Photodynamic Inactivation Effects Research Articles

Amino acids inhibit the photodynamic inactivation effect by hindering cellular oxidative stress

Photodynamic inactivation (PDI) could utilize light to activate reactive oxygen species (ROS) produced by photosensitizers to kill bacteria for preservation. To delve into the complex effects arising during the post-harvest PDI processing, we conducted experiments using Pseudomonas reinekei, a food spoilage bacteria extracted from rotten Pakchoi. Through analyzing the metabolomics results, we discovered that methionine (Met) and glutamate (Glu) exhibited significant inhibitory effects during the PDI process. The oxidative stress generated by light treatment resulted in a reduction of 30.31% and 36.37% in the levels of Met and Glu, respectively. The data also showed that exogenous Met and Glu reduced intracellular oxidative stress levels, increased peroxidase activity, and prevented the damage of intracellular material and cell membrane deformation. That amino acids could inhibit the effect of PDI by hindering oxidative stress. Therefore, the amino acid content should be considered when applying PDI to treat Met- or Glu-rich foods.

Erythrosine-Dialdehyde Cellulose Nanocrystal Coatings for Antibacterial Paper Packaging.

Though paper is an environmentally friendly alternative to plastic as a packaging material, it lacks antibacterial properties, and some papers have a low resistance to oil or water. In this study, a multifunctional paper-coating material was developed to reduce the use of plastic packaging and enhance paper performance. Natural cellulose nanocrystals (CNCs) with excellent properties were used as the base material for the coating. The CNCs were functionalized into dialdehyde CNCs (DACNCs) through periodate oxidation. The DACNCs were subsequently complexed using erythrosine as a photosensitizer to form an erythrosine-CNC composite (Ery-DACNCs) with photodynamic inactivation. The Ery-DACNCs achieved inactivations above 90% after 30 min of green light irradiation and above 85% after 60 min of white light irradiation (to simulate real-world lighting conditions), indicating photodynamic inactivation effects. The optimal parameters for a layer-by-layer dip coating of kraft paper with Ery-DACNCs were 4.5-wt% Ery-DACNCs and 15 coating layers. Compared to non-coated kraft paper and polyethylene-coated paper, the Ery-DACNC-coated paper exhibited enhanced mechanical properties (an increase of 28% in bursting strength). More than 90% of the bacteria were inactivated after 40 min of green light irradiation, and more than 80% were inactivated after 60 min of white light irradiation.

Photodynamic Eradication of Pseudomonas aeruginosa with Ru-Photosensitizers Encapsulated in Enzyme Degradable Nanocarriers.

The development of new approaches for the treatment of the increasingly antibiotic-resistant pathogen Pseudomonas aeruginosa was targeted by enhancing the effect of local antimicrobial photodynamic therapy (aPDT) using poly(ethylene glycol)-block-poly(lactic acid) (PEG114-block-PLAx) nanocarriers that were loaded with a ruthenium-based photosensitizer (PS). The action of tris(1,10-phenanthroline) ruthenium (II) bis(hexafluorophosphate) (RuPhen3) encapsulated in PEG114-block-PLAx micelles and vesicles was shown to result in an appreciable aPDT inactivation efficiency against planktonic Pseudomonas aeruginosa. In particular, the encapsulation of the PS, its release, and the efficiency of singlet oxygen (1O2) generation upon irradiation with blue light were studied spectroscopically. The antimicrobial effect was analyzed with two strains of Pseudomonas aeruginosa. Compared with PS-loaded micelles, formulations of the PS-loaded vesicles showed 10 times enhanced activity with a strong photodynamic inactivation effect of at least a 4.7 log reduction against both a Pseudomonas aeruginosa lab strain and a clinical isolate collected from the lung of a cystic fibrosis (CF) patient. This work lays the foundation for the targeted eradication of Pseudomonas aeruginosa using aPDT in various medical application areas.

Photodynamic inactivation of multidrug-resistant strains of Klebsiella pneumoniae and Pseudomonas aeruginosa in municipal wastewater by tetracationic porphyrin and violet-blue light: The impact of wastewater constituents.

