Remineralization of in vitro dental caries assessed with polarization-sensitive optical coherence tomography

Polarization-sensitive optical coherence tomography (PS-OCT) is potentially useful for imaging the nonsurgical remineralization of dental enamel. This study uses an all-fiber-based PS-OCT system operating at 1310 nm to image demineralized and fluoride-enhanced remineralized artificial lesions. PS-OCT images of lesions before and after remineralization are compared with the relative mineral loss DeltaZ (%vol x microm), obtained from high resolution digital microradiography (DM), and chemical composition changes by infrared spectroscopy. Severe early artificial caries show a significant increase in perpendicular-axis integrated reflectivity after remineralization. After sectioning the samples, DM demonstrates that the lesions remineralized with new mineral and the lesion surface zone show significant restoration of mineral volume. PS-OCT and DM both do not show a major change in lesion depth. For less severe artificial caries, the perpendicular-axis image resolves the scattering and depolarization of an outer growth layer after remineralization. This outer layer has a mineral volume close to that of sound enamel, and spectroscopic analysis indicates that the layer is a highly crystalline phase of apatite, without carbonate substitutions that increase the solubility of sound enamel. This study determines that PS-OCT can image the effects of fluoride-enhanced remineralization of mild and severe early artificial in vitro caries.     

Quantitative evaluation of the kinetics of human enamel simulated caries using photothermal radiometry and modulated luminescence

Photothermal radiometry and modulated luminescence (PTR-LUM) is an emerging nondestructive methodology applied toward the characterization and quantification of dental caries. We evaluate the efficacy of PTR-LUM in vitro to detect, monitor, and quantify human enamel caries. Artificial caries are created in extracted human molars (n = 15) using an acidified gel system (pH 4.5) for 10 or 40 days. PTR-LUM frequency scans (1 Hz-1 kHz) are performed before and during demineralization. Transverse microradiography (TMR) analysis, the current gold standard, follows at treatment conclusion to determine the mineral loss and depth of the artificially demineralized lesions. A theoretical model is applied to PTR experimental data to evaluate the changes in optothermophysical properties of demineralized enamel as a function of time. Higher optical scattering coefficients and poorer thermophysical properties are characteristic of the growing demineralized lesions, as verified by TMR, where the generated microporosities of the subsurface lesion confine the thermal-wave centroid. Enhanced optical scattering coefficients of demineralized lesions result in poorer luminescence yield due to scattering of both incident and converted luminescent photons. PTR-LUM sensitivity to changes in tooth mineralization coupled with opto-thermophysical property extraction illustrates the technique's potential for nondestructive quantification of enamel caries.     

Thermophotonic lock-in imaging of early demineralized and carious lesions in human teeth

As an extension of frequency-domain photothermal radiometry, a novel dental-imaging modality, thermophotonic lock-in imaging (TPLI), is introduced. This methodology uses photothermal wave principles and is capable of detecting early carious lesions and cracks on occlusal and approximal surfaces as well as early caries induced by artificial demineralizing solutions. The increased light scattering and absorption within early carious lesions increases the thermal-wave amplitude and shifts the thermal-wave centroid, producing contrast between the carious lesion and the intact enamel in both amplitude and phase images. Samples with artificial and natural occlusal and approximal caries were examined in this study. Thermophotonic effective detection depth is controlled by the modulation frequency according to the well-known concept of thermal diffusion length. TPLI phase images are emissivity normalized and therefore insensitive to the presence of stains. Amplitude images, on the other hand, provide integrated information from deeper enamel regions. It is concluded that the results of our noninvasive, noncontacting imaging methodology exhibit higher sensitivity to very early demineralization than dental radiographs and are in agreement with the destructive transverse microradiography mineral density profiles.     

Correlation of quantitative light-induced fluorescence and optical coherence tomography applied for detection and quantification of early dental caries

Fluorescence loss in enamel following demineralization has been correlated with the amount of mineral lost during the demineralization. The correlation between fluorescence loss measured by quantitative light-induced fluorescence (QLF) and the reflectivity loss measured by a versatile en face optical coherence tomography (OCT) system was investigated in a demineralization process to produce artificial dental caries. We used an OCT system that can collect A-scans (reflectivity versus depth), B-scans (longitudinal images), and C-scans (en face images). The power to the sample was 250 microW, the wavelength lambda = 850 nm, and the depth resolution in air 16 microm. A-scans, which show the profile of the reflectivity versus the depth of penetration into the tooth tissue, were used for quantitative analysis of the reflectivity loss. The results have shown that both the fluorescence radiance and reflectivity of the enamel decrease with increasing de- mineralization time. A linear correlation was observed between the percentage of fluorescence loss measured by QLF and the percentage of reflectivity loss measured by OCT. It was concluded that the decrease in reflectivity of the enamel during demineralization, measured by OCT, could be related to the amount of mineral lost during the de- mineralization process.     

