Spatial analysis of polarimetric images to enhance near-surface sampling sensitivity: feasibility in demineralized teeth and other tissue-like media.

Significance: Early tooth demineralization may be detectable through spatial analysis of polarized light images as demonstrated in this study. This may also prove useful in the early detection of epithelial tumors that comprise the majority of the cancer burden worldwide. Aim: The spatial properties of polarized light images have not been greatly exploited in biomedicine to improve sensitivity to superficial tissue regions; therefore, we investigate the optical sampling depth effects as a function of location in the backscattered polarimetric images. Approach: Backscattered linear polarization intensity distributions exhibit four-lobed patterns arising through single-scattering, multiple-scattering, and geometrical effects. These photon pathway dynamics are investigated through experimental imaging of microsphere suspensions along with corroborative computational polarization-sensitive Monte Carlo modeling. The studied sampling depth effects of linear and circular polarization images (explored in a previous study) are then evaluated on normal and demineralized human teeth, which are known to differ in their surface and sub-surface structures. Results: Backscattered linear polarization images exhibit enhanced sensitivity to near-surface properties of media (for example, surface roughness and turbidity) at specific locations within the four-lobed patterns. This yields improved differentiation of two tooth types when spatially selecting image regions in the direction perpendicular to the incident linear polarization vector. Circular polarimetric imaging also yields improved differentiation through spatial selection of regions close to the site of illumination. Improved sensitivity to superficial tissues is achieved through a combination of these linear and circular polarimetric imaging approaches. Conclusions: Heightened sampling sensitivity to tissue microstructure in the surface/near-surface region of turbid tissue-like media and dental tissue is achieved through a judicious spatial selection of specific regions in the resultant co-linear and cross-circular backscattered polarimetric images.

Quantitative analysis of the degree of demineralization for bleached enamel by optical coherence tomography.

BACKGROUND: Tooth bleaching imparts whitening effects along with adverse effects such as increased tooth sensitivity and enamel surface changes. Herein, we employed optical coherence tomography (OCT), a nondestructive optical detection technique, for evaluation of tooth enamel after treatment with peroxide-based bleaching agents. METHODS: Fifteen enamel samples were bleached using 38% acidic hydrogen peroxide-based bleach, subjected to OCT scanning, and then cross-sectioned and imaged under polarized light microscopy (PLM) and transverse microradiography (TMR). OCT cross-sectional images were compared with PLM and TMR. The depth and severity of demineralization produced in the bleached enamel were measured by OCT, PLM, and TMR. Comparison between the three techniques was performed using Kruskal-Wallis H non-parametric test and Pearson correlation. RESULTS: In comparison with PLM and TMR, OCT clearly detected the changes in the enamel surface after hydrogen peroxide bleaching. Significant correlations (p<0.05) were observed in lesion depth between OCT and PLM (r=0.820), OCT and TMR (r=0.822), and TMR and PLM (r=0.861). There was no statistically significant difference in demineralization depth values measured by OCT, PLM, and TMR (p>0.05). CONCLUSION: OCT can allow real-time, non-invasive imaging of artificially bleached tooth models and automatically measure the early changes in the enamel lesion structure upon exposure to hydrogen peroxide-based bleaching agents.

Detection of artificial enamel caries-like lesions with a blue hydroxyapatite-binding porosity probe.

