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

Abstract

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.

Candidalysin Is Required for Neutrophil Recruitment and Virulence During Systemic Candida albicans Infection

“…Zebrafish maintenance and experimental work was performed in accordance with UK Home Office regulations and the UK Animals (Scientific Procedures) Act 1986. London wild-type zebrafish larvae were maintained as previously described [34]. For infection, methylcellulose-embedded, tricaine-anesthetized dechorionated larvae were injected systemically by direct inoculation into the common cardinal vein at 2 days postfertilization with 500 colony-forming units of C albicans in PBS or PBS alone as control (n = 20 zebrafish injected per treatment group).…”

Mechanisms of vascular damage by systemic dissemination of the oral pathogen Porphyromonas gingivalis

“…Dysregulated adhesion molecule expression may initiate vascular pathology due to abnormally elevated vascular permeability leading to oedema, chronic inflammatory and vascular damage [26]. We recently showed that Pg can cause vascular damage in an in vivo zebrafish larvae model of systemic infection in a gingipain-dependent manner [27], suggesting that these bacterial proteases may be important factors in the development of CVD. Here, we hypothesised that gingipain-dependent cleavage of endothelial cell junction adhesion molecules is important in mediating the vascular damage.…”

Cold-Shock Domain Family Proteins (Csps) Are Involved in Regulation of Virulence, Cellular Aggregation, and Flagella-Based Motility in Listeria monocytogenes

“…Our aims in this study were to further assess such roles of Csps in L. monocytogenes virulence in vitro using a human macrophage cell line as well as in vivo using a zebrafish embryo based infection model. Zebrafish embryos are an attractive multicellular model for infection studies, that are easily accessible for microscopy due to their translucent nature, and have been widely used in microbial research with various bacteria including L. monocytogenes (Prajsnar et al, 2008 ; Levraud et al, 2009 ; Widziolek et al, 2016 ). Observations from these virulence studies also led us to examine the role of Csps in cellular aggregation as well as in extracellular motility and flagella production of L. monocytogenes .…”