Polyhexamethylene guanidine-phosphate enhances pro-coagulant activity of human erythrocytes and venous thrombosis in rats through phosphatidylserine externalization

Abstract

Polyhexamethylene guanidine-phosphate (PHMG-p) is a main compound used as a humidifier disinfectant, but the systemic health effects of PHMG-p still need to be explored. The circulatory and blood system is the organ that comes into contact with compounds absorbed into the body after inhalation exposure, resulting in various health problems, including cardiovascular diseases. This study examined the impact of PHMG-p on erythrocytes (red blood cells; RBCs), which are essential for sustaining circulatory health and are directly associated with thrombotic risks. We demonstrated that PHMG-p could enhance the thrombotic risk by promoting pro-coagulant activity and reducing erythrocyte deformability. In PHMG-p-exposed erythrocytes, phosphatidylserine externalization in the outer membrane and microvesicle generation were significantly increased under sub-hemolytic conditions, along with the morphological alterations in the erythrocytes. Exposure to PHMG-p induced erythrocyte phosphatidylserine externalization, leading to enhanced pro-coagulant activity, which was characterized by increased adhesion to vascular endothelial cells, elevated thrombin generation, and decreased deformability. Notably, calcium chelation effectively inhibited PS externalization and thrombin generation, highlighting the pivotal role of calcium influx in PHMG-p-induced thrombogenic alterations. Moreover, intratracheal instillation of PHMG-p promoted phosphatidylserine externalization and thrombin generation in rat erythrocytes, leading to a significant increase in thrombus formation, thereby corroborating the link between in vitro findings and the increased thrombotic risk observed in vivo. These findings suggest that PHMG-p may increase pro-thrombotic risk by promoting RBC pro-coagulant activity through calcium influx-driven PS externalization. © 2025 Elsevier B.V.