There is an increasing need to discover effective methods for treating municipal wastewater and addressing the threat of multidrug-resistant (MDR) strains of bacteria spreading into the environment and drinking water. Photodynamic inactivation (PDI) that combines a photosensitiser and light in the presence of oxygen to generate singlet oxygen and other reactive species, which in turn react with a range of biomolecules, including the oxidation of bacterial genetic material, may be a way to stop the spread of antibiotic-resistant genes. The effect of 5,10,15,20-(pyridinium-3-yl)porphyrin tetrachloride (TMPyP3) without light, and after activation with violet-blue light (VBL) (394 nm; 20 mW/cm2), on MDR strains of Pseudomonas aeruginosa, Klebsiella pneumoniae and K. pneumoniae OXA-48 in tap water and municipal wastewater was investigated. High toxicity (~2 μM) of TMPyP3 was shown in the dark on both strains of K. pneumoniae in tap water, while on P. aeruginosa toxicity in the dark was low (50 μM) and the PDI effect was significant (1.562 μM). However, in wastewater, the toxicity of TMPyP3 without photoactivation was much lower (12.5-100 μM), and the PDI effect was significant for all three bacterial strains, already after 10 min of irradiation with VBL (1.562-6.25 μM). In the same concentrations, or even lower, an anti-adhesion effect was shown, suggesting the possibility of application in biofilm control. By studying the kinetics of photoinactivation, it was found that with 1,562 μM of TMPyP3 it is possible to achieve the complete destruction of all three bacteria after 60 min of irradiation with VBL. This study confirmed the importance of studying the impact of water constituents on the properties and PDI effect of the applied photosensitiser, as well as checking the sensitivity of targeted bacteria to light of a certain wavelength, in conditions as close as possible to those in the intended application, to adjust all parameters and perfect the method.

High-Efficiency Photodynamic Control of Antibiotic-Resistant Bacteria and Antibiotic-Resistant Genes in Wastewater Using Hydrogel-Decorated Porous Polyurethane Sponges with Metal Nanoparticles

The overuse and misuse of antibiotics in hospitals and other healthcare settings can lead to the development and spread of multidrug resistance (MDR) bacteria and water contaminants. In this study, silver nanoparticles (AgNPs)-loaded chitosan (CS)-alginate (AA) hydrogels were prepared to control wastewater MDR bacterial contaminants. CS-AA/AgNP composites strongly inhibited a wastewater MDR Enterobacter tabaci biofilm on membrane surfaces. The effect of AgNP-loaded CS-AA composite hydrogels on the gelation time, swelling ratio, in vitro degradation, and in vitro release properties was investigated. Furthermore, due to their outstanding environmentally friendly superhydrophobicity/superoleophilicity, sponges have proven compelling advantages in the area of water contaminants. Therefore, CS-AA/AgNP composites were coated on polyurethane sponges (PUSs) for disinfection applications in wastewater. CS-AA/AgNP composites and CS-AA/AgNP-coated (5 wt %) PUS exhibited higher photodynamic inactivation (PDI) of E. tabaci under blue LED light. In particular, modified PUS exhibited almost complete inactivation (5.1 log cfu/mL) at 40 min. Furthermore, tetracycline antibiotic degradation efficiency was observed to be 72.91% when the modified PUSs were added to antibiotic-polluted water in the designed reactor. In correlation with the degradation of resistance bacteria and antibiotics, the modified PUS in the designed reactor with PDI effect completely degraded tetracycline antibiotic resistance genes (ARGs) tetA and tetB at 45 min and 60 min, respectively. Specifically, PDI action on the modified PUS effectively degraded ARGs such as sul1, tetA, and tetB by 4.2, 3.2, and 2.9 log reduction (copies/mL), respectively. Excellent removal of ARGs after blue LED treatment indicated that the release of MDR pathogens and free ARGs was effectively controlled by the modified PUS photodynamic reaction. Therefore, the modified PUS can be considered as an alternative approach to remove MDR bacteria and free ARGs in the large-scale photodynamic disinfection process.

