Raman spectroscopy with an integrated arrayed-waveguide grating

An integrated arrayed-waveguide grating fabricated in silicon-oxynitride technology is applied to Raman spectroscopy. After its validation by reproducing the well-known spectrum of cyclohexane, polarized Raman spectra are measured of extracted human teeth containing localized initial carious lesions. Excellent agreement is obtained between the spectra of healthy and carious tooth enamel measured with our integrated device and spectra recorded using a conventional Raman spectrometer. Our results represent a step toward the realization of compact, hand-held, integrated spectrometers, e.g. for the detection of dental caries at an early stage.     

Dental biothermophotonics: How photothermal methods are winning the race with X-rays for dental caries diagnostic needs of clinical dentistry

Recent trends in biothermophotonics of teeth are presented. The presentation is centered on the development of clinical-level frequency-domain photothermal radiometry and modulated luminescence to address issues associated with the early diagnosis of demineralization caries in human teeth. Biothermophotonic principles and applications to the detection of the carious state in human teeth as embodied by laser photothermal radiometry are presented and further supported by modulated luminescence. The emphasis is on recent developments with regard to abilities of these techniques to diagnose interproximal lesions between teeth, etching with phosphoric acid and with an artificial demineralization gel in order to simulate early demineralization, as well as demineralization and remineralization of dental crown enamel and root dentin. These are lesions which normally go undetected by X-ray radiographs. Comparisons with X rays, Micro-Computed Tomography (μ-CT) and Transverse Micro-Radiography (TMR) are discussed. A theoretical model involving coupled diffuse photon density and thermal-wave fields is developed and applied to frequency scans from demineralized artificial lesions to produce quantitative values for optical and thermophysical parameters of teeth as well as the thickness of the induced lesion.     

Local study of fissure caries by Fourier transform infrared microscopy and X‐ray diffraction using synchrotron radiation

Investigations of intact dental enamel as well as carious‐affected human dental enamel were performed using infrared spectromicroscopy and X‐ray diffraction applying synchrotron radiation. Caries of enamel was shown to be characterized by an increase in the number of deformation and valence vibrations for N—C—O, N—H and C=O bonds, a decrease of the crystallinity index, and by the absence of the preferable orientation of hydroxyapatite crystals within the affected enamel. This indicates the presence of destructive processes in the organic matrix of hard tooth tissues.     

Functional mapping of carious enamel in human teeth with Raman microspectroscopy

We employed Raman microspectroscopy to measure the Raman spectra of phosphate in sound and carious tooth substance. The peak intensity at 960 cm⁻¹ of the phosphate (PO₄³⁻) symmetric stretching vibrational mode (υ₁) in sound enamel was stronger than that of sound dentin, which indicated that sound enamel contained more phosphate than sound dentin. Furthermore, the element analysis of phosphate in sound teeth substance, measured using a scanning electron microscope (SEM) equipped with an energy dispersive X‐ray spectroscope (EDX), gave similar results to those of the Raman measurement. In addition, the border between sound enamel and dentin was clearly demonstrated by mapping the image of the Raman spectrum of phosphate. The mapping image of phosphate in the carious enamel region revealed a heterogeneous low Raman spectrum intensity of phosphate in the area surrounding carious enamel; this finding indicates that phosphate had dissolved from the tooth substance in such areas. In contrast with the decrease in the Raman spectrum intensity of phosphate, the intensity of amide I increased mainly in the low‐phosphate area. Although it remains very difficult to clinically identify the accurate border between sound and carious tooth substance, this distinction may be enabled by using the Raman spectrum of carious tooth substance. Copyright © 2008 John Wiley & Sons, Ltd.     

Broad‐beam PIXE and µ‐PIXE analysis of normal and in vitro demineralized dental enamel