OBJECTIVES: This in vitro study investigated the ability of a blue protein-based hydroxyapatite porosity probe to selectively detect artificial enamel caries-like lesions of varying severities. METHODS: Artificial caries-like lesions were formed in enamel specimens using a hydroxyethylcellulose-containing lactic acid gel for 4/12/24/72 or 168 h. One untreated group was used as a control. The probe was applied for 2 min and unbound probe rinsed off with deionized water. Surface color changes were determined spectrophotometrically (L*a*b* color space) and with digital photography. Lesions were characterized using quantitative light-induced fluorescence (QLF), Vickers surface microhardness, and transverse microradiography (TMR). Data were analyzed using one-way ANOVA. RESULTS: Digital photography did not reveal any discoloration in unaffected enamel. However, all lesions stained blue with color intensity positively correlated with demineralization times. The color data reflected similar trends: lesions became significantly darker (L* decreased) and bluer (b* decreased), while overall color differences (ΔE) increased significantly after probe application (4-h lesion, mean±standard deviation: ΔL*=-2.6 ± 4.1/Δb*=0.1 ± 0.8/ΔE=5.5 ± 1.3 vs. 168-h lesion: ΔL*=-17.3 ± 1.1/Δb*=-6.0 ± 0.6/ΔE=18.7 ± 1.1). TMR analysis revealed distinct differences in integrated mineral loss (ΔZ) and lesion depth (L) between demineralization times (4-h lesion: ΔZ=391±190 vol%min × µm/L = 18.1 ± 10.9 µm vs. 168-h lesion: ΔZ=3606±499 vol%min × µm/L = 111.9 ± 13.9 µm). QLF and microhardness were also able to differentiate between demineralization times. L and ΔZ strongly correlated (Pearson correlation coefficient [r]) with Δb* (L vs. Δb*: r=-0.90/ΔZ vs. Δb*: r=-0.90), ΔE (r = 0.85/r = 0.81), and ΔL* (r=-0.79/r=-0.73). CONCLUSION: Considering the limitations of this study, the blue protein-based hydroxyapatite-binding porosity probe appears to be sufficiently sensitive to distinguish between unaffected enamel and artificial caries-like lesions. CLINICAL SIGNIFICANCE: Early detection of enamel caries lesions remains one of the most critical aspects in the diagnosis and management of dental caries. This study highlighted the potential of a novel porosity probe in detecting artificial caries-like demineralization by objective means.

In vitro remineralization assessment of enamel subsurface lesions using different percentages of surface reaction-type pre-reacted glass-ionomer containing gum-based material.

OBJECTIVE: To identify the remineralization activity of enamel subsurface lesions using different percentages of surface pre-reacted glass-ionomer (S-PRG) filler containing gum-base material. METHODS: Gum extracts from gum-base materials containing 0wt%, 5wt%, and 10wt% S-PRG filler were prepared as GE0, GE5, and GE10, respectively. A total of 50 bovine enamel specimens were used, and the polished enamel surface of a 3 × 3 mm2 window area was exposed. The specimens were then subjected to a demineralization solution for seven days to create a subsurface enamel lesion. Remineralization was then conducted for seven days using a protocol whereby the specimens were immersed three times a day in prepared gum extracts (0wt%, 5wt%, and 10wt%) and artificial saliva of pH 7 (Control) for 20 min at 37 °C. Thereafter, remineralization assessment was performed by using Swept Source Optical Coherence Tomography (SS-OCT) and micro-computed tomography (µCT). Surface morphology and elemental analysis were conducted by scanning electron microscopy (SEM) and energy-dispersive X-ray spectrometry (EDS). RESULTS: The depths of the demineralized lesions in the GE5 and GE10 groups were significantly lower than those of the Control and the GE0 groups. SEM observations of the enamel surface morphology of the GE5 and GE10 groups indicated remineralization with S-PRG filler-related elements present. CONCLUSION: The GE5 and GE10 S-PRG filler containing gum-base materials showed significantly improved surface remineralization and reduced demineralization of the enamel lesions. EDS analysis suggested that the released ions from the S-PRG filler might be responsible for surface remineralization. CLINICAL SIGNIFICANCE: The S-PRG filler containing gum-base material may have a significant remineralization effect and improve the surface morphology of enamel subsurface lesions.

Monitoring lesion activity on primary teeth with CP-OCT and SWIR reflectance imaging.