The Synergic Effect of Erythrosine and Gold Nanoparticles in Photodynamic Inactivation

Photodynamic inactivation (PDI) is a process that uses photosensitizing substances to produce reactive oxygen species. This is achieved by exposing photosensitizers to specific wavelengths of light and causing oxidative damage in cells. This sterilization technique is commonly utilized and has been extensively investigated owing to its environmentally friendly and inert characteristics. In this study, erythrosine was selected as the photosensitizer and a green light-emitting diode was used as the light source. Due to their excellent biocompatibility, gold nanoparticles were added; these acted as a carrier for erythrosine, linking it to Escherichia coli (E. coli) cells. Colony-forming unit plate counting and LIVE/DEAD bacterial viability tests were performed. A synergic PDI effect of the photosensitizer, light, and gold nanoparticles was demonstrated. After irradiation for 9 min, a bacterial death rate higher than 97% was achieved. Finally, to study the mechanism of E. coli death, we conducted reactive oxygen species tests by adding different scavengers, and concluded that the bacterial death was due to the production of singlet oxygen (Type II reaction).

Nanoemulsions loaded with compound photosensitisers: synergistic photodynamic inactivation effects of curcumin and riboflavin tetra butyrate

SummaryAntimicrobial photodynamic therapy (aPDT) is an effective method in the field of sterilisation. Singlet oxygen (1O2) is an important substance in photodynamic sterilisation that needs to be detected. It involves energy transfer from an excited photosensitiser to surrounding oxygen molecules to produce cytotoxic 1O2 species, a process termed as type II reaction. Traditional photodynamic sterilisation for fresh produce generally involves only a single photosensitiser and uses nanoemulsions coating to make the photosensitiser work better. This allows the emulsion to produce only a limited amount of reactive oxygen species (ROS) in the presence of light, such that the bactericidal effect is limited. We have innovatively compounded two photosensitisers and stored them as nanoemulsions. The composite photosensitiser produces relatively higher ROS and the nanoemulsions have good stability as well as rheological properties, and the photosensitisers are well coated in the emulsions. In summary, nanoemulsions based on composite photosensitisers showed synergistic activity in controlling the growth of Staphylococcus aureus, which provides a novel approach for raw sterilisation to maintain food safety.

Photodynamic Inactivation Effectively Eradicates Candida auris Biofilm despite Its Interference with the Upregulation of CDR1 and MDR1 Efflux Genes.

Candida auris, in recent years, has emerged as a dangerous nosocomial pathogen. It represents a challenge for effective treatment because of its multiresistance. Photodynamic inactivation (PDI) is a promising way to solve problems with a wide range of resistant microorganisms. This study aimed to use PDI for the eradication of C. auris biofilms. Moreover, the regulation of the CDR1, CDR2, and MDR1 resistance genes was studied. Experiments were performed on 24 h biofilms formed by three clinical isolates of C. auris in vitro. PDI was performed in the presence of the photosensitizer methylene blue (0.25 mM) and samples were irradiated with a red laser (λ = 660 nm, 190 mW/cm2) for 79, 120, and 300 s. To confirm the PDI effect, confocal laser scanning microscopy was performed after treatment. Effective PDI was achieved in all strains. The highest inhibition was observed after 300 s irradiation, with over 90% inhibition compared with the non-irradiated control sample. PDI was observed to upregulate the expression of the CDR1 gene, but mainly the MDR1 gene. Despite this observation, PDI significantly decreased the survival of C. auris biofilm cells and proved to have great potential for the eradication of problematic resistant yeasts.

Photodynamic inactivation of pathogenic Gram-negative and Gram-positive bacteria mediated by Si(IV) phthalocyanines bearing axial ammonium units.

