Efficacy Of Photoinactivation Research Articles

Substantial improvement in photocatalysis performance of N-TiO2 immobilized on PMMA: Exemplified by inactivation of Staphylococcus aureus and Escherichia coli

Photocatalysis is an efficient process for degrading organic pollutants and inactivating pathogenic microorganisms. However, this process constantly suffers from turbidity shading and particle aggregation in a catalyst suspension system, thereby reducing its photocatalytic activity. Immobilizing the photocatalyst on the light-transmissible surface is a viable solution to the obstacles. So far, the photo-inactivation efficacy between the immobilized photocatalyst and suspension systems has yet to be compared and investigated. In this study, N-TiO2 (NT) immobilized on poly-methyl-methacrylate (PMMA) was fabricated via a dip-coating method, which has a high transmittance rate of 92 % - better than all of the previous works (50 %). By immobilizing N-TiO2 on PMMA, up to 60 % and 19 % improvements in inactivation efficiencies against Gram-positive Staphylococcus aureus (S. aureus) and Gram-negative Escherichia coli (E. coli) are achieved, respectively, relative to a photocatalyst suspension. Notably, reactive oxygen species (ROSs) detection results indicate that 5 g/L NT coated PMMA ((NT-PMMA)5) has higher intensities of singlet oxygen (1O2), hydroxyl radicals (HO•), and higher concentration of hydrogen peroxide (H2O2) than the NT suspension. The as-made NT-PMMA sustains a 99.99 % inactivation efficiency (5-log-inactivation) against S. aureus through five consecutive photocatalysis cycles of reuse. The inactivation kinetics of S. aureus and E. coli fit well with the modified Hom model. Atomic force microscopy observations indicate that the NT-PMMA inactivation causes more severe damage to S. aureus's cell wall than E. coli due to the different susceptibility of cell wall structure to ROSs. This study paves a substantial way for scaling up the immobilizing catalyst on PMMA for the effective photocatalytic inactivation of pathogens under visible light.

Synthesis and photodynamic antimicrobial chemotherapy against multi-drug resistant Proteus mirabilis of ornithine-porphyrin conjugates in vitro and in vivo.

For the treatment of bacterial infections, photodynamic antimicrobial chemotherapy (PACT) has the advantage of circumventing multi-drug resistance. In this work, new cationic photosensitizers against multi-drug resistant Proteus mirabilis (MRPM) were designed and synthesized by the conjugation of amino phenyl porphyrin with basic amino acid L-ornithine. Their photoinactivation efficacies against MRPM in vitro were reported and include the influence of laser energy, uptake, MIC and MBC, dose-dependent photoinactivation effects, membrane integrity, and fluorescence imaging. The PACT in vivo was evaluated using a wound mouse model infected by MRPM. Photosensitizer 4d displayed high photo inactivation efficacy against MRPM at 7.81 μM under illumination, and it could accelerate wound healing via bactericidal effect. These ornithine-porphyrin conjugates are potential photosensitizers for PACT in the treatment of MRPM infection.

Spectra of pathogens predict lethality of blue light photo-inactivation

Whole-cell spectra of human pathogens are examined to identify species-specific wavelengths of maximum light absorption. It is presented how these data relate to clinical efficacy for antimicrobial phototherapies. Twelve species of microorganisms were cultured to exponential growth then processed and suspended in a scattering solution. Diffuse reflectance from the sample was delivered to two spectrometers that covered a spectral range of 370nm to 1200nm. Reflectance spectra were converted to relative absorbance. Absorbance spectra from multiple trials were averaged to produce a representative spectrum for each species. All pathogens studied demonstrated an absorbance spectrum with a primary component in the range 405-426 nm and 1-4 secondary components in the range 445-636 nm. Antibiotic-resistant and antibiotic-sensitive strains of Pseudomonas aeruginosa (Pa) had identical absorbance spectra. Whole-cell spectra support the hypothesis that a pathogen’s absorbance is directly related to efficacy of photoinactivation. The spectral signatures in all pigmented bacteria tested appear to be that of various combinations of in situ porphyrins. The knowledge of the absorption spectrum of a given pathogen provides a logical start for selecting the appropriate light source for a clinical trial.

