Donald R. Schwass

A System of Calibrating Microtomography for Use in Caries Research

Desktop microcomputed tomography (micro-CT) offers a non-invasive 3-dimensional analysis of structures and their physical properties. To date, the use of micro-CT has mostly involved qualitative observations, with the extent of quantitative analysis relying on automated internal calibration by the micro-CT control software. However, the value of such calibration is limited by machine drift. For an accurate quantitative use of micro-CT, it is recognized that external means of calibration are needed. A novel system of calibration standards, also known as ‘phantoms’, is presented. A range of low mineral concentration phantoms involving triethylene glycol dimethacrylate/glycerolate dimethacrylate resin mixed with commercial pure hydroxyapatite (HAP), from 0.07 to 1.05 g/cm3, was fabricated. Sintered HAP was impregnated with the same resin, producing phantoms with medium-level mineral concentrations up to 1.90 g/cm3. These phantoms were easy to create, proved accurate and stable with repeated use, and were found to mimic the composite nature of dental enamel and dentine structures under investigation.

Evaluating the efficiency of caries removal using an Er:YAG laser driven by fluorescence feedback control

INTRODUCTION Caries lesions in dental hard tissues autofluoresce when exposed to light of certain wavelengths, whereas sound tissues do not, and this can be used as an in vitro histological marker for dental caries. Detection of autofluorescence is the basis of KaVo DIAGNOdent™ technology, and provides objective feedback control of laser-stimulated ablation of dental caries for the KaVo Key Laser 3™. This Er:YAG laser operates at 2940nm wavelength, and is effective at removal of infected dental hard tissues. Micro-computed tomography (micro-CT) allows the non-invasive investigation of three-dimensional structures and analysis of mineral density profiles of dentine following laser ablation. OBJECTIVE To evaluate removal of infected, demineralised dentine by Er:YAG irradiation with a laser feedback mechanism, using micro-CT. DESIGN 27 carious teeth (1 control) and 1 sound tooth, treated with the KaVo Key Laser 3™ using a KaVo™ non-contact 2060 handpiece at specific feedback settings, were examined using a Skyscan 1172 Micro-CT, to observe the efficiency of demineralised dentine removal. Grey scale images obtained were colour rendered to assist detection of demineralised tissue if present. RESULTS Complete removal of demineralised tissue occurred with laser-stimulated ablation under feedback control at values of 7 and 8 when measured by micro-CT. At greater values, removal of demineralised dentine was incomplete. CONCLUSION Examination of dental tissues by micro-CT allowed determination of the efficiency of Er:YAG laser-stimulated ablation. Feedback control of the KaVo Key Laser 3™ appeared to operate like a cut-off switch when infected dentine was eliminated, at a threshold of between 6 and 7.

Towards a tunable microemulsion method for nanoparticle synthesis

We present a new twist on a conventional nanoparticle preparation method, which allows the one-pot synthesis of two fractions of size-controlled, spherical, monodisperse silver nanoparticles (Ag NPs), one fraction of which can be suspended in organic solvent, and the other can be suspended in aqueous or other polar solvents, with no significant change to the core diameter of the suspended particles. Furthermore, we demonstrate that the careful selection of capping agents enables the tailoring of the NP surface and optical properties and controls the extent of partitioning between the organic and aqueous phases. This study can serve as a guide for the preparation of NPs with independent control over particle size, composition, surface coating and dispersing medium, thus allowing the contributions of each of these factors to the NP properties to be isolated and investigated individually.

Use of Micro-Computed Tomography for Dental Studies in Modern and Fossil Odontocetes: Potential Applications and Limitations

Teeth are important elements in studies of modern and fossil Cetacea (whales, dolphins), providing information on feeding habits, estimations of age and phylogenetic relationships. The growth layer groups (GLGs) recorded in dentine have demonstrated application for aging studies, but also have the potential to elucidate life history phenomena such as metabolic or physiologic events. Micro-Computed Tomography (Micro-CT) is a non-invasive and non-destructive technique that allows 3-dimensional study of mineralized tissues, such as human teeth, and their physical properties. Teeth from extant dolphins (Cetacea: Odontoceti) and some fossil odontocetes were scanned in a Skyscan 1172 Micro-CT desktop system. X-rays were generated at 100 kV and 100 µA for extant samples, and at 80kV and 124 µA for fossils. 0.5 mm thick aluminum and copper filters were used in the beam. Reconstructed images were informative for most extant species, showing a good resolution of the enamel layer, dentine and pulp cavity. Greyscale changes in the dentinal layers were not resolved enough to show GLGs. Visualization of the internal structure in fossil cetacean teeth depended on the degree of diagenetic alteration in the specimen; undifferentiated enamel and dentine regions probably reflect secondary mineralization. However, internal details were finely resolved for one fossil specimen, showing the enamel, internal layers of dentine and the pulp cavity. Micro-CT has been proven to be a useful tool for resolving the internal morphology of fossil and extant teeth of cetaceans before they are sectioned for other morphological analyses; however some methodological refinements are still necessary to allow better resolution of dentine for potential application in non-destructive age determination studies.

