differentiated into osteoblasts. However, mandible BMSCs demonstrated augmented alkaline phosphatase activity, mineralization, and osteoblast gene expression. Importantly, upon implantation into nude mice, mandible BMSCs formed 70% larger bone nodules containing three-fold more mineralized bone compared with long-bone BMSCs. Analysis of these data demonstrates an increased osteogenic potential and augmented capacity of mandible BMSCs to induce bone formation in vitro and in vivo. Our findings support differences in the mechanisms underlying mandible homeostasis and the pathophysiology of diseases unique to the jaws.

Although the pathogenesis of Streptococcus oralis may be different from that of other viridans group streptococci, S. oralis shares a high degree of DNA sequence similarity with these streptococci. As a result, discrimination of S. oralis from its close relatives has long been considered difficult. This study was conducted to find specific genes that allow for the in vitro identification of S. oralis, but not other oral commensals. Four hundred ninety S. oralis clones obtained by suppressive subtractive hybridization were used for Southern hybridization, and positive clones were sequenced. Of 5 S. oralis-specific clones, newly designed primer sets based on the glucosyltransferase regulatory gene amplified genomic DNA only from S. oralis strains, but not from any of the other 125 strains tested. Our findings may be useful for the future development of efficient diagnostic tools for the rapid identification and differentiation of S. oralis from other oral streptococci strains.

Epigallocatechin gallate (EGCg), the main antimicrobial tea catechin, has been reported to inhibit growth and virulence factors of oral pathogens in vitro. Although the mechanism is unclear, the potential of EGCg in reducing halitosis caused by volatile sulfur compounds (VSCs) has been suggested. This study tested the hypothesis that EGCg reduces VSCs by suppressing mgl, the gene encoding L-methionine-α-deamino-γ-mercaptomethane-lyase, responsible for methyl mercaptan (CH₃SH) production by oral anaerobes. In this study, the effect of EGCg on in vitro growth, CH₃SH production, and mgl gene expression in P. gingivalis W83 was investigated. EGCg inhibited growth of P. gingivalis W83 (MIC = 97.5 µg/mL) and was bactericidal (MBC = 187.5 µg/mL). At sub-MIC levels, EGCg inhibited CH₃SH production, and mgl mRNA and protein expression (p < 0.05). We conclude that EGCg may represent a natural and alternative agent to the antimicrobial chemicals currently available for halitosis control.

We have reported that mustard oil application to the rat dental pulp induces neuronal activation in the thalamus. To address the mechanisms involved in the thalamic changes, we performed neuronal responsiveness recording, immunohistochemistry, and molecular biological analysis. After mustard oil application, neuronal responsiveness was increased in the mediodorsal nucleus. When MK801 (an N-methyl-D-aspartate receptor antagonist) was applied to the mediodorsal nucleus, the enhanced responsiveness was decreased. N-methyl-D-aspartate receptor 2D, glial fibrillary acidic protein, and antigen-presenting cell-related gene mRNAs in the contralateral thalamus were up-regulated at 10 minutes after mustard oil application, but were down-regulated within 10 minutes after the antagonist application. OX6-expressing microglia and glial fibrillary acidic protein-expressing astrocytes did not increase until 60 minutes after mustard oil application. These results suggested that the thalamic neurons play some roles in regulating the glial cell activation in the mediodorsal nucleus via N-methyl-D-aspartate receptor 2D during pulp inflammation-induced central sensitization.

The underlying mechanism and the therapeutic regimen for the transition of reversible gingivitis to irreversible periodontitis are unclear. Since transforming growth factor (TGF)-β has been implicated in differentially regulated gene expression in gingival fibroblasts, we hypothesized that TGF-β signaling is activated in periodontitis-affected gingiva, along with enhanced collagen degradation, that is reversed by TGF-β inhibition. A novel three-dimensional (3D) gel-culture system consisting of primary human gingival fibroblasts (GF) and gingival epithelial (GE) cells in collagen gels was applied. GF populations from patients with severe periodontitis degraded collagen gels, which was reduced by TGF-β-receptor kinase inhibition. Up-regulation of TGF-β-responsive genes was evident in GF/GE co-cultures. Furthermore, the TGF-β downstream transducer Smad3C was highly phosphorylated in periodontitis-affected gingiva and 3D cultures. These results imply that TGF-β signaling is involved in fibroblast-epithelial cell interaction in periodontitis, and suggest that the 3D culture system is a useful in vitro model for therapeutic drug screening for periodontitis.