Metal oxide-doped elastomeric materials for amplifying visible light-based antimicrobial activity

Healthcare-associated infection through transmission of pathogenic bacteria poses a huge threat to public health. One of the main transmission routes is via contaminated surfaces, including those of medical devices, and therefore significant efforts are being invested in developing new surface decontamination strategies. This includes visible light-based approaches, which offer improved compatibility with mammalian cells but lower germicidal efficacy with respect to UV-light. This study investigates the potential to enhance the antimicrobial efficacy of 405 nm light for surface decontamination through use of a photocatalytic TiO2-doped elastomer, elastomers being selected due to their wide use in biomaterials. Poly(dimethylsiloxane) (PDMS) was doped with TiO2 nanoparticles and the surface elastomer etched to expose the embedded nanoparticles. As etching results in increased surface roughness, samples with control nanoparticles (SiO2 and Fe3O4) were also investigated to decouple the effects of roughness and photoinactivation upon bacterial attachment and inactivation. Characterisation by SEM, AFM and contact angle analysis confirmed that etching produced a rougher (39.3 ± 15.3 versus 5.11 ± 1.29 nm RMS roughness; etched versus unetched TiO2-PDMS), more hydrophobic surface (water contact angle of 120 ± 2.5° versus 110 ± 1.0°; etched TiO2-PDMS versus native PDMS). This surface, rich in exposed photocatalytic TiO2 nanoparticles, allows direct contact between contaminating bacteria and nanoparticles, enabling ROS generation in closer proximity to the bacteria and consequent enhancement of visible light treatment. Incorporating TiO2 into PDMS significantly improved the photoinactivation efficacy (mean bacterial count for light-treated samples normalised to untreated samples of 0.043 ± 0.0081) compared to PDMS alone (0.19 ± 0.036), when seeded with Staphylococcus aureus and exposed to 405 nm, 60 J cm−2 light. However, photoinactivation efficacy was significantly (p < 0.001) enhanced by etching the TiO2-PDMS surface (0.015 ± 0.0074), resulting in greater photoinactivation than that obtained for etched (47.0 ± 14.5 nm RMS roughness), non-photocatalytic SiO2-PDMS (0.10 ± 0.093). Results suggest this doping and etching strategy shows significant potential for facilitating decontamination of elastomer-based biomaterials.

Photodynamic Inactivation of Antibiotic-Resistant and Sensitive Aeromonas hydrophila with Peripheral Pd(II)- vs. Zn(II)-Phthalocyanines.

The antimicrobial multidrug resistance (AMR) of pathogenic bacteria towards currently used antibiotics has a remarkable impact on the quality and prolongation of human lives. An effective strategy to fight AMR is the method PhotoDynamic Therapy (PDT). PDT is based on a joint action of a photosensitizer, oxygen, and light within a specific spectrum. This results in the generation of singlet oxygen and other reactive oxygen species that can inactivate the pathogenic cells without further regrowth. This study presents the efficacy of a new Pd(II)- versus Zn(II)-phthalocyanine complexes with peripheral positions of methylpyridiloxy substitution groups (pPdPc and ZnPcMe) towards Gram-negative bacteria Aeromonas hydrophila (A. hydrophila). Zn(II)-phthalocyanine, ZnPcMe was used as a reference compound for in vitro studies, bacause it is well-known with a high photodynamic inactivation ability for different pathogenic microorganisms. The studied new isolates of A. hydrophila were antibiotic-resistant (R) and sensitive (S) strains. The photoinactivation results showed a full effect with 8 µM pPdPc for S strain and with 5 µM ZnPcMe for both R and S strains. Comparison between both new isolates of A. hydrophila (S and R) suggests that the uptakes and more likely photoinactivation efficacy of the applied phthalocyanines are independent of the drug sensitivity of the studied strains.

