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Abstract：
OBJECTIVE: Calcifying nanoparticles (CNPs), referred to as nanobacteria (NB), are recognized to be associated with ectopic calcification. This study aims to isolate and culture CNPs from the dental plaque of patients with periodontal disease and investigate their possible role in unravelling the aetiology of periodontal disease.
METHODS: Supragingival and subgingival plaques were sampled from 30 periodontitis patients for CNPs isolation and culture. Alkaline phosphatase (ALP) content changes were tracked over time. Positive samples underwent thorough morphological identification via hematoxylin and eosin (HE) staining, Alizarin red S (ARS), and transmission electron microscopy (TEM). The chemical composition of CNPs analysis involved calcium (Ca) and phosphorus (P) content determination, Fourier transform infrared spectroscopy (FTIR), and X-ray diffraction (XRD).
RESULTS: The subgingival plaque dental group exhibited a higher CNPs isolation rate at 36.67% (11/30) compared to the supragingival dental plaque group at 66.67% (20/30). ALP activity varied among the positive, negative and control groups. Morphological observation characterized the CNPs as round, oval, and ellipsoid particles with Ca deposits. Chemical analysis revealed the Ca/P ratio was 0.6753. Hydroxyl, methyl, carbonate, phosphate, hydrogen phosphate, and dihydrogen phosphate were detected by FTIR; the main chemical components detected by XRD were hydroxyapatite and tricalcium phosphate.
CONCLUSIONS: CNPs were found in periodontitis-related dental plaque and exhibited the potential to develop calcified structures resembling dental calculus. However, the potential involvement of ALP in CNPs formation requires deeper exploration, as does the precise nature of its role and the interrelation with periodontitis demand a further comprehensive investigation.
METHODS: Supragingival and subgingival plaques were sampled from 30 periodontitis patients for CNPs isolation and culture. Alkaline phosphatase (ALP) content changes were tracked over time. Positive samples underwent thorough morphological identification via hematoxylin and eosin (HE) staining, Alizarin red S (ARS), and transmission electron microscopy (TEM). The chemical composition of CNPs analysis involved calcium (Ca) and phosphorus (P) content determination, Fourier transform infrared spectroscopy (FTIR), and X-ray diffraction (XRD).
RESULTS: The subgingival plaque dental group exhibited a higher CNPs isolation rate at 36.67% (11/30) compared to the supragingival dental plaque group at 66.67% (20/30). ALP activity varied among the positive, negative and control groups. Morphological observation characterized the CNPs as round, oval, and ellipsoid particles with Ca deposits. Chemical analysis revealed the Ca/P ratio was 0.6753. Hydroxyl, methyl, carbonate, phosphate, hydrogen phosphate, and dihydrogen phosphate were detected by FTIR; the main chemical components detected by XRD were hydroxyapatite and tricalcium phosphate.
CONCLUSIONS: CNPs were found in periodontitis-related dental plaque and exhibited the potential to develop calcified structures resembling dental calculus. However, the potential involvement of ALP in CNPs formation requires deeper exploration, as does the precise nature of its role and the interrelation with periodontitis demand a further comprehensive investigation.
摘要：
目的：钙化纳米颗粒（CNPs），被称为纳米细菌（NB），被认为与异位钙化有关。本研究旨在从牙周病患者的牙菌斑中分离和培养CNP，并研究其在揭示牙周病病因中的可能作用。
方法：从30例牙周炎患者中取样进行CNP分离和培养。跟踪碱性磷酸酶(ALP)含量随时间的变化。阳性样品通过苏木精和伊红（HE）染色进行彻底的形态学鉴定，茜素红S(ARS)，和透射电子显微镜（TEM）。CNPs的化学成分分析涉及钙(Ca)和磷(P)含量的测定,傅里叶变换红外光谱(FTIR),和X射线衍射(XRD)。
结果：与66.67%（20/30）相比，龈下菌斑组的CNPs分离率为36.67%（11/30）。ALP活性在阳性之间变化，阴性组和对照组。形态学观察显示CNPs为圆形，椭圆形,和带有Ca沉积物的椭球颗粒。化学分析显示Ca/P比为0.6753。羟基,甲基，碳酸盐,磷酸盐,磷酸氢盐,用FTIR检测磷酸二氢盐和磷酸二氢盐;XRD检测的主要化学成分为羟基磷灰石和磷酸三钙。
结论：在与牙周炎相关的牙菌斑中发现了CNPs，并显示出形成类似牙结石的钙化结构的潜力。然而,ALP在CNPs形成中的潜在参与需要更深入的探索，其作用的确切性质以及与牙周炎的相互关系也需要进一步全面的研究。
方法：从30例牙周炎患者中取样进行CNP分离和培养。跟踪碱性磷酸酶(ALP)含量随时间的变化。阳性样品通过苏木精和伊红（HE）染色进行彻底的形态学鉴定，茜素红S(ARS)，和透射电子显微镜（TEM）。CNPs的化学成分分析涉及钙(Ca)和磷(P)含量的测定,傅里叶变换红外光谱(FTIR),和X射线衍射(XRD)。
结果：与66.67%（20/30）相比，龈下菌斑组的CNPs分离率为36.67%（11/30）。ALP活性在阳性之间变化，阴性组和对照组。形态学观察显示CNPs为圆形，椭圆形,和带有Ca沉积物的椭球颗粒。化学分析显示Ca/P比为0.6753。羟基,甲基，碳酸盐,磷酸盐,磷酸氢盐,用FTIR检测磷酸二氢盐和磷酸二氢盐;XRD检测的主要化学成分为羟基磷灰石和磷酸三钙。
结论：在与牙周炎相关的牙菌斑中发现了CNPs，并显示出形成类似牙结石的钙化结构的潜力。然而,ALP在CNPs形成中的潜在参与需要更深入的探索，其作用的确切性质以及与牙周炎的相互关系也需要进一步全面的研究。
