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Porphyromonas gingivalis (Pg ) is a keystone pathogen in the aetiology of chronic periodontitis. However, recent evidence suggests that the bacterium is also able to enter the bloodstream, interact with host cells and tissues, and ultimately contribute to the pathogenesis of cardiovascular disease (CVD). Here we established a novel zebrafish larvae systemic infection model showing that  Pg  rapidly adheres to and penetrates the zebrafish vascular endothelium causing a dose- and time-dependent mortality with associated development of pericardial oedemas and cardiac damage. The  in vivo  model was then used to probe the role of  Pg  expressed gingipain proteases using systemically delivered gingipain-deficient  Pg  mutants, which displayed significantly reduced zebrafish morbidity and mortality compared to wild-type bacteria. In addition, we used the zebrafish model to show efficacy of a gingipain inhibitor (KYT) on  Pg -mediated systemic disease, suggesting its potential use therapeutically. Our data reveal the first real-time  in vivo  evidence of intracellular  Pg  within the endothelium of an infection model and establishes that gingipains are crucially linked to systemic disease and potentially contribute to CVD.

Zebrafish as a new model to study effects of periodontal pathogens on cardiovascular diseases