Quantification of pain or discomfort induced by pressure is essential for understanding human responses to physical stimuli and improving user interfaces. Pain research has been conducted to investigate physiological signals associated with discomfort and pain perception. This study analyzed changes in electrodermal activity (EDA), tissue oxygen saturation (StO2), heart rate variability (HRV), and Visual Analog Scale (VAS) under pressures of 10, 20, and 30 kPa applied for 3 min to the thigh, knee, and calf in a seated position. Twenty participants were tested, and relationships between biosignals, pressure intensity, and pain levels were evaluated using Friedman tests and post-hoc analyses. Multiple linear regression models were used to predict VAS and pressure, and five machine learning models (SVM, Logistic Regression, Random Forest, MLP, KNN) were applied to classify pain levels (no pain: VAS 0, low: VAS 1–3, moderate: VAS 4–6, high: VAS 7–10) and pressure intensity. The results showed that higher pressure intensity and pain levels affected sympathetic nervous system responses and tissue oxygen saturation. Most EDA features and StO2 significantly changed according to pressure intensity and pain levels, while NN interval and HF among HRV features showed significant differences based on pressure intensity or pain level. Regression analysis combining biosignal features achieved a maximum R2 of 0.668 in predicting VAS and pressure intensity. The four-level classification model reached an accuracy of 88.2% for pain levels and 81.3% for pressure intensity. These results demonstrated the potential of EDA, StO2, HRV signals, and combinations of biosignal features for pain quantification and prediction.
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