Antimicrobial Activity of a Colloidal AgNP Suspension Demonstrated In Vitro against Monoculture Biofilms: Toward a Novel Tooth Disinfectant for Treating Dental Caries

A novel silver nanoparticle (AgNP) formulation was developed as a targeted application for the disinfection of carious dentine. Silver nitrate (AgNO3) was chemically reduced using sodium borohydrate (NaBH4) in the presence of sodium dodecyl sulfate (SDS) to form micelle aggregate structures containing monodisperse 6.7- to 9.2-nm stabilized AgNPs. AgNPs were characterized by measurement of electrical conductivity and dynamic light scattering, scanning electron microscopy, transmission electron microscopy, and inductively coupled plasma mass spectrometry. Antimicrobial activity of AgNPs was tested against planktonic cultures of representative gram-positive and gram-negative oral bacteria using well diffusion assays on tryptic soy broth media and monoculture biofilms grown with brain heart infusion ± sucrose anaerobically at 37°C on microtiter plates. Biofilm mass was measured by crystal violet assay. Effects were compared to silver diamine fluoride and chlorhexidine (negative controls) and 70% isopropanol (positive control) exposed cultures. In the presence of AgNPs, triplicate testing against Streptococcus gordonii DL1, C219, G102, and ATCC10558 strains; Streptococcus mutans UA159; Streptococcus mitis I18; and Enterococcus faecalis JH22 for planktonic bacteria, the minimum inhibitory concentrations were as low as 7.6 µg mL–1 and the minimum bacteriocidal concentrations as low as 19.2 µg mL–1 silver concentration. Microplate readings detecting crystal violet light absorption at 590 nm showed statistically significant differences between AgNP-exposed biofilms and where no antimicrobial agents were used. The presence of sucrose did not influence the sensitivity of any of the bacteria. By preventing in vitro biofilm formation for several Streptococcus spp. and E. faecalis, this AgNP formulation demonstrates potential for clinical application inhibiting biofilms.

Surface Characteristics and Biofilm Development on Selected Dental Ceramic Materials

Background Intraoral adjustment and polishing of dental ceramics often affect their surface characteristics, promoting increased roughness and consequent biofilm growth. This study correlated surface roughness to biofilm development with four commercially available ceramic materials. Methods Four ceramic materials (Vita Enamic®, Lava™ Ultimate, Vitablocs Mark II, and Wieland Reflex®) were prepared as per manufacturer instructions. Seventeen specimens of each material were adjusted and polished to simulate clinical intraoral procedures and another seventeen remained unaltered. Specimens were analysed by SEM imaging, confocal microscopy, and crystal violet assay. Results SEM images showed more irregular surface topography in adjusted specimens than their respective controls. Surface roughness (Ra) values were greater in all materials following adjustments. All adjusted materials with the exception of Vitablocs Mark II promoted significantly greater biofilm growth relative to controls. Conclusion Simulated intraoral polishing methods resulted in greater surface roughness and increased biofilm accumulation.

Beam hardening correction tool improves the diagnosis of incipient caries lesions in Micro-CT images

OBJECTIVE To evaluate the effect of Beam Hardening Correction tool (BHC) on Micro-CT images used in the diagnosis of incipient caries lesions. METHODS Six posterior teeth, with 6 sound proximal surfaces and 6 natural white spot lesions, were scanned using a SkyScan1174 device. Each tooth was imaged 16 times in various acquisition parameters, and reconstructed 3 times applying different levels of BHC (0, 15 and 30%). The 288 datasets were analyzed by three examiners. Gold-standard was obtained by microscopic analyses under polarized light. Area under the ROC curve was obtained for each protocol and the BHC parameter was isolated to individually observe the influence of such parameters of reconstruction. RESULTS Microtomographic images were accurate for the diagnosis of incipient caries lesions even when the BHC tool was not applied. Accuracy was perfect in 50 % of the protocols without BHC. When adding the artefact correction, the number of protocols that reached perfect accuracy increased to 81.25%. No difference was found between the protocols that used 15 or 30% BHC. Statistically significant differences were found only in four comparisons, and on those the images that had BHC were more accurate. Two non-BHC protocols reached accuracy scores of 0.653 and 0.881, and were improved to perfect accuracy with BHC application. CONCLUSION The use of the BHC tool should be encouraged when using Micro-CT to detect incipient caries lesions, especially when low-quality parameters aiming less acquisition and processing time are used.

