Two-photon Excited Photodynamic Therapy Research Articles

Antitumor Cream: Transdermal Hydrogel Containing Liposome-Encapsulated Ruthenium Complex for Infrared-Controlled Multimodal Synergistic Therapy.

A transdermal drug delivery cream, which is non-invasive and painless, containing a liposome-encapsulated Ru(II) complex (LipoRu) is created for the treatment of skin cancer. This formulation capitalizes on the synergistic antitumor effects of two-photon excited photodynamic therapy (PDT), photothermal therapy (PTT), and chemotherapy. LipoRu exhibits effective tumor accumulation, efficient cellular uptake, pH-sensitive and infrared-accelerated release, and dual localization to the nucleus and mitochondria. The released Ru(II) complexes within cells exert multiple antitumor mechanisms, such as DNA topoisomerase and RNA polymerase inhibition, Type I and II PDT, PTT, DNA photodamage, and apoptosis and ferroptosis induction. The biodistribution and therapeutic efficacy of LipoRu in vivo are systematically compared via three distinct administration routes: intratumoral injection, intravenous injection, and transdermal delivery through topical cream application. The positive therapeutic effects of the LipoRu cream fabricated here in subcutaneous tumor-bearing mice offer optimistic potential for the painless and non-invasive treatment of both early-stage and advanced skin cancers, as well as superficially located solid tumors.

Iridium(III)-Based Infrared Two-Photon Photosensitizers: Systematic Regulation of Their Photodynamic Therapy Efficacy.

Cyclometalated iridium(III) complexes are of significant importance in the field of antitumor photodynamic therapy (PDT), whether they exist as single molecules or are incorporated into nanomaterials. Nevertheless, a comprehensive examination of the relationship between their molecular structure and PDT effectiveness remains awaited. The influencing factors of two-photon excited PDT can be anticipated to be further multiplied, particularly in relation to intricate nonlinear optical properties. At present, a comprehensive body of research on this topic is lacking, and few discernible patterns have been identified. In this study, through systematic structure regulation, the nitro-substituted styryl group and 1-phenylisoquinoline ligand containing YQ2 was found to be the most potent infrared two-photon excitable photosensitizer in a 4 × 3 combination library of cyclometalated Ir(III) complexes. YQ2 could enter cells via an energy-dependent and caveolae-mediated pathway, bind specifically to mitochondria, produce 1O2 in response to 808 nm LPL irradiation, activate caspases, and induce apoptosis. In vitro, YQ2 displayed a remarkable phototherapy index for both malignant melanoma (>885) and non-small-cell lung cancer (>1234) based on these functions and was minimally deleterious to human normal liver and kidney cells. In in vivo antitumor phototherapy, YQ2 inhibited tumor growth by an impressive 85% and could be eliminated from the bodies of mice with a half-life as short as 43 h. This study has the potential to contribute significantly to the development of phototherapeutic drugs that are extremely effective in treating large, profoundly located solid tumors as well as the understanding of the structure-activity relationship of Ir(III)-based PSs in PDT.

Emodin-Based Nanoarchitectonics with Giant Two-Photon Absorption for Enhanced Photodynamic Therapy.

Two-photon-excited photodynamic therapy (TPE-PDT) has significant advantages over conventional photodynamic therapy (PDT). However, obtaining easily accessible TPE photosensitizers (PSs) with high efficiency remains a challenge. Herein, we demonstrate that emodin (Emo), a natural anthraquinone (NA) derivative, is a promising TPE PS with a large two-photon absorption cross-section (TPAC: 380.9 GM) and high singlet oxygen (1 O2 ) quantum yield (31.9 %). When co-assembled with human serum albumin (HSA), the formed Emo/HSA nanoparticles (E/H NPs) possess a giant TPAC (4.02×107 GM) and desirable 1 O2 generation capability, thus showing outstanding TPE-PDT properties against cancer cells. In vivo experiments reveal that E/H NPs exhibit improved retention time in tumors and can ablate tumors at an ultra-low dosage (0.2 mg/kg) under an 800 nm femtosecond pulsed laser irradiation. This work is beneficial for the use of natural extracts NAs for high-efficiency TPE-PDT.

