Search Results (241)

This study reports the development of chitosan-based (CS) films incorporating riboflavin (RF) as a natural photosensitizer to create sustainable, light-activated antimicrobial packaging materials. The films were prepared by solvent casting, and their photochemical behavior under blue LED light (450 nm) was investigated, including RF photodegradation kinetics and structural changes in the film-forming solution analyzed by ¹H NMR spectroscopy. Mechanical, thermal, optical, and barrier properties were also characterized to assess packaging suitability. Upon illumination, CS/RF films generated reactive oxygen species, particularly singlet oxygen (¹O₂), leading to visible color changes and significant antimicrobial activity against Pseudomonas fluorescens. Bacterial growth was reduced by up to 97% after 120 min of irradiation (0.92 J cm⁻²), with efficacy observed at both room temperature and 4 °C. The incorporation of RF did not alter the films’ mechanical properties, while thermal stability was preserved, optical transparency was modulated, and excellent oxygen barrier performance was maintained, although water vapor permeability remained moderate. These findings demonstrate that CS/RF films combine functionality and sustainability, offering a promising strategy for extending food shelf life through light-activated antimicrobial action. Validation under real storage conditions is recommended to confirm their potential in diverse food systems. Full article

Photodynamic therapy (PDT) is a promising antimicrobial strategy whose efficacy depends largely on the photosensitizers (PSs) used. While conventional PDT relies on a single PS, recent studies suggest that combining different PSs may improve outcomes by introducing complementary mechanisms. However, such combinations also add complexity, as timing, composition, and PS interactions must be considered alongside bacterial structures, uptake pathways, and light dosimetry. This study investigated the effects of PSs, methylene blue (MB), Photodithazine (PDZ), and their combinations on the PDT of Gram-negative bacterium Klebsiella pneumoniae. MB-mediated PDT demonstrated greater antibacterial effectiveness than PDZ-PDT. The combination of MB and PDZ produced varying results. When applied simultaneously, PDZ dose-dependently decreased MB’s antibacterial activity. Sequential treatment with PDZ followed by MB showed only slight antagonism compared to MB alone, while the reverse order (MB → PDZ) nearly abolished MB’s activity. Since both PSs are activated at the same wavelength (660 nm), their combined use was not additive. Photobleaching was performed on individuals and combined PSs to compare inactivation results with changes in chemical properties under red light (660 nm). This study highlights the limitations of using two photosensitizers together in antimicrobial photodynamic therapy and emphasizes the need for further optimization of combination protocols. Full article

Clavibacter michiganensis causes significant crop losses in tomatoes, and the disease may be transferred by plant seeds. This study evaluates the efficacy of Photodynamic Inactivation (PDI) with a water-soluble hypericin derivative, developed as a complex with polyvinylpyrrolidone (high hypericin-loaded PVP, HHL-PVP), as a decontamination strategy for tomato seeds. HHL-PVP was chosen for its overall stability, as the complex remains stable in solution for over 950 days, maintains its absorption capacity after illumination with 200 J·cm⁻², and produces reactive oxygen species (ROS) even at concentrations as low as 1 µM. PDI against C. michiganensis with 5 μM HHL-PVP, 10 min drug to light interval (DLI), and illumination with red light (600–700 nm, 100 J·cm⁻²) exceeded the antimicrobial effect of a 99.9% reduction in liquid culture. Increasing the DLI to 24 h did not alter the photokilling effect. A 14 h light/10 h dark cycle in white light (118 J·cm⁻²) with 0.3 µM HHL-PVP inhibited the growth of C. michiganensis by more than 6 log₁₀ steps, indicating that HHL-PVP has a stable and long-lasting photokilling effect. The combination of HHL-PVP with potassium iodide (KI, 100 mM) completely eradicated C. michiganensis in liquid culture with red and white light, indicating KI’s role in enhancing phototoxicity. Tomato seed photodynamic decontamination using 1.0 µM HHL-PVP activated by 200 J·cm⁻² white light inactivated >5 log₁₀ of C. michiganensis, without diminishing sprouting. An addition of 100 mM KI increased the percentage of sprouted seedlings and inactivated 100% of bacteria. These results demonstrate that HHL-PVP-mediated PDI combined with KI could be highly effective as a preventative strategy in tomato protection against C. michiganensis. Full article