Dental enamel has been widely studied by particle‐induced x‐ray emission (PIXE), but less attention was paid to its demineralization, which leads to caries formation. Using broad‐beam PIXE and µ‐PIXE, we investigated normal enamel and the in vitro formation of pre‐carious lesion in lactic acid solution, aiming also to evaluate intercusp differences within the same tooth. Broad‐beam PIXE was performed using 3.0 MeV protons, and µ‐PIXE maps of Ca, Fe and Zn were collected with 3.1 MeV protons at ～4 µm resolution. In normal enamel a differentiated Ca‐rich surface layer was observed, where Fe and Zn reached their highest levels. In deeper layers, Fe and Zn evidenced quasiperiodic patterns of maxima, possibly due to coupled diffusion‐reaction catalytic processes involved in the enamel growth. Both Fe and Zn appeared to be located in a few distinct types of pools. Near the surface, demineralization induced an increase of Fe, Cu, Zn, Sr and Pb with respect to Ca, attributed to partial hydroxyapatite dissolution and/or to chelate extraction and concentration of trace metals. Ca maps revealed limited changes in the surface layer and a massive loss in the inner enamel; here, Fe was almost depleted and Zn partially removed. The maps of Ca, Fe and Zn demonstrated major intercusp variations in both normal and altered enamel. Thus, broad‐beam PIXE and µ‐PIXE, which do not require (semi)thin sectioning of the tooth as the conventional methods, provide compositional and structural insight of normal dental enamel, of its intercusp variability and of the alterations produced by in vitro demineralization, largely not accessible to the current techniques, and highly relevant for understanding the incipient caries formation. Copyright © 2008 John Wiley & Sons, Ltd.     

电感耦合等离子体-原子发射光谱法测定龋齿中微量元素

牙齿样品经HNO3-HClO4体系消解,采用电感耦合等离子体-原子发射光谱法(ICP-AES)测定龋齿和健康齿的牙釉质与牙本质中Zn、Fe、Cu、Sr、Mg、Cr、Al和Pb等微量元素。方法相对标准偏差在1.1%—2.7%之间,标准回收率在90.0%—106.7%之间。实验测得龋齿牙釉质与牙本质中Zn、Fe、Cu和Sr含量均明显低于健康齿,而Mg、Cr、Al和Pb含量则均高于健康齿,说明牙齿中的微量元素是指示龋病的一种因子。本实验为口腔医学研究提出了一个简单、准确和可行的研究方法,也为医院口腔科预防和治疗龋齿,提供参考数据。     

Preparation of fluoride substituted apatite cements as the building blocks for tooth enamel restoration

Fluoride substituted apatite cement (fs-AC) was synthesized by using the cement powders of tetracalcium phosphate (TTCP) and sodium fluoride (NaF), and the cement powders were mixed with diluted phosphoric acid (H₃PO₄) as cement liquid to form fs-AC paste. The fs-AC paste could be directly filled into the carious cavities to repair damaged dental enamel. The results indicated that the fs-AC paste was changed into fluorapatite crystals with the atom molar ratio for calcium to phosphorus of 1.66 and the F ion amount of 3 wt% after self-hardening for 2 days. The solubility of fs-AC in Tris-HCl solution (pH 6) was slightly lower than hydroxyapatite cement (HAC) that was similar to the apatite in enamel, indicating the fs-AC was much insensitive to the weakly acidic solution than the apatite in enamel. The fs-AC was tightly combined with the enamel surface because of the chemical reaction between the fs-AC and the apatite in enamel after the caries cavities was filled with fs-AC. The extracts of fs-AC caused no cytotoxicity on L929 cells, which satisfied the relevant criterion on dental biomaterials, revealing good cytocompatibility. The fs-AC had potential prospect for the reconstitution of carious lesion of dental enamel.     

牙釉质中稀土和微量元素的ICP-MS方法检测

主要研究应用电感耦合等离子体质谱法测定牙釉质中稀土和微量元素的实验方法。实验证明牙釉质样品经硝酸-过氧化氢体系消解,样品溶解完全,可以满足ICP-MS检测方法的要求。方法的精密度用相对标准偏差表示在0.50%～6.79%之间,方法准确度以标准回收率度量在94.45%～104.78%之间,完全能满足生物样品的测试要求。本实验为口腔医学研究提出一个简便、准确和可行的研究方法的同时,也为龋齿病防治提供了有龋齿和无龋齿中样品中稀土微量元素测试数据。     

FT‐Raman investigation of human dental enamel surfaces

FT‐Raman spectra of human enamel surfaces from sound, affected (with 1 cavity) and highly affected (with at least 3 cavities) tooth samples were analyzed by principal component analysis (PCA). Major differences between the unaffected and affected tooth samples seem to arise from the structural changes along the c‐axis of hydroxyapatite, the chief crystalline component of human dental enamel. Based on Fisher index calculations, the most discriminative value was obtained for the intensity of the only Raman active ν₂PO₄³⁻ (E₁) symmetric deformation mode at 428 cm⁻¹. Moreover, these changes can be observed through the whole tooth enamel surface, establishing a predisposition to caries correlated to chemical and structural composition of tooth enamel. No spectral changes regarding the CO₃²⁻ substitution were detected by both nondestructive FT‐Raman and FTIR (Fourier transform infrared) spectroscopy of the powdered teeth samples. Copyright © 2009 John Wiley & Sons, Ltd.     