Photoinactivation of bacteriophage MS2, Tulane virus and Vibrio parahaemolyticus in oysters by microencapsulated rose bengal

Abstract Bivalve molluscan shellfish such as oysters are important vectors for the transmission of foodborne pathogens including both viruses and bacteria. Photoinactivation provides a cold-sterilization option against the contamination as excited photosensitizers could transfer electronic energy to oxygen molecules producing reactive oxygen species such as singlet oxygen, leading to oxidative damage and death of the pathogens. However, the efficacy of photoinactivation is very often compromised by the presence of food matrix due to the nonselective reactions of short-lived singlet oxygen with organic matter other than the target pathogens. In order to address this issue, we encapsulated a food-grade photosensitizer rose bengal (RB) in alginate microbeads. An extra coating of chitosan effectively prevented the release of RB from the microbeads in seawater, and more importantly, enhanced the selectivity of the photoinactivation via the electrostatic interactions between cationic chitosan and anionic charge of the virus particles (bacteriophage MS2 and Tulane virus) and the Gram-negative bacteria Vibrio parahaemolyticus (V. parahaemolyticus). The treatment of oysters with microencapsulated RB resulted in significantly higher reductions of MS2 phage, Tulane virus and V. parahaemolyticus than free RB and non-RB carrying microbeads (P&amp;lt;0.05) tested with both in vitro and in vivo experimental set-ups. This study demonstrated a new strategy in delivering comprehensively formulated biochemical sanitizers in bivalve shellfish through their natural filter-feeding activity and thereby enhancing the mitigation efficiency of foodborne pathogen contamination.

Mapping of UV-C dose and SARS-CoV-2 viral inactivation across N95 respirators during decontamination

During public health crises like the COVID-19 pandemic, ultraviolet-C (UV-C) decontamination of N95 respirators for emergency reuse has been implemented to mitigate shortages. Pathogen photoinactivation efficacy depends critically on UV-C dose, which is distance- and angle-dependent and thus varies substantially across N95 surfaces within a decontamination system. Due to nonuniform and system-dependent UV-C dose distributions, characterizing UV-C dose and resulting pathogen inactivation with sufficient spatial resolution on-N95 is key to designing and validating UV-C decontamination protocols. However, robust quantification of UV-C dose across N95 facepieces presents challenges, as few UV-C measurement tools have sufficient (1) small, flexible form factor, and (2) angular response. To address this gap, we combine optical modeling and quantitative photochromic indicator (PCI) dosimetry with viral inactivation assays to generate high-resolution maps of “on-N95” UV-C dose and concomitant SARS-CoV-2 viral inactivation across N95 facepieces within a commercial decontamination chamber. Using modeling to rapidly identify on-N95 locations of interest, in-situ measurements report a 17.4 ± 5.0-fold dose difference across N95 facepieces in the chamber, yielding 2.9 ± 0.2-log variation in SARS-CoV-2 inactivation. UV-C dose at several on-N95 locations was lower than the lowest-dose locations on the chamber floor, highlighting the importance of on-N95 dose validation. Overall, we integrate optical simulation with in-situ PCI dosimetry to relate UV-C dose and viral inactivation at specific on-N95 locations, establishing a versatile approach to characterize UV-C photoinactivation of pathogens contaminating complex substrates such as N95s.

Use of Micellar Delivery Systems to Enhance Curcumin’s Stability and Microbial Photoinactivation Capacity

Microbial photoinactivation using ultraviolet (UV) or visible light can be enhanced by photosensitizers. This study assessed the efficacy of encapsulating a food-grade photosensitizer (curcumin) in surfactant micelles on its water dispersibility, chemical stability, and antimicrobial activity. Stock curcumin-surfactant solutions were prepared with Surfynol 465 (S465) or Tween 80 (T80) (5 mM sodium citrate buffer). The antimicrobial activity of curcumin-loaded surfactant solutions was determined by monitoring the inactivation of Escherichia coli O157: H7 and Listeria innocua after 5-min irradiation with UV-A light (λ = 365 nm). The solutions mixed with the bacterial suspensions contained 1 µM curcumin and each surfactant below, near, and above their critical micelle concentrations (CMCs). The addition of surfactants at any level to the curcumin solution enhanced its dispersibility, stability, and efficacy as a photosensitizer, thereby enhancing its antimicrobial activity. Gram-positive bacteria were more susceptible than Gram-negative bacteria when curcumin-loaded micelles were used against them. The photoinactivation efficacy of curcumin-surfactant solutions depended on the pH of the solution (low > high), surfactant type (S465 > T80), and the amount of surfactant present (below CMC ≥ near CMC > above CMC = unencapsulated curcumin). This result suggests that excessive partitioning of curcumin into micelles reduced its ability to interact with microbial cells. Synergistic antimicrobial activity was observed when S465 was present below or near the CMC with curcumin at pH 3.5, which could be attributed to a more effective interaction of the photosensitizer with the cell membranes as supported by the fluorescence lifetime micrographs. The use of a micelle-based delivery system facilitates adsorption and generation of reactive oxygen species in the immediate environment of the microbial cell, enhancing photoinactivation.

