Subgingival lipid A profile and endotoxin activity in periodontal health and disease
Regulation of lipopolysaccharide (LPS) chemical composition, particularly its lipid A domain, is an important, naturally occurring mechanism that drives bacteria-host immune system interactions into either a symbiotic or pathogenic relationship. Members of the subgingival oral microbiota can critically modulate host immuno-inflammatory responses by synthesizing different LPS isoforms. The objectives of this study were to analyze subgingival lipid A profiles and endotoxin activities in periodontal health and disease and to evaluate the use of the recombinant factor C assay as a new, lipid A-based biosensor for personalized, point-of-care periodontal therapy.
Materials and methods
Subgingival plaque samples were collected from healthy individuals and chronic periodontitis patients before and after periodontal therapy. Chemical composition of subgingival lipid A moieties was determined by ESI-Mass Spectrometry. Endotoxin activity of subgingival LPS extracts was assessed using the recombinant factor C assay, and their inflammatory potential was examined in THP-1-derived macrophages by measuring TNF-α and IL-8 production.
Characteristic lipid A molecular signatures, corresponding to over-acylated, bi-phosphorylated lipid A isoforms, were observed in diseased samples. Healthy and post-treatment samples were characterized by lower m/z peaks, related to under-acylated, hypo-phosphorylated lipid A structures. Endotoxin activity levels and inflammatory potentials of subgingival LPS extracts from periodontitis patients were significantly higher compared to healthy and post-treatment samples.
This is the first study to consider structure-function-clinical implications of different lipid A isoforms present in the subgingival niche and sheds new light on molecular pathogenic mechanisms of subgingival biofilm communities.
Subgingival endotoxin activity (determined by lipid A chemical composition) could be a reliable, bacterially derived biomarker and a risk assessment tool for personalized periodontal care.
KeywordsLipopolysaccharide Lipid A Subgingival microbiota Biomarker Periodontal diseases
This study was supported by the Oral and Dental Research Trust’s GSK Research Grants Programme, the Dowager Countess Eleanor Peel Trust (219), and the New National Excellence Program of the Hungarian Ministry of Human Capacities (ÚNKP-17-4-III and NKFIH K-125275). Plymouth University Peninsula Schools of Medicine and Dentistry also acknowledges the support of the National Institute of Health Research Clinical Research Network (NIHR CRN).
Compliance with ethical standards
Conflict of interest
SZ has a patent on periodontitis biomarkers, publication number GB2549712. Author AS declares that he has no conflict of interest. Author ZH declares that she has no conflict of interest. Author CM declares that she has no conflict of interest. Author MJ declares that he has no conflict of interest. Author LB declares that she has no conflict of interest. Author AK declares that she has no conflict of interest. Author AF declares that he has no conflict of interest. Author SJ declares that he has no conflict of interest.
All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.
Informed consent was obtained from all individual participants included in the study.
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