Three/four-dimensional (3D/4D) microscopic imaging and processing in clinical dental research

Yu, Hong; Houshmandi, Mojgan

doi:10.1186/s12903-016-0282-0

Cited by 6 publications

(6 citation statements)

References 28 publications

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“…K2 also disrupts epithelial barrier function. 33 Impairment of epithelial barrier function leading to enhanced penetration of microbial products is central to the concept that destructive periodontitis is an immuno-pathological response to microbial products. 27 …”

Section: Discussionmentioning

confidence: 99%

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An early report: a modified porphyrin-linked metronidazole targeting intracellular Porphyromonas gingivalis in cultured oral epithelial cells

2017

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Porphyromonas gingivalis (P. gingivalis) has a strong association with the pathogenesis of periodontal disease. Recurrence of periodontal disease following therapy is attributed to numerous factors, and of growing interest is the potential problem of intracellular bacteria that are able to persist and multiply within the host cell, thereby facilitating relapse of infection. The effect of antibiotic therapy in controlling P. gingivalis is questionable. Accordingly, while metronidazole is very effective against anaerobic extracellular P. gingivalis by disrupting the DNA of anaerobic microbial cells, this antibiotic does not effectively penetrate into mammalian cells to inhibit intracellular bacteria. Therefore in the present study, a modified porphyrin-linked metronidazole adducts, developed in our laboratory, was used to kill intracellular P. gingivalis. A series of experiments were performed, including cytotoxicity assays and cellular uptake of adducts by flow cytometry coupled with live cell imaging analysis, P. gingivalis invasion and elimination assays, and the analysis of colocalization of P. gingivalis and porphyrin-linked metronidazole by confocal laser scanning microscopy. Findings indicated that P. gingivalis and porphyrin-linked metronidazole were colocalized in the cytoplasm, and this compound was able to kill P. gingivalis intracellular with a sufficient culture time. This is a novel antimicrobial approach in the elimination of P. gingivalis from the oral cavity.

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Section: Discussionmentioning

confidence: 99%

“…Live cells without fixation were to determine the penetration/localization of this compound (auto-fluorescence in red) for up to 4 h. 3D reconstructions were built up by z -stack images using the 3D Olympus Fluoview software (Shinjuku-ku, Tokyo, Japan). 33 …”

Section: Methodsmentioning

confidence: 99%

An early report: a modified porphyrin-linked metronidazole targeting intracellular Porphyromonas gingivalis in cultured oral epithelial cells

2017

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“…73 This feature allows to image for example cells, tissues, 3D scaffolds/sponges, which showed promise in copious application, including microuidics and exible electronics, [74][75][76][77] or nanostructures and subsequently illustrate them 3-dimensional. [78][79][80] Nowadays, the research is shiing towards nanotechnology (for example nanocontainers) and fundamental biology of cells, which are in a size-range of tens to few hundred nm. 81 Here, optical microscopy methods lack sufficient resolution due to the diffraction limit described by Abbe.…”

Section: Combination Of Afm With Optical Surface Sensitive Methodsmentioning

confidence: 99%

Recent advances in hybrid measurement methods based on atomic force microscopy and surface sensitive measurement techniques

2017

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“…Considering the technical challenges required for traditional serial thin sectioning approaches, especially for composite soft/hard tissues, alternative workflows avoiding sample decalcification have been developed ( Figure 1 ). Recently, various fluorescence microscopy techniques, including confocal scanning or light sheet fluorescence microscopy (LSFM), combined or not with clearing methods, have been proposed for composite structures (including dental and bone tissue) imaging ( Kabasawa et al, 1995 ; Ye et al, 2016 ; Hong et al, 2019 ). Confocal microscopy, useful for high-resolution analysis, generates 3D images for about a few hundred microns (100–200 μm) of sample depth.…”

Section: Introductionmentioning

confidence: 99%

A Spectral Principal Component Analysis-Based Framework for Composite Hard/Soft Tissue Fluorescence Image Investigation

et al. 2022

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Traditional thin sectioning microscopy of large bone and dental tissue samples using demineralization may disrupt structure morphologies and even damage soft tissues, thus compromising the histopathological investigation. Here, we developed a synergistic and original framework on thick sections based on wide-field multi-fluorescence imaging and spectral Principal Component Analysis (sPCA) as an alternative, fast, versatile, and reliable solution, suitable for highly mineralized tissue structure sustain and visualization. Periodontal 2-mm thick sections were stained with a solution containing five fluorescent dyes chosen for their ability to discriminate close tissues, and acquisitions were performed with a multi-zoom macroscope for blue, green, red, and NIR (near-infrared) emissions. Eigen-images derived from both standard scaler (Std) and Contrast Limited Adaptive Histogram Equalization (Clahe) pre-preprocessing significantly enhanced tissue contrasts, highly suitable for histopathological investigation with an in-depth detail for sub-tissue structure discrimination. Using this method, it is possible to preserve and delineate accurately the different anatomical/morphological features of the periodontium, a complex tooth-supporting multi-tissue. Indeed, we achieve characterization of gingiva, alveolar bone, cementum, and periodontal ligament tissues. The ease and adaptability of this approach make it an effective method for providing high-contrast features that are not usually available in standard staining histology. Beyond periodontal investigations, this first proof of concept of an sPCA solution for optical microscopy of complex structures, especially including mineralized tissues opens new perspectives to deal with other chronic diseases involving complex tissue and organ defects. Overall, such an imaging framework appears to be a novel and convenient strategy for optical microscopy investigation.

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Three/four-dimensional (3D/4D) microscopic imaging and processing in clinical dental research

Cited by 6 publications

References 28 publications

An early report: a modified porphyrin-linked metronidazole targeting intracellular Porphyromonas gingivalis in cultured oral epithelial cells

An early report: a modified porphyrin-linked metronidazole targeting intracellular Porphyromonas gingivalis in cultured oral epithelial cells

Recent advances in hybrid measurement methods based on atomic force microscopy and surface sensitive measurement techniques

A Spectral Principal Component Analysis-Based Framework for Composite Hard/Soft Tissue Fluorescence Image Investigation

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