Microcomputed Tomography Evaluation of Dentine Mineral Concentration in Primary Molars Managed by Three Treatment Protocols

The objectives of the study were to quantify the dentine mineral concentration (DMC) in teeth restored conventionally, according to the atraumatic restorative treatment (ART) and ultraconservative (UCT) protocols (open cavities and small ART restorations), and the DMC underneath the open cavities of teeth managed by UCT versus nontreated, open cavities. We studied 50 teeth with restorations/open cavities, 39 restored teeth (9 by conventional restorative treatment [CRT], 17 by ART, and 13 by UCT) and 16 teeth with open cavities. Each restoration/open cavity was scanned using microcomputed tomography, with 3 hydroxyapatite disks with respective densities of 1.24, 1.33, and 1.57 g/cm3 as a reference. Images were reconstructed and the greyscale images were converted into DMC values. For each restoration/open cavity, 15 measurements of dentine immediately underneath and from the corresponding area in sound dentine were taken. DMC was expressed as a percentage of the DMC of sound dentine. ANOVA and the Student t test were used for statistical analysis. The mean DMC underneath restorations of the ART protocol group (98.93%) was statistically significantly higher than that of the UCT protocol group (91.98%), but not of the CRT protocol group (91.33%). On multiple surfaces, mean DMC in the axial area (94.32%) was statistically significantly higher than in the gingival area (92.80%). The mean DMC of open cavities managed by UCT protocol (89.05%) was statistically significantly higher than in nontreated open cavities (83.90%). In conclusion, a dentine-hypermineralized area underneath ART restorations was observed. Managing open cavities with a toothbrush and fluoride toothpaste (the UCT protocol) resulted in higher mineralized dentine underneath the cavity than in nontreated open cavities.

Resin Infiltration of Caries in Primary Molars in a Community Setting: 24-Month Randomized Controlled Trial Findings

This study aimed to assess whether resin infiltration of primary molar proximal lesions is more effective than noninvasive measures in radiographically controlling carious lesion progression into the dentin. A split-mouth randomized controlled trial included 90 children, each with 2 proximal lesions confined to the inner half of the enamel or ≤0.5 mm into the dentin. For each child, lesions were randomly allocated to test (infiltration: DMG Icon preproduct and fluoride varnish) or control (fluoride varnish) status. The primary outcome measure was 24-mo radiographic lesion progression. Placement of a restoration during the study period was counted as lesion progression. Proportions of teeth with progressed lesions were compared using the McNemar test. Children also reported on the treatment’s acceptability to them. Children (46% female) ranged in age from 6 to 9 y. Their mean number of decayed, missing, and filled teeth (d3mft) was 2.8 (SD 2.6). At baseline, 58% and 42% of children were at moderate and low risk, respectively. Test and control lesions presented with similar radiographic lesions at baseline. At the 24-mo follow-up, 6 children had moved and 30 teeth had exfoliated. In the test and control groups, 15 of 66 lesions (22.7%) and 30 of 69 lesions (43.5%) had progressed, respectively (P < 0.05). The 2-y therapeutic effect (based on pairwise radiographic readings) of infiltration over fluoride varnish was 20.8% (95% confidence interval, 10.6%−30.2%). Nearly all children (96.7%) had enjoyed their visit to the clinic, and more than two-thirds (72.2%) were not worried about returning for treatment. Infiltration is more efficacious than fluoride varnish for controlling carious lesion progression in proximal lesions in primary molars, and most children find the treatment acceptable (Australian New Zealand Clinical Trials Registry ANZCTR.org.au ACTRN12611000827932). Knowledge Transfer Statement: These study findings can help clinicians decide which caries management approach they wish to use to prevent progression of proximal lesions in primary molars. With consideration of cost and patient preference, this information could lead to more appropriate therapeutic decisions.