Triplet-Triplet Annihilation Upconverting Liposomes: Mechanistic Insights into the Role of Membranes in Two-Dimensional TTA-UC.

Triplet-triplet annihilation upconversion (TTA-UC) implemented in nanoparticle assemblies is of emerging interest in biomedical applications, including in drug delivery and imaging. As it is a bimolecular process, ensuring sufficient mobility of the sensitizer and annihilator to facilitate effective collision in the nanoparticle is key. Liposomes can provide the benefits of two-dimensional confinement and condensed concentration of the sensitizer and annihilator along with superior fluidity compared to other nanoparticle assemblies. They are also biocompatible and widely applied across drug delivery modalities. However, there are relatively few liposomal TTA-UC systems reported to date, so systematic studies of the influence of the liposomal environment on TTA-UC are currently lacking. Here, we report the first example of a BODIPY-based sensitizer TTA-UC system within liposomes and use this system to study TTA-UC generation and compare the relative intensity of the anti-Stokes signal for this system as a function of liposome composition and membrane fluidity. We report for the first time on time-resolved spectroscopic studies of TTA-UC in membranes. Nanosecond transient absorption data reveal the BODIPY-perylene dyad sensitizer has a long triplet lifetime in liposome with contributions from three triplet excited states, whose lifetimes are reduced upon coinclusion of the annihilator due to triplet-triplet energy transfer, to a greater extent than in solution. This indicates triplet energy transfer between the sensitizer and the annihilator is enhanced in the membrane system. Molecular dynamics simulations of the sensitizer and annihilator TTA collision complex are modeled in the membrane and confirm the co-orientation of the pair within the membrane structure and that the persistence time of the bound complex exceeds the TTA kinetics. Modeling also reliably predicted the diffusion coefficient for the sensitizer which matches closely with the experimental values from fluorescence correlation spectroscopy. The relative intensity of the TTA-UC output across nine liposomal systems of different lipid compositions was explored to examine the influence of membrane viscosity on upconversion (UC). UC showed the highest relative intensity for the most fluidic membranes and the weakest intensity for highly viscous membrane compositions, including a phase separation membrane. Overall, our study reveals that the co-orientation of the UC pair within the membrane is crucial for effective TTA-UC within a biomembrane and that the intensity of the TTA-UC output can be tuned in liposomal nanoparticles by modifying the phase and fluidity of the liposome. These new insights will aid in the design of liposomal TTA-UC systems for biomedical applications.

BODIPY-Perylene Charge Transfer Compounds; Sensitizers for Triplet-Triplet Annihilation Up-conversion.

BODIPY heterochromophores, asymmetrically substituted with perylene and/or iodine at the 2 and 6 positions were prepared and investigated as sensitizers for triplet-triplet annihilation up conversion (TTA-UC). Single-crystal X-ray crystallographic analyses show that the torsion angle between BODIPY and perylene units lie between 73.54 and 74.51, though they are not orthogonal. Both compounds show intense, charge transfer absorption and emission profiles, confirmed by resonance Raman spectroscopy and consistent with DFT calculations. The emission quantum yield was solvent dependent but the emission profile remained characteristic of CT transition across all solvents explored. Both BODIPY derivatives were found to be effective sensitizers of TTA-UC with perylene annihilator in dioxane and DMSO. Intense anti-Stokes emission was observed, and visible by eye from these solvents. Conversely, no TTA-UC was observed from the other solvents explored, including from non-polar solvents such as toluene and hexane that yielded brightest fluorescence from the BODIPY derivatives. In dioxane, the power density plots obtained were strongly consistent with TTA-UC and the power density threshold, the Ith value (the photon flux at which 50 % of ΦTTAUC is achieved), for B2PI was observed to be 2.5x lower than of B2P under optimal conditions, an effect ascribed to the combined influence of spin-orbit charge transfer intersystem crossing (SOCT-ISC) and heavy metal on the triplet state formation for B2PI.

In Cellulo Light-Induced Dynamics in a BODIPY-Perylene Dyad.

BODIPY-based donor-acceptor dyads are widely used as sensors and probes in life science. Thus, their biophysical properties are well established in solution, while their photophysical properties in cellulo, i. e., in the environment, in which the dyes are designed to function, are generally understood less. To address this issue, we present a sub-ns time-resolved transient absorption study of the excited-state dynamics of a BODIPY-perylene dyad designed as a twisted intramolecular charge transfer (TICT) probe of the local viscosity in live cells.

A biphasic response of polymerized Type 1 collagen architectures to dermatan sulfate.