Near-infrared light activated photosensitizer with specific imaging of lipid droplets enables two-photon excited photodynamic therapy.

Two-photon excited phototherapy has attracted considerable attention due to its advantages such as deeper penetration depth and higher spatial resolution. The lack of a high-performance photosensitizer with large two-photon absorption cross-sections and specific targeting ability makes the efficacy of phototherapy in the treatment of cancer unsatisfactory. Here, a new BODIPY-derived photosensitizer 6DBF2 is designed with two-photon photosensitization for two-photon excited photodynamic therapy in vivo. 6DBF2 possesses good two-photon absorption and efficient 1O2 generation upon near-infrared laser excitation. Excellent targeting specificities to lipid droplets of 6DBF2 without any encapsulation or modification at a low working concentration of 0.1 μM is in favor of efficient photodynamic therapy. In vitro cancer cell ablation and in vivo tumor ablation inside mice models upon two-photon irradiation in NIR demonstrate the outstanding therapeutic performance of 6DBF2 in two-photon excited photodynamic therapy. This work thus discusses a rare example of lipid droplets targeting two-photon excited photodynamic therapy for deep cancer tissue imaging and treatment under near-infrared light irradiation.

Photosensitivity of Different Nanodiamond-PMO Nanoparticles in Two-Photon-Excited Photodynamic Therapy.

In addition to their great optical properties, nanodiamonds (NDs) have recently proved useful for two-photon-excited photodynamic therapy (TPE-PDT) applications. Indeed, they are able to produce reactive oxygen species (ROS) directly upon two-photon excitation but not with one-photon excitation; Methods: Fluorescent NDs (FNDs) with a 100 nm diameter and detonation NDs (DNDs) of 30 nm were compared. In order to use the gems for cancer-cell theranostics, they were encapsulated in a bis(triethoxysilyl)ethylene-based (ENE) periodic mesoporous organosilica (PMO) shell, and the surface of the formed nanoparticles (NPs) was modified by the direct grafting of polyethylene glycol (PEG) and amino groups using PEG-hexyltriethoxysilane and aminoundecyltriethoxysilane during the sol-gel process. The NPs' phototoxicity and interaction with MDA-MB-231 breast cancer cells were evaluated afterwards; Results: Transmission electronic microscopy images showed the formation of core-shell NPs. Infrared spectra and zeta-potential measurements confirmed the grafting of PEG and NH2 groups. The encapsulation of the NDs allowed for the imaging of cancer cells with NDs and for the performance of TPE-PDT of MDA-MB-231 cancer cells with significant mortality. Multifunctional ND@PMO core-shell nanosystems were successfully prepared. The NPs demonstrated high biocompatibility and TPE-PDT efficiency in vitro in the cancer cell model. Such systems hold good potential for two-photon-excited PDT applications.

Photon Characteristics and Behavior under Refractive Index

The effective mass, energy and momentum of photon in medium is described. The photon has both kinetic and potential energy and displays an effective mass when travelling through a medium. The photon is a type of elementary particle that serves as the quantum of the electromagnetic field and is explained through equations to have an elastic constant, volume, momentum, force, power and zero rest mass. The theory that photons a massless only applies to rest mass. A photon undergoes different dynamics such as strain and changes in effective mass under the influence of the refractive index that gives rise to a refractive force. This study synthesizes some properties of energy such as elastic constant, weight, and volume in order to explain the nature of photons and photon dynamics under refractive index. In this study, a helical spring model has been used to describe the behavior of a photon in the medium. The model connects together the exhibited properties such as energy, effective mass, spin, elastic constant and volume. Further explanation is given to the effect of gravitational force on wavelength and refractive index of air, and how gravitational force causes red and blue shifts in photon propagation. The quantization of energy has been extended to the quantization of space, time and matter. This study is consistent and consolidates the existing classical and quantum theories. There are further potential applications in optical communication, quantum cryptography, quantum optics, and medicine, especially in the use of two-photon excitation microscopy and photodynamic therapy.

