Charles R. Parkinson

Scanning Electrochemical Microscopy as a Quantitative Probe of Acid-Induced Dissolution: Theory and Application to Dental Enamel

This Article reports the use of scanning electrochemical microscopy (SECM) for the quantitative study of acid-induced dissolution. An ultramicroelectrode (UME) is used to generate a flux of protons galvanostatically just above a sample surface, creating controlled acid challenges relevant to acid erosion. The electrochemical technique produces etch features in the sample, which are characterized by white light interferometry (WLI). The technique has been applied to bovine enamel where understanding the kinetics of dissolution is important in the context of acid erosion. Dissolution has been observed as a fast process, but the high rates of mass transport in SECM allow the surface kinetics of dissolution to be evaluated. Key attributes of SECM for these studies are the ability to deliver high, controllable, and local acid challenges in a defined way and that multiple dissolution measurements can be performed on one sample, eliminating intersample variability effects. A novel moving boundary finite element model has been designed to describe the etching process, which allows the etch kinetics to be evaluated quantitatively, simply by measuring the size and shape of etch features over time.

Initial stages of enamel erosion: An in situ atomic force microscopy study

Time-resolved in situ atomic force microscopy has been employed to examine erosion progression in untreated and fluoride-treated enamel specimens during exposure to citric acid. Contact with the acidic reaction solution initiated the emergence and growth of dissolution pits in both the native and the fluoride-treated enamel. In native enamel, pits are first observed after 90 min exposure to the reaction solution, compared to 250 min in the case of the fluoride-treated enamel. These findings indicate that, within the constraints of this study, a single application of fluoride solution (1000 mg/L, 2 min) confers protection to the enamel surface against acid-mediated erosion. This paper also highlights the potential role intrinsic defects may have on the susceptibility of enamel to erosion and, in part, may explain why some people are more susceptible to acid erosion.

Strontium effects on root dentin tubule occlusion and nanomechanical properties

OBJECTIVES Dentin hypersensitivity often is treated by promotion of dentin tubule occlusion. In this in vitro study we evaluated nanomechanical properties and degree of tubule occlusion conferred to sound and demineralized human root dentin following treatment with a 10% (w/w) strontium acetate solution and its relation to the treatment duration and delivery method. METHODS 24 human cervical root dentin disks (8 groups of 3) were polished through 0.25 μm. 12 disks were subjected to an acid challenge (1% citric acid, pH 3.8) for 2 min. The specimens were incubated in artificial saliva, treated by soaking or brushing with deionized (DI) water or a solution of 10% strontium acetate for 2 min twice a day for 28 days. The occlusion percent and nanomechanical properties were determined at the baseline, 5, 14 and 28 days. Cross-sectioned specimens were prepared to evaluate the depth affected by strontium acetate / dentin interaction by SEM. Statistical analysis was performed using linear mixed effects models. RESULTS A 10% strontium acetate treatment over 5-28 days significantly increased tubule occlusion for normal root dentin and to a lesser extent for demineralized dentin and increased the AFM based nanomechanical properties of demineralized dentin. Brushing was more effective than soaking in recovery of properties of demineralized dentin when treated with strontium. No difference in tubuleocclusion was found between the two delivery methods. SIGNIFICANCE Strontium acetate itself proved to have the ability to occlude dentin tubules and result in small changes in the mechanical properties of dentin.

A randomised clinical trial investigating calcium sodium phosphosilicate as a dentine mineralising agent in the oral environment

OBJECTIVE The ability of a dentifrice containing the bioactive material calcium sodium phosphosilicate (CSPS) to remineralise the surface of dentine and physically occlude patent tubules was investigated in a 20 day in situ randomised clinical study. METHODS Changes in surface microhardness and surface topography of dentine specimens treated for 5, 10, 15 and 20 days, twice daily with either a dentifrice containing 5% CSPS or a fluoride-only containing placebo dentifrice were compared. The substantivity of any mineral deposits formed on the surface of dentine were investigated by the application of an intra-oral dietary acid challenge twice daily during the final 10 days of treatment. RESULTS After 5 and 10 days of treatment, the dentine samples in both treatment groups demonstrated an increase in surface microhardness. After 10 days of treatment the increase in surface hardness was directionally greater for the specimens treated with 5% CSPS dentifrice. Introducing an intra-oral acid exposure resulted in a reduction in surface microhardness which was significantly greater for the specimens treated with the placebo dentifrice compared to the dentifrice containing 5% CSPS, at day 20. Occlusion of the patent tubules was evident at each time-point and was significantly greater for the 5% CSPS containing dentifrice on days 5 and 10. On day 15 both dentifrices demonstrated the same degree of occlusion. CONCLUSION This in situ study demonstrated that dentifrice containing 5% CSPS may have potential to mineralise and occlude the dentine in the oral environment. CLINICAL SIGNIFICANCE This work provides evidence of potential agents that can be used to reduce the pain of dentine hypersensitivity when formulated into dentifrice and applied as part of a normal oral hygiene routine.

