ORASPORT Study Protocol: A Prospective Observational Study on the Impact of Endurance Physical Activity on Oral Microbiology and Immunology

Morgane Ortis^1,2*Margaux Dubois^1,2Valerie Bougault³Kevin Legueult⁴Julien Fassy⁵Bernard Mari⁵Marie-France Bertrand^1,2Laurence Lupi^1,2Alain DOGLIO^1,6

• ¹Laboratoire MICORALIS, Université Côte d'Azur, Nice, France
• ²Institut de médecine bucco-dentaire, Centre Hospitalier Universitaire de Nice, Pôle Odontologie, Nice, France
• ³Universite Cote d'Azur Laboratoire Motricite Humaine Expertise Sport Sante, Nice, France
• ⁴Département de Santé Publique, Université Côte d'Azur, Centre Hospitalier Universitaire de l’Archet, Nice, France
• ⁵Institut de Pharmacologie Moleculaire et Cellulaire, Valbonne, France
• ⁶Unité de Thérapie Cellulaire et Génique (UTCG), Centre Hospitalier Universitaire de Nice, Nice, France

The oral cavity offers a non-invasive window into systemic health. Elite athletes often present with poor oral health, potentially linked to high physiological stress, psychological strain, and specific dietary habits. This study explores the relationship between physical activity and oral health, with a particular focus on identifying key changes in the oral microbiota and salivary immune profiles. The main objective is to compare the composition of oral microbiota and the levels of salivary cytokines among young healthy individuals with varying levels of endurance physical activity. The ORASPORT study is a prospective cross-sectional non-interventional study involving 200 participants recruited in a single center. The participants are healthy young individuals aged 18–30, with or without regular endurance physical activity. They will be divided into three groups according to their physical activity level, following international guidelines set by the World Health Organization (WHO). For each consenting participant, a standardized oral health assessment will be conducted by a dentist to establish a comprehensive oral health profile. Participants will also complete a detailed questionnaire to document individual data on physical activity, dietary habits, and overall health status. Additionally, oral biological samples, including saliva and gingival smears, will be collected for microbiological analysis and cytokine detection. Microbiological analysis of all oral samples will be performed using high-throughput qPCR with Biomark HD™ technology, to simultaneously detect 40 oral bacterial species, selected to represent both commensal and pathogenic oral flora, as well as the presence of 8 Human Herpesviruses (HHVs). Concurrently, the salivary concentrations of 9 inflammatory cytokines will be quantified using Meso Scale DiscoveryⓇ multiplex immunoassays. Statistical analysis will include both univariate and multivariate models to identify correlations between physical activity, oral health parameters, and microbial-immune profiles. This study protocol integrates clinical, microbiological, and immunological analysis to identify non-invasive biomarkers of oral health and physical performance. This study complies with French bioethics regulations (Jardé Law) and with data protection under European GDPR requirements. It received French ethical approval from the Committee for the Protection of Persons in February 2023. Findings will be shared through publications in peer-reviewed journals and presentations at international conferences. Clinical trial number: NCT05765422