Original Articles
Indocyanine green-loaded perfluorocarbon nanoemulsions for bimodal 19F-magnetic resonance/nearinfrared fluorescence imaging and subsequent phototherapy
Abstract
We have developed an indocyanine green-loaded perfluorocarbon (ICG/PFCE) nanoemulsion as a multifunctional theranostic nanomedicine which enables not only 19F magnetic resonance (MR)/near-infrared fluorescence (NIRF) bimodal imaging but also subsequent photodynamic/photothermal dual therapy of cancer. The hydrodynamic size of ICG/PFCE nanoemulsions was 164.2 nm. The stability of indocyanine green (ICG) in aqueous solution was significantly improved when loaded on perfluorocarbon nanoemulsions. In addition, ICG/PFCE nanoemulsions showed good dispersion stability in aqueous media containing 10% fetal bovine serum, for at least 14 days.
19F-MRI of ICG/PFCE nanoemulsions showed that the signal intensity increased with increasing nanoemulsion concentration with no signal observed from the surrounding background. Using NIRF imaging with perfluorocarbon nanoemulsion alone, without ICG, did not produce NIRF, while clear and bright fluorescent images were obtained with ICG/PFCE nanoemulsions at 10-μM ICG equivalent. The capacity of ICG-loaded nanoemulsions to generate heat following light irradiation by using an 810-nm laser was comparable to that of free ICG, while singlet oxygen generation of ICG-loaded nanoemulsions was significantly better than that of free ICG.
In vitro cytotoxicity tests and fluorescence microscopy confirmed biocompatibility of the nanoemulsion. Upon light irradiation, U87MG glioblastoma cells incubated with ICG/PFCE nanoemulsions underwent necrotic cell death. The therapeutic mechanism during light illumination appears to be mainly due to the photodynamic effect at lower ICG concentrations, whilst the photothermal effect became more obvious at increased ICG concentrations, enabling combined photodynamic/photothermal therapy of cancer cells.
19F-MRI of ICG/PFCE nanoemulsions showed that the signal intensity increased with increasing nanoemulsion concentration with no signal observed from the surrounding background. Using NIRF imaging with perfluorocarbon nanoemulsion alone, without ICG, did not produce NIRF, while clear and bright fluorescent images were obtained with ICG/PFCE nanoemulsions at 10-μM ICG equivalent. The capacity of ICG-loaded nanoemulsions to generate heat following light irradiation by using an 810-nm laser was comparable to that of free ICG, while singlet oxygen generation of ICG-loaded nanoemulsions was significantly better than that of free ICG.
In vitro cytotoxicity tests and fluorescence microscopy confirmed biocompatibility of the nanoemulsion. Upon light irradiation, U87MG glioblastoma cells incubated with ICG/PFCE nanoemulsions underwent necrotic cell death. The therapeutic mechanism during light illumination appears to be mainly due to the photodynamic effect at lower ICG concentrations, whilst the photothermal effect became more obvious at increased ICG concentrations, enabling combined photodynamic/photothermal therapy of cancer cells.