Conjugated Polymers with Aggregation-Induced Emission Characteristics for Fluorescence Imaging and Photodynamic Therapy.

Accurate diagnosis and treatment have been extensively developed in the field of biomedicine, which put forward higher requirements for the development of biomedical materials with high efficiency and selectivity. Among them, conjugated polymers featuring aggregation-induced emission (AIE) characteristics (AIE conjugated polymers) have stood out in recent years owing to their unique properties, such as intense solid emission, high light-harvesting ability, efficient energy transfer, and high 1 O2 generation ability, which empower them with effective biomedical functions in fluorescence imaging (FLI), photodynamic therapy (PDT), FLI-guided PDT, two-photon excited photodynamic therapy (2PE-PDT), etc. In this review, we highlight recent progress in AIE conjugated polymers and their applications in anticancer and antibacterial areas based on FLI and PDT, and summarize the mechanism of color-tuned fluorescence emission and efficient 1 O2 generation ability. The challenges and perspectives for the future development of AIE conjugated polymers are also discussed.

Biological Assessment of Laser-Synthesized Silicon Nanoparticles Effect in Two-Photon Photodynamic Therapy on Breast Cancer MCF-7 Cells.

Driven by their distinctive physiological activities, biological properties and unique theranostic modalities, silicon nanoparticles (SiNPs) are one of the promising materials for the development of novel multifunctional nanoplatforms for biomedical applications. In this work, we assessed the possibility to use laser-synthesized Si NPs as photosensitizers in two-photon excited photodynamic therapy (TPE-PDT) modality. Herein, we used an easy strategy to synthesize ultraclean and monodispersed SiNPs using laser ablation and fragmentation sequences of silicon wafer in aqueous solution, which prevent any specific purification step. Structural analysis revealed the spherical shape of the nanoparticles with a narrow size distribution centered at the mean size diameter of 62 nm ± 0.42 nm, while the negative surface charge of −40 ± 0.3 mV ensured a great stability without sedimentation over a long period of time. In vitro studies on human cancer cell lines (breast and liver) and healthy cells revealed their low cytotoxicity without any light stimulus and their therapeutic potential under TPE-PDT mode at 900 nm with a promising cell death of 45% in case of MCF-7 breast cancer cells, as a consequence of intracellular reactive oxygen species release. Their luminescence emission inside the cells was clearly observed at UV-Vis region. Compared to Si nanoparticles synthesized via chemical routes, which are often linked to additional modules with photochemical and photobiological properties to boost photodynamic effect, laser-synthesized SiNPs exhibit promising intrinsic therapeutic and imaging properties to develop advanced strategy in nanomedicine field.

Design and synthesis of efficient heavy-atom-free photosensitizers for photodynamic therapy of cancer.

Novel thiocarbonyl derivatives (NIS and CRNS) with excellent ROS generation abilities are synthesized and studied as potential photosensitizers for one- and two-photon excited photodynamic therapy. In particular, NIS-Me and CRNS display outstanding phototoxicity toward HeLa cells under two-photon excitation (800 nm) with negligible dark toxicity.

Highly efficient singlet oxygen generation, two-photon photodynamic therapy and melanoma ablation by rationally designed mitochondria-specific near-infrared AIEgens.