Comparison of the effect of ethanol and potassium iodide in antibacterial photodynamic therapy on gram negative pathogens

Objectives: Antibiotics is the most common treatment for bacterial infections. However, bacteria can change their genetic material, develop antibiotic resistance and cannot be treated. This brings the need for new treatment methods. Antibacterial Photodynamic Therapy is becoming a promising approach to treat bacterial infections. It is based on the use of photosensitizer to be activated by light with an appropriate wavelength and it will result in reactive oxygen species which are responsible for the destruction of pathogens. In this study, Antibacterial Photodynamic Therapy was examined for the treatment of Pseudomonas aeruginosa with the help of ethanol and potassium iodide. Methods: The effect of Chlorin e6-based photoinactivation was studied on Multidrug resistant P. aeruginosa upon irradiation with 655-nm diode laser. Then ethanol and potassium iodide was added to the mechanism separately to increase the efficacy of photoinactivation. After each application, serial dilution method was used for the determination of viable cells. Results: Outcomes showed that only Antibacterial Photodynamic Therapy causes a mortality rate of 75%. Addition of ethanol causes a mortality rate of 93% and addition of potassium iodide causes a mortality rate of 99.9% with less amount of Chlorin e6 and light dose. Conclusions: Chlorin-e6 based photoinactivation did not provide high mortality rate on P. aeruginosa. The use of ethanol and potassium iodide increased the effect of photoinactivation. The highest mortality rate was obtained with potassium iodide. It was understood that potassium iodide was a better concomitant agent to increase the bactericidal effect of Antibacterial Photodynamic Therapy on P. aeruginosa.

Photoinactivation of Staphylococci with 405 nm Light in a Trachea Model with Saliva Substitute at 37 °C.

The globally observed rise in bacterial resistance against antibiotics has increased the need for alternatives to antibiotic treatments. The most prominent and important pathogen bacteria are the ESKAPE pathogens, which include among others Staphylococcus aureus, Klebsiella pneumoniae and Acinetobacter baumannii. These species cause ventilator-associated pneumonia (VAP), which accounts for 24% of all nosocomial infections. In this study we tested the efficacy of photoinactivation with 405 nm violet light under conditions comparable to an intubated patient with artificial saliva for bacterial suspension at 37 °C. A technical trachea model was developed to investigate the visible light photoinactivation of Staphylococcus carnosus as a non-pathogen surrogate of the ESKAPE pathogen S. aureus (MRSA). The violet light was coupled into the tube with a fiber optic setup. The performed tests proved, that photoinactivation at 37 °C is more effective with a reduction of almost 3 log levels (99.8%) compared to 25 °C with a reduction of 1.2 log levels. The substitution of phosphate buffered saline (PBS) by artificial saliva solution slightly increased the efficiency during the experimental course. The increased efficiency might be caused by a less favorable environment for bacteria due to for example the ionic composition.

Effect of TiO2 Loading, Water Depth and Light Intensity on Photo-Disinfection Efficacy of Escherichia Coli O157:H7 Using TiO2 NP-Embedded Cellulose Acetate Film in Water

