During apoptosis, a form of programmed cell death, cells fragment into small (~1-5μm), membrane-bound extracellular vesicles (EVs) known as apoptotic bodies (ApoBDs), in a process termed apoptotic cell disassembly. During both normal physiology as well as under pathological conditions, ApoBDs are increasingly recognised as potential mediators of intercellular communication, e.g. via the transfer of miRNA, proteins and other factors, to target cells. Atherosclerosis is a chronic inflammatory condition in which the formation of arterial plaques leads to hardening and narrowing of blood vessels, placing patients at risk of cardiovascular disease. During pathogenesis of atherosclerosis, the death of endothelial cells (ECs) contributes to loss of barrier function and promotes the formation of plaques in regions of EC damage. Although ECs generate an abundance of ApoBDs during apoptosis, a specific role for EC-derived ApoBDs (EC-ApoBDs) in the progression of atherosclerosis remains poorly defined. In the present study, a detailed characterisation of EC disassembly was performed, revealing that ApoBD formation in ECs is regulated by ROCK-1 and can be pharmacologically inhibited, providing tools for functional in vivo studies. To elucidate the functional significance of ApoBD generation by ECs under conditions of atherosclerosis-associated inflammation, the specific cargo and role in clearance of EC-ApoBDs were investigated. Profiling of EC-ApoBDs was performed via cytokine antibody array to reveal that EC-ApoBDs generated under inflammatory conditions contain high levels of pro-inflammatory cytokines including MCP-1 and IL-8. Furthermore, the ability of EC-ApoBDs to be cleared from the vasculature via phagocytosis was investigated using an in vivo murine clearance model, revealing that EC-ApoBDs may travel to distal regions to undergo clearance. These findings provide important insights into the potential functions of EC-ApoBDs and may contribute to future studies involving the therapeutic targeting of EC disassembly for the treatment of atherosclerosis.