Photosensitizers (PSs) with multiple characteristics, including efficient singlet oxygen (1O2) generation, cancer cell-selective accumulation and subsequent mitochondrial localization as well as near-infrared (NIR) excitation and bright NIR emission, are promising candidates for imaging-guided photodynamic therapy (PDT) but rarely concerned. Herein, a simple rational strategy, namely modulation of donor–acceptor (D–A) strength, for molecular engineering of mitochondria-targeting aggregation-induced emission (AIE) PSs with desirable characteristics including highly improved 1O2 generation efficiency, NIR emission (736 nm), high specificity to mitochondria, good biocompatibility, high brightness and superior photostability is demonstrated. Impressively, upon light irradiation, the optimal NIR AIE PS (DCQu) can generate 1O2 with efficiency much higher than those of commercially available PSs. The excellent two-photon absorption properties of DCQu allow two-photon fluorescence imaging of mitochondria and subsequent two-photon excited PDT. DCQu can selectively differentiate cancer cells from normal cells without the aid of extra targeting ligands. Upon ultralow-power light irradiation at 4.2 mW cm−2, in situ mitochondrial photodynamic activation to specifically damage cancer cells and efficient in vivo melanoma ablation are demonstrated, suggesting superior potency of the AIE PS in imaging-guided PDT with minimal side effects, which is promising for future precision medicine.

Engineered g-C3N4 Quantum Dots for Tunable Two-Photon Imaging and Photodynamic Therapy.

Near-infrared (NIR)-light-triggered diagnostic and therapeutic systems normally are the integration of several components, whose complex structures and low reproducibility restrict their further applications. Herein, we proposed a single component of defect graphitic phase carbon nitride quantum dots (g-C3N4 QDs) as a dual-functional nanoplatform, which could synchronously achieve two-photon imaging (TPI) and two-photon excited photodynamic therapy (TPE-PDT) under an 800 nm NIR laser. In order to regulate the competitive capability between TPI and PDT, three kinds of engineered g-C3N4 QDs (CN, CN-DPT, and CN-THDT QDs) with different two-photon catalytic capabilities to generate reactive oxygen species (ROS) were prepared through copolymerization melamine with selected monomers. In the evaluation of in vitro cytotoxicity, three g-C3N4 QDs possessed very good biocompatibilities, which could be delivered to tumor cells for imaging and therapy. Under an 800 nm NIR light, CN-DPT QDs exhibited a bright TPI effect and produced an efficient ROS to achieve TPE-PDT, thus suggesting that the single component of CN-DPT QDs could perform as an appropriate dual-functional TPI and TPE-PDT probe triggered by an 800 nm NIR light. The engineered g-C3N4 QDs with a high stability and excellent biocompatibility have contributed to the method of TPI and TPE-PDT, which may lead to g-C3N4-based nanomaterials as novel imaging and therapeutic agents in cancer treatment.

Rational Design of Organic Probes for Turn-On Two-Photon Excited Fluorescence Imaging and Photodynamic Therapy

Summary The rational design of two-photon absorption (2PA) organic dyes with 2PA cross-section (δ) values tunable by mild stimuli under physiological conditions remains a challenge. Using a simulation-assisted structure screening method, we have designed an acetal terminated distyrylbenzene derivative (Ace-DSB), which can be readily converted into aldehyde terminated molecules (Ald-DSB) to exhibit a significant increase in δ values at 760 nm. The conversion reaction, which dramatically turns on the 2PA absorption capability of the molecules, can occur in an intracellular environment. Furthermore, Ald-DSB exhibits 40 times higher efficiency of two-photon generation of singlet oxygen than Ace-DSB. These turn-on features allow enhanced two-photon laser confocal scanning microscopic imaging and two-photon excited photodynamic therapy (2PE-PDT) for MCF-7 cancer cells and melanoma tumors. This work opens a new avenue toward the design of turn-on-type two-photon absorption molecules for both in vitro and in vivo tumor imaging and 2PA-PDT.