OBJECTIVE: The purpose of this study was to use cross polarization optical coherence tomography (CP-OCT) and short wavelength infrared imaging (SWIR) reflectance imaging to monitor changes in the structure and activity of early occlusal caries on primary teeth over a period of 6 months during intervention with fluoride. METHODS: Participants (n = 29) aged 6-10 each with two suspected active occlusal lesions on primary teeth completed the study. Fluoride varnish was applied to tooth surfaces every 3-months and participants were instructed to brush twice daily with a fluoride toothpaste. Images were acquired using CP-OCT every 3 months for 6 months. SWIR reflectance images were acquired during forced air-drying of the lesions for 30 s at 0 and 6-months. RESULTS: Most of the 42 lesions appeared initially active at baseline. Only 6 lesions appeared arrested at baseline based on the presence of a highly mineralized transparent surface layer (TSL) in CP-OCT images. At 6 months, 14 of the lesions appeared arrested including the 6 initially arrested lesions and the TSL thickness increased significantly (p < 0.0001). The mean lesion depth (Ld) and the integrated reflectivity over the lesion depth (ΔR) increased significantly (p < 0.05) after 6 months for the 42 lesions analyzed. SWIR reflectance images showed that there was a significantly higher (p < 0.05) delay before changes in intensity were measured for active lesions versus arrested lesions during lesion drying. CONCLUSION: CP-OCT was able to monitor changes in lesion structure and activity including the formation of a highly mineralized TSL indicative of lesion arrest during nonsurgical intervention. Time-resolved SWIR reflectance imaging also shows that there are differences in the dehydration kinetics between active and arrested lesions. This study demonstrates two independent imaging methods that can be used to monitor changes in lesion activity over time.

Demonstration of an optical dentin hardness measuring device using bovine dentin with different demineralization times.

Significance: The increase in root caries is a serious problem as society ages. Root caries is diagnosed by inspection and palpation, which are qualitative. A method to objectively and quantitatively evaluate the progress of root caries in a clinical setting is strongly desired. The root caries could be diagnosed by measuring hardness because dentin becomes softer as the caries progresses. Vickers hardness has been customarily used as an indicator of tooth hardness. However, this method cannot be used to in vivo teeth because the teeth must be dried prior to measurement to make the indentation. A hardness meter using an indenter with light for tooth monitoring (HAMILTOM) is proposed as an optical device. HAMILTOM could measure hardness of teeth in wet condition as a dark area while applying a load to dentins without drying. Therefore, HAMILTOM may realize hardness measurements of in vivo teeth in a clinical setting quantitatively. Aim: The aim of our study is to demonstrate the optical dentin hardness measuring device HAMILTOM using bovine dentin with different demineralization times and to evaluate the correlation between the dark areas measured by HAMILTOM and the Vickers hardness measured by the Vickers hardness tester. Approach: The samples were 20 bovine dentins. They were demineralized by a lactic acid solution with different times and divided into groups 1 and 2 of 10 samples each. In both groups, the dark areas and Vickers hardness were measured for each sample. Group 1 was used to obtain a calibration curve to calculate Vickers hardness from the dark area. Group 2 was used to validate the calibration curve obtained from the dentin samples of group 1. Results: The areas appearing black without a total internal reflection of the indenter measured by HAMILTOM increased as the demineralization time increased. Additionally, the Vickers hardness of group 2 calculated by the dark areas of group 2 and the calibration curve obtained in group 1 and the Vickers hardness of group 2 measured by the Vickers hardness tester were strongly correlated with a determination coefficient of 0.99. Conclusions: The results demonstrate that HAMILTOM may be a suitable alternative to the conventional method. Unlike the conventional method, which cannot be used for in vivo teeth, HAMILTOM holds potential to quantitatively evaluate the progress of caries in in vivo teeth.

Evaluation of alveolar bone hypomineralization in pediatric hypophosphatasia using orthopantomography.