Measurement of bone mineral density via light scattering

In this study we have investigated the potential of optical techniques to monitor changes in bone mineral density (BMD) via changes in scattering coefficient. For each of five bone samples, diffuse reflection and transmission coefficients were measured over the wavelength range 520-960 nm using an integrating sphere and CCD spectrometer. These were converted into optical absorption and scattering coefficients using a Monte Carlo inversion procedure. Measurements were made on samples immersed in formic acid solution for different lengths of time in order to investigate the effect of reduction in BMD on the optical properties. After full demineralization, the optical scattering coefficient fell by a factor 4. From the observed degree of fluctuation of the measurements, we estimate that BMD could be measured with an accuracy of 7% if optical scattering can be measured with an accuracy of 10%. We also report preliminary measurements of bone scattering using optical coherence tomography (OCT). An inter-side variability of 3% is obtained on dry samples with and without overlying periosteum. These results suggest that minimally invasive techniques for measuring optical scattering, such as OCT, may have a role in monitoring regional changes in BMD. This could be an important advance in our understanding of bone remodelling and its relationship to osteoarthritis. Both the integrating sphere and OCT measurements also suggest that light transport in bone is spatially anisotropic. OCT was used to assess probability of obtaining results in vivo.     

Imaging caries lesions and lesion progression with polarization sensitive optical coherence tomography

New diagnostic tools are needed for the characterization of dental caries in the early stages of development. If carious lesions are detected early enough, they can be arrested without the need for surgical intervention. The objective of this study was to demonstrate that polarization sensitive optical coherence tomography (PS-OCT) can be used for the imaging of early caries lesions and for the monitoring of lesion progression over time. High-resolution polarization resolved images were acquired of natural caries lesions and simulated caries lesions of varying severity created over time periods of 1 to 14 days. Linearly polarized light was incident on the tooth samples and the reflected intensity in both orthogonal polarizations was measured. PS-OCT was invaluable for removing the confounding influence of surface reflections and native birefringence necessary for the enhanced resolution of the surface structure of caries lesions. This study demonstrated that PS-OCT is well suited for the imaging of interproximal and occlusal caries, early root caries, and for imaging decay under composite fillings. Longitudinal measurements of the reflected light intensity in the orthogonal polarization state from the area of simulated caries lesions linearly correlated with the square root of time of demineralization indicating that PS-OCT is well suited for monitoring changes in enamel mineralization over time.     

基于光学相干断层成像量化评估平滑面早期牙釉质龋的研究进展

背景：平滑面早期牙釉质龋虽然已发生脱矿改变但无肉眼可见的龋洞形成,目前临床采用的常规检测方法难以对其准确诊断。 目的：应用光学相干断层成像技术对平滑面早期牙釉质龋的量化研究进展进行综述。 方法：由第一作者检索CNKI和EBSCO HOST数据库中1979/2011发表的关于光学相干断层成像技术定量研究平滑面早期龋的相关文献,以“光学相干断层成像,早期龋,平滑面,量化评估”和“optical coherence tomography, early caries, smooth surface,quantitative assessment”为检索词进行检索。纳入31篇进行综述。 结果与结论：新兴的光学相干断层成像技术能够检测龋变牙釉质的背向散射信号使脱矿牙釉质成像,并应用深度、反射率、衰减系数等定量指标对平滑面早期牙釉质龋进行量化评估,为牙釉质早期龋的诊断提供了新的方法。     

Ability of laser fluorescence device associated with fluorescent dyes in detecting and quantifying early smooth surface caries lesions

A laser fluorescence (LF) device is a portable tool, but it does not measure minor mineral changes. Our in vitro study aim is to propose the association of an LF with two fluorescent dyes and to evaluate the performance in detecting and quantifying early demineralization. Artificial caries lesions are created in 40 primary canine teeth using a demineralizing solution (pH=4.8) for 12, 24, 48, and 96 h. LF measurements are performed with DIAGNOdent after demineralization in these samples and in 20 sound primary teeth. Measurements with LF with 0.2-mM tetrakis(N-methylpyridyl)porphyrin (LF TMPyP) and with 4-mM protoporphyrin IX (LF PPIX) are made. The amount of calcium loss is determined by atomic emission spectrometry. A correlation between LF and LF with dyes and mineral loss and receiver operating characteristics analysis are performed, as well as comparisons of sensitivity, specificity, and accuracy values. Significant correlation is obtained with LF TMPyP and mineral loss of lesions demineralized for 24, 48, and 96 h. Better performance is achieved with LF TMPyP for all parameters than with LF alone. LF PPIX does not present good results. In conclusion, LF TMPyP provides good performance in detecting and quantifying very early enamel caries lesions.     