HighlightsPhoto-disinfection using TiO2 NP-embedded CA film showed great potential.TiO2 loading, water depth, and light intensity affected TiO2 photo-disinfection.Bacterial reduction can be predicted based on water depth and light intensity.Durability of the TiO2 NP-embedded CA film has been demonstrated.Abstract. Photocatalysis disinfection has great potential for food and water safety applications. TiO2 NP-embedded cellulose acetate (CA) film has shown effectiveness in inactivating Escherichia coli (E. coli) O157:H7 in water. This study evaluated the effect of several operational parameters such as TiO2 load, light intensity, and water depth on photo-disinfection efficacy of TiO2 NP-embedded CA film. Effect of TiO2 load on photocatalysis disinfection efficacy was investigated by assessing the disinfection rate of the films at different TiO2 levels in inoculated water at a weak UV-A light intensity. The individual and interaction effects of light intensity and water depth on photocatalysis disinfection were evaluated using the response surface methodology (RSM). Based on a central composite design, 13 runs of experiments were conducted with the combination of the two factors at three levels. The effect of film size on photo-disinfection efficacy was also investigated. Results showed that CA film with a TiO2 load of 0.82 mg/cm2 yielded the highest disinfection rate. An empirical model was established to predict the effect of light intensity and water depth on bacterial reduction with a high coefficient of determination (R2 = 0.98). An increased water depth or a decreased light intensity reduced photo-inactivation efficacy. A 5 log reduction was achieved during a 3-h treatment at a light intensity of 1 mW/cm2 and water depth at 1.6 cm. The photo-disinfection efficacy of the film was not significantly (p&amp;gt;0.05) affected by film size. The CA film was able to maintain antimicrobial efficacy after four repeated uses. Results from this study demonstrated that UV-A assisted photo-disinfection using TiO2 NP-embedded CA film has great potential to inactivate pathogens in water. Keywords: Cellulose Acetate, E. coli O157:H7, Photocatalysis, TiO2 nanoparticles, Water Treatment.

Modulation of aminolevulinic acid-based photoinactivation efficacy by iron in vitro is cell type dependent

Iron availability to cells may be modified in the tumour microenvironment, which may be involved in treatment response. Iron availability affects the conversion of protoporphyrin IX to heme, which likely determines the efficacy of aminolevulinic acid-based photodynamic therapy (ALA-based PDT). We compared photoinactivation efficacy in three oesophageal cell lines in culture media differing in iron content, DMEM and RPMI 1640, and in RPMI 1640 supplemented with iron to understand the importance of iron presence for ALA-based PDT outcome. ALA-based PDT was more efficacious in DMEM than in RPMI 1640 in all tested cell lines. Consistently, the highest protoporphyrin IX fluorescence signals, indicating the highest level of protoporphyrin IX production, were detected from cell colonies incubated in DMEM compared to those incubated in RPMI 1640 irrespective of iron presence. Components in the culture media other than iron ions are likely to be responsible for the observed differences in two culture media. Nevertheless, iron supplementation to RPMI 1640 showed that the presence of ferric ions in the concentration range 0–8 mg/l affected ALA-based PDT efficacy in a cell type-dependent manner. In poorly differentiated carcinoma cells, the increased efficacy of ALA-induced photoinactivation in the presence of 0.1 mg/l of supplemented iron was found. At the same iron concentration, the slightly different mitochondrial potential at no modifications of the iron labile pool was observed. The efficacy of ALA-based PDT in vitro depends on the choice of culture medium and the presence of iron ions in culture medium depending on intrinsic properties of cells.

Photodynamic Inactivation of Methicillin-Resistant Staphylococcus aureus by a Natural Food Colorant (E-141ii).

This study evaluates the photosensitizing effectiveness of sodium copper chlorophyllin, a natural green colorant commonly used as a food additive (E-141ii), to inactivate methicillin-sensitive and methicillin-resistant Staphylococcus aureus under red-light illumination. Antimicrobial photodynamic inactivation (aPDI) was tested on a methicillin-sensitive reference strain (ATCC 25923) and a methicillin-resistant Staphylococcus aureus strain (GenBank accession number Mh087437) isolated from a clinical sample. The photoinactivation efficacy was investigated by exposing the bacterial strains to different E-141ii concentrations (0.0, 1.0, 2.5, 5.0, 10.0, and 20.0 µM) and to red light (625 nm) at 30 J cm−2. The results showed that E-141ii itself did not prevent bacterial growth for all tested concentrations when cultures were placed in the dark. By contrast, E-141ii photoinactivated both methicillin-sensitive Staphylococcus aureus (MSSA) and methicillin-resistant Staphylococcus aureus (MRSA) under red-light illumination. However, different dose responses were observed for MSSA and MRSA. Whilst the MSSA growth was inhibited to the detection limit of the method with E-141ii at 2.5 µM, >10 µM concentrations were required to inhibit the growth of MRSA. The data also suggest that E-141ii can produce reactive oxygen species (ROS) via Type I reaction by electron transfer from its first excited singlet state to oxygen molecules. Our findings demonstrate that the tested food colorant has great potential to be used in aPDI of MRSA.