EPR-dosimetry with carious teeth

The effect of caries in EPR dosimetry of tooth enamel (in the dose range of 0–1 Gy) was investigated. The enamel of each tooth was divided into carious, non-carious and intermediate portions. The EPR signals of enamel at g = 2.0018 (dosimetric) and g = 2.0045 (native) were examined. The intensity of the dosimetric signal was the same for all three portions, while that of the native signal was higher for carious portions than for non-carious and intermediate portions. Reconstruction of the laboratory applied doses was done using all portions. Reasonable correlation between nominal and reconstructed doses was found in most cases. The effect of alkali treatment on the native and dosimetric signals of enamel was also tested. Reduction of the native signal intensity, particularly in the carious portions, was found to be the only significant effect. This resulted in a slight improvement in the accuracy of the reconstructed doses.     

Phase-resolved photoacoustic spectroscopy to monitor dental plaque on tooth enamel

Dental plaque is a biofilm on the enamel surface of the teeth, formed by colonizing bacteria. It is essential to monitor the biofilm presence on the surface of the enamel of the teeth, since plaque is a reason for periodontal disease and dental caries. This paper proposes applying photoacoustic spectroscopy to monitor and the phase-resolved photoacoustic method to analyze the biofilm plaque on samples of human dental enamel facets. The biofilm growth occurred in the oral cavity environment at deposition time of 48 and 72?h. Photoacoustic ex vivo monitoring found that: (1) Photoacoustic spectroscopy is a helpful tool to identify the bactericidal effect on dental plaque; (2) the biofilm deposition provides an optical absorption of the Soret band at 410?nm and increase of hydroxyl absorption band; (3) the biofilm thickness by phase-resolved photoacoustic method is in agreement with the literature in situ measurements. These new knowledges may help in development of a greater practicality and safety equipment able to identify and quantify dental biofilm, which may be achieved with low-cost Soret-band light emitting diode as light excitation.     

Absorption and thermal study of dental enamel when irradiated with Nd:YAG laser with the aim of caries prevention

It is widely recognized that Nd:YAG can increase enamel resistance to demineralization; however, the safe parameters and conditions that enable the application of Nd:YAG laser irradiation in vivo are still unknown. The aim of this study was to determine a dye as a photoabsorber for Nd:YAG laser and to verify in vitro a safe condition of Nd:YAG irradiation for caries prevention. Fifty-eight human teeth were selected. In a first morphological study, four dyes (waterproof India ink., iron oxide, caries indicator and coal paste) were tested before Nd:YAG laser irradiation, under two different irradiation conditions: 60 mJ/pulse and 10 Hz (84.9 J/cm²); 80 mJ/pulse and 10 Hz (113.1 J/cm²). In a second study, the enamel surface and pulp chamber temperatures were evaluated during laser irradiations. All dyes produced enamel surface melting, with the exception of the caries indicator, and coal paste was the only dye that could be completely removed. All irradiation conditions produced temperature increases of up to 615.08°C on the enamel surface. Nd:YAG laser irradiation at 60 mJ/pulse, 10 Hz and 84.9 J/cm² promoted no harmful temperature increase in the pulp chamber (ANOVA, p < 0.05). Among all dyes tested, the coal paste was an efficient photoabsorber for Nd:YAG irradiation, considered feasible for clinical practice. Nd:YAG laser at 84.9 J/cm² can be indicated as a safe parameter for use in caries prevention.     

