Induction of Immunogenic Cell Death by Photodynamic Therapy Mediated by Aluminum-Phthalocyanine in Nanoemulsion

Photodynamic therapy (PDT) has been clinically employed to treat mainly superficial cancer, such as basal cell carcinoma. This approach can eliminate tumors by direct cytotoxicity, tumor ischemia, or by triggering an immune response against tumor cells. Among the immune-related mechanisms of PDT, the induction of immunogenic cell death (ICD) in target cells is to be cited. ICD is an apoptosis modality distinguished by the emission of damage-associated molecular patterns (DAMP). Therefore, this study aimed to analyze the immunogenicity of CT26 and 4T1 treated with PDT mediated by aluminum-phthalocyanine in nanoemulsion (PDT-AlPc-NE). Different PDT-AlPc-NE protocols with varying doses of energy and AlPc concentrations were tested. The death mechanism and the emission of DAMPs–CRT, HSP70, HSP90, HMGB1, and IL-1β–were analyzed in cells treated in vitro with PDT. Then, the immunogenicity of these cells was assessed in an in vivo vaccination-challenge model with BALB/c mice. CT26 and 4T1 cells treated in vitro with PDT mediated by AlPc IC50 and a light dose of 25 J/cm² exhibited the hallmarks of ICD, i.e., these cells died by apoptosis and exposed DAMPs. Mice injected with these IC50 PDT-treated cells showed, in comparison to the control, increased resistance to the development of tumors in a subsequent challenge with viable cells. Mice injected with 4T1 and CT26 cells treated with higher or lower concentrations of photosensitizer and light doses exhibited a significantly lower resistance to tumor development than those injected with IC50 PDT-treated cells. The results presented in this study suggest that both the photosensitizer concentration and light dose affect the immunogenicity of the PDT-treated cells. This event can affect the therapy outcomes in vivo.

On the Synergism of Biogenic Gold Nanoparticles and Hydroxyaluminum Phthalocyanines in the Photoeradication of Staphylococcus aureus

Due to the unusual properties of gold nanoparticles, these structures are widely used in medicine and biology. This paper describes for the first time the synthesis of colloidal gold nanoparticles by the cell-free filtrate obtained from the Coriolus versicolor biomass and the use of these biogenic nanostructures to increase the photosensitizing efficiency of di- (AlPcS2) and tetrasulfonated (AlPcS4) hydroxyaluminum phthalocyanines in antibacterial photodynamic therapy. The obtained monodisperse particles were extremely stable, and this remarkable stability was due to the presence of phosphoprotein as a capping agent. The studied gold nanoparticles had a spherical shape, were uniformly distributed, and were characterized by a single plasmon band at wavelength of 514–517 nm. Almost 60% of the gold particles were found to be in the range of 13 to 15 nm. In accordance with the regulations of the American Microbiological Society, indicating that any antimicrobial technique must kill at least 3 log CFU (99.9%) to be accepted as “antimicrobial”, this mortality of Staphylococcus aureus was shown to be achieved in the presence of AlPcS4 + AuNPs mixture and 4.8 J cm−2 light dose compared to AlPcS4 alone, which required a light dose of 24 J cm−2. The best effect of increasing the effectiveness of combating this pathogen was observed in the case of AlPcS2 + AuNPs as a photosensitizing mixture. The light dose of 24 J cm−2 caused a lethal effect of the studied coccus in the planktonic culture.

Temperature-responded tunable metalenses based on phase transition materials

Once the metalenses are fabricated, the functions of most are invariable. The tunability and reconfigurability are useful and cost-saving for metalenses in realistic applications. We demonstrated this tunability here via a novel hybrid metalens with the strategic placement of an ultra-thin VO2 layer. The hybrid metalens is capable of dynamically modulating the focusing intensity of transmitted light at 1550 nm, and demonstrated a 42.28% focusing efficiency of the incident light and 70.01% modulation efficiency. The hybrid metalens’ optothermal simulations show the optothermal conversion progress of dynamic focusing, and the maximum laser density of 1.76×103 W/cm2 can be handled at an ambient temperature lower than 330 K. The hybrid metalens proposed in this work, a light-dose sensitive tunable smart metalens that can protect other instruments/systems or materials from damage, has its specific applications such as anti-satellite blinding, bio-imaging, etc.

An adaptive microscope for the imaging of biological surfaces

Scanning fluorescence microscopes are now able to image large biological samples at high spatial and temporal resolution. This comes at the expense of an increased light dose which is detrimental to fluorophore stability and cell physiology. To highly reduce the light dose, we designed an adaptive scanning fluorescence microscope with a scanning scheme optimized for the unsupervised imaging of cell sheets, which underly the shape of many embryos and organs. The surface of the tissue is first delineated from the acquisition of a very small subset (~0.1%) of sample space, using a robust estimation strategy. Two alternative scanning strategies are then proposed to image the tissue with an improved photon budget, without loss in resolution. The first strategy consists in scanning only a thin shell around the estimated surface of interest, allowing high reduction of light dose when the tissue is curved. The second strategy applies when structures of interest lie at the cell periphery (e.g. adherens junctions). An iterative approach is then used to propagate scanning along cell contours. We demonstrate the benefit of our approach imaging live epithelia from Drosophila melanogaster. On the examples shown, both approaches yield more than a 20-fold reduction in light dose -and up to more than 80-fold- compared to a full scan of the volume. These smart-scanning strategies can be easily implemented on most scanning fluorescent imaging modality. The dramatic reduction in light exposure of the sample should allow prolonged imaging of the live processes under investigation.

