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During delivery, nanoparticles must sequentially traverse multiple biological barriers, from systemic circulation and tumor accumulation to vascular extravasation, basement\u2010membrane crossing, interstitial penetration, and cellular internalization. In this review, we distinguish the sheet\u2010like perivascular basement membrane from the fibrillar, hydrated interstitial extracellular matrix and discuss how intertumoral, intratumoral, and stage\u2010specific heterogeneity alters the dominant transport barriers across different tumor contexts. We then summarize three complementary strategies for engineering nanoparticles to improve deep tumor penetration: 1) Nanocarrier design, which tunes particle shape, size, surface charge, mechanical stiffness, and ligand presentation to balance cellular interactions and interstitial mobility; 2) Vascular and Stromal Modulation, which alleviates transport barriers through vascular normalization, transient basement\u2010membrane modulation, controlled extracellular\u2010matrix remodeling, and regulation of matrix production by cancer\u2010associated fibroblasts; 3) Active and Assisted Transport, which promotes trans\u2010barrier delivery through transcytosis, self\u2010propelled nanomotors, and externally applied physical energy. We discuss the associated challenges and optimization strategies, particularly those related to spatial penetration, tumor accumulation, model\u2010dependent efficacy, and safety. Finally, we highlight the translational limitations and future opportunities of tumor\u2010penetrating nanomedicines from a clinical perspective.<\/jats:p>","DOI":"10.1002\/smll.75972","type":"journal-article","created":{"date-parts":[[2026,10,4]],"date-time":"2026-10-04T13:35:51Z","timestamp":1791120951000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Engineering Nanoparticles to Navigate Biological Barriers for Deep Tumor Penetration"],"prefix":"10.1002","author":[{"given":"Lincan","family":"Zeng","sequence":"first","affiliation":[{"name":"The Second Affiliated Hospital Guangdong Provincial Key Laboratory of Allergy and Clinical Immunology Guangzhou Medical University  Guangzhou P. R. 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