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New Molecular Insights into Antimicrobial Photo-Treatments

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A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Molecular Microbiology".

Deadline for manuscript submissions: closed (20 October 2025) | Viewed by 16716

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Special Issue Editor

Dr. Pietro Delcanale

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Guest Editor

Department of Mathematical, Physical and Computer Sciences, University of Parma, 43124 Parma, Italy
Interests: photo-active molecules; fluorescence microscopy; nanomedicine; photodynamic inactivation

Special Issue Information

Dear Colleagues,

The recent COVID-19 pandemic highlighted the need for the identification of new and effective treatments against emerging health threats. The insurgence of new microbial infections, including the growing number of antibiotic-resistant bacterial species, calls for a renewed effort towards the development of antimicrobials that do not induce resistance in microorganisms. Antimicrobial photo-treatments, such as photodynamic inactivation (PDI), antimicrobial photo-thermal therapy (PTT) or blue/near UV light-induced inactivation, are regarded as a viable alternative to antibiotics in a large variety of cases.

We invite investigators who are active in the field to submit original research papers as well as review articles to this Special Issue, to highlight the realization and the use of molecular and nano-systems for antimicrobial photo-treaments, to provide new insights into the fundamental mechanisms of microbial photo-inactivation or to propose stimulating ideas and novel applications to expand the use of the methodology. Scientific contributions are expected to provide molecular-level information about the systems and/or the processes under study. The issue will focus on, but will not be limited to, the following aspects:

Fundamental mechanisms of PDI, PTT or UV light-based antimicrobial treatments;
New photo-activated materials, including molecules and nanomaterials;
New formulations of existing photo-activated agents;
Targeted photo-treatments against bacteria and viruses;
Photo-treatments exploiting endogenous molecules;
Visualization of functional photo-activated agents in situ with advanced microscopy;
Combination treatments with, e.g., antibiotics or antibodies;
Photo-activated molecular systems for industrial and large-scale applications.

Dr. Pietro Delcanale
Guest Editor

Manuscript Submission Information

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Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. International Journal of Molecular Sciences is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. There is an Article Processing Charge (APC) for publication in this open access journal. For details about the APC please see here. Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

photodynamic inactivation
photo-thermal therapy
photosensitization
antimicrobials
reactive oxygen species
photo-treatments
antibiotic resistance
singlet oxygen
antivirals

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Published Papers (10 papers)

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Research

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27 pages, 8214 KB

Open AccessArticle

The Immunostimulatory Effect of MIL-101(Al)-NH₂ In Vivo and Its Potential to Overcome Bacterial Resistance to Penicillin Enhanced by Hypericin-Induced Photodynamic Therapy

by Mariana Máčajová, Ľuboš Ambro, Majlinda Meta, Ľuboš Zauška, Terézia Gulyásová, Boris Bilčík, Ivan Čavarga, Gabriela Zelenková, Erik Sedlák, Miroslav Almáši and Veronika Huntošová

Int. J. Mol. Sci. 2025, 26(23), 11681; https://doi.org/10.3390/ijms262311681 - 2 Dec 2025

Cited by 1 | Viewed by 428

Abstract

The increasing prevalence of multidrug-resistant bacteria necessitates alternative therapeutic strategies that combine antimicrobial efficacy with immunomodulatory properties. Here, we report the immunostimulatory activity and antibacterial potential of the amino-functionalized metal–organic framework MIL-101(Al)-NH₂ as a carrier for penicillin (PEN) and hypericin (Hyp), a photodynamically active compound. Structural and physicochemical characterization confirmed successful encapsulation of PEN, Hyp, and their combination within MIL-101(Al)-NH₂, with distinct effects on porosity, release kinetics, and thermal stability. Drug release studies revealed rapid Hyp liberation triggered by serum components, whereas PEN exhibited a biphasic, diffusion-controlled profile. Using a quail chorioallantoic membrane (CAM) model, we demonstrated that MIL-101(Al)-NH₂ enhances interferon-α expression, indicating intrinsic immunostimulatory activity, and that Hyp-loaded systems promote angiogenic responses. In a bacterial infection CAM model, MIL-101(Al)-NH₂ carriers loaded with Hyp or Hyp/PEN induced immunomodulatory changes and, upon photodynamic activation, inhibited bacterial growth. While Gram-negative Escherichia coli remained resistant, Gram-positive Staphylococcus epidermidis was effectively suppressed by photodynamic therapy (PDT), and Hyp/PEN co-delivery overcame bacterial resistance to PEN. These results highlight MIL-101(Al)-NH₂ as a multifunctional nanoplatform with immunostimulatory capacity and PDT-enhanced antibacterial activity, offering a promising strategy to combat antibiotic resistance and infections associated with medical implants. Full article