Quantitative light-induced fluorescence: a potential tool for general dental assessment

Current dental diagnostic methods can detect caries but cannot quantify the mineral status of a lesion. Quantitative light-induced fluorescence (QLF) measures the percentage of fluorescence change of demineralized enamel with respect to surrounding sound enamel, and relates it directly to the amount of mineral lost during demineralization. Development of caries-like lesions and subsequent remineralization of the lesions were monitored by QLF. The results showed that the percentage of fluorescence change (Delta Q) increased linearly with the demineralization time and decreased with increased remineralization time. Stained teeth were whitened with a bleaching agent and the change in stain intensity (Delta E) was quantified using QLF. The results showed that Delta E decreased linearly as the tooth regained its natural color. Factors that might affect the use of QLF to detect and quantify caries were also examined. It was concluded that QLF could be used to detect and longitudinally monitor the progression or remineralization of incipient caries, however lesion detection may be limited by the presence of saliva or plaque and enhanced by staining. The change in shade of discolored teeth by whitening agents could be quantitatively measured by QLF.     

Ex vivo detection and characterization of early dental caries by optical coherence tomography and Raman spectroscopy

Early dental caries detection will facilitate implementation of nonsurgical methods for arresting caries progression and promoting tooth remineralization. We present a method that combines optical coherence tomography (OCT) and Raman spectroscopy to provide morphological information and biochemical specificity for detecting and characterizing incipient carious lesions found in extracted human teeth. OCT imaging of tooth samples demonstrated increased light backscattering intensity at sites of carious lesions as compared to the sound enamel. The observed lesion depth on an OCT image was approximately 290 microm matching those previously documented for incipient caries. Using Raman microspectroscopy and fiber-optic-based Raman spectroscopy to characterize the caries further, spectral changes were observed in PO4 (3-) vibrations arising from hydroxyapatite of mineralized tooth tissue. Examination of various ratios of PO4 (3-) nu2, nu3, nu4 vibrations against the nu1 vibration showed consistent increases in carious lesions compared to sound enamel. The changes were attributed to demineralization-induced alterations of enamel crystallite morphology and/or orientation. OCT imaging is useful for screening carious sites and determining lesion depth, with Raman spectroscopy providing biochemical confirmation of caries. The combination has potential for development into a new fiber-optic diagnostic tool enabling dentists to identify early caries lesions with greater sensitivity and specificity.     

生物活性玻璃对早期人工釉质龋的再矿化

背景：生物活性玻璃具有良好的生物相容性,且具有抑制口腔致龋细菌和牙周相关细菌及抗牙本质过敏的作用。 目的：评价生物活性玻璃促进早期人工釉质龋再矿化的作用。 方法：将新鲜拔除的30颗牛切牙制成人工龋模型,将标本在37 ℃人工脱矿液内脱矿72 h,用扫描电镜观察脱矿后釉质表面的平滑情况,用显微硬度仪测量脱矿后釉质的显微硬度。然后随机分为3组,每组10个。采用pH循环法模拟人口腔环境,将3组标本分别浸泡在生物活性玻璃溶液、氟化钠溶液及人工唾液内,3次/d,10 min/次,循环浸泡20 d,扫描电镜检测标本脱矿及再矿化情况,用显微硬度计检查牙釉质显微硬度。 结果与结论：浸泡在生物活性玻璃溶液中的牙釉质表面较浸泡在其他两溶液中的牙釉质表面光滑平整,无空隙存在;浸泡在生物活性玻璃溶液中的牙釉质表面显微硬度高于浸泡在其他两溶液中的牙釉质表面显微硬度(P < 0.05)。说明生物活性玻璃在体外实验中能促进早期釉质龋的再矿化。     

体外动物组织热变过程中的光学特性变化

目的 研究热变过程中生物组织的光学特性变化。方法 采用犬肝、犬肾、犬肌肉和鼠肌肉4种动物组织作为样本,使用积分球系统测量650 nm波长激光照射下的样本在热风枪从30℃升温到100℃过程中的准直透射光、漫透射光和漫反射光光强随温度变化的数据,根据Kubelka-Munk模型计算得出吸收系数、散射系数和散射各向异性因子变化曲线。结果 温度变化使组织光学特性改变,热变过程中样本透射减弱,吸收系数和散射系数增大,散射各向异性因子减小,不同组织的变化有一定差别。结论 热变性使组织透射减弱,吸收和散射增强。     

