Comparative study on the detection of early dental caries using thermo-photonic lock-in imaging and optical coherence tomography.

Early detection of dental caries is known to be the key to the effectiveness of therapeutic and preventive approaches in dentistry. However, existing clinical detection techniques, such as radiographs, are not sufficiently sensitive to detect and monitor the progression of caries at early stages. As such, in recent years, several optics-based imaging modalities have been proposed for the early detection of caries. The majority of these techniques rely on the enhancement of light scattering in early carious lesions, while a few of them are based on the enhancement of light absorption at early caries sites. In this paper, we report on a systemic comparative study on the detection performances of optical coherence tomography (OCT) and thermophotonic lock-in imaging (TPLI) as representative early caries detection modalities based on light scattering and absorption, respectively. Through controlled demineralization studies on extracted human teeth and µCT validation experiments, several detection performance parameters of the two modalities such as detection threshold, sensitivity and specificity have been qualitatively analyzed and discussed. Our experiment results suggests that both modalities have sufficient sensitivity for the detection of well-developed early caries on occlusal and smooth surfaces; however, TPLI provides better sensitivity and detection threshold for detecting very early stages of caries formation, which is deemed to be critical for the effectiveness of therapeutic and preventive approaches in dentistry. Moreover, due to the more specific nature of the light absorption contrast mechanism over light scattering, TPLI exhibits better detection specificity, which results in less false positive readings and thus allows for the proper differentiation of early caries regions from the surrounding intact areas. The major shortcoming of TPLI is its inherent depth-integrated nature, prohibiting the production of depth-resolved/B-mode like images. The outcomes of this research justify the need for a light-absorption based imaging modality with the ability to produce tomographic and depth-resolved images, combining the key advantages of OCT and TPLI.

Longitudinal correlation of 3D OCT to detect early stage erosion in bovine enamel.

Erosive tissue-loss in dental enamel is of significant clinical concern because the net loss of enamel is irreversible, however, initial erosion is reversible. Micro-hardness testing is a standard method for measuring initial erosion, but its invasive nature has led to the investigation of alternative measurement techniques. Optical coherence tomography (OCT) is an attractive alternative because of its ability to non-invasively image three-dimensional volumes. In this study, a four-dimensional OCT system is used to longitudinally measure bovine enamel undergoing a continuous erosive challenge. A new method of analyzing 3D OCT volumes is introduced that compares intensity projections of the specimen surface by calculating the slope of a linear regression line between corresponding pixel intensities and the associated correlation coefficient. The OCT correlation measurements are compared to micro-hardness data and found to exhibit a linear relationship. The results show that this method is a sensitive technique for the investigation of the formation of early stage erosive lesions.

Optical diagnosis and characterization of dental caries with polarization-resolved hyperspectral stimulated Raman scattering microscopy.

We report the utility of a rapid polarization-resolved hyperspectral stimulated Raman scattering (SRS) imaging technique developed for optical diagnosis and characterization of dental caries in the tooth. Hyperspectral SRS images (512 × 512 pixels) of the tooth covering both the fingerprint (800-1800 cm(-1)) and high-wavenumber (2800-3600 cm(-1)) regions can be acquired within 15 minutes, which is at least 10(3) faster in imaging speed than confocal Raman mapping. Hyperspectral SRS imaging uncovers the biochemical distributions and variations across the carious enamel in the tooth. SRS imaging shows that compared to the sound enamel, the mineral content in the body of lesion decreases by 55%; while increasing up to 110% in the surface zone, indicating the formation of a hyper-mineralized layer due to the remineralization process. Further polarized SRS imaging shows that the depolarization ratios of hydroxyapatite crystals (ν 1-PO4 (3-) of SRS at 959 cm(-1)) of the tooth in the sound enamel, translucent zone, body of lesion and the surface zone are 0.035 ± 0.01, 0.052 ± 0.02, 0.314 ± 0.1, 0.038 ± 0.02, respectively, providing a new diagnostic criterion for discriminating carious lesions from sound enamel in the teeth. This work demonstrates for the first time that the polarization-resolved hyperspectral SRS imaging technique can be used for quantitatively determining tooth mineralization levels and discriminating carious lesions from sound enamel in a rapid fashion, proving its promising potential of early detection and diagnosis of dental caries without labeling.

