Accurate diagnosis of pulpal health is crucial to identify the most effective therapeutic approach. However, differentiating pulpal conditions, which may require different treatment approaches, remains a challenge. This study aimed to address this gap by investigating the protein levels of 17 inflammatory biomarkers simultaneously in the dental pulp with different clinical diagnoses. This study employed a cross-sectional exploratory design, enrolling 64 adult patients. After obtaining ethical approval, researchers conducted clinical and radiographic examinations to categorise teeth into four diagnostic groups: normal pulp, reversible pulpitis, symptomatic irreversible pulpitis and asymptomatic irreversible pulpitis. Pulpal blood samples were then collected and analysed using Luminex technology to measure the levels of inflammatory proteins and matrix metalloproteinases (MMPs). Statistical analyses, including the Mann-Whitney U test and Spearman's rank correlation, were used to compare the levels of these markers across the different diagnoses and to assess their correlation with patient symptoms. The study revealed significant increases in several inflammatory proteins, including IL-4, IL-8, MCP-1, MIP-1α, RANTES and MMP-9, in both types of irreversible pulpitis cases compared with other diagnostic categories p < 0.05. These elevated levels exhibited positive correlations with patient-reported pain scores, instances of spontaneous pain and bleeding times. Notably, only IL-4 and IL-7 exhibited correlations with prolonged bleeding times (over 10 min) p < 0.05, while IL-1α and MMP-2 were associated with shorter bleeding times (under 10 min) p < 0.05. Additionally, IL-8 and MCP-1 levels were significantly associated with positive palpation findings p < 0.05, whereas MIP-1α and MMP-1 were correlated with positive percussion results p < 0.05. Differential specific inflammatory potential biomarker levels may differentiate pulpal disease states. Integrating molecular diagnostics into longitudinal clinical trials and eventually into routine endodontic practice, could revolutionise treatment decisions.