蛋清热变过程中的光学特性变化

目的为解决手术中激光切割能力下降的问题,以鸡蛋蛋清为样本,研究热变过程中生物组织的光学特性变化。方法使用积分球系统测量650nm激光照射下的样本在热风枪加热下,从26％升温到99℃过程中的准直透射光、漫透射光和反射光光强随温度变化的数据,计算吸收系数和散射系数变化曲线。结果样本热变前光学特性受温度影响较小,热变过程中透射率降低,吸收系数和散射系数增大。结论手术中低强度激光照射组织发生热变使吸收能力下降,应保持较高激光强度以达到消融阈值。     

Effect of light losses of sample between two integrating spheres on optical properties estimation

A Monte Carlo algorithm is applied to simulate the measurements of a sample with glass slides sandwiched between the double integrating sphere (DIS) setup. The effects caused by various parameters, such as the sample port of integrating sphere, thicknesses, and optical properties of the sample, on light losses and optical properties estimated by the inverse adding-doubling method (IAD) have been investigated. The results show that the light loss greatly increases the estimated error of absorption coefficient and slightly affects the estimated scattering coefficient. When the increase of apparent absorption of the sample induced by the light loss is 59%, the relative error of the scattering coefficient is less than 2% and that of the absorption coefficient reaches 28%. Enhancing the sample port diameter or decreasing the thickness of the sample can reduce the error effectively, and the effect of the former is much greater than that of the latter. In addition, the IAD method is proved to be valid for estimating the optical properties of a highly scattering or highly absorbing sample. This study can not only evaluate the error of optical properties estimation, but also provide optimal ways for the design of DIS and a scheme for acquiring accurate optical properties of tissue.     

Effects of fluoride release from bis-GMA/TEGDMA resin regulated by gamma-methacryloxypropyltrimethoxysilane on demineralization of bovine enamel

We previously demonstrated that fluoride release from resins could be regulated by the polysiloxane coating of the fluoride additives. The present study investigated the effects of regulated fluoride release from resin on enamel demineralization in vitro. Bovine enamel cavities were restored with bis-GMA/TEGDMA resins containing 50 wt% NaF powders treated with or without gamma-methacryloxypropyltrimethoxysilane. Specimens were immersed in distilled water that was changed daily to measure the amount of fluoride released over 40 days, and thereafter subjected to pH-cycling. Microradiographic observations were performed to determine total mineral loss (AZ) and lesion depth (Ld) on the enamel. In addition, fluorine distribution was analyzed using EPMA. The resin containing untreated NaF exhibited high-rate and short-term fluoride release, whereas the resin containing treated NaF released low concentrations of fluoride over a longer period. The former showed high fluorine uptake in the adjacent enamel. In contrast, the latter showed high fluorine uptake not only in the adjacent enamel, but also in a wider area of enamel surface. The latter also showed lower AZ and Ld values in the surrounding enamel, indicating a high inhibitory effect on caries formation. Therefore, it is suggested that regulated fluoride release from the resin based on polysiloxane coating is effective in preventing caries formation.     

Determination of the optical properties of the human uterus using frequency-domain photon migration and steady-state techniques

The optical properties (absorption and transport scattering coefficients) of freshly excised, bulk human uterine tissues were measured at 630 nm using frequency-domain and steady-state photon migration techniques. Measurements were made on both normal (pre- and post-menopausal) and non-neoplastic fibrotic tissues. The absorption coefficient of normal post-menopausal tissue (approximately 0.06 mm(-1)) was found to be significantly greater than that of normal pre-menopausal tissue (0.02-0.03 mm(-1)) and pre-menopausal fibrotic tissue (0.008 mm(-1)). The transport scattering coefficient was similar in all three tissue types considered (0.6-0.9 mm(-1)). From the preliminary results presented here, we conclude that optical properties can be reliably calculated either from the frequency-dependent behaviour of diffusely propagating photon density waves or by combining the frequency-independent photon density wave phase velocity with steady-state light penetration depth measurements. Instrument bandwidth and tissue absorption relaxation time ultimately determine the useful frequency range necessary for frequency-domain photon migration (FDPM) measurements. Based on the optical properties measured in this study, we estimate that non-invasive FDPM measurements of normal uterine tissue require modulation frequencies in excess of 350 MHz.     