Wide-field in vivo oral OCT imaging.

We have built a polarization-sensitive swept source Optical Coherence Tomography (OCT) instrument capable of wide-field in vivo imaging in the oral cavity. This instrument uses a hand-held side-looking fiber-optic rotary pullback catheter that can cover two dimensional tissue imaging fields approximately 2.5 mm wide by up to 90 mm length in a single image acquisition. The catheter spins at 100 Hz with pullback speeds up to 15 mm/s allowing imaging of areas up to 225 mm(2) field-of-view in seconds. A catheter sheath and two optional catheter sheath holders have been designed to allow imaging at all locations within the oral cavity. Image quality of 2-dimensional image slices through the data can be greatly enhanced by averaging over the orthogonal dimension to reduce speckle. Initial in vivo imaging results reveal a wide-field view of features such as epithelial thickness and continuity of the basement membrane that may be useful in clinic for chair-side management of oral lesions.

Decrease in fluorescence lifetime by glycation of collagen and its application in determining advanced glycation end-products in human dentin.

Advanced Glycation End-products (AGEs) are produced by the Maillard reaction, which causes cross-linking of collagen and results in changes in the mechanical properties of collagen tissues. Several types of AGE fluoresce, and measurement of this fluorescence is effective for determining the presence of AGEs. Because fluorescence intensity by steady-state fluorometry is affected by sample surface condition and light source, we focused on fluorescence lifetime measurement (FLM). We found that fluorescence lifetime of collagen gel decreased with glycation progress. In vivo application of FLM for determination of AGEs was confirmed in human dentin.

Automated assessment of the remineralization of artificial enamel lesions with polarization-sensitive optical coherence tomography.

Accurate measurement of the highly mineralized transparent surface layer that forms on caries lesions is important for diagnosis of the lesion activity because chemical intervention can slow or reverse the caries process via remineralization. Previous in-vitro and in-vivo studies have demonstrated that polarization-sensitive optical coherence tomography (PS-OCT) can nondestructively image the subsurface lesion structure and the highly mineralized transparent surface zone of caries lesions. The purpose of this study was to develop an approach to automatically process 3-dimensional PS-OCT images and to accurately assess the remineralization process in simulated enamel lesions. Artificial enamel lesions were prepared on twenty bovine enamel blocks using two models to produce varying degree of demineralization and remineralization. The thickness of the transparent surface layer and the integrated reflectivity of the subsurface lesion were measured using PS-OCT. The automated transparent surface layer detection algorithm was able to successfully detect the transparent surface layers with high sensitivity ( = 0.92) and high specificity ( = 0.97). The estimated thickness of the transparent surface layer showed a strong correlation with polarized light microscopy (PLM) measurements of all regions (R(2) = 0.90). The integrated reflectivity, ΔR, and the integrated mineral loss, ΔZ, showed a moderate correlation (R(2) = 0.32). This study demonstrates that PS-OCT can automatically measure the changes in artificial enamel lesion structure and severity upon exposure to remineralization solutions.

Biochemical characterization of human gingival crevicular fluid during orthodontic tooth movement using Raman spectroscopy.

This study used Raman spectroscopy to report the first human gingival crevicular fluid (GCF) biochemical characterization during the early phase of orthodontic tooth movement. This technique allows for label-free and noninvasive biochemical change monitoring in GCF during orthodontic tooth movement. Ten orthodontic patients (20.8 ± 2.5 years) participated in the study. GCF samples were obtained before (baseline, 0 days) and during orthodontic treatment at 1, 7 and 28 days. For Raman spectroscopic measurement, GCF samples (5 µl) were deposited onto a gold-coated substrate, then dried at room temperature. Raman spectra GCF analysis during orthodontic treatment indicated that the hydroxyapatite to primarily collagen-dominated matrix band (phosphate 984 cm(-1)/amide I 1667 cm(-1)) intensity ratio decreased at day 7 (P < 0.05). The carbonate apatite to hydroxyapatite ratio (carbonate 1088 cm(-1)/phosphate 984 cm(-1)) was significantly higher on day 7 compared to day 0 (P < 0.05). These results indicate that demineralization occurs during the alveolar bone remodeling process. We also found notable peak shifts in the amide I range during orthodontic tooth movement. The 1658 cm(-1) in baseline red shifted to 1667 cm(-1) at orthodontic treatment day 7. Curve fitting in the amide I (1615-1725 cm(-1)) range demonstrated that increased random coil conformation was accompanied by a decrease in ß-sheet structure during orthodontic tooth movement. Thus, we suggest Raman spectroscopy could be used for label-free, non-invasive GCF quality assessment during orthodontic tooth movement. Furthermore, this method may prove to be a powerful diagnostic and prognostic tool for monitoring orthodontic tooth movement in a clinical setting.

