ABSTRACT

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 “releasewindow”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.

Figure 1. Conceptual framework and operational principle of the MORPHEUS platform. (a) A schematic illustration of the MORPHEUS platform based on an aptamer-functionalized AuNI chip and programmable thermoplasmonic regulation. Local thermoplasmonic modulation enables reversible capture-to-release of CD63-positive EVs through the conformational switching of a controlled aptamer. (b) Multicyclic manipulations of EV-CD63 at varying concentrations, demonstrating the reusability and regeneration capability of the aptamer sensor. (c) Enhanced target association and dissociation kinetics by leveraging the thermoplasmonic MORPHEUS and the energy landscape of the biomolecular binding.

Figure 2. Optimization of the thermoplasmonic MORPHEUS operating conditions for thermodynamic- and kinetic-enhanced biomolecular detection and regeneration. (a) Structural characteristics of the selected CD63-specific aptamers at varying temperatures. (b) Quantitative CD63 detection sensorgrams with Aptamer 2 at 25 °C. (c) Quantitative regressions of the two aptamers characterized at 25 °C. (d) Temperature-dependent MORPHEUS bioassay performance for Aptamer 1. (e) Thermoplasmonic temperature dependence of Aptamer 1 for detecting 100 pg/mL CD63. (f) Identification of the optimal regeneration temperature for Aptamer 2. (g) Cycling stability of the MORPHEUS platform over 30 consecutive thermoplasmonic capture-release cycles. (h) Regeneration efficiency and recovery performance during repeated thermoplasmonic cycling. For panels (c), (d), and (e), data are shown as mean ± s.d. from three independent measurements (n = 3).

https://doi.org/10.1021/acsnano.6c05601