The photodynamic inactivation (PDI) of microorganisms has gained interest as an efficient option for conventional antibiotic treatments. Recently, Si(IV) phthalocyanines (SiPcs) have been highlighted as promising photosensitizers (PSs) to the PDI of microorganisms due to their remarkable absorption and emission features. To increase the potential of cationic SiPcs as PS drugs, one novel (1a) and two previously described (2a and 3a) axially substituted PSs with di-, tetra-, and hexa-ammonium units, respectively, were synthesized and characterized. Their PDI effect was evaluated for the first time against Escherichia coli and Staphylococcus aureus, a Gram-negative and a Gram-positive bacterium, respectively. The photodynamic treatments were conducted with PS concentrations of 3.0 and 6.0μM under 60min of white light irradiation (150mW.cm-2). The biological results show high photodynamic efficiency for di- and tetra-cationic PSs 1a and 2a (6.0μM), reducing the E. coli viability in 5.2 and 3.9 log, respectively (after 15min of dark incubation before irradiation). For PS 3a, a similar bacterial reduction (3.6 log) was achieved but only with an extended dark incubation period (30min). Under the same experimental conditions, the photodynamic effect of cationic PSs 1a-3a on S. aureus was even more promising, with abundance reductions of ca. 8.0 log after 45-60min of PDI treatment. These results reveal the high PDI efficiency of PSs bearing ammonium groups and suggest their promising application as a broad-spectrum antimicrobial to control infections caused by Gram-negative and Gram-positive bacteria.

Photodynamic inactivation effects on foodborne pathogens and its application in food products

Photodynamic inactivation effects on foodborne pathogens and its application in food products

Selective photodynamic inactivation of Helicobacter pylori by a cationic benzylidene cyclopentanone photosensitizer - an in vitro and ex vivo study.

The rise in the antibiotic resistance rate of Helicobacter pylori has led to an increasing eradication failure of this carcinogenic bacterial pathogen worldwide. This underlines the need for alternative antibacterial strategies against H. pylori infection. Antimicrobial photodynamic therapy (aPDT) is a promising non-pharmacological antibacterial technology. In this study, the selective killing activities of three benzylidene cyclopentanone (BCP) photosensitizers (Y1, P1 and P3) towards H. pylori over normal human gastric epithelial GES-1 cells were evaluated and the ex vivo photodynamic inactivation effect was preliminarily assessed on twelve H. Pylor-infected mice. Results showed that under the irradiation of 24J/cm2 532nm laser, Y1, P1 and P3 at 2.5μM induced a 3-log10 reduction of H. pylori CFU (99.9% killing). Confocal images showed that P3, unlike Y1 and P1, could not be uptaken by GES-1 cells. P3 at 2.5 to 20μM showed not significant (p>0.05) phototoxicity to GES-1 cells, nevertheless, Y1 and P1 under the same concentrations exhibited remarkable phototoxicity to GES-1 cells. In the co-culture of H. pylori and GES-1 cells, P3 at 2.5μM led to a complete eradication of H. pylori under the irradiation of 24J/cm2 532nm laser. While for the GES-1 cells, no significant (p>0.05) phototoxicity was observed under the same aPDT dosage. The ex vivo experiments showed that P3 mediated aPDT resulted in 82.4% to 100% reduction of H. pylori CFU without damaging the gastric mucosa. To sum up, P3 is a promising anti-H. pylori photosensitizer with the ability to selectively photo-inactivate H. pylori while sparing normal gastric tissues.

Fabrics Attached with Highly Efficient Aggregation-Induced Emission Photosensitizer: Toward Self-Antiviral Personal Protective Equipment.

Personal protective equipment (PPE) is vital for the prevention and control of SARS-CoV-2. However, conventional PPEs lack virucidal capabilities and arbitrarily discarding used PPEs may cause a high risk for cross-contamination and environmental pollution. Recently reported photothermal or photodynamic-mediated self-sterilizing masks show bactericidal-virucidal abilities but have some inherent disadvantages, such as generating unbearable heat during the photothermal process or requiring additional ultraviolet light irradiation to inactivate pathogens, which limit their practical applications. Here, we report the fabrication of a series of fabrics (derived from various PPEs) with real-time self-antiviral capabilities, on the basis of a highly efficient aggregation-induced emission photosensitizer (namely, ASCP-TPA). ASCP-TPA possesses facile synthesis, excellent biocompatibility, and extremely high reactive oxygen species generation capacity, which significantly outperforms the traditional photosensitizers. Meanwhile, the ASCP-TPA-attached fabrics (ATaFs) show tremendous photodynamic inactivation effects against MHV-A59, a surrogate coronavirus of SARS-CoV-2. Upon ultralow-power white light irradiation (3.0 mW cm-2), >99.999% virions (5 log) on the ATaFs are eliminated within 10 min. Such ultralow-power requirement and rapid virus-killing ability enable ATaFs-based PPEs to provide real-time protection for the wearers under indoor light irradiation. ATaFs' virucidal abilities are retained after 100 washings or continuous exposure to office light for 2 weeks, which offers the benefits of reusability and long-term usability. Furthermore, ATaFs show no toxicity to normal skin, even upon continuous high-power light illumination. This self-antiviral ATaFs-based strategy may also be applied to fight against other airborne pathogens and holds huge potential to alleviate global PPE supply shortages.

