Development of an Integrated Dental Plaque Bioanalytical Platform for Point-of-Care Diagnostics of Oral Diseases - ABSTRACT Dental caries poses a significant financial burden to the oral healthcare system in the US. Hence, early detection and preventive measures for primary caries are essential for reducing treatment costs. It is well-established that biofilm metabolic activity (lactic acid production) in the oral cavity, particularly close to the teeth, plays a pivotal role in the progression of dental caries, and hydrogen peroxide (H2O2) has been identified as a crucial factor in controlling lactic acid producing bacteria. We therefore propose to develop a quantitative and easy-to-read electrochemical biosensing platform that will use live dental plaque samples for potential point-of-care diagnostics to test the following hypothesis: Can the capacity of dental plaque to produce metabolites—such as H2O2, associated with oral health, and lactic acid, linked to caries development—be used to establish threshold values for the early detection of caries? AIM 1: Detect and quantify hydrogen peroxide and lactate produced by varying ratios of the bacterial species S. sanguinis (Ss) and S. mutans (Sm). Our innovative electrochemical sensors will be used along with microliter volume droplet hydrogel electrochemical cells, which can trap and analyze the picomoles of metabolites (H2O2 and lactate) produced by different ratios of beneficial bacteria (Ss) and pathogenic bacteria (Sm). AIM 2: Establish a correlation between oral caries conditions and untreated dental plaque metabolic activity. We will establish the correlation between the ratio of H2O2 to lactate produced by dental plaque from patients and determine the threshold value that distinguishes non-caries from early caries conditions. We will also investigate the time-dependent metabolic activity of dental plaque in the presence of various sugars such as glucose and sucrose to gain insights into the cariogenic potential of dental plaque samples. Our analytical platform will eliminate the need for sample pretreatment, separation, or preconcentration and thus will enable the rapid and accurate analysis of metabolites produced by dental plaque. This study will validate dental plaque as a noninvasive bio-sample for point-of-care diagnostics in oral health, paving the way for early detection of dental caries and personalized prevention plans. The need for invasive treatments will thereby be reduced, ultimately lowering healthcare costs.