Search Results (25)

Evolving antibiotic resistance of bacteria is a prevailing global challenge in health care and requires the development of safe and efficient alternatives to classic antibiotics. Photodynamic Inactivation (PDI) has proven to be a promising alternative for treatment of a broad range of microorganisms. Photodynamic Inactivation uses photoactive molecules that generate reactive oxygen species (ROS) upon illumination and in the presence of oxygen, which immediately kill pathogenic target organisms. Relevant photoactive properties are provided by berberine. Originally extracted from Barberry (Berberis vulgaris), it is a natural compound widely used in Traditional Chinese Medicine for its antimicrobial and anti-inflammatory effects. With this study, we demonstrated the potential of berberine chloride hydrate (Ber) as a photosensitizer for PDI of important human pathogens, Gram(+) Staphylococcus capitis subsp. capitis, Gram(+) Staphylococcus aureus, and Gram(−) Escherichia coli. In vitro experiments on planktonic and biofilm cultures were conducted focusing on Ber activated with visible light in the blue wavelength range. The number of planktonic S. capitis cells was reduced by 7 log₁₀ steps using 100 µM Ber (5 min incubation, illumination with 435 nm LED array, radiant exposure 25 J/cm²). For an antibacterial effect of 4 log₁₀ steps, static S. capitis biofilms required 1 mM Ber, a drug-to-light interval of 60 min, and illumination with 100 J/cm². Almost all planktonic cells of Staphylococcus aureus could be photokilled using 100 µM Ber (drug-to-light interval of 30 min, radiant exposure 25 J/cm²). Biofilms of S. aureus could be phototreated (3 log₁₀ steps inactivation) when using 1 mM Ber incubated for 5 min and photoactivated with 100 J/cm². The study is highlighted by the proof that PDI treatment using Ber showed an antibacterial effect on Gram(−) E. coli. Planktonic cells could be reduced by 3 log₁₀ steps with 100 µM Ber (5 min incubation, 435 nm, 25 J/cm²). With 5 mM ethylenediamine tetraacetic acid disodium salt dihydrate (Na₂EDTA) or 1.2% polyaspartic acid (PASA) in addition, a relative inactivation of 4 log₁₀ steps and 7 log₁₀ steps, respectively, was detectable. Furthermore, we showed that an antibacterial effect of 3.4 log₁₀ towards E. coli biofilms was given when using 1 mM Ber (5 min incubation, 435 nm, 100 J/cm²). These results underscore the significance of PDI-treatment with Ber as a natural compound in combination with blue light as valuable antimicrobial application. Full article

Photodynamic inactivation (PDI) is a highly effective treatment that can eliminate harmful microorganisms in a variety of settings. This study explored the efficacy of a curcumin-rich extract, Curcuma L., (Cur)- and essential oil component, trans-cinnamaldehyde, (Ca)-mediated PDI against Listeria monocytogenes ATCC 15313 (Lm) including planktonic cells and established biofilms on silicone rubber (Si), polytetrafluoroethylene (PTFE), stainless steel 316 (SS), and polyethylene terephthalate (PET). Applying Ca- and Cur-mediated PDI resulted in planktonic cell reductions of 2.7 and 6.4 log CFU/cm², respectively. Flow cytometric measurements (FCMs) coupled with CFDA/PI and TOTO^®-1 staining evidenced that Ca- doubled and Cur-mediated PDI quadrupled the cell damage. Moreover, the enzymatic activity of Lm cells was considerably reduced by Cur-mediated PDI, indicating its superior efficacy. Photosensitization also affected Lm biofilms, but their reduction did not exceed 3.7 log CFU/cm². Cur-mediated PDI effectively impaired cells on PET and PTFE, while Ca-mediated PDI caused no (TOTO^®-1) or only slight (PI) cell damage, sparing the activity of cells. In turn, applying Ca-mediate PDI to Si largely diminished the enzymatic activity in Lm. SS contained 20% dead cells, suggesting that SS itself impacts Lm viability. In addition, the efficacy of Ca-mediated PDI was enhanced on the SS, leading to increased damage to the cells. The weakened viability of Lm on Si and SS could be linked to unfavorable interactions with the surfaces, resulting in a better effect of Ca against Lm. In conclusion, Cur demonstrated excellent photosensitizing properties against Lm in both planktonic and biofilm states. The efficacy of Ca was lower than that of Cur. However, Ca bears potent antibiofilm effects, which vary depending on the surface on which Lm resides. Therefore, this study may help identify more effective plant-based compounds to combat L. monocytogenes in an environmentally sustainable manner. Full article

