Enhanced Photodynamic Therapy

The following sections are included:

• Preface
• About the Editors
• Contents

The current pandemic SARS-CoV-2 (also known as 2019-nCoV and COVID-19) viral infection is growing globally and has created a disastrous situation all over the world. One of the biggest challenges is that no drugs are available to treat this life-threatening disease. As no drugs are available for definitive treatment of this disease and the mortality rate is very high, there is an utmost need to cure the infection using novel technologies. This study will point out some new antimicrobial technologies that have great potentials for eradicating and preventing emerging infections. They can be considered as treatments of choice for viral infections in the future.

This study is aimed at the chemical synthesis of light-activated cobalt-doped zinc oxide and its further doping on reduced graphene oxide (RGO) and assessment of its antibacterial activity on antibiotic-resistant waterborne pathogens including Enterococcus faecalis, Staphylococcus aureus, Klebsiella pneumonia, and Pseudomonas aeruginosa. The synthesized nanoparticles were characterized via UV–vis spectroscopy, scanning electron microscopy (SEM), and energy-dispersive X-ray spectroscopy (EDS). The minimal inhibitory concentration (MIC) of nanoparticles portrayed a significant killing of both Gram-positive and Gram-negative bacteria. The synthesized nanoparticles were further found as active killers of bacteria in drinking water. Further, these nanoparticles were found photothermally active alongside ROS generators. The photokilling activity makes them ideal replacement candidates for traditional water filters.

Bacterial resistance is today a matter of great medical concern, so it is urgent to investigate alternatives to alleviate it. Photodynamic inactivation (PDI) is a method that has been endorsed to inactivate different pathogens, including bacteria, fungi and viruses. PDI is achieved by using a photosensitizer (PS) molecule which generates reactive oxygen species under visible or UV radiation. We use visible light and UV-A radiation to excite four commercial PSs (methylene blue, rose bengal, riboflavin and curcumin), and nanoparticles synthesized in our laboratory. Despite these PSs having been thoroughly studied in the past by other research groups, in order to compare their effects in an appropriate way, we matched the number of photons they absorb. We found that methylene blue leads to the major inactivation of Escherichia coli. Furthermore, we evaluated the production of singlet oxygen and hydroxyl radicals in the photoinactivation process.

Surgical tumor resection is a common approach to cancer treatment. India Ink tattoos are widely used to aid tumor resection by localizing and mapping the tumor edge at the surface. However, India Ink tattoos are easily obscured during electrosurgical resection, and fade in intensity over time. In this work, a novel near-infrared (NIR) fluorescent marker is introduced as an alternative. The NIR marker was made by mixing indocyanine green (ICG), biocompatible cyanoacrylate, and acetone. The marking strategy was evaluated in a chronic ex vivo feasibility study using porcine tissues, followed by a chronic in vivo mouse study while compared with India Ink. In both studies, signal-to-noise (SNR) ratios and dimensions of the NIR markers and/or India Ink over the study period were calculated and reported. Electrocautery was performed on the last day of the mouse study after mice were euthanized, and SNR ratios and dimensions were quantified and compared. Biopsy was performed at all injection sites and slides were examined by a pathologist. The proposed NIR marker achieved (i) consistent visibility in the 26-day feasibility study and (ii) improved durability, visibility, and biocompatibility when compared to traditional India Ink over the six-week period in an in vivo mouse model. These effects persist after electrocautery whereas the India Ink markers were obscured. The use of a NIR fluorescent presurgical marking strategy has the potential for intraoperative tracking during long-term treatment protocols.