Quantitative physiology and immunohistochemistry of oral lesions.

Angiogenesis and hypoxia are reported to correlate with tumor aggressiveness. In this study, we investigated the potential of optically measured total hemoglobin concentration (THC) and blood oxygen saturation (StO2) as a quantitative measure of angiogenesis and hypoxia in oral lesions with an immunohistochemical comparison. 12 normal subjects and 40 oral patients (22 oral squamous cell carcinoma (SCC), 18 benign/premalignant lesions including 11 verrucous hyperplasia (VH) and 7 hyperkeratosis/parakeratosis (HK)) were studied. The results showed that the THC measurement was consistent with vascular endothelial growth factor (VEGF) and microvessel staining in the stromal area, but StO2 was not associated with HIF-1α. We observed inflammation induced neovascular formation in the stromal area of VH and HK that were likely attributed to higher-than-control THC and StO2 and resulted in no difference in optical measurements between all lesions. However, we found that in majority of SCC, the ratio of THC and StO2 levels between lesions and the surrounding tissues provide potential distinguishing characteristics from VH, which are not visually differentiable from SCC, with a sensitivity, specificity, and accuracy of 91%, 68%, and 76%, respectively.

Towards a bimodal proximity sensor for in situ neurovascular bundle detection during dental implant surgery.

Proof of concept results are presented towards an in situ bimodal proximity sensor for neurovascular bundle detection during dental implant surgery using combined near infrared absorption (NIR) and optical coherence tomography (OCT) techniques. These modalities are shown to have different sensitivity to the proximity of optical contrast from neurovascular bundles. NIR AC and DC signals from the pulsing of an artery enable qualitative ranging of the bundle in the millimeter range, with best sensitivity around 0.5-3mm distance in a custom phantom setup. OCT provides structural mapping of the neurovascular bundle at sub-millimeter distances in an ex vivo human jaw bone. Combining the two techniques suggests a novel ranging system for the surgeon that could be implemented in a "smart drill." The proximity to the neurovascular bundle can be tracked in real time in the range of a few millimeters with NIR signals, after which higher resolution imaging OCT to provide finer ranging in the sub-millimeter distances.

Zirconia dental implants degradation by confocal Raman microspectroscopy: analytical simulation and experiments.

Subsurface hydrothermal degradation of yttria stabilized tetragonal zirconia polycrystals (3Y-TZP) is presented. Evaluation of low temperature degradation (LTD) phase transformation induced by aging in 3Y-TZP is experimentally studied by Raman confocal microspectroscopy. A non-linear distribution of monoclinic volume fraction is determined in depth by using different pinhole sizes. A theoretical simulation is proposed based on the convolution of the excitation intensity profile and the Beer-Lambert law (optical properties of zirconia) to compare between experiment and theory. The calculated theoretical degradation curves matche closely to the experimental ones. Surface transformation (V0) and transformation factor in depth (T) are obtained by comparing simulation and experience for each sample with nondestructive optical sectioning.

Accelerated measurement of perikymata by an optical instrument.

The proposed device considerably reduces the measuring time of important microscopic features of tooth crown surfaces. The instrumentation is accompanied by a computer program to analyse the results. Tooth enamel is formed by ameloblasts, which demonstrate daily secretory rhythms developing tissue-specific structures known as cross striations, and longer period markings that are referred as striae of Retzius. These striae correspond to linear structures on the enamel surface. This newly developed optical measuring instrument can automatically, precisely and accurately record the number and periodicity of perikymata on the dental crown. Furthermore it can characterize the variability in periodicity of perikymata in hominids. The depth of field can be extended as desired by taking several images with different focus positions and combining them into a single composite image that contains all regions fully focused.