Backround: The agr (accessory gene regulator) quorum sensing (QS) system of Staphylococcus aureus participates significantly in its virulence and biofilm formation—either through its activation or suppression. The aim of this study was to investigate the impact of photoactive nanomaterials that have been functionalized with erythrosine B (EryB) on the modulation of this agr QS system on three methicillin-resistant S. aureus (MRSA). Methods: The functionality of the agr system was determined by the CAMP test and by quantitative PCR (qPCR) to analyze the expression of the hld gene, which is located within the RNAIII and encodes δ-hemolysin. The biofilm was evaluated by crystal violet assay and fluorescence microscopy. The anti-biofilm activity was determined by calculating the colony-forming units. The relative expression of the hld gene, determined by qPCR. Results: Using the CAMP test, S66 and S68 strains were found to be agr-positive, and strain S73 was agr-negative. The relative expression of the hld gene increased only in the agr-positive strains (600- and 1000-fold). In these strains, the biofilm was less compact compared to the dense biofilm formed by the agr-negative strain. The anti-biofilm effectiveness on the nanocomposite with EryB after irradiation reduced the growth of biofilm cells by 100- to 1000-fold compared to the biofilm on polyurethane alone. The qPCR results showed a significant decrease in the relative expression of the hld gene in the agr-positive strains after irradiation compared to the non-irradiated samples. Conclusions: These results suggest that photoactive nanocomposites with EryB can significantly reduce biofilm formed by MRSA strains, regardless of the functionality of the agr QS system. Full article

Food spoilage and contamination are major global challenges, reducing food quality, safety, and availability, causing significant economic losses. This study evaluates the photodynamic and sonodynamic antibacterial activities of grape leaf extracts from Beer and Hanut Orcha varieties. The extracts were tested against Staphylococcus aureus and Escherichia coli under illumination and ultrasonic activation. The results demonstrated that the photodynamic and sonodynamic treatments significantly enhanced the antibacterial efficacy of the extracts when higher concentrations of the extracts and prolonged exposure led to complete bacterial eradication. Separation of the extracts using RP-18 cartridges (Yicozoo Energy Technology Co., Ltd., Xi’an, China) enabled us to get an active fraction containing components responsible for antimicrobial effects. Singlet oxygen generation measurements confirmed the involvement of reactive oxygen species in bacterial inactivation under illumination. Using HPLC/MS, the active components responsible for the photodynamic properties of the extracts were identified as quercetin 3’-O-glucuronide and pheophorbide a. The findings suggest that these natural extracts, in combination with photodynamic and sonodynamic activation, represent promising alternatives to conventional antibiotics. Further studies should focus on the isolation of active individual compounds, the improvement of treatment parameters, and the investigation of molecular mechanisms to facilitate the development of practical applications in medicine and food preservation. Full article

This study evaluated the effectiveness of photodynamic treatment (PDT) using riboflavin (Rbf) and blue light-emitting diode (BL) irradiation for microbial inactivation and quality preservation in fresh betel leaves (Piper betle L.). Non-pathogenic surrogates Escherichia coli K-12 and Listeria innocua were used to model Gram-negative and Gram-positive bacteria. The combined Rbf-BL treatment significantly reduced microbial populations by up to 5.3 log CFU/g for E. coli and 6.2 log CFU/g for L. innocua on leaf surfaces (p < 0.05) and 1.3–1.5 log CFU/mL in broth cultures. Treated samples showed significantly higher total soluble solids (12.0 ± 0.0 °Brix), total phenolic content (0.17 ± 0.02 mmol GAE/g, p < 0.05), and antioxidant activity (62.0 ± 3.1% DPPH inhibition, p < 0.05), with minimal color alteration after treatment (ΔE = 4.68). The total fluence measured at the leaf surface was approximately 11.72 J/cm². As a mild thermal treatment utilizing a GRAS photosensitizer, riboflavin-assisted PDT presents a promising strategy for enhancing microbial safety and promoting phytochemical quality in betel leaves. Full article