To date, the clinical use of functional near-infrared spectroscopy (NIRS) to detect cerebral ischemia has been largely limited to surgical settings, where motion artifacts are minimal. In this study, we present novel techniques to address the challenges of using NIRS to monitor ambulatory patients with kidney disease during approximately eight hours of hemodialysis (HD) treatment. People with end-stage kidney disease who require HD are at higher risk for cognitive impairment and dementia than age-matched controls. Recent studies have suggested that HD-related declines in cerebral blood flow might explain some of the adverse outcomes of HD treatment. However, there are currently no established paradigms for monitoring cerebral perfusion in real-time during HD treatment. In this study, we used NIRS to assess cerebral hemodynamic responses among 95 prevalent HD patients during two consecutive HD treatments. We observed substantial signal attenuation in our predominantly Black patient cohort that required probe modifications. We also observed consistent motion artifacts that we addressed by developing a novel NIRS methodology, called the HD cerebral oxygen demand algorithm (HD-CODA), to identify episodes when cerebral oxygen demand might be outpacing supply during HD treatment. We then examined the association between a summary measure of time spent in cerebral deoxygenation, derived using the HD-CODA, and hemodynamic and treatment-related variables. We found that this summary measure was associated with intradialytic mean arterial pressure, heart rate, and volume removal. Future studies should use the HD-CODA to implement studies of real-time NIRS monitoring for incident dialysis patients, over longer time frames, and in other dialysis modalities.

The automatic and accurate identification of apoptosis facilitates large-scale cell analysis. Most identification approaches using nucleus fluorescence imaging are based on specific morphological parameters. However, these parameters cannot completely describe nuclear morphology, thus limiting the identification accuracy of models. This paper proposes a new feature extraction method to improve the performance of the model for apoptosis identification. The proposed method uses a histogram of oriented gradient (HOG) of high-frequency wavelet coefficients to extract internal and edge texture information. The HOG vectors are classified using support vector machine. The experimental results demonstrate that the proposed feature extraction method well performs apoptosis identification, attaining 95:7% accuracy with low cost in terms of time. We confirmed that our method has potential applications to cell biology research.

Photobiomodulation (PBM) promoting wound healing has been demonstrated by many studies. Currently, 630 nm and 810 nm light-emitting diodes (LEDs), as light sources, are frequently used in the treatment of diabetic foot ulcers (DFUs) in clinics. However, the dose–effect relationship of LED-mediated PBM is not fully understood. Furthermore, among the 630 nm and 810 nm LEDs, which one gets a better effect on accelerating the wound healing of diabetic ulcers is not clear. The aim of this study is to evaluate and compare the effects of 630 nm and 810 nm LED-mediated PBM in wound healing both in vitro and in vivo. Our results showed that both 630 nm and 810 nm LED irradiation significantly promoted the proliferation of mouse fibroblast cells (L929) at different light irradiances (1, 5, and 10 mW/cm². The cell proliferation rate increased with the extension of irradiation time (100, 200, and 500 s), but it decreased when the irradiation time was over 500 s. Both 630 nm and 810 nm LED irradiation (5 mW/cm² significantly improved the migration capability of L929 cells. No difference between 630 nm and 810 nm LED-mediated PBM in promoting cell proliferation and migration was detected. In vivo results presented that both 630 nm and 810 nm LED irradiation promoted the wound healing and the expression of the vascular endothelial growth factor (VEGF) and transforming growth factor (TGF) in the wounded skin of type 2 diabetic mice. Overall, these results suggested that LED-mediated PBM promotes wound healing of diabetic mice through promoting fibroblast cell proliferation, migration, and the expression of growth factors in the wounded skin. LEDs (630 nm and 810 nm) have a similar outcome in promoting wound healing of type 2 diabetic mice.

Reduced nicotinamide adenine dinucleotide (NADH) plays a crucial role in many biochemical reactions in human metabolism. In this work, a flow-mediated skin fluorescence (FMSF)–post-occlusion reactive hyperaemia (PORH) system was developed for noninvasive and in vivo measurement of NADH fluorescence and its real-time dynamical changes in human skin tissue. The real-time dynamical changes of NADH fluorescence were analyzed with the changes of skin blood flow measured by laser speckle contrast imaging (LSCI) experiments simultaneously with FMSF-PORH measurements, which suggests that the dynamical changes of NADH fluorescence would be mainly correlated with the intrinsic changes of NADH level in the skin tissue. In addition, Monte Carlo simulations were applied to understand the impact of optical property changes on the dynamical changes of NADH fluorescence during the PORH process, which further supports that the dynamical changes of NADH fluorescence measured in our system would be intrinsic changes of NADH level in the skin tissue.