Hypervirulent and hypermucoviscous strains of Klebsiella pneumoniae challenged by antimicrobial strategies using visible light

Introduction: Infections caused by hypervirulent and/or hypermucoviscous Klebsiella pneumoniae (K. pneumoniae) strains are frequently reported worldwide. Since convergence of hypervirulence and drug-resistance emerged as a serious clinical problem, novel therapeutic strategies are worthy of investigation. In this regard, antimicrobial photodynamic therapy and blue light have proven to be effective against a broad-spectrum of clinically relevant pathogens but have never been tested for hypervirulent/hypermucoviscous strains. Thus, this study investigated the influence of hypermucoviscosity and hypervirulence over the photoinactivation efficacy of blue light alone or antimicrobial photodynamic therapy mediated by methylene blue and red light.Methods: Five clinical isolates of K. pneumoniae were screened for hypermucoviscosity by string test and for hypervirulence by a Galleria mellonella model of systemic infection. Strains were then challenged by both photoinactivation methods performed in vitro. All tests also included a non-hypervirulent/hypermucoviscous control strain for comparison.Results: All K. pneumoniae strains were effectively inactivated by both light-based antimicrobial strategies. Hypervirulent/hypermucoviscous strains exposed to photodynamic therapy presented rapid and consistent inactivation kinetics, while blue light led to slower and more variable inactivation kinetics.Conclusion: Hypermucoviscosity and hypervirulence does not confer tolerance in K. pneumoniae against photoinactivation. Antimicrobial photodynamic therapy represents an interesting alternative to treat localised infections because it is a fast procedure with high effectiveness. On the other hand, antimicrobial blue light could be used to decontaminate hospital environments since no photosensitiser administration is required and harmful effects of ultraviolet light are avoided. Therefore, visible light-based strategies present great potential for the development of safe and effective antimicrobial technologies against such aggressive pathogens.

Determining and unravelling origins of reduced photoinactivation efficacy of bacteria in milk

Bovine mastitis is an endemic disease of dairy cattle that is considered to be one of the most frequent and costly diseases in veterinary medicine. An increase in the incidence of disease results in the increased use of antibiotics, which in turn increases the potential of bacterial resistance. This study aimed to investigate the effectiveness of antimicrobial photodynamic therapy (aPDT) in the treatment of bovine mastitis, as an alternative to systemic antibiotics. To identify the key factors affecting photoinactivation efficacy, realistic experiments in view of the end-use were conducted in milk samples using two different photosensitizers: methylene blue (MB) and silicon (IV) phthalocyanine derivative (SiPc). We explored the effects of divalent ions and fat content on the aPDT outcome and determined influence of different proteins on aPDT efficacy. Levels of bacterial sensitivity to PSs varied depending on the type of bacteria (Gram-positive vs. Gram-negative) and light exposure time. Critical interrelated factors affecting aPDT in milk were identified and an efficient combination of treatment conditions that can lead to a full photodynamic inactivation of bacteria was determined.

Evaluation of the photodynamic therapy as a potential technique for blood decontamination

Evaluation of the photodynamic therapy as a potential technique for blood decontamination

Synthesis, characterization and in vitro photodynamic antimicrobial activity of basic amino acid–porphyrin conjugates

Photodynamic antimicrobial chemotherapy (PACT), as a novel and effective modality for the treatment of infection with the advantage of circumventing multidrug resistance, receives great attention in recent years. The photosensitizer is the crucial element in PACT, and cationic porphyrins have been demonstrated to usually be more efficient than neutral and negatively charged analogues towards bacteria in PACT. In this work, three native basic amino acids, l-lysine, l-histidine and l-arginine, were conjugated with amino porphyrins as cationic auxiliary groups, and 13 target compounds were synthesized. This paper reports their syntheses, structural characterizations, oil-water partition coefficients, singlet oxygen generation yields, photo-stability, as well as their photo inactivation efficacies against methicillin-resistant Staphylococcus aureus (MRSA), Escherichia coli and Pseudomonas aeruginosa in vitro. The preliminary structure-activity relationship was discussed. Compound 4i, with porphyrin bearing four lysine moieties, displays the highest photo inactivation efficacy against the tested bacterial strains at 3.91 μM with a low light dose (6 J/cm(2)), and it is stable in serum and lower cytotoxicity to A929 cells. These basic amino acid-porphyrin conjugates are potential photosensitizers for PACT.