Opportunistic premise plumbing pathogens (OPPPs) form a group of microorganisms that normally live in water supply systems and have adapted especially well to the conditions in premise plumbing systems, and as such pose a threat to human health. Since the beginning of the 21st century, this threat has been escalating, and it is becoming increasingly evident that current water disinfection methods fall short in effectively controlling these pathogens. In researching new approaches to this emergency, phototherapy looks promising, especially one that combines photosensitizers, light, and oxygen, which is known as photodynamic inactivation (PDI). This review describes the main characteristics of the recognized (Pseudomonas aeruginosa, Legionella pneumophila, and Mycobacterium avium) and most important emerging OPPPs, and it offers a brief overview of current disinfection methods and their limitations in the fight against OPPPs. The principle and outcomes of PDI with endogenous and, in particular, exogenous photosensitizers are then explained and described through representative examples of PDI on recognized and emerging OPPPs and their biofilms. Finally, the prospects and future directions of PDI research in water disinfection and control of OPPPs are discussed. Full article

In this work we pursued research involving the microwave-assisted N-alkylation of a NH pyrrolidine-fused chlorin with methyl 4-(bromomethyl) benzoate and subsequent ester hydrolysis as a straightforward strategy to obtain carboxylic acid functionality in the pyrrolidine-fused chlorin, as a single reaction product. We studied the reaction’s scope by extending the N-alkylation of the free-base chlorin and its corresponding Zn(II) complex to other alkyl halides, including 1,4-diiodobutane, N-(2-bromoethyl)phthalimide, and 2-bromoethanaminium bromide. In addition, two new chlorin–dansyl dyads were synthesized by reacting dansyl chloride with the 2-aminoethyl pyrrolidine-fused chlorin (dyad 6) and NH pyrrolidine-fused chlorin (dyad 7). According to spectral studies, the linker length between the two fluorophores influences the response of the dyads to the solvent polarity. Because of the simplicity of these approaches, we believe it will enable access to a vast library of custom-tailored N-functionalized chlorins while preserving their important absorption and emission spectra as photosensitizers in photodynamic therapy (PDT) of cancer and photodynamic inactivation (PDI) of microorganisms. Full article

The climate crisis is one of the biggest challenges for humanity in the 21st century. Production and consumption of meat contributes to global warming by causing emissions of climate-relevant gases. Freshly grown sprouts are part of an alternative, as they are less polluting but still a nutritious food. However, warm humid sprouting conditions may cause pathogenic microorganisms to thrive. Decontamination methods for raw sprouts are therefore relevant. Photodynamic Inactivation (PDI) is a novel approach that uses photoactivatable molecules (photosensitisers, PS) and visible or near-infrared light to produce reactive oxygen species (ROS). These ROS kill microorganisms by oxidative processes. Here, we test the application of PDI based on sodium-magnesium-chlorophyllin (Chl, approved as food additive E140) for photo-decontamination of mung bean, radish, and buckwheat seeds. Seeds were contaminated with Listeria innocua, serving as a model system for Listeria monocytogenes, subjected to PDI using an LED array with 395 nm and tested for remaining bacterial contamination by CFU counting. PDI based on 100 µM Chl reduces the bacterial load of mung bean and radish seeds by 99.9% (radiant exposure 56.4 J/cm² and 28.2 J/cm², respectively), and of buckwheat seeds by <90% reduction after illumination with 28.2 J/cm². Neither weight nor the germination rates of seeds are affected by PDI. Interestingly, the effect of PDI on seeds is partially maintained on stored sprouts after germination: The bacterial load on mung bean sprouts is reduced by more than 99.9% after phototreatment of seeds with 100 µM Chl and illumination at 56.4 J/cm². In conclusion, we suggest PDI based on Chl as an effective and biocompatible method for the decontamination of seeds and sprouts for human consumption from Listeria. Full article