Background/Objectives: Covalent conjugation of an antibiotic vancomycin (VCM) moiety and a photosensitizing mesochlorin (mChl_Pd) unit into one molecular entity may present the potential to produce the combinatorial effect of both antibacterial photodynamic therapeutic (aPDT) and antibiotic activities. Our recent study indicated that a short linkage of <4 (C−C/or C−N) bond distances between these two moieties resulted in significant steric hindrance due to the bulky VCM, which greatly reduces the accessibility of the agent to the cell surface of methicillin-resistant Staphylococcus aureus (MRSA). The observed aPDT efficacy was found to be minimal. Here, we report that the revision of this linkage, via an EG₁₀ unit using identical synthetic procedures, was able to resolve the issue. Methods: Accordingly, the corresponding combinatorial aPDT−antibiotic compound, consisting of two covalently bonded quaternary ammonium pentacationic arms on the mesochlorin chromophore core, designated as VCMe-mChl_Pd-N₁₀⁺ (LC40e⁺), was prepared for applications in antibacterial photodynamic inactivation (aPDI) activity. It was selected to investigate its enhanced binding and targeting ability to the surface of Gram-positive MRSA cells. Subsequent antibacterial photodynamic therapeutic (aPDT) activity to inactivate MRSA was investigated to substantiate the corresponding cell-surface binding effect on the efficacy of aPDT. Results: We found that the covalent combination of 10 positive charges and an MRSA-targeting vancomycin (VCM) moiety in a conjugated structure, functioning as an antibiotic–decacationic photosensitizing agent (Abx-dcPS), was capable of largely improving the MRSA cell-targeting efficiency. Importantly, variation in the chain length of the oligo(ethylene glycol) linker of VCMe-mChl_Pd-N₁₀⁺, which was sufficiently long enough to properly separate the photoactive mesochlorin ring moiety from the VCM moiety within the molecular structure, resulted in significantly enhanced aPDT activity. The new conjugate provided nearly complete eradication (>6.5-log₁₀ colony-forming units (CFU) reduction) of MRSA cells in vitro. The aPDT efficacy followed the order Abx-dcPS (combinatorial decacationic) > dcPS (decacationic) >> nPS (nonionic). This order was also verified by the relative physical binding trend of these PSs using either nPS-, dcPS-, or Abx-dcPS-pretreated and pre-fixed MRSA cells in investigations of fluorescent confocal microscopy, UV–vis fluorescence spectroscopy, and transmission electron microscopy (TEM). Conclusions: Furthermore, the molecular conjugate of Abx-dcPS may provide covalent co-delivery of two drug components concurrently, which might also serve as an effective antibiotic agent after aPDT and potentially prevent the reoccurrence of MRSA-induced infection. Full article