Antibacterial Photodynamic Inactivation of Fagopyrin F from Tartary Buckwheat (Fagopyrum tataricum) Flower against Streptococcus mutans and Its Biofilm.

The objective of this study was to determine reactive oxygen species (ROS) produced by fagopyrin F-rich fraction (FFF) separated from Tartary buckwheat flower extract exposed to lights and to investigate its antibacterial photodynamic inactivation (PDI) against Streptococcus mutans and its biofilm. ROS producing mechanisms involving FFF with light exposure were determined using a spectrophotometer and a fluorometer. S. mutans and its biofilm inactivation after PDI treatment of FFF using blue light (BL; 450 nm) were determined by plate count method and crystal violet assay, respectively. The biofilm destruction by ROS produced from FFF after exposure to BL was visualized using confocal laser scanning microscopy (CLSM) and field emission scanning electron microscope (FE-SEM). BL among 3 light sources produced type 1 ROS the most when applying FFF as a photosensitizer. FFF exposed to BL (5 and 10 J/cm2) significantly more inhibited S. mutans viability and biofilm formation than FFF without the light exposure (p < 0.05). In the PDI of FFF exposed to BL (10 J/cm2), an apparent destruction of S. mutans and its biofilm were observed by the CLSM and FE-SEM. Antibacterial PDI effect of FFF was determined for the first time in this study.

Photodynamic inactivation of Staphylococcus aureus using tetraethylene glycol-substituted Zn(II) phthalocyanine

ABSTRACT Methicillin resistant Staphylococcus aureus infections are increasing, especially in intensive care units. A new method for photodynamic inactivation (PDI) generates reactive oxygen species by photosensitization to kill bacteria. We investigated the PDI effect of tetraethylene glycol-substituted Zn(II) phthalocyanine (TEG-P) on S. aureus strains including two standards (ATCC 25923 and ATCC 43400) and 20 clinically isolated methicillin sensitive and 20 methicillin resistance strains. We also investigated three treated groups: 650 nm laser only, TEG-P only and TEG-P + laser, plus one control group. Treatments included 0.5, 1, 2, 4, 8, 16, 32 µg/ml concentrations of TEG-P. No suppression of bacterial growth was observed in the control, laser only and TEG-P only groups whether or not S. aureus was methicillin resistant. Bacterial growth was suppressed by 85% using 8 µg/ml TEG-P and completely suppressed by 32 µg/ml TEG-P in the TEG-P + laser group. A combination of TEG-P + laser treatment may be an alternative to conventional antibiotics for routine treatment of S. aureus infections, although further investigation of the effect at the tissue level is required.

Antibacterial activity against Staphylococcus aureus of curcumin-loaded chitosan spray coupled with photodynamic treatment