Red emitting conjugated polymer based nanophotosensitizers for selectively targeted two-photon excitation imaging guided photodynamic therapy

Two-photon excitation (2PE) photodynamic therapy (PDT) is a non-invasive technique for the treatment of cancer. However, its clinical applications are limited by small two-photon absorption cross section values of conventional photosensitizers. Here we designed multifunctional conjugated polymer based nanoparticles consisting of a conjugated polymer, a photosensitizer and a red-emitting dye, which can realize simultaneous 2PE red emission imaging and 2PE-PDT activities. The working principle is based on a 2PE fluorescence resonance energy transfer strategy from the conjugated polymer to photosensitizing and imaging agents. In these nanoparticles (NPs), the conjugated polymer, PPBF, was chosen as a two-photon light-harvesting material while the photosensitizer (tetraphenylporphyrin, TPP) and the red-emitting dye (TPD) were chosen as energy acceptors. The 2PE emission of TPP and TPD was enhanced by up to ∼161 and ∼23 times, respectively. The 2PE-PDT activity of these NPs was significantly improved compared with those NPs without PPBF by up to ∼149 times. Further surface-functionalization with folic acid (FA) groups allows these nanoparticles to exhibit selective affinity toward KB cancer cells. These NPs could act as novel 2PE conjugated polymer based nanoparticles combined with the advantages of low dark cytotoxicity, selective targeting and imaging-guided 2PE-PDT activities.

SHG-enhanced NIR-excited in vitro photodynamic therapy using composite nanoparticles of barium titanate and rose Bengal.

Near infrared (NIR) light excited photodynamic therapy (PDT) has been considered as a possible way to increase the therapy depth. Besides the traditional two-photon excited PDT and upconversion PDT by rare-earth ion materials, SHG has drawn much attention recently to act as an additional choice to achieve NIR light excited PDT. Herein, by using the electrostatic absorption method, barium titanate and rose Bengal composite nanoparticles (BT@PAH/RB/PAH, BT–RB) were synthesized. Compared with rose Bengal (RB) molecules and a mixture of barium titanate nanoparticles and RB (BT + RB), BT–RB nanoparticles were shown to be able to produce more reactive oxygen species (ROS) ex vivo and in vitro. Afterwards, the SHG-enhanced localized PDT was applied on Hela cells, in which BT–RB nanoparticles showed a better performance than BT + RB. Our work has shown that the SHG-enhanced PDT has good prospects and the close combination of harmonic nanoparticles and photosensitizers may facilitate the development of novel reagents for NIR light excited PDT.

A Porphyrin Dimer-GdDOTA Conjugate as a Theranostic Agent for One- and Two-Photon Photodynamic Therapy and MRI.

A molecular theranostic agent designed for photodynamic therapy (PDT) treatment in the near-infrared and for imaging tissue tumors with magnetic resonance imaging (MRI) is reported. It consists of a linear π-conjugated Zn(II) porphyrin dimer linked at each extremity to a GdDOTA-type complex. This agent has shown very promising potential for PDT applications with good singlet oxygen generation in DMSO and high linear absorption in the near-infrared (λmax = 746 nm, ε ≈ 105 M-1 cm-1). Moreover, this molecule has a propensity for two-photon excited PDT with high two-photon cross sections (∼8000 GM in 880-930 nm range), which should allow for deeper tumor treatments and higher spatial precision as compared to conventional one-photon PDT. Regarding the MRI contrast agent properties, the molecule has shown superior relaxivity (14.4 mM-1 s-1 at 40 MHz, 298 K) in comparison to clinical contrast agents and the ability to be internalized in cells, thanks to its amphiphilic character. Irradiation of HeLa cells using either one-photon (740 nm) or two-photon excitation (910 nm) has led in both cases to important cell death.

Fluorescence Resonance Energy Transfer Based Highly Efficient Theranostic Nanoplatform for Two-Photon Bioimaging and Two-Photon Excited Photodynamic Therapy of Multiple Drug Resistance Bacteria.