Photodynamic inactivation and antimicrobial photodynamic therapy (PDI/APDT) based on the toxic properties of reactive oxygen species (ROS), which are generated by a number of photoexcited dyes, are promising for preventing and treating infections, especially those associated with drug-resistant pathogens. The negatively charged bacterial cell surface attracts polycationic photosensitizers, which contribute to the vulnerability of the bacterial plasma membrane to ROS. The integrity of the plasma membrane is critical for the viability of the bacterial cell. Polycationic phthalocyanines are regarded as promising photosensitizers due to their high quantum yields of ROS generation (mainly singlet oxygen), high extinction coefficients in the far-red spectral range, and low dark toxicity. For application in PDI/APDT, the wide range of possibilities of modifying the chemical structure of phthalocyanines is particularly valuable, especially by introducing various peripheral and non-peripheral substituents into the benzene rings. Depending on the type and location of such substituents, it is possible to obtain photosensitizers with different photophysical properties, photochemical activity, solubility in an aqueous medium, biocompatibility, and tropism for certain structures of photoinactivation targets. In this study, we tested novel water-soluble Zn (II) phthalocyanines bearing four 4-((diethylmethylammonium)methyl)phenoxy substituents with symmetric and asymmetric charge distributions for photodynamic antibacterial activity and compared them with those of water-soluble octacationic zinc octakis(cholinyl)phthalocyanine. The obtained results allow us to conclude that the studied tetracationic aryloxy-substituted Zn(II) phthalocyanines effectively bind to the oppositely charged cell wall of the Gram-negative bacteria E. coli. This finding is supported by data on bacteria’s zeta potential neutralization in the presence of phthalocyanine derivatives and fluorescence microscopy images of stained bacterial cells. Asymmetric substitution influences the aggregation and fluorescent characteristics but has little effect on the ability of the studied tetracationic phthalocyanines to sensitize the bioluminescent E. coli K12 TG1 strain. Both symmetric and asymmetric aryloxy-substituted phthalocyanines are no less effective in PDI than the water-soluble zinc octakis(cholinyl)phthalocyanine, a photosensitizer with proven antibacterial activity, and have significant potential for further studies as antibacterial photosensitizers. Full article

Fermented vegetables, which are valued for their distinctive organoleptic properties and nutritional profile, are susceptible to quality deterioration during processing and storage because microorganisms inhabit vegetable raw materials. The metabolic processes of these microorganisms may induce texture degradation, chromatic alterations, flavor diminution, and spoilage. Conventional inactivation methods employing thermal sterilization or chemical preservatives achieve microbial control through nonselective inactivation, inevitably compromising the regional sensory characteristics conferred by indigenous fermentative microbiota. Recent advances in existing antimicrobial technologies offer promising alternatives for selective microbial management in fermented vegetable matrices. Existing modalities, including cold plasma, electromagnetic wave-based inactivation (e.g., photodynamic inactivation, pulsed light, catalytic infrared radiation, microwave, and radio frequency), natural essential oils, and lactic acid bacterial metabolites, demonstrate targeted pathogen inactivation while maintaining beneficial microbial consortia essential for quality preservation when properly optimized. This paper explores the applications, mechanisms, and targeted microbes of these technologies in fermented vegetable ingredients, aiming to provide a robust theoretical and practical framework for the use of selective inactivation strategies to manage the fermentation process. By assessing their impact on the initial microbial community, this review aims to guide the development of methods that ensure product safety while safeguarding the characteristic flavor and quality of fermented vegetables. Full article

The increasing reliance on light-based antimicrobial technologies, such as antimicrobial blue light (aBL) and antimicrobial photodynamic inactivation (aPDI), underscores the urgent need to comprehend bacterial survival strategies beyond conventional resistance. Two key phenotypes—tolerance and resilience—have emerged as critical but often conflated mechanisms by which bacteria withstand oxidative and photodynamic stress. While tolerance refers to delayed bacterial killing without changes in MIC, resilience encompasses the active restoration of cellular function after transient stress exposure. Both phenomena may impair treatment outcomes and contribute to long-term persistence, even in the absence of genetic resistance. This review dissects the molecular mechanisms underlying tolerance and resilience, with a focus on their relevance to bacterial responses to reactive oxygen species generated by light-based or chemical stressors. The regulatory and effector overlap between these phenotypes is examined, including antioxidant defense systems, DNA repair pathways, and metabolic rewiring. Furthermore, the role of phenotypic heterogeneity and cross-stress protection in blurring the boundary between survival and recovery is discussed, highlighting challenges in experimental interpretation. Finally, the implications of these adaptive strategies are evaluated in the context of antimicrobial efficacy and safety, with an emphasis on kinetic assays and multidimensional profiling as tools to capture complex treatment outcomes. Clarifying the distinction between tolerance and resilience may help guide the development of robust and evolutionarily stable antimicrobial phototherapies. Full article