Hypophosphatasia (HPP) is a metabolic disease characterized by impaired bone mineralization and early exfoliation of primary teeth. This study was performed to develop a method for quantitatively evaluating alveolar bone hypomineralization using orthopantomographic images. Alveolar bone density was defined according to the pixel values and corrected by brightness shown by an indicator applied to the orthopantomographic device. Images of 200 healthy subjects (aged 2-15 years) were classified into five age groups. The corrected pixel values were significantly lower in the younger group than in those aged 14-15 years (2-4, 5-7, and 8-10 years versus 14-15 years: P < 0.0001, 11-13 years versus 14-15 years: P < 0.01). Orthopantomographic images of 17 patients with HPP were evaluated. The corrected pixel values of three-fourths of the patients with odonto type HPP were lower than the mean values of the healthy group. One-third of patients treated with enzyme replacement therapy showed higher corrected pixel values than the healthy group. Our results suggest that odonto type HPP without skeletal problems is occasionally accompanied by hypomineralization of alveolar bone and that alveolar bone hypomineralization in patients with severe HPP is possibly improved by enzyme replacement therapy.

Quantifying the demineralisation of enamel using a hyperspectral camera measuring fluorescence loss.

BACKGROUND: The gold standard for quantifying mineral loss of enamel is transverse microradiography (TMR) and is complimented by the non-destructive quantitative light induced fluorescence (QLF) which measures changes in autofluorescence. Fluorescence loss has been shown to correlate with mineral loss. Building upon the established method, the use of hyperspectral fluorescence imaging (HI) allows the capture of a broader range of wavelengths to quantify fluorescence changes more accurately. METHODS: Bovine Enamel was demineralised within the dual constant depth film fermenter over 14 days and analysed using TMR, QLF and HI. The mineral change values were compared using Pearson's Correlation Coefficient. RESULTS: The analysis showed a statistically significant correlation that was equal between TMR and HI (r = 0.844) and TMR and QLF (r = 0.844), but weaker between QLF and HI (r = 0.811). CONCLUSIONS: The correlations indicate that HI is a promising valid non-destructive method for quantifying mineral loss from bovine enamel that is as accurate as QLF and complements TMR.

Effect of ion-releasing filler-containing gel application on dentin remineralization using optical coherent tomography.

PURPOSE: To examine the effects of an ion-releasing filler-containing gel on the remineralization of dentin using optical coherence tomography (OCT). METHODS: Dentin slabs of bovine teeth were sliced and shaped into a rectangular form. Specimens were treated with undersaturated 0.1 M lactic acid buffer solution (pH 4.75) for 10 minutes and then placed in artificial saliva (pH 7.0). This procedure was repeated three times a day for 28 days. The dentin remineralization effects of a fluoride/S-PRG filler-containing gel (PRG) and a 38% SDF solution (SDF) on dentin slabs of bovine teeth were compared. After treatment, the dentin slabs were immersed in a 0.1 M lactic acid buffer solution and then placed in artificial saliva. This procedure was repeated three times a day for 28 days. OCT imaging was conducted on the selected location of the dentin surface. The peak intensity and width at 1/e² were recorded in each of the six areas on the sample and averaged. Each group had a sample size of 10. Knoop hardness number (KHN) measurements and scanning electron microscopy (SEM) observations were also conducted. The data for each group were subjected to a one-way repeated-measures ANOVA and Tukey tests (α= 0.05). The samples were also observed using SEM. RESULTS: The peak signal intensities of SDF and PRG decreased on day 7 and then slightly increased during the experimental period for the one-off application groups and then decreased for frequent-time application groups. Although the width at 1/e² in the untreated specimens decreased over the test period, SDF and PRG for the one-off application groups exhibited an increase in widths on day 7 followed by a slight decrease, whereas it increased for the frequent-time application groups. The average KHN of the dentin samples exhibited the same tendency as the width at 1/e². Closure of the dentin tubules and crystal precipitation were detected on the surface of both SDF and PRG groups. CLINICAL SIGNIFICANCE: S-PRG filler-containing gel might have the ability to prevent dentin demineralization and could be useful for the prevention of hard-to-access lesions. This material achieved remineralization of the demineralized root dentin and had the same remineralization ability as SDF in vitro.

Enamel demineralization model in primary teeth: Micro-CT and SEM assessments of artificial incipient lesion.