With the aim of contributing to the development of more efficient materials for wound care, new topical formulations based on bacterial nanocellulose (BNC) hydrogels containing propolis were produced. Characterizations confirmed the incorporation of propolis into the BNC matrix, maintaining its structure and properties. Rheological analysis confirmed that the hydrogels showed thixotropic behavior appropriate for topical application. Chromatographic profiles showed sustained release of propolis biomarkers for at least 20 h. The formulations did not present mutagenicity. For application in photodynamic inactivation (PDI), BNC/propolis hydrogels were prepared with the photosensitizers methylene blue (MB). Spectroscopy and confocal fluorescence microscopy confirmed the interaction of MB and propolis in BNC hydrogels, as well as the formation of a new composite material. In the antibacterial assays, formulations containing MB and propolis significantly reduced Staphylococcus aureus growth. In the presence of light, BNC/MB hydrogels completely inhibited the microorganism. Therefore, the results suggest potential materials for the prevention or treatment of Staphylococcus aureus infections in wounds. Full article

The increase in the antibiotic resistance of bacteria is a serious threat to public health. Photodynamic inactivation (PDI) of micro-organisms is a reliable antimicrobial therapy to treat a broad spectrum of complex infections. The development of new photosensitizers with suitable properties is a key factor to consider in the optimization of this therapy. In this sense, four corroles were designed to study how the number of cationic centers can influence the efficacy of antibacterial photodynamic treatments. First, 5,10,15-Tris(pentafluorophenyl)corrole (Co) and 5,15-bis(pentafluorophenyl)-10-(4-(trifluoromethyl)phenyl)corrole (Co-CF₃) were synthesized, and then derivatized by nucleophilic aromatic substitution with 2-dimethylaminoethanol and 2-(dimethylamino)ethylamine, obtaining corroles Co-3NMe₂ and Co-CF₃-2NMe₂, respectively. The straightforward synthetic strategy gave rise to macrocycles with different numbers of tertiary amines that can acquire positive charges in an aqueous medium by protonation at physiological pH. Spectroscopic and photodynamic studies demonstrated that their properties as chromophores and photosensitizers were unaffected, regardless of the substituent groups on the periphery. All tetrapyrrolic macrocycles were able to produce reactive oxygen species (ROS) by both photodynamic mechanisms. Uptake experiments, the level of ROS produced in vitro, and PDI treatments mediated by these compounds were assessed against clinical strains: methicillin-resistant Staphylococcus aureus and Klebsiella pneumoniae. In vitro experiments indicated that the peripheral substitution significantly affected the uptake of the photosensitizers by microbes and, consequently, the photoinactivation performance. Co-3NMe₂ was the most effective in killing both Gram-positive and Gram-negative bacteria (inactivation > 99.99%). This work lays the foundations for the development of new corrole derivatives having pH-activable cationic groups and with plausible applications as effective broad-spectrum antimicrobial photosensitizers. Full article

Photodynamic inactivation (PDI) is a therapeutic approach based on combined use of light, oxygen, and a photosensitizing agent (PS). These three components interact to generate reactive oxygen species, which are cytotoxic and irreversibly damage vital components of microbial cells, leading to death. However, this methodology has not managed to be completely specific in its mode of action since the photosensitizer can bind to both pathogenic and commensal microorganisms and even to host cells. Since subsequent irradiation of such cells could lead to their destruction, it is desirable to direct the photodynamic activity to the target cell. Therefore, the objective of this work was to direct the destruction of pathogenic microorganisms without affecting the normal flora. This could be achieved by binding the photosensitizing molecule to an antibody against the surface of the target organism. Therefore, a TCPP-IgG conjugate was synthesized using 4,4′,4″,4‴-(porphine-5,10,15,20-tetrayl)tetrakis(benzoic acid) (TCPP) and the antibody anti-protein A of Staphylococcus aureus. The UV-visible spectra of TCPP-IgG showed the typical Soret and Q bands characteristic of porphyrin derivatives and, additionally, a new band was observed, corresponding to the absorbance of the protein. However, the results indicated that the conjugation reaction affects the photochemical properties of fluorescent emission and the production of reactive oxygen species compared to TCPP free base. As a consequence, a lower cytotoxicity was observed in planktonic cells of S. aureus. PDI can become a promising therapeutic alternative, having as a strategy the specific control of bacterial death for an efficient eradication. Full article