Diffusive transport within dentinal tubules: An X-ray microtomographic study

The hydrodynamic theory of dentine hypersensitivity proposes that external stimuli cause dentinal fluid movement within dentinal tubules thereby triggering mechanosensitive nerves and eliciting a pain response. The aim of this study was to employ X-ray microtomography (XMT) to monitor diffusion of caesium acetate through dentine to investigate the extent to which transport occurs within the primary tubules compared to that through branched microtubules believed to run perpendicular to the direction of the primary dentinal tubules. 2.0-mm thick coronal dentine disks masked to leave half of the upper surface exposed were imaged by XMT, initially in water, which was then replaced with an aqueous solution of 0.50 mol l(-1) caesium acetate. Further XMT images were acquired after 1 and 6 days immersion. The XMT images were used to measure the change in the X-ray linear attenuation coefficient resulting from caesium acetate ingress into dentine. There was clearly considerable ingress of caesium acetate into the dentine lying below the exposed surface, but considerably less beneath the sealed surface, suggesting that diffusive transport occurs predominantly in the direction of the primary dentinal tubules, with no significant lateral transport. Primary tubules are clearly the dominant transmission route for triggering the mechanosensitive nerves present at the dentine-pulp interface, and for delivery of nerve desensitising agents.

Measurement of surface roughness changes of unpolished and polished enamel following erosion

Objectives To determine if Sa roughness data from measuring one central location of unpolished and polished enamel were representative of the overall surfaces before and after erosion. Methods Twenty human enamel sections (4x4 mm) were embedded in bis-acryl composite and randomised to either a native or polishing enamel preparation protocol. Enamel samples were subjected to an acid challenge (15 minutes 100 mL orange juice, pH 3.2, titratable acidity 41.3mmol OH/L, 62.5 rpm agitation, repeated for three cycles). Median (IQR) surface roughness [Sa] was measured at baseline and after erosion from both a centralised cluster and four peripheral clusters. Within each cluster, five smaller areas (0.04 mm2) provided the Sa roughness data. Results For both unpolished and polished enamel samples there were no significant differences between measuring one central cluster or four peripheral clusters, before and after erosion. For unpolished enamel the single central cluster had a median (IQR) Sa roughness of 1.45 (2.58) μm and the four peripheral clusters had a median (IQR) of 1.32 (4.86) μm before erosion; after erosion there were statistically significant reductions to 0.38 (0.35) μm and 0.34 (0.49) μm respectively (p<0.0001). Polished enamel had a median (IQR) Sa roughness 0.04 (0.17) μm for the single central cluster and 0.05 (0.15) μm for the four peripheral clusters which statistically significantly increased after erosion to 0.27 (0.08) μm for both (p<0.0001). Conclusion Measuring one central cluster of unpolished and polished enamel was representative of the overall enamel surface roughness, before and after erosion.

Clinical enamel surface changes following an intra-oral acidic challenge

OBJECTIVES Investigation of early enamel erosion using replica impressions to compare changes in enamel surface topography in vivo prior to and over a 24 h period following acid challenge. METHOD A single treatment, blinded, enamel replica clinical study was undertaken in 20 healthy subjects. Replica tooth impressions were taken at baseline, following acid challenge and 2, 4, 7 and 24 h post challenge. Subjects consumed 500 ml of acidic soft drink over 30 min. Scanning electron microscopy of surface tomography was characterised with a descriptive 5 point scale by four judges. Duplicate impressions were taken to assess reproducibility. RESULTS 18 subjects had scorable sequences. Descriptive analyses showed erosive changes following acid consumption and reparative changes in the subsequent 24 h period. Comparing baseline replica to the 24 h replica, there were no significant differences (p=0.26) in tooth surface characteristics. Comparing the replica taken immediately following acidic challenge with the subsequent replicas at 2, 4, 7 and 24 h, showed clear reduction of erosive effects on the enamel surface at 2 h (p=0.02) and a highly significant reduction at 4, 7 and 24 h (p<0.001). CONCLUSION This methodology demonstrated the ability to follow the progression and recovery of early erosive enamel lesions over 24 h being accurate and reproducible. This study suggests enamel repair commences within 2 h following a substantial acidic challenge and is completed 4-24 h later. After 24 h, the tooth surface appeared visibly indistinguishable from the original tooth surface, suggestive of a recovery process occurring. CLINICAL SIGNIFICANCE Healthy erosive lifestyles often culminate in tooth wear. The time taken for enamel remineralisation following acidic challenge is unknown however, this study suggests the repair process is relatively slow following a substantial acidic challenge, and at least 4-24 h should elapse prior to further acidic consumption to allow for recovery.