Many studies have analyzed different tooth demineralization models, which generate artificial incipient lesions; however, most of them are complex, slow, not clear and results could not be employed in both primary and permanent teeth because of chemical content differences among them. This study evaluates a demineralization model on primary enamel, under three incubation periods; quantifying artificial incipient lesions formation, and depth by micro-CT, complementing with SEM for morphological characterization. Sixteen healthy human anterior primary teeth extracted for prolonged retention and orthopedic/orthodontic reasons were included in this study, previous informed consent. The sample was randomly assigned to four groups n = 4: G_Control, G_2D, G_4D, and G_7D. Micro-CT and SEM were performed during two stages: before demineralization (BD) and after demineralization (AD). A t-student test was carried out to determine differences among groups (p ≤ .05). No incipient lesions were observed in control group. Artificial lesion depth was similar among experimental groups; values were from 38.16 ± 05.40 µm to 42.61 ± 04.75 µm. An amount of 14 to 17 artificial incipient lesions were formed per group, the extension and distribution were different for each incubation period. Five erosive lesions were produced in G_7D. All experimental groups were able to form incipient artificial lesions in primary enamel. SEM characterization revealed more pronounced changes on the enamel surface, as the days of immersion in the demineralization solution increased. The 4-day incubation period is the most recommended for the demineralization model, due to the formation of incipient lesions only and its extension, which facilitates their assessment.

Initial investigations of a novel bioluminescence method for imaging dental demineralization.

OBJECTIVES: In this in vitro study, a bioluminescent marker was investigated for its potential to illuminate the assessment of dental caries and dental erosion, which are significant clinical and public health problems, through its binding of those ions, notably Ca2+ , known to be released during the process of demineralization. MATERIALS AND METHODS: The light output from the selected bioluminescent marker was investigated in several experiments, including: (a)contact with a range of Ca2+ ion concentrations; (b) treatment of extracted teeth with solutions of differing pH, followed by application of the bioluminescence marker to assess Ca2+ ion release; and (c) application of the marker to freshly extracted teeth with natural and artificially created caries lesions on occlusal and smooth surfaces to image the Ca2+ ion distribution. RESULTS: The results of: experiment (a) showed that the light output from the marker increases with increasing Ca2+ concentration and of experiment (b) showed increases in light being observed as increasingly acidic solutions were applied. The results of experiment (c) showed the bioluminescence images of the extracted teeth produced "demineralization maps" of the imaged surfaces. CONCLUSIONS: These results demonstrate the ability of a novel bioluminescence technology to image Ca2+ ions on tooth enamel surfaces which has potential in dental caries and dental erosion applications and provides the scientific basis for the ongoing development of that novel technology.

High Contrast Reflectance Imaging of Enamel Demineralization and Remineralization at 1950-nm for the Assessment of Lesion Activity.

BACKGROUND AND OBJECTIVES: Previous studies have shown that large changes in the diffuse reflectivity of caries lesions during drying with air can be used to assess lesion activity. The largest changes occur at short wavelength infrared (SWIR) wavelengths coincident with high water absorption. The strongest water absorption in the SWIR occurs at 1950 nm. In this study changes in the reflectivity of simulated lesions with varying degrees of remineralization was measured at 1500-2340 nm and at 1950 nm as the samples were dried with air. STUDY DESIGN/MATERIALS AND METHODS: Twenty bovine enamel surfaces each with five treatment windows were exposed to two demineralization/remineralization regimens to produce simulated lesions of varying depth, severity, and mineral gradients. An extended range tungsten-halogen lamp with a long pass filter (1500-2340 nm) and a broadband amplified spontaneous emission source centered near the peak of the water-absorption band at 1950-nm were used as light sources and an extended range InGaAs camera (1000-2340 nm) was used to acquire reflected light images as the samples were dried with air. Lesions were also assessed using digital microscopy, polarized light microscopy, optical coherence tomography, and transverse microradiography. RESULTS: Both wavelength ranges showed extremely high lesion contrast (>0.9) for all six lesion treatment windows in both models. The change in contrast (ΔI) was significantly higher for the 1950 nm broadband source for all the intact lesion windows compared with the 1500-2340 nm wavelength range. CONCLUSION: SWIR light at 1950 nm yields extremely high contrast of demineralization and appears to be the optimum wavelength for the assessment of lesion activity on tooth coronal surfaces. Lasers Surg. Med. 00:00-00, 2020. © 2020 Wiley Periodicals LLC.