Near-infrared (NIR) light between 700 and 2500 nm, which is in the range of the first, second, and third biological windows, has the capability to penetrate biological tissues and blood, which provides a huge advantages of higher penetration depth. However, because of the lack of available biocompatible single photon probes in NIR window, there is an urgent need for new theranostic material, which could be used for two-photon bioimaging as well as for two-photon photodynamic therapy (PDT) in biological window. Driven by the need, the current manuscript reports gold nanoclusters (GNCs) attached graphene quantum dot (GQD) based two-photon excited theranostic nanoplatform with high two-photon absorption, very strong two-photon luminescence, as well as two-photon stability in NIR region. Experimental result shows strong two-photon luminescence and two-photon-induced PDT, which is based on fluorescence resonance energy transfer (FRET) mechanism, where graphene quantum dots with very high two-photon absorption act as two-photon donors and gold nanoclusters act as acceptors. Reported data indicate that 1O2 generation efficiency enhances tremendously due to the FRET process, which increases the two-photon excited PDT efficiency for multiple drug resistance bacteria (MDRB). Reported data indicate that the nanoplatform has the capability for bright two-photon bioimaging and two-photon photodynamic therapy for MRSA and carbapenem-resistant (CRE) Escherichia coli. Reported nanoplatform is a promising candidate to serve as a contrast agent for multiphoton imaging as well as for two-photon excited PDT agent to eliminate multidrug-resistant strains.

A simple mitochondrial targeting AIEgen for image-guided two-photon excited photodynamic therapy.

Two-photon excited photodynamic therapy (TP-PDT) is not only able to offer deeper penetration depth but also much more precise 3D treatment than traditional one-photon excited PDT. However, the achievement of TP-PDT requires photosensitizers with large two-photon absorption cross sections, efficient generation of reactive oxygen species, and bright two-photon fluorescence. In this work, we present a simple AIE luminogen (AIEgen), IQ-TPA, with mitochondrial targeting and susceptible two-photon excitation for image-guided photodynamic therapy in cancer cells. This feasibility of utilizing small molecular multifunctional AIEgens for TP-PDT was demonstrated together with the merits of tiny size, good cell permeability, low dark cytotoxicity and easy synthesis, showing great potential for the development of future theranostic systems.

Ru(II) polypyridyl complexes as photosensitizers in (two-photon excitation) photodynamic therapy

Ru(II) polypyridyl complexes as photosensitizers in (two-photon excitation) photodynamic therapy

Porphyrin- or phthalocyanine-bridged silsesquioxane nanoparticles for two-photon photodynamic therapy or photoacoustic imaging.

Porphyrin- or phthalocyanine-bridged silsesquioxane nanoparticles (BSPOR and BSPHT) were prepared. Their endocytosis in MCF-7 cancer cells was shown with two-photon excited fluorescence (TPEF) imaging. With two-photon excited photodynamic therapy (TPE-PDT), BSPOR was more phototoxic than BSPHT, which in contrast displayed a very high signal for photoacoustic imaging in mice.

Lysosome-targetable polythiophene nanoparticles for two-photon excitation photodynamic therapy and deep tissue imaging.

Two-photon excitation (TPE) photodynamic therapy (PDT) has attracted great interest due to its distinctive properties, e.g., good penetration ability of biological tissues and less damage to healthy tissues. However, the conventional photosensitizers (PSs) for PDT have poor subcellular location capability and low two-photon absorption (TPA) cross section in the phototherapeutic window. Herein, we report a fascinating multi-functional TPE PS, polythiophene nanoparticles (PT NPs), for simultaneous lysosome-targetable fluorescence imaging and PDT. PT NPs show bright yellow fluorescence, good water solubility as well as excellent photo- and pH-stability. Moreover, PT NPs exhibit high singlet oxygen generation quantum yield (∼42%) and large TPA cross section (∼3420 GM). A fluorescence imaging penetration depth of 1800 μm could be reached in the tissue phantom under the TPE mode. Due to these outstanding merits and their unique clathrin- and caveola-independent intracellular uptake pathway, PT NPs have great potential for application in TPE fluorescence imaging and photodynamic therapy.