龋病相关微生物及龋损组织自体荧光表征研究

发布时间：2019-05-17 19:10

【摘要】：目的：龋病是世界范围内广泛存在的口腔常见疾病,我国患龋率高达90%以上。随着口腔保健意识的增强,龋齿风险评估和早期检测成为口腔临床医学领域面临的重要问题。本论文从自体荧光光谱技术角度展开研究,为无创、准确、可定量的光学诊断技术应用于龋病精确化防控奠定实验基础。 方法及结果： ①应用荧光分光光度计对6种龋病相关微生物逐一进行光谱表征参数分析,发现6种龋病相关微生物在随着激发波长的递增,发射波出现了红移的现象；6种龋病相关微生物在350nm激发波长下,均存在436+4nm的特征荧光峰位值,提示此特征荧光来自于非卟啉物质。加血培养促进了龋病相关微生物的自体荧光代谢环境,微生物浓度与荧光强度之间存在线性正相关。 ②通过模拟口腔环境,形成不同致龋强度的人工菌斑,测试获得菌斑荧光光谱表征参数。结果显示培养第8天,1%蔗糖浓度组的菌斑溶液荧光强度高于其他蔗糖浓度组(P0.05)。部分实验组荧光强度第8天高于第4天(P0.05)；采用逐步回归分析方法设立了牙釉质片微硬度下降百分比、细菌粘附度与荧光参数的函数关系。 ③采用荧光分光光度计获得不同龋损程度离体牙在405nm波长激发下的荧光光谱分布特征,结果显示荧光光谱面积比率值P与龋齿检测仪DD值之间具有较好的线性关系(R=-0.9085)。分析了不同激光功率和不同检测部位对荧光光谱表征参数的影响。 ④在健康及不同龋损程度离体牙的荧光光谱特征参数基础上,分析了龋损等级与荧光面积比率值的关系(P0.01),龋损等级与主波长的关系(P0.01),荧光面积比率值α与荧光图像分量比率p的关系(P0.01),根据ROC分析方法获得了荧光图像分量比率β的龋损分级标准。 结论：本文探索了龋病相关微生物、人工菌斑、健康及不同龋损程度牙体组织的自体荧光光谱特征；通过荧光强度对口腔微生物进行定量化分析是可行的；研究了人工牙菌斑的荧光光谱表征性参数与其致龋性的关系；设立了基于龋损组织自体荧光光谱参数的荧光光谱面积比率值、荧光图像分量比率值,为基于自体荧光技术的龋活跃性评价和龋损分级提供了理论依据,为后期的龋齿检测仪研发提供必要的实验基础。
[Abstract]:Objective: caries is a common oral disease in the world. The incidence of caries in China is as high as 90%. With the increasing awareness of oral health care, dental caries risk assessment and early detection have become an important problem in the field of oral clinical medicine. In this paper, the study was carried out from the point of view of autofluorescence spectroscopy, which laid an experimental foundation for the application of noninvasive, accurate and quantitative optical diagnosis technology in the accurate prevention and control of caries. Methods and results: (1) the spectral characterization parameters of six kinds of caries related microorganisms were analyzed by fluorescence spectrophotometer one by one. It was found that the emission waves of 6 kinds of caries related microorganisms shifted red with the increase of excitation wavelength. The characteristic fluorescence peaks of 436 4nm were found in all six caries-related microorganisms at 350nm excitation wavelength, suggesting that the characteristic fluorescence came from non-porphyrin substances. Blood culture promoted the autofluorescence metabolic environment of caries-related microorganisms, and there was a linear positive correlation between microbial concentration and fluorescence intensity. (2) artificial plaque with different caries intensity was formed by simulating oral environment, and the fluorescence spectrum characterization parameters of plaque were obtained. The results showed that on the 8th day of culture, the fluorescence intensity of plaque solution in 1% sucrose concentration group was higher than that in other sucrose concentration groups (P 0.05). The fluorescence intensity of some experimental groups on the 8th day was higher than that on the 4th day (P 0.05). The percentage of microhardness decrease and the functional relationship between bacterial adhesion and fluorescence parameters were established by stepwise regression analysis. (3) the distribution characteristics of fluorescence spectra of isolated teeth with different degrees of caries excited by 405nm wavelength were obtained by fluorescence spectrophotometer. The results showed that there was a good linear relationship between fluorescence spectral area ratio P and DD value of dental caries detector (R 鈮，

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