Sub-lethal photodynamic inactivation renders Staphylococcus aureus susceptible to silver nanoparticles

Staphylococcus aureus is a common etiological factor in infections of burns and other chronic wounds. The development of an effective and fast-acting treatment would be enormously beneficial and is highly desired. We focused on testing the bactericidal efficacy of photoinactivation using a known photosensitizer (protoporphyrin IX, PPIX) in sequential combination with silver nanoparticles against S. aureus. Using PPIX-based photoinactivation followed by silver nanoparticles we obtained a high bactericidal effect (7 log10 units reduction) with limited harmful effects on human epidermal keratinocytes. Moreover, we observed that the use of silver nanoparticles prevents bacterial re-growth 24 h post-PDI treatment. A sequential combination of photoinactivation and silver nanoparticles represents a potentially effective antibacterial approach.

Silencing of ALA dehydratase affects ALA-photodynamic therapy efficacy in K562 erythroleukemic cells

Synthesis of protoporphyrin IX (PpIX) by malignant cells is essential for the success of ALA-based photodynamic therapy (PDT). Two key enzymes that were described as affecting PpIX accumulation during ALA treatment are porphobilinogen deaminase (PBGD) and ferrochelatase. Here, we show that down regulation of ALA dehydratase (ALAD) expression and activity by specific shRNA induced a marked decrease in PpIX synthesis in K562 erythroleukemic cells. Photo-inactivation efficacy following ALA-PDT was directly correlated with ALAD-silencing and cellular levels of PpIX. MTT metabolism following ALA-PDT was shown to be 60% higher in ALAD-silenced cells in comparison to control cells, indicating that mitochondria were protected in the silenced cells. Morphological analysis by scanning electron microscopy (SEM) of cells treated by ALA-PDT showed no morphological changes in ALAD-silenced cells, in contrast to controls exhibiting cell deformations and lysis. Membrane integrity following ALA-PDT was kept intact and undamaged in ALAD-silenced cells as examined by Annexin V-FITC/PI staining and LDH-L leakage. We conclude that ALAD, although it is present in the cell at abundant levels, has a major and limiting role in regulating PpIX synthesis and ALA-PDT outcome.

Syntheses and photodynamic activities of novel trisulfonated zinc phthalocyanine derivatives.

The synthesis of water-soluble, unsymmetrical, trisulfonated zinc phthalocyanines (ZnPcS3) as single products of the ring expansion of boron tri(4-sulfo)subphthalocyanine (SubPc) is reported. The novel, water-soluble trisulfo-SubPcB(OH) was prepared via hydrolysis of the tris(4-chlorosulfonyl)SubPcB(Br) which in turn was obtained from the condensation of 4-(chlorosulfonyl)phthalonitrile with BBr3 in 1-chlorobenzene. A number of ZnPcS3 analogues were prepared via the reaction of S3SubPcB (OH) with different diiminoisoindoline derivatives of increasing hydrophobicity. The reaction proceeds at relative low temperature with acceptable yields. Metalation of free base Pc's with zinc acetate dihydrate afforded the corresponding zinc complexes. Photodynamic activities were measured against the EMT-6 mouse mammary tumor cell line and compared to those of the known ZnPcS3 and ZnPcS4. Added (t-Bu)benzo and (t-Bu)naphtho groups increased the in vitro cell photoinactivation efficacy of the ZnPcS3, whereas addition of a fourth sulfobenzo or bulky diphenylpyrazino group decreased the activity of the parent molecule. The (t-Bu)naphthotrisulfobenzoporphyrazine induced the best in vivo photodynamic tumor control which, combined with its good solubility and broad absorption spectrum, renders this compound an interesting dye for photodynamic applications in medicine.