Photodynamic therapy (PDT) not only destroys tumor cells directly but also induced anti-tumor immune response through damage-associated molecular patterns (DAMPs). It is reported that anti-tumor response was associated with light dose and photosensitizer used in PDT. In this study, 4T1 tumor cells were implanted on both the right and left flanks of mice. Only the right tumor was treated by HpD-PDT, while the left tumor was not irradiated. The anti-tumor immune response induced by HpD-PDT was investigated. The expression of DAMPs and co-stimulatory molecules induced by HpD-PDT were tested by immunofluorescence and flow cytometry in vivo. Different light doses of PDT were designed to treat 4T1 cells. The killing effect was assessed by CCK-8 kit and apoptosis kit. The expression of DAMPs on 4T1 cells after HpD-PDT were evaluated by flow cytometry, western blot and ATP kit. This study showed that CD4⁺T, CD8⁺T and the production of IFN-γ were increased significantly on day 10 in right-tumor after PDT treatment compared with control group. HpD-PDT enhanced the expression of calreticulin (CRT) on tumor tissue. Importantly, co-stimulatory molecular OX-40 and 4-1BB were elevated on CD8⁺T cells. In vitro, immunogenic death of 4T1 cells was induced after PDT. Besides, the expression of DAMPs increased with the increasing of energy density. This study indicates that anti-tumor immune effect was induced by HpD-PDT. The knowledge of the involvement of CRT, ATP and costimulatory molecules uncovers important mechanistic insight into the anti-tumor immunogenicity. It was the first time that co-stimulatory molecules were investigated and found to elevate after PDT.

Cerebral edema is a severe complication of acute ischemic stroke with high mortality but limited treatment. Although parameters such as brain water content and intracranial pressure may represent the global assessment of edema, optical properties can appear heterogeneously throughout the cerebral tissue relative to the site of injury. In this study, we have monitored the edema formation and progression in both permanent and transient middle cerebral artery occlusion models in rats. Edema was reflected by the decrease of optical attenuation coefficient (OAC) value in OCT system. By utilizing swept-source optical coherence tomography (SS-OCT), we found that in photochemically induced permanent focal stroke model, both the edema size and edema index, steadily developed until the end of monitor (7 h). Comparatively, when transient ischemia was introduced with endothelin-1 (ET-1), the edema was detected as early as 15 min, and began to recover after 30 min until monitor was finished (3 h). Despite the majority of the edema being recovered to some extent, the condition of a small region within the edema kept deteriorating, presumably due to the reperfusion damage which might result in serious clinical outcomes. Our study has compared the edema characteristics from two different acute ischemic stroke situations. This work not only confirms the capability of OCT to temporal and spatial monitor of edema but is also able to locate focal conditions at some areas that might highly determine the prognosis and treatment decisions.

Fluorescence imaging is very useful for skin cancer lesions detection because of its properties of noninvasion and fast imaging. However, conventional fluorescence imaging devices’ excitation light source and camera are usually separated, which will cause problems such as complicated structure, large volume, and poor illumination homogeneity. In this paper, we introduce a miniature portable fluorescence imaging device to diagnose skin cancer. A coaxial design has been introduced to combine the exciting light source and fluorescence receiver as an integral part, which significantly reduces the size of the device and ensures illumination homogeneity. The volume of the device is less than 3:5 × 3:5 × 9:5 cm³ with weight of 150 g, and the total power (including the excitation lamp) is only 1.5 W. It is used to detect the squamous cell carcinoma mice for demonstration. The results show that the location of the cancer lesions can be easily distinguished from the images captured by the device. It can be efficiently used to detect early skin tumors with noninvasion. It also has prospects to be integrated with other diagnostic methods such as ultrasound probe, for multiple diagnose of skin tumors thanks to its miniature size.