Photodynamic treatment is a nonthermal disinfection technology, which uses photosensitizers under specific light conditions to inactivate microorganisms. Curcumin, a component of turmeric and with many medicinal properties, has been used as food additive in many countries. In the study, curcumin was used as a photosensitizer. A controllable LED photodynamic apparatus with a wavelength of 460–465 nm was prepared from a 96-well plate, an LED dimmer and a transformer. The photodynamic activity of curcumin-loaded chitosan against Staphylococcus aureus and its biofilms on stainless-steel were investigated. The results showed that photodynamic inactivation effects relied on curcumin concentration, co-incubation time and illumination time. When 25 μmol/L curcumin-loaded chitosan was co-incubated with Staphylococcus aureus suspension (approximately 6.0 log CFU/mL) for 10 min, the viable count was reduced by 5.0 log CFU/mL, after 10 min exposure to light. On the surface of stainless-steel coupons coated with S. aureus biofilm, the viable count was significantly reduced by more than 5.0 log CFU/cm2 (initial number 5.6 logCFU/m2), after spraying with curcumin (50 μmol/L)-loaded chitosan and 10 min light treatment. So, curcumin photodynamic technology is expected to show great potential application for the inactivation of microbials on the surface of food equipment and fresh food products.

Enhancement of photodynamic inactivation of planktonic cultures of Staphylococcus aureus by DMMB-AuNPs.

Photodynamic inactivation is a promising method for the treatment of infectious diseases. Nanotechnology through gold nanoparticles, as a tool to improve the delivery of photosensitizer is an attractive approach to enhance photodynamic inactivation of bacteria. Moreover, gold nanoparticles enchance the absorption of light due to their plasmon resonance. The aim of this study was to evaluate in vitro photodynamic inactivation effects of 1.9-Dimethyl-Methylene Blue (DMMB)-AuNPs associated with the red LED (λ630 ηm ± 20 ηm, 125 mW, 12 J / cm², 192 s) on S. aureus strain. Eight experimental groups were studied: Control, LED, AuNPs, AuNPs + LED, DMMB, DMMB + LED, DMMB + AuNPs, DMMB + AuNPs + LED. After incubation, the number of bacteria surviving each treatment was determined and then enumerated by viable counting (CFU / mL). The logarithm of CFU / mL (CFU/mL log10) was calculated. All experiments realized in triplicate. The statistical analyses included one-way ANOVA tests, Tukey's multiple comparisons and nonlinear regression, p values <0.05 were considered statistically significant. According to results, the photodynamic inactivation of S. aureus on groups DMMB + LED and DMMB-AuNPs + LED, showed a significant reduction of the microbial load (p < 0.0001) when compared to the Control group. The decimal reduction (RD) of these groups were 99.96 % (RD = 3) and 99.994 % (RD = 4) respectively. In conclusion, these findings demonstrated that photodynamic inactivation is enhanced by using DMMB-AuNPs on S. aureus.

Photoeradication of bacteria with porphycenes: Substituent effects on the efficiency

Considering the world-wide problem of growing antibiotic resistance of bacteria, photodynamic inactivation (PDI) has a potential to become the treatment approach against some infectious diseases. In our study, four differently substituted porphycenes were compared in terms of their bactericidal activity against E. faecalis. All tested compounds had a similar photophysical characteristics, i.e., there were no significant differences in the location of absorption bands or molar absorption coefficients. Also, singlet oxygen generation quantum yields were very similar. Surprisingly, differently substituted porphycenes caused very diverse PDI effects. Special attention was drawn to the tert-butyl moieties. Our studies demonstrated that the presence of these substituents lowers the bactericidal potential significantly and can completely block the activity when more than one moiety is introduced to the molecule. The porphycenes lacking tert-butyl groups exhibited much higher PDI potential and we assign this effect to different interactions of the differently substituted porphycenes with the bacterial cells. Most likely, the presence of tert-butyls impairs cell penetration by the photosensitizer. These results remind that the favorable photophysical characteristics do not ensure that the compound considered as a potential PDI agent can reach the microbial cells.

Photodynamic inactivation of S. aureus with a water-soluble curcumin salt and an application to cheese decontamination.

