Thermodynamic-Kinetic Tailored Photothermal-Responsive Molecular Switching for Extracellular Vesicle Manipulation

Extracellular vesicles (EVs) are central mediators of intercellular communication and promising carriers for oncology diagnosis and therapeutic delivery, yet their isolation remains challenged by inadequate specificity, irreversible capture, and vesicle structural damage. Here, we present the Multifunctional Optically Regulated Plasmonic Heating-Enhanced Ultrasensitive Sensing (MORPHEUS) system for programmable and reversible EV manipulation with thermoplasmonics-regulated aptamer switches. Through spatiotemporal control of localized thermoplasmonic heating, MORPHEUS established a temperature-defined "capture window" with enhanced binding kinetics, followed by a mild "release window" enabling 98.94% nondestructive EV recovery. The localized thermoplasmonic heating fine-tuned the conformational dynamics of the CD63 aptamer and free-energy landscape, accelerating association and dissociation kinetics by 3-fold and 415-fold, respectively, for precise binding-to-releasing control over intact EVs. Thermodynamically, the programmed photothermal heating conditions reduced the energy barriers for molecular desolvation and structural rearrangement, enhancing conformational sampling and facilitating interfacial water molecule release, resulting in entropic gains. This dual kinetic-thermodynamic regulation enabled reversible aptasensing regeneration while preserving EV structural integrity, membrane protein activity, and nucleic acid cargo. The platform maintained stable operation over 30 consecutive capture-release cycles with a capture-signal coefficient of variation (CV) of 3.24%. Benefiting from the preserved biological integrity, MORPHEUS-enriched EVs retained efficient drug-loading capability and therapeutic activity in cellular models, highlighting the potential of programmable thermoplasmonic regulation for EV-based diagnostic and therapeutic engineering.