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. Full article

In this study, we describe the semisynthesis of cost-effective photosensitizers (PSs) derived from chlorophyll a containing different substituents and using previously described methods from the literature. We compared their structures when used in photodynamic inactivation (PDI) against Staphylococcus aureus, Escherichia coli, and Candida albicans under different conditions. The PSs containing carboxylic acids and butyl groups were highly effective against S. aureus and C. albicans following our PDI protocol. Overall, our results indicate that these nature-inspired PSs are a promising alternative to selectively inactivate microorganisms using PDI. Full article

The aim of this study was to evaluate the effect of chlorophyllin-based photodynamic inactivation (Chl-PDI) on biofilm formation and fresh-cut pakchoi quality during storage. Firstly, Chl-based PDI reduced the amount of biofilm in an in vivo experiment and inactivated the food spoilage bacteria. Antibacterial mechanism analysis indicated that the bacterial extracellular polysaccharides and extracellular proteins were vulnerable targets for attacks by the Chl-based PDI. Then, the food spoilage microorganisms (Pseudomonas reinekei and Pseudomonas palleroniana) were inoculated onto the surface of fresh-cut pakchoi. We used chlorophyllin (1 × 10⁻⁵ mol/L) and 405 nm light (22.27 J/cm² per day) to investigate the effect of Chl-based PDI treatment on fresh-cut pakchoi quality during storage. The results showed that Chl-based PDI increased the visual quality and the content of chlorophyll, VC, total soluble solids, and SOD activity and decreased the occurrence of leaf yellowing and POD activity. These suggest that Chl-based PDI can be used for the preservation of fresh-cut pakchoi and has the potential to inhibit biofilm formation of food spoilage bacteria. It is of great significance for the effective processing and traditional vegetable preservation. Full article

The past 2 years have been marked by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic. This virus is found in the intestinal tract and reaches the wastewater system, and, consequently, the natural receiving water bodies, and inappropriate or/and inefficient WW treatment is a means of contamination. In the present work, we used a SARS-CoV-2 model—the phage Phi6—to evaluate its survival under different environmental conditions (pH, temperature, salinity, solar, and UV-B irradiation). Then, we tested the efficiency of photodynamic inactivation (PDI) as a WW disinfection alternative method, and, additionally, the impact on the cultivable native marine microorganisms of the PDI-treated WW was evaluated. Full article

Global crop production depends on strategies to counteract the ever-increasing spread of plant pathogens. Antibiotics are often used for large-scale treatments. As a result, Erwinia amylovora, causal agent of the contagious fire blight disease, has already evolved resistance to streptomycin (Sm). Photodynamic Inactivation (PDI) of microorganisms has been introduced as innovative method for plant protection. The aim of this study is to demonstrate that E. amylovora resistant to Sm (E. amylovora^SmR) can be killed by PDI. Two photosensitizers, the synthetic B17-0024, and the natural derived anionic sodium magnesium chlorophyllin (Chl) with cell-wall-permeabilizing agents are compared in terms of their photo-killing efficiency in liquid culture with or without 100 µg/mL Sm. In vitro experiments were performed at photosensitizer concentrations of 1, 10 or 100 µM and 5 or 30 min incubation in the dark, followed by illumination at 395 nm (radiant exposure 26.6 J/cm²). The highest inactivation of seven log steps was achieved at 100 µM B17-0024 after 30 min incubation. Shorter incubation (5 min), likely to represent field conditions, reduced the photo-killing to 5 log steps. Chlorophyllin at 100 µM in combination with 1.2% polyaspartic acid (PASA) reduced the number of bacteria by 6 log steps. While PASA itself caused some light independent toxicity, an antibacterial effect (3 log reduction) was achieved only in combination with Chl, even at concentrations as low as 10 µM. Addition of 100 µg/mL Sm to media did not significantly increase the efficacy of the photodynamic treatment. This study proves principle that PDI can be used to treat plant diseases even if causative bacteria are resistant to conventional treatment. Therefore, PDI based on natural photosensitizers might represent an eco-friendly treatment strategy especially in organic farming. Full article