Collagen I, the most abundant extracellular matrix (ECM) protein in vertebrate tissues provides mechanical durability to tissue microenvironments and regulates cell function. Its fibrillogenesis in biological milieu is predominantly regulated by dermatan sulfate proteoglycans, proteins conjugated with iduronic acid-containing dermatan sulfate (DS) glycosaminoglycans (GAG). Although DS is known to regulate tissue function through its modulation of Coll I architecture, a precise understanding of the latter remains elusive. We investigated this problem by visualizing the fibrillar pattern of fixed Coll I gels polymerized in the presence of varying concentrations of DS using second harmonic generation microscopy. Measuring mean second harmonic generation signal (which estimates the ordering of the fibrils), and surface occupancy (which estimates the space occupied by fibrils) supported by confocal reflectance microscopy, our observations indicated that the effect on fibril pattern of DS is contextual upon the latter's concentrations: Lower levels of DS resulted in sparse disorganized fibrils; higher levels restore organization, with fibrils occupying greater space. An appropriate change in elasticity as a result of DS levels was also observed through atomic force microscopy. Examination of dye-based GAG staining and scanning electron microscopy suggested distinct constitutions of Coll I gels when polymerized with higher and lower levels of DS. We observed that adhesion of the invasive ovarian cancer cells SKOV3 decreased for lower DS levels but was partially restored at higher DS levels. Our study shows how the Coll I gel pattern-tuning of DS is of relevance for understanding its biomaterial applications and possibly, pathophysiological functions.

Third-harmonic generation in multilayer Tin Diselenide under the influence of Fabry-Perot interference effects.

Two-dimensional layered materials are in general known to exhibit strong layer dependent nonlinear optical response owing to the crystal symmetry and associated phase matching considerations. Here we report up-conversion of 1550 nm incident light using third-harmonic generation (THG) in multilayered tin di-selenide (SnSe2) and study its thickness dependence by simultaneously acquiring spatially-resolved images in the forward and backward propagation direction. We find good agreement between the experimental measurements and a coupled-wave equation model we have developed when including the effect of Fabry-Perot interference between the SnSe2 layer and the surrounding medium. We extract the magnitude of the third order electronic nonlinear optical susceptibility of SnSe2, for the first time to our knowledge, by comparing its nonlinear response with a glass substrate and find this to be ∼1500 times higher than that of glass. We also study the polarization dependence and find good agreement with the expected angular dependence of nonlinear polarization considering the crystal symmetry of SnSe2. The large nonlinear optical susceptibility of multi-layer SnSe2 makes it a promising material for studying nonlinear optical effects. This work demonstrates that in addition to the large inherent nonlinear optical susceptibility, the high refractive index of these materials and optical absorption above the bandgap strongly influence the overall nonlinear optical response and its thickness dependence characteristics.

Iduronate-2-Sulfatase-Regulated Dermatan Sulfate Levels Potentiate the Invasion of Breast Cancer Epithelia through Collagen Matrix.

Cancer epithelia show elevation in levels of sulfated proteoglycans including dermatan sulfates (DS). The effect of increased DS on cancer cell behavior is still unclear. We hypothesized that decreased expression of the enzyme Iduronate-2-sulfatase (IDS) can lead to increased DS levels, which would enhance the invasion of cancer cells. Breast cancer sections shows depleted IDS levels in tumor epithelia, when compared with adjacent untransformed breast tissues. IDS signals showed a progressive decrease in the non-transformed HMLE, transformed but non-invasive MCF-7 and transformed and invasive MDA-MB-231 cells, respectively, when cultured on Type 1 collagen scaffolds. DS levels measured by ELISA increased in an inverse-association with IDS levels. Knockdown of IDS in MCF-7 epithelia also increased the levels of DS. MCF-7 cells with depleted IDS expression, when imaged using two photon-excited fluorescence and second harmonic generation microscopy, exhibited a mesenchymal morphology with multiple cytoplasmic projections compared with epithelioid control cells, interacted with their surrounding matrix, and showed increased invasion through Type 1 collagen matrices. Both these traits were phenocopied when control MCF-7 cells were cultivated on Type 1 collagen gels polymerized in the presence of DS. In monolayer cultures, DS had no effect on MCF-7 migration. In the context of our demonstration that DS enhances the elastic modulus of Type 1 collagen gels, we propose that a decrease of IDS expression leads to accumulation within cancer epithelia of DS: the latter remodels the collagen around cancer cells leading to changes in cell shape and invasiveness through fibrillar matrix milieu.

Microscopic study of resonant third-harmonic generation from amorphous silicon nanodisk arrays.

A detailed microscopic study of third-harmonic generation (THG) from two-dimensional arrays of sub-wavelength spaced amorphous silicon nanodisks is reported. The arrays are designed to support broadband, minimally angle-sensitive resonances for the fundamental excitation wavelength in the 1500 nm region. This results in resonantly enhanced visible THG in the green spectral range with ∼500-fold enhancement on-resonance, compared to the unpatterned a-Si thin-film. THG multispectral microscopic imaging reveals individual nanodisks with enhanced nonlinear signal on-resonance. For increasing pump intensities, spatially dependent saturation effects are observed for the first time, to the best of our knowledge, in such dielectric nanostructure arrays with THG images showing a reversal of contrast.