Chiral nanomaterials hold significant promise for enhancing clinical therapeutic outcomes, thanks to their unique chiral selectivity in biological systems. Despite this potential, the innovative approach has garnered limited focus in the realm of cancer therapy. In this work, we synthesized multi-shelled mesoporous silica nanocarriers by a biphasic stratification reaction system. Owing to a lamellar micelle spiral self-assembly happens at the water/oil bio-interface, the fabricated silica nanosphere has a unique chirality which is different from conventional dendritic mesoporous silica nanoparticles. On one hand, after loading with the clinically approved dye molecule indocyanine green (ICG), the long-off peak fluorescence at second near-infrared window (NIR II, 1000–1700 nm) exhibits excellent photostability, with a significant enhancement under continuous exposure to daylight or an 808 nm laser, which is highly beneficial for long-duration surgical navigation in clinical settings. On the other hand, due to its unique chiral architecture, chiral nanostructured mesoporous silica (cSi) nanocarriers demonstrate unparalleled cellular uptake capability compared to traditional mesoporous silica. Furthermore, thanks to cSi's targeted accumulation in tumor tissues and its ability to synergistically promote photothermal augmented chiral-specific tumor cell killing, the malignant tissue is effectively suppressed, heralding a new way in cancer-fighting.