Cell Damage–Like Proteomic Signatures Distinguish Cell‐Free and Concentrated Ascites Reinfusion Therapy ( CART ) Membranes in Malignant Ascites: A Mechanistic Study

Introduction Cell‐free and concentrated ascites reinfusion therapy (CART) alleviates the symptoms of malignant ascites but may trigger inflammation. We investigated why AHF‐MO filter (A‐A)–processed concentrates showed weaker bioactivity than EC20W filter (A‐E)–processed concentrates. Methods Ascites from three patients with ovarian cancer were processed into untreated ascites (UA), A‐A, or A‐E. Whole ascites, extracellular vesicles (EVs), and EV‐depleted supernatants were injected intraperitoneally into mice. Serum cytokines were measured, and label‐free proteomics was used to identify inflammatory pathways. Results A‐E induced the strongest proinflammatory and Th17/type I interferon responses compared to UA. A‐A produced intermediate activation. EV‐depleted A‐E supernatants remained highly bioactive. A‐E was enriched in proteins associated with cellular damage pathways and proinflammatory signaling. A‐A exhibited attenuated changes and preserved regulatory proteins. Conclusion A‐A yields a more balanced inflammatory and proteomic profile than A‐E. Our findings provide a preliminary mechanistic rationale for its use in CART and offer targets for membrane optimization.