Photodynamic priming with triple-receptor targeted nanoconjugates that trigger T cell-mediated immune responses in a 3D in vitro heterocellular model of pancreatic cancer

Photodynamic priming (PDP), a collateral effect of photodynamic therapy, can transiently alter the tumor microenvironment (TME) beyond the cytotoxic zone. Studies have demonstrated that PDP increases tumor permeability and modulates immune-stimulatory effects by inducing immunogenic cell death, via the release of damage-associated molecular patterns and tumor-associated antigens. Pancreatic ductal adenocarcinoma (PDAC) is one of the deadliest of cancers with a stubborn immunosuppressive TME and a dense stroma, representing a challenge for current molecular targeted therapies often involving macromolecules. We, therefore, tested the hypothesis that PDP’s TME modulation will enable targeted therapy and result in immune stimulation. Using triple-receptor-targeted photoimmuno-nanoconjugate (TR-PINs)-mediated PDP, targeting epidermal growth factor receptor, transferrin receptor, and human epidermal growth factor receptor 2 we show light dose-dependent TR-PINs mediated cytotoxicity in human PDAC cells (MIA PaCa-2), co-cultured with human pancreatic cancer-associated fibroblasts (PCAFs) in spheroids. Furthermore, TR-PINs induced the expression of heat shock proteins (Hsp60, Hsp70), Calreticulin, and high mobility group box 1 in a light dose and time-dependent manner. TR-PINs-mediated T cell activation was observed in co-cultures of immune cells with the MIA PaCa-2-PCAF spheroids. Both CD4+ T and CD8+ T cells showed light dose and time-dependant antitumor reactivity by upregulating degranulation marker CD107a and interferon-gamma post-PDP. Substantial tumor cell death in immune cell-spheroid co-cultures by day 3 shows the augmentation by antitumor T cell activation and their ability to recognize tumors for a light dose-dependent kill. These data confirm enhanced destruction of heterogeneous pancreatic spheroids mediated by PDP-induced phototoxicity, TME modulation and increased immunogenicity with targeted nanoconstructs.

Seasonal Efficiency of Supplemental LED Lighting on Growth and Photomorphogenesis of Sweet Basil

For decisions on supplemental lighting a quantitative knowledge of the plants' responses to light under varying conditions is fundamental. In this study, we developed light dose-response curves of growth and morphological traits for Ocimum basilicum L. and examined the effects of light color (blue, red, and white plus far-red) and natural environment (season) on these curves. Four greenhouse experiments were conducted throughout the year to determine the efficiencies of the light regimes on growth and their effects on plant morphology. A special aspect was the photosynthetic efficiency of far-red light. Linear and monomolecular relationships were found for the relationships between plant traits and supplemental light dose. Traits related to biomass productivity increased linearly with light dose whereas some morphological characters showed a saturation behavior. Red light and white plus far-red light were more efficient in plant dry weight production than blue light, and the plants adapted differently to the light qualities: higher biomass under red light was related to a plant architecture more favorable for light capture, i.e., taller plants and bigger leaves. White plus far-red light, on the other hand, increased leaf mass per area (LMA) and light use efficiency (LUE). Blue light resulted in lowest plant light interception and LUE. Considering photosynthetic effects of near-infrared light (PPFD800, 400–800 nm) instead of photosynthetic photon flux density (PPFD700, 400–700 nm) led to strongly reduced efficiencies. Traits related to photosynthesis such as dry weight, LMA and LUE were particularly affected by PPFD800. There were no interactions between the efficiencies of the different light colors and the seasons. Efficiencies of all light regimes were significantly lower during summer compared to spring and winter. Higher dry weight production during summer compared to winter and spring were a consequence of increased light interception rather than changes in LUE. The observed differences in seasonal efficiencies were directly linked to the amount of natural light present as indicated by changes in the ratio of supplemental to natural light.

PHOTODYNAMIC THERAPY AS A COMPONENT OF PREOPERATIVE PREPARATION FOR PATIENTS WITH STOMACH CANCER

The study and analysis of the treatment outcomes were carried out in 69 patients aged from 40 to 85 years (mean age 53.0±1.0) who had tumours of the body or cardiac part of the stomach T3-4 (according to the UICC classification (ninth edition, 2014)). The distribution of patients according to the localization of tumour lesions in the stomach was as follows: 43 (62.3%) cases found in the body region and 22 (31.9%) found in the proximal section (of which 14 (20 3%) were in the cardia (Type III according to Siewert J.R.), 8 (11.6%) patients had the transition to the abdominal oesophagus (type II according to Siewert JR), 4 (5.8%) patients had the tumour located in the stump of the resected organ. Histologically, the vast majority of neoplasms were represented by adenocarcinoma (94.8%). All other types of tumours made up for 5.2%. The patients received photodynamic therapy with the Photolon photosensitizer and subsequent irradiation with a laser with a wavelength λ = 0.67 μm crossed irradiation fields, which was introduced through the biopsy channel of the endoscope. The light dose and the number of irradiation sessions were determined depending on the degree of damage to the oesophagus and stomach (from 4 to 7 sessions). The light dose of the session was up to 200 J. The power density of the laser radiation is 25 mW/cm2. Endoscopic photodynamic therapy can be used at the stage of combined treatment in combination with chemotherapy and surgery and is an effective means of preventing the failure of the oesophageal-small intestinal anastomosis in the early postoperative period. Due to photodynamic therapy in the preoperative period, the number of complications in the early postoperative period has significantly decreased. The leakage of the oesophageal-intestinal anastomosis developed in 4 cases, which make up 5.8%.