Photodynamic action has been used for diverse biomedical applications, such as treating a broad range of bacterial infections. Based on the combination of light, dioxygen, and photosensitizer (PS), the photodynamic inactivation (PDI) approach led to the formation of reactive oxygen species (ROS) and represented a non-invasive, non-toxic, repeatable procedure for pathogen photoinactivation. To this end, different tetrapyrrolic macrocycles, such as porphyrin (Por) dyes, have been used as PSs for PDI against microorganisms, mainly bacteria. Still, there is significant room for improvement, especially new PS molecules. Herein, unsymmetrical new pyridinone (3–5) and thiopyridyl Pors (7) were prepared with α-, β-, or γ-cyclodextrin (CD) units, following their quaternization to perform the corresponding free-base Pors (3a–5a and 7a), and were compared with the already-known Pors 6a and 8a, both bearing thiopyridinium and CD units. These water-soluble porphyrins were evaluated as PSs, and their photophysical and photochemical properties and photodynamic effects on E. coli were assessed. The presence of one CD unit and three positive charges on the Por structure (3a–5a and 7a) enhanced their aqueous solubility. The photoactivity of the cationic Pors 3a–5a and 6a–8a ensured their potential against the Gram-negative bacterium E. coli. Within each series of methoxypyridinium vs thiopyridinium dyes, the best PDI efficiency was achieved for 5a with a bacterial viability reduction of 3.5 log₁₀ (50 mW cm⁻², 60 min of light irradiation) and for 8a with a total bacterial viability reduction (>8 log₁₀, 25 mW cm⁻², 30 min of light irradiation). Here, the presence of the methoxypyridinium units is less effective against E. coli when compared with the thiopyridinium moieties. This study allows for the conclusion that the peripheral charge position, quaternized substituent type/CD unit, and affinity to the outer bacterial structures play an important role in the photoinactivation efficiency of E. coli, evidencing that these features should be further addressed in the pursuit for optimised PS for the antimicrobial PDI of pathogenic microorganisms. Full article

The last two years have been marked by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic. This virus is found in the intestinal tract; it reaches wastewater systems and, consequently, the natural receiving water bodies. As such, inefficiently treated wastewater (WW) can be a means of contamination. The currently used methods for the disinfection of WW can lead to the formation of toxic compounds and can be expensive or inefficient. As such, new and alternative approaches must be considered, namely, photodynamic inactivation (PDI). In this work, the bacteriophage φ6 (or, simply, phage φ6), which has been used as a suitable model for enveloped RNA viruses, such as coronaviruses (CoVs), was used as a model of SARS-CoV-2. Firstly, to understand the virus’s survival in the environment, phage φ6 was subjected to different laboratory-controlled environmental conditions (temperature, pH, salinity, and solar and UV-B irradiation), and its persistence over time was assessed. Second, to assess the efficiency of PDI towards the virus, assays were performed in both phosphate-buffered saline (PBS), a commonly used aqueous matrix, and a secondarily treated WW (a real WW matrix). Third, as WW is generally discharged into the marine environment after treatment, the safety of PDI-treated WW was assessed through the determination of the viability of native marine water microorganisms after their contact with the PDI-treated effluent. Overall, the results showed that, when used as a surrogate for SARS-CoV-2, phage φ6 remains viable in different environmental conditions for a considerable period. Moreover, PDI proved to be an efficient approach in the inactivation of the viruses, and the PDI-treated effluent showed no toxicity to native aquatic microorganisms under realistic dilution conditions, thus endorsing PDI as an efficient and safe tertiary WW disinfection method. Although all studies were performed with phage φ6, which is considered a suitable model of SARS-CoV-2, further studies using SARS-CoV-2 are necessary; nevertheless, the findings show the potential of PDI for controlling SARS-CoV-2 in WW. Full article