In this study, the optimal parameters for the photodynamic inactivation (PDI) of Staphylococcus aureus in bacterial suspensions and in cheese were assessed using a water-soluble curcumin salt as the photosensitizer (PS). The in vitro study aimed at finding the optimal concentration and light dose to promote S. aureus photokilling. Four main groups were proposed: CONTROL (L-C-), LIGHT (L+C-), CUR (L-C+) and PDI (L+C+). A fixed light dose (LED, 450 ± 10 nm, 10 J cm-2) was applied using four different PS concentrations (0.75, 1.0, 1.5 and 3.0 mg mL-1). The dose also varied from 10-100 J cm-2 for a fixed concentration. High inactivation rates were observed for all light doses, with a maximum reduction of 7.58 log10 at 100 J cm-2 (p ≪ 0.05). Saturation of the PDI effect was observed after a 10 minute illumination time, as well as a slight decrease in the S. aureus population for increasing illumination times in the L+C- group. As an application, the concentration showing the best decontamination performance in vitro (0.75 mg mL-1) was applied to decontaminate cheese in loco. PDI in two types of coalho cheese, a rennet-coagulated cheese commonly consumed in Brazil, was investigated. The results showed no significant inactivation in unpasteurized cheese, but a 4.34 log10 reduction for t > 5 min in pasteurized specimens. In conclusion, the present PDI-catalyzed curcumin photosensitizer inactivated S. aureus at statistically significant levels in vitro, in pasteurized cheese, but not in unpasteurized specimens.

In vitro photodynamic inactivation effects of hypocrellin B on azole-sensitive and resistant Candida albicans.

The extensive use of antifungal drugs has led to resistance from Candida albicans. The search for alternative treatment against drug-resistant C. albicans is highly desirable. Antimicrobial photodynamic therapy (aPDT) is an emerging and promising approach for treating localized and superficial C. albicans infections. The aim of this study was to investigate the photodynamic inactivation (PDI) effects of hypocrellin B (HB) on azole-sensitive and resistant C. albicans in vitro. The PDI efficacies of HB on standard C. albicans strain (ATCC 10231), azole-sensitive clinical isolate of C. albicans, and azole-resistant clinical isolate of C. albicans were assessed. The uptake of HB in C. albicans cells was investigated by confocal laser scanning microscopy (CLSM). The PDI effects on cellular structure and surface characteristics were investigated by transmission electron microscopy (TEM) and scanning electron microscopy (SEM). HB exhibited no significant dark toxicity, but inactivated the azole-sensitive and resistant C. albicans in a light-dose and PS concentration-dependent manner. CLSM images indicated that PDI treated C. albicans cells showed stronger fluorescence compared to untreated cells. TEM images suggested that significant damage to the cell wall, membrane, and cytoplasm were induced by HB-mediated PDI. SEM analysis revealed that the surface of C. albicans cells became twisted and ruptured after PDI treatment. Azole-sensitive and resistant C. albicans could be effectively inactivated by HB in the presence of light, and HB-mediated aPDT shows promise as an antifungal treatment for C. albicans.

Smart Hybrid Nanocomposite for Photodynamic Inactivation of Cancer Cells with Selectivity.

Photodynamic therapy has been efficiently applied for cancer therapy. Here, we have fabricated the folic acid (FA)- and pheophorbide A (PA)-conjugated FA/PA@Fe3O4 nanoparticle (smart hybrid nanocomposite, SHN) to enhance the photodynamic inactivation (PDI) of specific cancer cells. SHN coated with the PDI agent is designed to have selectivity for the folate receptor (FR) expressed on cancer cells. Structural characteristics and morphology of the fabricated MNPs were studied with X-ray diffraction and scanning electron microscopy. The photophysical properties of SHN were investigated with absorption, emission spectroscopies, and Fourier transform infrared spectroscopy. In addition, the magnetic property of Fe3O4 nanoparticle (MNP) can be utilized for the collection of SHNs by an external magnetic field. The photofunctionality was given by the photosensitizer, PA, which generates reactive oxygen species by irradiation of visible light. Generation of singlet oxygen was directly evaluated with time-resolved phosphorescence spectroscopy. Biocompatibility and cellular interaction of SHN were also analyzed by using various cancer cells, such as KB, HeLa, and MCF-7 cells which express different levels of FR on the surface. Cellular adsorption and the PDI effect of SHN on the various cancer cells in vitro were correlated well with the surface expression levels of FR, suggesting potential applicability of SHN on specific targeting and PDI of FR-positive cancers.