牙齿龋齿损伤可见-近红外光谱偏振检测研究

针对不同龋齿损伤程度有效表征问题,探究一种光谱偏振无损低耗的检测方法,作为对常规检测方法如化学分析法、表面轮廓测定法、显微放射照相术的有益补充。牙齿龋齿损伤由脱矿溶解牙釉质与牙本质的钙与磷酸盐造成,牙齿组织表面微观结构发生了不同程度的变化,结构的变化导致其对光波散射特性与相位改变呈现不同程度的差异。基于偏振光谱对观测样本表面微观结构敏感的特征,不同龋齿损伤样本其表面微观结构变化与偏振信息强烈关联,提出一种光谱偏振有效表征牙齿龋齿损伤度的方法。分别选取450,550,670和860 nm四个不同观测光谱研究波段,选择六个不同龋坏程度的实验样本,选取偏振度参量描述不同牙齿样本的光谱偏振特性。实验结果表明,同一观测光谱波段随着牙齿龋坏程度的加深其偏振度表征参量呈现增加趋势,同时,对于同一观测样本450 nm光谱波段的偏振特性强于其他波段,呈现观测光谱波段与其偏振特性负相关关系。为进一步定量刻画偏振光谱与牙齿龋坏损伤等级的表征关系,构建光谱偏振特征参量与不同损伤等级牙齿样本的指数关联数学模型,通过数学模型解译其内在耦合关联。为有效验证构建模型的稳健性,对模型仿真预测结果与实测数据进行定量化验证,选取和方差（SSE）、模型确定系数（R-square）、均方根（RMSE）等评价因子对构建的模型进行定量评判。结果表明,四个不同光谱波段的模型确定系数均接近于1,和方差与均方根数值均较小,且450nm观测光谱波段接近于0,定量评价结果验证了构建模型的稳健性与有效性,构建模型可有效解译不同龋齿样本与光谱偏振特征的耦合关联。该研究内容有效扩展了牙齿龋齿检测方式,揭示了光谱偏振可有效表征龋齿损伤,发展了一种无损低耗的新型偏振光谱检测技术手段。     

Elemental and chemical compounds study of enamel and dentine of porcine primary molar teeth

For the enamel and dentin of porcine primary molar teeth, the spatial variability of the elements and chemical groups was studied with the aim of identifying the chemistry of porcine teeth, and it was then compared with results for human primary teeth. Porcine teeth were taken from specially bred animals. An electron microprobe was applied for the elemental analyses and a Raman microscope for the determination of chemical groups. The distribution of the main elements (Ca, P) was similar for both porcine and human teeth, whereas major differences were found in the distributions of minor elements (Mg, Sr, Cl, C). Carbonates were present in porcine dentin mainly as A-substituted species, whereas the B-substituted ones prevailed in enamel. The mechanism of asymmetric enrichment of Ca and P was quantitatively calculated as a consequence of Mg release. The layer close to the surface of the enamel with air had a unique composition. Cl involvement in enamel close to the boundary with air is a complex process, coupled with the involvement of Sr and release of Mg. The discoveries shed light on the role of elements in apatites, especially the role of carbonates and magnesium in porcine teeth, and put in doubt the potential use of pig apatite as a substitute for human apatite in filling materials.     

A Raman spectroscopic study of teeth affected with molar–incisor hypomineralisation

Raman spectroscopy was used to chemically map lesions associated with molar–incisor hypomineralisation in human teeth. Three teeth with hypomineralised lesions of differing severity, described as white, yellow or brown, were mapped using integral ratios of major component bands (hydroxyapatite, amide I and b‐type carbonate) and principal component analysis scores values. These lesions were found to contain depleted levels of mineral (hydroxyapatite) compared with those of healthy enamel. Principal component analysis also highlighted changes in the phosphate structure and variations in various organic constituents. These variations were consistent with increased disorder in the mineral component of the hypomineralised tooth lesions. Scanning electron microscopy–energy dispersive X‐ray spectroscopy supported the findings based on Raman spectroscopy. Copyright © 2015 John Wiley & Sons, Ltd.     

Numerical and experimental study of excited light and auto-fluorescence diffusion in teeth

Excited light and corresponding intrinsic fluorescence diffusion inside teeth tissue are an essential problem for light-based carious lesion detection. Based on finite element numerical analysis of diffusion equation, the photon density distribution of both excited light and autofluorescence of 2D premolar teeth model is obtained. The dependence of excited light and autofluorescence density distribution inside the teeth model on the scattering coefficient of enamel (5–25 mm⁻¹) and dentine (100–140 mm⁻¹) is numerically simulated and analyzed. The fitted results reveal that fluorescence intensity decreases exponentially. Optical penetration depth and fluorescence relative depth declined with the increment of scattering coefficient of enamel. And the dentine had the opposite effect. Finally, the experiment of measurement of fluorescence intensity on the teeth surface is conducted and the result is compared with the numerical computation.     