An in-situ pilot study to investigate the native clinical resistance of enamel to erosion

OBJECTIVES To investigate the differences in susceptibility of the surface of native and polished enamel to dietary erosion using an in-situ model. METHODS Thirty healthy volunteers (n = 10 per group) wore mandibular appliances containing 2 native and 2 polished enamel samples for 30 min after which, the samples were exposed to either an ex-vivo or in-vivo immersion in orange juice for 5, 10 or 15 min and the cycle repeated twice with an hours interval between them. Samples were scanned with a non-contacting laser profilometer and surface roughness was extracted from the data, together with step height and microhardness change on the polished enamel samples. RESULTS All volunteers completed the study. For native enamel there were no statistical difference between baseline roughness values versus post erosion. Polished enamel significantly increased mean (SD) Sa roughness from baseline for each group resulting in roughness change of 0.04 (0.03), 0.06 (0.04), 0.04 (0.03), 0.06 (0.03), 0.08 (0.05) and 0.09 (0.05) μm respectively. With statistical differences between roughness change 45 min in-vivo versus 45 min ex-vivo (p < 0.05). Microhardness significantly decreased for each polished group, with statistical differences in hardness change between 30 min in-vivo versus 30 min ex-vivo (p < 0.05), 45 min in-vivo versus 30 min ex-vivo (p < 0.01), 45 min in-vivo versus 45 min ex-vivo (p < 0.01). CONCLUSIONS The native resistance to erosion provided clinically is a combination of the ultrastructure of outer enamel, protection from the salivary pellicle and the overall effects of the oral environment. CLINICALTRIALS. GOV IDENTIFIER NCT03178968. CLINICAL SIGNIFICANCE This study demonstrates that outer enamel is innately more resistant to erosion which is clinically relevant as once there has been structural breakdown at this level the effects of erosive wear will be accelerated.

Precision of 655 nm Confocal Laser Profilometry for 3D surface texture characterisation of natural human enamel undergoing dietary acid mediated erosive wear

OBJECTIVES To assess the precision of optical profilometry for characterising the 3D surface roughness of natural and polished human enamel in order to reliably quantify acid mediated surface roughness changes in human enamel. METHODS Forty-two enamel samples were prepared from extracted human molars and either polished flat or left unmodified. To investigate precision, the variability of thirty repeated measurements of five areas of one polished and one natural enamel sample was assessed using 655nm Confocal Laser Profilometry. Remaining samples were subjected to forty-five minutes orange juice erosion and microstructural changes were analysed using Sa roughness change (μm) and qualitatively using surface/subsurface confocal microscopy. RESULTS Enamel surface profilometry from the selected areas revealed maximal precision of 5nm for polished enamel and 23nm for natural enamel. After erosion, the polished enamel revealed a 48% increase in mean (SD) Sa roughness of 0.10 (0.07)μm (P<0.05), whereas in contrast the natural enamel revealed a 45% decrease in mean (SD) roughness of -0.32 (0.42)μm (P<0.05). These data were supported by qualitative confocal images of the surface/subsurface enamel. SIGNIFICANCE This study demonstrates a method for precise surface texture measurement of natural human enamel. Measurement precision was superior for polished flat enamel in contrast to natural enamel however, natural enamel responds very differently to polished enamel when exposed to erosion challenges. Therefore, thus future studies characterising enamel surface changes following erosion on natural enamel may provide more clinically relevant responses in comparison to polished enamel.

Novel Air Stimulation MR-Device for Intraoral Quantitative Sensory Cold Testing

The advent of neuroimaging in dental research provides exciting opportunities for relating excitation of trigeminal neurons to human somatosensory perceptions. Cold air sensitivity is one of the most frequent causes of dental discomfort or pain. Up to date, devices capable of delivering controlled cold air in an MR-environment are unavailable for quantitative sensory testing. This study therefore aimed at constructing and evaluating a novel MR-compatible, computer-controlled cold air stimulation apparatus (CASA) that produces graded air puffs. CASA consisted of a multi-injector air jet delivery system (AJS), a cold exchanger, a cooling agent, and a stimulus application construction. Its feasibility was tested by performing an fMRI stimulation experiment on a single subject experiencing dentine cold sensitivity. The novel device delivered repetitive, stable air stimuli ranging from room temperature (24.5°C ± 2°C) to −35°C, at flow rates between 5 and 17 liters per minute (l/min). These cold air puffs evoked perceptions similar to natural stimuli. Single-subject fMRI-analysis yielded brain activations typically associated with acute pain processing including thalamus, insular and cingulate cortices, somatosensory, cerebellar, and frontal brain regions. Thus, the novel CASA allowed for controlled, repetitive quantitative sensory testing by using air stimuli at graded temperatures (room temperature down to −35°C) while simultaneously recording brain responses. No MR-compatible stimulation device currently exists that is capable of providing non-contact natural-like stimuli at a wide temperature range to tissues in spatially restricted areas such as the mouth. The physical characteristics of this novel device thus holds promise for advancing the field of trigeminal and spinal somatosensory research, namely with respect to comparing therapeutic interventions for dentine hypersensitivity.