Estimation of lesion progress in artificial root caries by swept source optical coherence tomography in comparison to transverse microradiography

This study aimed to investigate whether swept source optical coherence tomography (SS-OCT) could estimate the lesion depth and mineral loss quantitatively without the use of polarization sensitivity, and to examine a relationship between OCT data and transverse microradiography (TMR) lesion parameters. Twenty-four bovine root dentin specimens were allocated to three groups of 4-, 7-, and 14-day demineralization. Cross-sectional images of the specimens before and after the demineralization were captured by OCT at 1319 nm center wavelength. Following the demineralization, these specimens were cut into sections for TMR analysis. Correlations between the OCT data and TMR lesion parameters were examined. TMR images of the specimens showed cavitated lesions (lesion depth or LD(TMR): 200 to 500 μm, ΔZ or mineral loss: 10,000 to 30,000 vol % μm). The OCT images showed "boundaries," suggesting the lesion front. Integrated dB values before and after the demineralization and their difference (R(D), R(S), and ΔR, respectively) were calculated from the lesion surface to the corrected depth of boundary (LD(OCT)). A statistically significant correlation was found between LD(OCT) and LD(TMR) (p < 0.05, r = 0.68). Similarly, statistically significant correlations were found between ΔZ and R(D) or ΔR. The OCT showed a potential for quantitative estimation of lesion depth and mineral loss with cavitated dentin lesions in vitro.     

Measurements of angular distributions of Rayleigh and Mie scattering events in biological models

Remission spectroscopy in living tissues can only be performed because scattering processes induce a pronounced amount of backscattered light. New types of scattering chambers were constructed in order to investigate the angular distribution of light intensities caused by scattering events typical for living tissues. Different solutions and suspensions containing microspheres, haemoglobin molecules, erythrocytes and liver homogenate were used in order to gain basic information applicable to remission spectrophotometry in tissue. Reflection spectroscopy in biological tissues is only possible because of the scattering properties of the material under observation. Light which penetrates the sample is remitted from the tissue. The remission is caused by the scattering of the light by different biological structures and particles. This scattered light gives information about the optical properties of the particles and structures in an integral form. A scattering chamber for small sample volumes has been developed for measurements of the optical properties of various particles in suspension. Measurements of scattering diagrams from microspheres with an average diameter of 7 microns, artificial particles, haemoglobin solutions, mixtures of microspheres and haemoglobin as well as erythrocytes and mitochondria have been performed. The different optical behaviour of the measured signals is qualitatively discussed.     

Optical properties of porcine skin dermis between 900 nm and 1500 nm

The weak absorption of shortwave infrared light by skin tissues between 700 and 1500 nm offers an important window for diagnosis by optical means. The strong scattering of shortwave infrared light by the skin, however, presents a challenge to the modelling of light propagation through the skin and the understanding of skin optics. We have measured the collimated and diffuse transmittance and diffuse reflectance of porcine skin dermis samples within 30 h post-mortem. Monte Carlo simulations have been performed to inversely determine the absorption coefficient, scattering coefficient and anisotropy factor of the dermis samples in the spectral range from 900 to 1500 nm. We further analyse the sensitivity of the values of the parameters to the experimental errors and inverse calculation procedures. The state of the cellular integrity of the skin samples following optical measurements was verified using transmission electron microscopy. These results were correlated to study post-mortem effects on the in vitro optical properties of porcine dermis. We concluded that for samples stored within crushed ice for up to 30 h post-mortem the wavelength dependence of optical properties of the dermis remains unchanged while the values of the parameters vary moderately due to modification of the water content of the tissue.     

Nanobacteria’s potential involvement in enamel repair in caries

Dental caries is the accumulation of numerous episodes of demineralization and remineralization, rather than a unidirectional demineralization process. Demineralization and remineralization occur constantly either simultaneously or alternately and whether a lesion will progress or be repaired depends upon the predominant process over periods. Even if fluoride has demonstrated the anti-caries effect by shifting the demineralization/remineralization balance favorably, little is known about non-fluoride action in favor of the balance and the effect of fluoride could not fully explain enamel repair in caries. Recently, in vitro experiments demonstrated enamel repair by synthetic apatite nanocrystals which showed the strong affinity, excellent biocompatibility, mechanical improvement, and a higher resistance to acids than apatite from teeth. This reminds us of a controversial microorganism called nanobacteria (NB) which form nanocrystalline apatite around themselves. Although NB have been detected in some pathological calcifications, epidemiologic literature suggests that they are widespread present in the healthy people blood. Considering the similarity of synthetic nanocrystalline apatite to that of NB and blood circulation communicating with saliva, we put forward a hypothesis that NB may act in enamel surface just like what the synthetic nanocrystalline apatite does in vitro to repair enamel in caries.