Wavelength-Resolved 3-Dimensional Fluorescence Lifetime Imaging

We have developed a compact system for wide-field fluorescence imaging, resolved in three spatial dimensions, lifetime and wavelength, that is based on a gated optical intensifier and an all-solid-state diode pumped Cr:LiSAF oscillator-amplifier system. Exploiting spectral separation, the system has been applied to human teeth, obtaining good lifetime contrast between enamel, dentin and caries. Exploiting spectral separation combined with depth resolution, the study of fluorescent microspheres led to an enhancement in both lifetime contrast and lateral resolution.     

Changes in dental enamel oven heated or irradiated with Er,Cr:YSGG laser. Analysis by FTIR

This study evaluated the change that occurs in dental enamel under action of oven heating or Er,Cr:YSGG laser irradiation aiming to obtain a structure more resistant to demineralization. Enamel powder was obtained from bovine teeth. Samples were subjected to oven heating at temperatures of 200, 400, 600, 800, and 1000°C or during laser irradiation with energy densities of 7.53, 10.95, and 13.74 J/cm². The infrared thermography was used to measure the surface temperature generated in the solid samples of enamel during lasers irradiation. The samples were analyzed by Fourier transform infrared spectroscopy (FTIR), which shows changes on enamel oven heated or laser irradiated, due to treatments, related to carbonates, adsorbed water and hydroxyl content. These compositional effects were more evident in lased samples. These changes may alter the material properties such as its solubility, and decrese of demineralization that is important for caries prevention.     

离体龋齿的光热锁相成像检测试验

牙齿龋损是人类一种广泛存在的口腔疾病,如何能够尽早的发现早期龋齿,对患者牙齿龋损的预防与治疗具有重要意义。介绍了基于光热效应的光热锁相热成像技术,光热锁相热成像技术具有无损伤、高效率及探测面积大等优势被广泛应用于各类材料的无损检测。基于光热锁相热成像技术原理对人为模拟龋损的人类离体牙齿组织进行成像检测试验研究。首先开展了对不同模拟龋损位置离体牙齿的光热锁相成像试验,采用防酸指甲油对牙齿组织测试面进行开窗受控龋损(开窗大小为5 mm×5 mm),其中模拟龋损部位分别选择在牙齿组织的邻接面及牙颌面。试验结果显示,牙齿组织模拟龋损位置热波幅值增大,相位滞后增大且光热锁相成像的幅值图与相位图与X射线成像相比对模拟龋损部位表现出高敏感及高特异性;光热锁相成像技术对邻接面龋损比牙颌面龋损具有较高特异性。其次开展了对平滑面牙齿组织样本的多天(0,1,2,4及6 d)模拟龋损光热锁相成像跟踪试验研究,牙齿组织龋损程度采用X射线方法进行量化。试验结果表明,当龋损时间小于6 d时,X射线方法无法对龋损位置进行有效识别,而采用光热锁相热成像技术检测模拟龋损1 d的牙齿组织时,幅值差为3.82,相位差为10.57°,证明光热锁相热成像方法对牙齿组织龋损具有较高敏感性。提取的光热锁相成像幅值与相位与龋损时间具有良好的相关性。     

Evaluation of enamel crystallites in subsurface lesion by microbeam X‐ray diffraction

Early caries lesion is a demineralization process that takes place in the top 0.1 mm layer of tooth enamel. In this study, X‐ray microbeam diffraction was used to evaluate the hydroxyapatite crystallites in the subsurface lesion of a bovine enamel section and the results are compared with those obtained by transversal microradiography, a method commonly used for evaluation of tooth mineral. Synchrotron radiation from SPring‐8 was used to obtain a microbeam with a diameter of 6 µm. Wide‐angle X‐ray diffraction reports the amount of hydroxyapatite crystals, and small‐angle X‐ray scattering reports that of voids in crystallites. All three methods showed a marked decrease in the enamel density in the subsurface region after demineralization. As these diffraction methods provide structural information in the nanometre range, they are useful for investigating the mechanism of the mineral loss in early caries lesion at a nanometre level.