RADAR-Renewable and Safe Aromatic Compounds as Replacements for Substances of Concern

Bisphenol A (BPA) is a widely used industrial chemical with well-documented endocrine-disrupting and immunotoxic effects. Bisguaiacol (BG) has been proposed as a safer alternative due to its structural similarity but potentially lower toxicity. This project aims to conduct an experimental risk assessment of BG by focusing on its immunotoxicological effects using in vitro models and developing New approach methodologies (NAM)s to improve the experimental risk assessment. Objectives 1. Evaluate the cytotoxicity of BG on immune cell lines (e.g., THP-1 monocytes, Jurkat T cells) using MTT and LDH assays. 2. Investigate BG’s effects on immune cell activation and cytokine production through ELISA and flow cytometry. 3. Assess oxidative stress induction via ROS detection assays and glutathione depletion measurements. 4. Compare BG’s immunomodulatory properties with BPA to determine its relative safety. 5. Develop a microfluidic system based NAMs to improve experimental risk assessment. Methodology Human-derived immune cells will be exposed to environmentally relevant concentrations of BG and BPA. Cell viability assays will determine cytotoxic thresholds. Changes in cytokine secretion (e.g., IL-6, TNF-α, IL-10) will be quantified using ELISA. Flow cytometry will assess changes in immune cell markers (CD69, CD86). Oxidative stress will be measured using DCFH-DA fluorescence for ROS generation and GSH/GSSG assays for antioxidant capacity. Comparative analyses with BPA will identify whether BG presents lower immunotoxic risks. Expected Outcomes This study will provide critical data on BG’s immunotoxicological profile, helping determine its suitability as a BPA alternative. If BG demonstrates reduced immune system disruption, it may serve as a safer material in consumer products. Impact The findings will contribute to regulatory assessments of BPA alternatives, supporting the development of safer industrial chemicals.