Yeasts of the genus Candida (C.) and the bacterium Staphylococcus aureus (S. aureus) are among the most common pathogens responsible for infections that are difficult to treat, including those resistant to standard therapy. In recent decades, this has become an increasing clinical problem. In response to the limitations of traditional procedures, antimicrobial photodynamic therapy (aPDT), which combines light, a photosensitizer, and oxygen, is gaining growing interest. The aim of this study was to evaluate the in vitro effectiveness of aPDT using a 635 nm diode laser in combination with toluidine blue O (TBO) against Candida spp. and S. aureus. Reference strains of C. albicans, C. glabrata, C. krusei, and S. aureus were subjected to aPDT. In phase I of this study, the optimal TBO incubation time was assessed with constant laser parameters. In phase II, the impact of the physical parameters of the laser, irradiation time, and output power, was analyzed, with the TBO incubation time set based on the phase I results, to evaluate the degree of microbial reduction (CFU/mL). Statistical analyses were then conducted to assess significance. TBO-mediated aPDT significantly reduced microbial viability, depending on incubation time and laser settings. The minimal effective incubation times were 10 min for Candida spp. and 5 min for S. aureus. The highest pathogen inactivation efficacy was observed at an output power of 400 mW and an irradiation time of 120 s. The use of the photosensitizer or laser alone did not result in significant antimicrobial effects. TBO-mediated aPDT may serve as an effective complement to conventional antimicrobial therapy and, in selected cases (e.g., drug resistance), has the potential to partially or fully replace it. The observed minimal effective incubation times provide a practical baseline, but further statistical comparisons are required to determine whether these durations are truly optimal. Full article

Hydrogel materials have become an efficient, bioactive, and multifunctional alternative with great potential for biomedical applications. In this work, photoactive films were successfully designed for optical processing, and their photoactivity was tested in photodynamic therapy (PDT), such as antimicrobial patches. The stimulus-response hydrogel films are made of a hydrophilic polymer based on vinyl monomers, specifically 2-hydroxyethyl methacrylate (HEMA) and acrylamide (AAm), in a 1:1 molar ratio, along with the photochromic agent, 3,3-dimethylindolin-6′-nitrobenzoespiropirano (BSP), and a crosslinking agent, N,N’-methylenebisacrylamide (MBA). These hydrogel films were successfully created using the photoinitiator 2-hydroxy-4′-(2-hydroxyethoxy)-2-methylpropiophenone (IRGACURE 2959), MBA, and BSP in different concentrations (0.1, 0.3, and 0.5 mol%), which were later tested in photodynamic therapy (PDT) with the photosensitizer Ru(bpy)₂²⁺ against Staphylococcus aureus. The results showed that, while free Ru(bpy)₂²⁺ needed concentrations of 4–8 µg/mL to eliminate methicillin-sensitive (MSSA) strains, only partial inactivation was achieved for methicillin-resistant (MRSA) strains. The addition of the hydrogel films with BSP improved their effectiveness, lowering the minimum inhibitory concentration (MIC) to 2 µg/mL to fully inactivate MSSA and MRSA strains. These findings demonstrate that the combined use of hydrogel films containing BSP and Ru(bpy)₂²⁺ within a hydrogel matrix not only boosts antimicrobial activity but also highlights the potential of these photoactive films as innovative photosensitive antimicrobial coatings. This synergistic effect of BSP and Ru(bpy)₂²⁺ indicates that these materials are promising candidates for next-generation antimicrobial coatings and creative photosensitive materials. Full article

