In Situ Raman Study of Neurodegenerated Human Neuroblastoma Cells Exposed to Outer-Membrane Vesicles Isolated from <i>Porphyromonas gingivalis</i>

In Situ Raman Study of Neurodegenerated Human Neuroblastoma Cells Exposed to Outer-Membrane Vesicles Isolated from <i>Porphyromonas gingivalis</i>

The aim of this study was to elucidate the chemistry of cellular degeneration in human neuroblastoma cells upon exposure to outer-membrane vesicles (OMVs) produced by <i>Porphyromonas gingivalis</i> (<i>Pg</i>) oral bacteria by monitoring their metabolomic evolution using in...

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Bibliographic Details
Main Authors: Giuseppe Pezzotti, Tetsuya Adachi, Hayata Imamura, Davide Redolfi Bristol, Keiji Adachi, Toshiro Yamamoto, Narisato Kanamura, Elia Marin, Wenliang Zhu, Toshihisa Kawai, Osam Mazda, Toru Kariu, Tomonori Waku, Frank C. Nichols, Pietro Riello, Flavio Rizzolio, Tania Limongi, Kazu Okuma
Format: Article
Language:English
Published: MDPI AG 2023-08-01
Series:International Journal of Molecular Sciences
Subjects:
outer-membrane vesicles
<i>Porphyromonas gingivalis</i>
Raman spectroscopy
phosphorylated dihydroceramides
cysteine proteases
amyloid β
Online Access:https://www.mdpi.com/1422-0067/24/17/13351
Description
Summary:The aim of this study was to elucidate the chemistry of cellular degeneration in human neuroblastoma cells upon exposure to outer-membrane vesicles (OMVs) produced by <i>Porphyromonas gingivalis</i> (<i>Pg</i>) oral bacteria by monitoring their metabolomic evolution using in situ Raman spectroscopy. <i>Pg</i>-OMVs are a key factor in Alzheimer’s disease (AD) pathogenesis, as they act as efficient vectors for the delivery of toxins promoting neuronal damage. However, the chemical mechanisms underlying the direct impact of <i>Pg</i>-OMVs on cell metabolites at the molecular scale still remain conspicuously unclear. A widely used in vitro model employing neuroblastoma SH-SY5Y cells (a sub-line of the SK-N-SH cell line) was spectroscopically analyzed in situ before and 6 h after <i>Pg</i>-OMV contamination. Concurrently, Raman characterizations were also performed on isolated <i>Pg</i>-OMVs, which included phosphorylated dihydroceramide (PDHC) lipids and lipopolysaccharide (LPS), the latter in turn being contaminated with a highly pathogenic class of cysteine proteases, a key factor in neuronal cell degradation. Raman characterizations located lipopolysaccharide fingerprints in the vesicle structure and unveiled so far unproved aspects of the chemistry behind protein degradation induced by <i>Pg</i>-OMV contamination of SH-SY5Y cells. The observed alterations of cells’ Raman profiles were then discussed in view of key factors including the formation of amyloid β (Aβ) plaques and hyperphosphorylated Tau neurofibrillary tangles, and the formation of cholesterol agglomerates that exacerbate AD pathologies.
ISSN:1661-6596
1422-0067

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