Abstract
Periodontal infections cause inflammatory destruction of the tooth supporting tissues. We recently developed a dynamic, in vitro periodontal organotypic tissue model in a perfusion bioreactor system, in co-culture with an 11-species subgingival biofilm, which may recapitulate early events during the establishment of periodontal infections. This study aimed to characterize the global proteome regulations in this host-biofilm interaction model. Semi-quantitative shotgun proteomics were applied for protein identification and quantification in the co-culture supernatants (human and bacterial) and the biofilm lysates (bacterial). A total of 896 and 3363 proteins were identified as secreted in the supernatant and expressed in the biofilm lysate, respectively. Enriched gene ontology analysis revealed that the regulated secreted human tissue proteins were related to processes of cytoskeletal rearrangement, stress responses, apoptosis, and antigen presentation, all of which are commensurate with deregulated host responses. Most secreted bacterial biofilm proteins derived from their cytoplasmic domain. In the presence of the tissue, the levels of Fusobacterium nucleatum, Actinomyces oris and Campylobacter rectus proteins were significantly regulated. The functions of the up-regulated intracellular (biofilm lysate) proteins were associated with cytokinesis. In conclusion, the proteomic overview of regulated pathways in this host-biofilm interaction model provides insights to the early events of periodontal pathogenesis.
Highlights
The periodontium is a syncytium of specialised tissues that surround and support the teeth
We recently developed a complex periodontal infection model[13] that includes an 11-species biofilm used to challenge a generated organotypic tissue consisting of gingival epithelial, gingival fibroblast and monocytic cells grown on collagen sponges
Histology and electron microscopy revealed that the tissue consisted of a superficial epithelial-like layer and an underlying collagen-supported connective tissue rich in gingival fibroblasts and monocytic cells
Summary
The periodontium is a syncytium of specialised tissues that surround and support the teeth. The study of periodontal disease on the proteomic level has become increasingly popular in recent years, as a result of improvements in mass spectrometry-based technologies[14]. The present study utilised semi-quantitative proteomics with the aim to characterise changes in the proteome that take place in our recently established in vitro periodontal infection model[13]. This approach aspires to unravel in more detail the intricate interactions between the gingival tissue and subgingival biofilms during the early stages of the establishment of periodontal inflammation
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