The alarming increase in infections caused by antimicrobial-resistant bacteria is increasingly posing a critical threat to public health. For this reason, the scientific community is focusing on alternative therapeutic strategies, such as antimicrobial photodynamic therapy (aPDT). This review examined the use of natural photosensitizers (PSs) in aPDT, emphasizing how they may produce high yields of reactive oxygen species when activated by light and consequently inactivate a wide range of pathogens, including bacteria embedded in biofilms, efficiently. The main methodologies and several strategies of incorporation into cutting-edge nanotechnological delivery systems of the most prevalent natural PSs (curcuminoids, perylenequinones, tetrapyrrolic macrocycles, and flavins) have been analyzed. Although natural PSs have benefits in terms of environmental sustainability and biocompatibility, their clinical use is frequently constrained by low bioavailability and solubility, issues that are being addressed more and more through novel formulations and dual-mode treatments. Studies conducted both in vitro and in vivo highlight these compounds’ strong antibacterial and wound-healing properties. In conclusion, natural molecule-based aPDT is a flexible and successful strategy for combating antimicrobial resistance, deserving of more translational study and clinical advancement. Full article

Photodynamic inactivation (PDI) is an innovative non-thermal sterilization and preservation method that has recently emerged as a safe, effective, cost-effective and environmentally sustainable alternative for biomedical applications. Curcumin (Cur), a commonly used food additive, possesses photosensitizing properties. In this study, we investigated the effect of curcumin-mediated photodynamic treatment (Cur-PDT) on the preservation of fresh wolfberries. Our experimental data revealed that a Cur-PDT treatment using a cur concentration of 500 μmol/L for 30 min, with 20 W irradiation, achieved the best preservation effect on fresh wolfberries. This intervention significantly slowed the decline in post-harvest hardness and delayed the progression of decay. It also reduced the accumulation of malondialdehyde (MDA), hydrogen peroxide (H₂O₂) and superoxide anion (•O₂⁻). Notably, at day 3, the enzymatic activities of catalase (CAT) and ascorbate peroxidase (APX) in Cur-PDT-treated wolfberries were 1.12 and 1.88 times higher, respectively, than those in the control group. These elevated enzyme activities promoted the biosynthesis and recycling of ascorbic acid (AsA) and glutathione (GSH), leading to their substantial accumulation under oxidative stress conditions. By modulating the antioxidant defense system, Cur-PDT has the potential to extend the shelf-life of post-harvest wolfberries and enhance their overall quality attributes, thereby maintaining physiological homeostasis during storage. Full article

Municipal wastewaters pose a severe risk to the environment and human health when discharged untreated. This is due to their high content of pathogens, such as viruses and bacteria, which can cause diseases like cholera. Herein, the research and development of porphyrin-modified polyethersulfone (PES) ultrafiltration (UF) membranes was conducted to improve bacterial inactivation in complex municipal wastewater and enhance the fouling resistance and filtration performance. The synthesis and fabrication of porphyrin nanofillers and the resultant membrane characteristics were studied. The incorporation of porphyrin-based nanofillers improved the membrane’s hydrophilicity, morphology, and flux (247 Lm⁻² h⁻¹), with the membrane contact angle (CA) decreasing from 90° to ranging between 58° and 50°. The membrane performance was monitored for its flux, antifouling properties, reusability potential, municipal wastewater, and humic acid. The modified membranes demonstrated an effective application in wastewater treatment, achieving notable antibacterial activity, particularly under light exposure. The In-BP@SW/PES membrane demonstrated effective antimicrobial photodynamic effects against both Gram-positive S. aureus and Gram-negative E. coli. It achieved at least a 3-log reduction in bacterial viability, meeting Food and Drug Administration (FDA) standards for efficient antimicrobial materials. Among the variants tested, membranes modified with In-PB@SW nanofillers exhibited superior antifouling properties with flux recovery ratios (FRRs) of 78.9% for the humic acid (HA) solution and 85% for the municipal wastewater (MWW), suggesting a strong potential for long-term filtration use. These results highlight the promise of porphyrin-functionalized membranes as multifunctional tools in advanced water treatment technologies. Full article