Terahertz time-domain spectroscopy for monitoring the curing of dental composites.

We apply terahertz (THz) time-domain spectroscopy for monitoring the curing process of three different light-curing dental composites. Exact knowledge of the sample thickness is required for a precise determination of the THz dielectric parameters, as the materials exhibit shrinkage when they are cured. We find very small but significant changes of the THz refractive index and absorption coefficient during stepwise light exposure. The changes in the refractive index are correlated with changes in the density of the materials. Furthermore, the refractive index and the sample thickness are found to give the most reliable result for monitoring the curing process of the dental composites.

Multispectral near-IR reflectance and transillumination imaging of teeth.

NIR imaging methods do not require ionizing radiation and have great potential for detecting caries lesions (tooth decay) on high-risk proximal and occlusal tooth surfaces and at the earliest stages of development. Previous in vitro and in vivo studies at 1300-nm demonstrated that high contrast reflectance and transillumination images could be acquired of caries lesions on tooth proximal and occlusal surfaces where most new decay is found. Water absorption varies markedly between 1200 and 1600-nm and the scattering properties of enamel and the underlying dentin have not been characterized in this region. Hyperspectral reflectance studies show lower reflectivity from sound enamel and dentin at NIR wavelengths with higher water absorption. The purpose of this imaging study was to determine which NIR wavelengths between 1200 and 1600-nm provide the highest contrast of demineralization or caries lesions for each of the different modes of NIR imaging, including transillumination of proximal and occlusal surfaces along with cross polarization reflectance measurements. A tungsten halogen lamp with several spectral filters and a Ge-enhanced CMOS focal plane array (FPA) sensitive from 400 to 1600-nm were used to acquire the images of caries lesions on extracted teeth. Artificial interproximal lesions were created on twelve tooth sections of 5 & 6-mm thickness that were used for transillumination imaging. Fifty-four extracted teeth with suspected occlusal lesions were also examined in both occlusal transillumination and reflectance imaging modes. Cavity preparations were also cut into whole teeth and filled with composite and used to compare the contrast between composite and enamel at NIR wavelengths. NIR wavelengths longer than 1400-nm are likely to have better performance for the transillumination of occlusal caries lesions while 1300-nm appears best for the transillumination of proximal surfaces. Loss of mobile water in enamel markedly reduced the transparency of the enamel at all NIR wavelengths. Significantly higher contrast was attained for reflectance measurements at wavelengths that have higher water absorption, namely 1460-nm. Wavelengths with higher water absorption also provided higher contrast of composite restorations.

Laser ultrasonic evaluation of human dental enamel during remineralization treatment.

In this work a non-destructive laser ultrasonic technique is used to quantitatively evaluate the progressive change in the elastic response of human dental enamel during a remineralization treatment. The condition of the enamel was measured during two weeks treatment using laser generated and detected surface acoustic waves in sound and demineralized enamel. Analysis of the acoustic velocity dispersion confirms the efficacy, as well as illuminating the progress, of the treatment.

Imaging carious dental tissues with multiphoton fluorescence lifetime imaging microscopy.

In this study, multiphoton excitation was utilized to image normal and carious dental tissues noninvasively. Unique structures in dental tissues were identified using the available multimodality (second harmonic, autofluorescence, and fluorescence lifetime analysis) without labeling. The collagen in dentin exhibits a strong second harmonic response. Both dentin and enamel emit strong autofluorescence that reveals in detail morphological features (such as dentinal tubules and enamel rods) and, despite their very similar spectral profiles, can be differentiated by lifetime analysis. Specifically, the carious dental tissue exhibits a greatly reduced autofluorescence lifetime, which result is consistent with the degree of demineralization, determined by micro-computed tomography. Our findings suggest that two-photon excited fluorescence lifetime imaging may be a promising tool for diagnosing and monitoring dental caries.