Determination of the precise location and the degree of the Choroidal neovascularization (CNV) lesion is essential for diagnosation Neovascular age-related macular degeneration (AMD) and evaluation the efficacy of treatment. Noninvasive imaging techniques with specific contrast for CNV evaluation are demanded. In this paper, two noninvasive imaging techniques, namely Optical coherence tomography (OCT) and Photoacoustic microscopy (PAM), are combined to provide specific detection of CNV for their complimentary contrast mechanisms. In vivo time-serial evaluation of Laser-induced CNV in rats is present at days 1, 3, 5, 7, 14, 21 after laser photocoagulation is applied to the rat fundus. Both OCT and PAM show that the CNV increases to its maximum at day 7 and decreases at day 14. Quantification of CNV area and CNV thickness is given. The dual-modal information of CNV is consistent with the histologic evaluation by hematoxylin and eosin (H&E) staining.

Exosomes are lipid bilayer vesicles released by cells and serve as natural carriers for cell–cell communication. Exosomes provide a promising approach to the diagnosis and treatment of diseases and are considered as an alternative to cell therapy. However, one main restriction in their clinical application is that the current understanding of these vesicles, especially their in vivo behaviors and distributions, remains inadequate. Here, we reviewed the current and emerging methods for in vivo imaging and tracking of exosomes, including fluorescence imaging, bioluminescence imaging, nuclear imaging, X-ray imaging, magnetic resonance imaging, photoacoustic imaging, and multimodal imaging. In vivo imaging and tracking of exosomes by these methods can help researchers further understand their uptake mechanism, biodistribution, migration, function, and therapeutic performance. The pioneering studies in this field can elucidate many unknown exosomal behaviors at different levels. We discussed the advantages and limitations of each labeling and imaging strategy. The advances in labeling and in vivo imaging will expand our understanding of exosomes and promote their clinical application. We finally provide a perspective and discuss several important issues that need to be explored in future research. This review highlights the values of efficient, sensitive, and biocompatible exosome labeling and imaging techniques in disease theranostics.

Apoptosis is very important for the maintenance of cellular homeostasis and is closely related to the occurrence and treatment of many diseases. Mitochondria in cells play a crucial role in programmed cell death and redox processes. Nicotinamide adenine dinucleotide (NAD(P)H) is the primary producer of energy in mitochondria, changing NAD(P)H can directly reflect the physiological state of mitochondria. Therefore, NAD(P)H can be used to evaluate metabolic response. In this paper, we propose a noninvasive detection method that uses two-photon fluorescence lifetime imaging microscopy (TP-FLIM) to characterize apoptosis by observing the binding kinetics of cellular endogenous NAD(P)H. The result shows that the average fluorescence lifetime of NAD(P)H and the fluorescence lifetime of protein-bound NAD(P)H will be affected by the changing pH, serum content, and oxygen concentration in the cell culture environment, and by the treatment with reagents such as H₂O₂ and paclitaxel. Taxol (PTX). This noninvasive detection method realized the dynamic detection of cellular endogenous substances and the assessment of apoptosis.

Photodynamic therapy (PDT) dosimetry, including light dose, photosensitizer dose and tissue oxygen, has been a research focus in PDT. In this work, we present a three-dimensional (3D) quantification of protoporphyrin IX (PpIX) using combined spatial frequency domain imaging (SFDI) and diffuse fluorescence tomography (DFT). The SFDI maps both the distributions of tissue absorption and scattering properties at three wavelengths and accordingly provides the optical background for DFT and extracts the tissue oxygenation for assessing the therapeutic outcomes, while DFT dynamically monitors the 3D distribution of PpIX dose from measured fluorescence signals for the procedure optimization. A pilot in vivo application in tumor nude models showed that the proposed SFDI/DFT is able to dynamically trace changes in the PpIX concentration and tissue oxygen during the treatment, rendering it a potentially powerful tool for PDT to improve clinical efficacy.