Impact of early biofilm on the assessment of initial enamel erosion with swept-source optical coherence tomography.

Objective: This study examined the impact of early biofilm on the tooth surface, during the assessment of initial enamel erosion using swept-source optical coherence tomography (SS-OCT). Method and materials: Forty-five enamel windows of 2 × 4 mm2 were prepared on 23 extracted human teeth. The specimens were exposed to citric acid (pH 3.2) for 30 minutes and randomly divided into three groups (n = 15): Group 1, no biofilm; Group 2, 1-day-old biofilm; and Group 3, 3-day-old biofilm. Specimens in Groups 2 and 3 were inoculated with oral bacteria (Streptococcus sanguinis, Streptococcus mitis, and Actinomyces naeslundii) to produce early laboratory-cultivated biofilms for 1 and 3 days respectively. Surface microhardness (SMH) measurements were taken at pre- (t1) and post-erosion (t2); and SS-OCT scans were done at t1, t2, and post-biofilm cultivation (t3). Integrated reflectivity (IR) of the tooth-air interface (IRsurface) and enamel (IRenamel) were computed from the mean A-scans. Statistical analysis was performed using paired t tests and one-way ANOVA (α = .05). Results: A significant increase in IRenamel was observed at t2 (P < .05). At t3, IRsurface between Group 1 (control) and Group 2 (P = .012) as well as Group 3 (P = .001) were significantly different. Significant variances in IRenamel were perceived between t2 and t3 for Groups 2 and 3 but not for Group 1. Conclusion: As early biofilm affected SS-OCT assessment of initial enamel erosion, they should be removed from the tooth surface prior to OCT procedures..

Acid-induced demineralisation of human enamel as a function of time and pH observed using X-ray and polarised light imaging.

Acid-induced enamel demineralisation affects many individuals either by exposure to acidic diets, acidic gas pollution (dental erosion) or to dental plaque acids (dental caries). This study aimed to develop in situ X-ray and light imaging methods to determine progression of enamel demineralisation and the dynamic relationship between acid pH and mineral density. Hourly digital microradiograph time-lapse sequences showed the depth of enamel demineralisation in 500 µm thick sections progressed with time from the surface towards the dentine following a power-law function, which was 21% faster than the lateral demineralisation progression after exposure for 85 h to lactic acid (10%, pH 2.2). The minimum greyscale remaining (mineral content) within the induced enamel lesion followed an exponential decay, while the accumulated total greyscale loss with time was linear, which showed a constant anisotropic mineral release within the enamel architecture. This 85 h demineralisation method studied by polarised light microscopy time-lapse sequences showed that once the demineralisation front reached the enamel Hunter-Schreger bands, there was preferential demineralisation along those bands. Mineral density loss was linear with increasing pH acidity between pH 5.2 and pH 4.0 (with 0.4 pH increments) when incubated over a 3-week period exposed to 0.5% lactic acid. At pH 4.0, there was complete mineral loss in the centre of the demineralised area after the 3-week period and the linear function intercepted the x-axis at ~ pH 5.5, near the critical pH for hydroxyapatite (HAp). These observations showed how intrinsic enamel structure and pH affected the progression of demineralisation. STATEMENT OF SIGNIFICANCE: Hydroxyapatite crystallites (HAp) in human enamel dissolve when exposed to an acidic environment but little is known about how the intrinsic structures in enamel and pH influence the demineralisation kinetics. We have developed a time-lapse in situ microradiography method to quantify microscopic anisotropic mineral loss dynamics in response to an acid-only caries model. Correlation with polarised light microscopy time-lapse sequences showed that larger structures in enamel also influence demineralisation progression as demineralisation occurred preferentially along the Hunter-Schreger bands (decussating prismatic enamel). The pH-controlled enamel mineral release in a linear manner quantifying the relationship between HAp orientation and acid solubility. These findings should direct the development of improved anti-demineralisation/ remineralisation treatments to retain/ restore the natural intrinsic enamel structure.

