Background: Calcium disodium Ethylene diamine tetra-acetic acid (EDTA) is a common food additive and an ingredient in cosmetic and industrial products. It's used in food to preserve flavor, color, and texture. However, like many food additives, it has become quite controversial.Aim of the study: assessment of the potential hepatic, renal, testicular, and hematological toxicities of chronic daily oral administration of EDTA (E385) in adult male albino rats and the involved role of p53 expression. Materials and methods: twenty four adult male albino rats were partitioned into two distinct groups: Control groups (Subdivided into negative and positive control groups) (8 rats each); group Ia was negative control groups received nothing, group Ib a received 0.5 ml saline via gastric gavage daily for 90 days; and groupⅡ(8rats) received EDTA by gavage at a dosage of 20 mg/kg/day for 90 days, dose equivalent to 1/100th LD50. Upon completion of the experiment, blood and tissue samples were obtained from the liver, kidney, and testis for conducting biochemical, hematological, and histopathological analyses. Results: Hematological analyses revealed significant decreases in white blood cells (WBCs), red blood cells (RBCs), platelet count (PT), hemoglobin level (Hb), and hematocrit value (HCT%), in the EDTA intoxicated group compared to the control groups. Biochemical tests revealed significant changes in liver and kidney functions in the EDTA intoxicated group compared to the control groups. The EDTA intoxicated group exhibited multiple histopathological changes in the liver, kidney, and testis through immunohistochemical alterations in P53 and androgen receptor expressions. Conclusion: Chronic oral administration of EDTA for consecutive 90 days exhibits toxic effects on the tissue of the liver, kidney, and testis through a mechanism that involves p53 expression in addition to its toxic effects on hematological parameters in male albino rats.