Pro-CanDy (Probiotic-Loaded Hydrogels for the Prevention and Management of Oral Candidiasis and Dysbiosis) - Contact PD/PI: MYLONAKIS, ELEFTHERIOS PROJECT SUMMARY: Disruptions in microbial homeostasis due to age, antibiotic use, chronic illness, or immunosuppression promote overgrowth of opportunistic pathogens such as Candida albicans and Streptococcus mutans. This contributes to oral conditions like candidiasis, caries, and mucosal inflammation, and is linked to cardiovascular disease, diabetes, autoimmune disorders, adverse pregnancy outcomes, and gut-brain axis disturbances. Conventional antifungal therapies often fail to resolve biofilm-associated infections and may drive resistance, underscoring the need for safe, locally-acting alternatives. We have developed Lactobacillus paracasei 28.4-based probiotic strategies to restore mucosal homeostasis and target fungal and bacterial pathogens. Originally isolated from a caries-free individual, L. paracasei 28.4 inhibits C. albicans and S. mutans by reducing adhesion, biofilm formation, and virulence gene expression. Delivered via gellan gum hydrogels, it significantly lowers fungal burden and inflammation in murine oropharyngeal candidiasis models. Its postbiotic components also suppress Candida auris and drug-tolerant persister cells. Preliminary studies further enhanced efficacy by integrating L. paracasei 28.4 with caffeic acid phenethyl ester (CAPE), a natural antifungal and immunomodulator, in biocompatible hydrogels enabling controlled release and targeted mucosal delivery. We propose to advance gellan-based hydrogels with L. paracasei 28.4, with or without CAPE, to inhibit virulence, disrupt C. albicans and S. mutans biofilms, enhance epithelial barrier function, and modulate innate immunity. Aim 1 will optimize formulation for stability, probiotic viability, controlled release, and epithelial compatibility. Aim 2 will test safety, microbiome effects, and efficacy in murine models of candidiasis and dysbiosis. Aim 3 focuses on translational development, including in vitro safety testing, evaluation in immunocompromised hosts, and GMP-aligned formulation. This project introduces a novel, multi-modal strategy to manage oral dysbiosis via localized delivery of probiotics and natural antifungals—limiting pathogen overgrowth, restoring microbial balance, and reducing reliance on conventional drugs. Our gellan-based hydrogel co-delivers L. paracasei 28.4 ± CAPE directly to oral tissues, disrupting C. albicans and S. mutans biofilms, preventing recolonization, and preserving mucosal integrity. By maintaining probiotic viability and enabling targeted CAPE release, the system achieves strong efficacy in murine models. This biocompatible platform integrates pathogen control, immune modulation, and microbiome restoration, offering a transformative, resistance-limiting approach for difficult oral infections. References Cited Page 1