Ultrasonic Monitoring of Dentin Demineralization.

Demineralization is a process of loss of minerals in the dental hard tissue that affects seriously the health of the patients, as it diminishes the tooth resistance, generating chewing problems by altering the occlusal structure, hypersensitivity, and pulpal problems. Demineralization can be produced by pathological processes as erosion or caries, or by surgical processes as etching. Due to the complexity of natural demineralization processes, it is mandatory to provide quantitative and standardized tests to allow their study in controlled laboratory conditions. Ultrasonic techniques are suitable for this purpose as they are nondestructive, quick, and provide localized mechanical information about the tissue, which is related with its degree of demineralization. In the present work, we evaluate the complete process of demineralization of the human dentin under controlled laboratory conditions using a pulse-echo ultrasonic technique. Up to 15 human dentin teeth have been demineralized with phosphoric acid at 10%. The time-of-flight measurements using the pulse-echo system allows to obtain the speed of sound in healthy (3415 m/s) and demineralized dentin tissue (1710 m/s), as well as to characterize the dynamical process of the acid penetration, which generates well-defined boundaries between two media (demineralized and mineralized dentin), showing very different mechanical properties. These boundaries advance in depth at an initial rate of [Formula: see text]/min, decelerating at -9.3 nm/min2 until the whole demineralization of the sample is achieved. In addition, the technique allows to measure the relevance of the demineralization produced by the acid residues inside the tooth once it has been removed from the acidic solution. Beyond the assessment of artificial demineralization lesions under laboratory conditions, as demonstrated in this article, the proposed technique opens new approaches to the assessment of demineralization caused by natural caries in vivo.

Relationship between Changes in Chemical Composition of Enamel Subsurface Lesions and the Emitted Nonlinear Optical Signals: An in vitro Study.

The development of new diagnostic technologies based on the light scattering and autofluorescence properties of dental tissues is required to improve the diagnostic ability of initial caries lesions earlier than previously done and promoting the potential of treatment without surgical intervention. The aim of this study is to correlate fluorescence-based results provided by multiphoton microscopy (MPM) with confocal Raman microscopy records using phosphate level at 960 cm-1 and the organic matrix at ∼2,931 cm-1 in healthy and demineralized human enamel. Measurements on 14 teeth were made using two incident lights of different wavelengths, released by confocal Raman microscopy and MPM. Raman phosphate peak intensity at 960 cm-1 along with organic to mineral ratio at (2,931/430 cm-1) and nonlinear optical signals (second harmonic generation [SHG] and intrinsic two-photon excited fluorescence [I2PEF]) were recorded from the demineralized and healthy enamel sites. Raman spectral maps showed that the higher the organic/mineral ratio in the demineralized enamel, the lower the intensity of mineral component in the same zone. MPM revealed new optical indicators of carious lesion as shown by the presence of a red-shifted fluorescence peak in the 650- to 750-nm area of the fluorescence spectrum of demineralized enamel. Moreover, on sample regions with insignificant autofluorescence, the emergence of the SHG signal could be noted. By comparing I2PEF images with the structural motifs observed by the confocal Raman imaging system, the morphological similarity of the acquired images was quite evident. Any change in the I2PEF spectra reflects alterations in the chemical composition of enamel. These findings may provide an important basis for potentially valuable applications of photonic tools in the clinical diagnosis of tooth pathological conditions, besides exposing the fundamental role of organic matrix in enamel integrity and reparation.

New missense variants in RELT causing hypomineralised amelogenesis imperfecta.

