Small extracellular vesicles (EVs) were isolated from a model of normal breast fibroblasts— immortalized cells derived from healthy reduction mammoplasty tissue—treated with γ-radiation to study the effects of radiation therapy on intercellular communication within the breast stroma. EVs were isolated using the gold-standard differential speed centrifugation protocol. Proteins were subsequently extracted, and a quantitative examination of the alterations in the EV proteome was pursued. Both label-free and isobaric label-based methodologies were conducted to comprehensively characterize proteomic changes.