Abstract
A thermosensitive polymeric nanoformulation (NF) was fabricated for thermally enhanced near-infrared (NIR) fluorescence imaging (FLI). It comprised core-shell nanoparticles (NPs) with a polystyrene core and a thermosensitive shell of a co-polymer of N-isopropylacrylamide and acrylamide [poly(NIPAM-co-AA)], which underwent a reversible conformational transition at 38-40 °C (corresponding to a lower critical solution temperature, LCST), leading to a reversible shrinkage of NPs from ∼250 nm to ∼140 nm for temperatures above LCST. The NIR dye 3782SL or photosensitizer HPPH were loaded to the NP shells. While the fluorescence of 3782SL and HPPH was quenched in water, it recovered in the NPs dispersion as a result of adsorption by NPs. Fluorescence for 3782SL and HPPH in NF increased when the temperature increased above LCST. Heating of HPPH-loaded NFs led to the elongation of the HPPH fluorescence lifetime and increased the generation of singlet oxygen ((1)O(2)). This occurred as a result of the NP shrinkage, corresponding shell compaction and NP aggregation, which hindered the internal conversion for photoexcited molecules adsorbed by NPs, and resulted in an increase in other deactivation pathways, namely fluorescence emission and intersystem crossing. The latter led to an increase in the triplet yield and, consequently, in singlet oxygen generation. Fluorescence microscopy revealed a 2-3-fold increase in the 3782SL or HPPH fluorescence signal from the NF-treated cells after they were heated up to 40 °C. Comparable results were obtained for the FLI of mice in vivo, after subcutaneous, intravenous, or intratumoral NF injections and localized heating by NIR (1.3 μm) laser irradiation. The developed NF holds immense potential for thermally enhanced FLI and photodynamic therapy.