Amelogenesis imperfecta (AI) is a heterogeneous group of genetic diseases characterised by dental enamel malformation. Pathogenic variants in at least 33 genes cause syndromic or non-syndromic AI. Recently variants in RELT, encoding an orphan receptor in the tumour necrosis factor (TNF) superfamily, were found to cause recessive AI, as part of a syndrome encompassing small stature and severe childhood infections. Here we describe four additional families with autosomal recessive hypomineralised AI due to previously unreported homozygous mutations in RELT. Three families carried a homozygous missense variant in the fourth exon (c.164C>T, p.(T55I)) and a fourth family carried a homozygous missense variant in the 11th exon (c.1264C>T, p.(R422W)). We found no evidence of additional syndromic symptoms in affected individuals. Analyses of tooth microstructure with computerised tomography and scanning electron microscopy suggest a role for RELT in ameloblasts' coordination and interaction with the enamel matrix. Microsatellite genotyping in families segregating the T55I variant reveals a shared founder haplotype. These findings extend the RELT pathogenic variant spectrum, reveal a founder mutation in the UK Pakistani population and provide detailed analysis of human teeth affected by this hypomineralised phenotype, but do not support a possible syndromic presentation in all those with RELT-variant associated AI.

Caries detection enhancement using texture feature maps of intraoral radiographs.

OBJECTIVES: Dental caries are caused by tooth demineralization due to bacterial plaque formation. However, the resulting lesions are often discrete and thus barely recognizable in intraoral radiography images. Therefore, more advanced detection techniques are in great demand among dentists and radiographers. This study was performed to evaluate the performance of texture feature maps in the recognition of discrete demineralization related to caries plaque formation. METHODS: Digital intraoral radiology image analysis protocols incorporating first-order features (FOF), co-occurrence matrices, gray tone difference matrices, run-length matrices (RLM), local binary patterns (LBP), and k-means clustering (CLU) were used to transform the digital intraoral radiology images of 10 patients with confirmed caries, which were retrospectively reviewed in a dental clinic. The performance of the resulting texture feature maps was compared with that of radiographic images by radiologists and dental specialists. RESULTS: Significantly improved detection of caries spots was achieved by employing the CLU and FOF texture feature maps. The caries-affected area with sharp margins was well defined using the CLU approach. A pseudo-three-dimensional effect was observed in outlining the demineralization zones inside the cavity with the FOF 5 protocol. In contrast, the LBP and RLM techniques produced less satisfactory results with unsharp edges and less detailed depiction of the lesions. CONCLUSIONS: This study illustrated the applicability of texture feature maps to the recognition of demineralized spots on the tooth surface debilitated by caries and identified the best performing techniques.

Early detection of enamel demineralization by optical coherence tomography.

Enamel is the outermost layer of the tooth that protects it from invasion. In general, an acidic environment accelerates tooth demineralization, leading to the formation of cavities. Scanning electron microscopy (SEM) is conventionally used as an in vitro tool for the observation of tooth morphology changes with acid attacks. Yet, SEM has intrinsic limitations for the potential application of in vivo detection in the early demineralization process. In this study, a high-resolution optical coherence tomography (OCT) system with the axial and transverse resolutions of 2.0 and 2.7 µm in teeth has been utilized for characterizing the effect of the acidic environment (simulated by phosphoric acid) on the enamel topology. The scattering coefficient and the surface roughness of enamel can be directly derived from the OCT results, enabling a quantitative evaluation of the topology changes with demineralization. The dynamic process induced by the acid application is also recorded and analyzed with OCT, depicting the evolution of the demineralization process on enamel. Notably, the estimated enamel scattering coefficient and surface roughness significantly increase with the application time of acid and the results illustrate that the values of both parameters after demineralization are significantly larger than those obtained before the demineralization, illustrating both parameters could be effective to differentiate the healthy and demineralized teeth and determine the severity. The obtained results unambiguously illustrate that demineralization of the tooth surface can be successfully detected by OCT and further used as an indicator of early-stage cavity formation.