(This article belongs to the Special Issue New Molecular Insights into Antimicrobial Photo-Treatments)

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20 pages, 3070 KB

Open AccessArticle

Rose Bengal-Incorporated Supramolecular Gels as a Topical Platform for Localized Antimicrobial Photodynamic Therapy

by Kavya Anguluri, Saman Bagherpour, Ana C. Calpena, Lyda Halbaut, Alba Espargaró, Raimon Sabate and Lluïsa Pérez-García

Int. J. Mol. Sci. 2025, 26(23), 11455; https://doi.org/10.3390/ijms262311455 - 26 Nov 2025

Viewed by 550

Abstract

Efficient and localized singlet oxygen (SO) generation is essential for improving antimicrobial photodynamic therapy (aPDT). In this study, a bis-imidazolium-based amphiphilic gelator is used, which self-assembles into a supramolecular gel in a water–ethanol medium and incorporates Rose Bengal (RB) as a photosensitizer. The gel network provides a confined environment that promotes SO formation under light irradiation. RB@Gel was characterized with respect to its morphology, degradation behavior, and swelling properties. Biopharmaceutical assessment included in vitro release, ex vivo permeation studies and Hen’s Egg Test–Chorioallantoic Membrane (HET-CAM) assay. Rheological measurements confirmed a viscoelastic profile, indicating structural stability and suitability for localized therapeutic applications. SO production within the gel was quantified using tetrasodium 9,10-anthracenediyl-bis(methylene)dimalonate (NaABMA), showing higher efficiency than that of RB in solution. The RB@Gel exhibited significant aPDT against E. coli in a direct-surface contact assay. Overall, the RB@Gel provides a stable, suitable platform capable of efficient SO generation and potent antibacterial activity, highlighting its promise for localized aPDT applications. Full article

(This article belongs to the Special Issue New Molecular Insights into Antimicrobial Photo-Treatments)

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16 pages, 1960 KB

Open AccessArticle

Photodynamic Inactivation Enhances Antibiotic Efficacy Without Affecting Drug Stability: Insights into Photosensitizer–Antibiotic Combination Therapies

by Rocío B. Acosta, Edgardo N. Durantini and Mariana B. Spesia

Int. J. Mol. Sci. 2025, 26(23), 11267; https://doi.org/10.3390/ijms262311267 - 21 Nov 2025

Viewed by 569

Abstract

Photodynamic inactivation (PDI) represents a promising strategy to overcome bacterial resistance by combining light, oxygen, and a photosensitizer (PS) to generate reactive oxygen species (ROS) that damage essential cellular components. Combining PDI with conventional antibiotics (ATBs) may further enhance bacterial eradication through complementary mechanisms. In this study, the tetracationic 5,10,15,20-tetra(4-N,N,N-trimethylammoniophenyl)porphyrin (TMAP⁴⁺) was evaluated in combination with ATBs: ampicillin (AMP) and rifampicin (RIF) against Staphylococcus aureus and cephalexin (CFX) against Escherichia coli. The photostability of all agents was assessed under the experimental irradiation conditions, and no evidence of physical interaction between TMAP⁴⁺ and the ATBs was detected. AMP and CFX remained photostable, while RIF exhibited only minimal photodegradation under white light, confirming its stability during PDI treatments. The antimicrobial assays revealed that irradiation significantly enhanced the bactericidal activity of TMAP⁴⁺. When combined with ATBs, photoactivated TMAP⁴⁺ led to a pronounced reduction in the minimum inhibitory concentration (MIC) values of AMP and RIF for S. aureus and of CFX for E. coli, indicating additive effects. Growth curve analyses corroborated these results, showing delayed bacterial growth and decreased maximal optical densities in the combined treatments compared to single agents. Overall, these findings demonstrate that the photodynamic process can potentiate the antimicrobial effect of conventional ATBs without compromising their stability, supporting the potential of PS–ATB combination therapies as a valuable approach to improve antibacterial efficacy and mitigate ATB resistance. Full article

(This article belongs to the Special Issue New Molecular Insights into Antimicrobial Photo-Treatments)

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14 pages, 1814 KB

Open AccessArticle

The Combined Photosensitizers in Antimicrobial Photodynamic Therapy: The Case of Methylene Blue and Photodithazine Against Klebsiella pneumoniae

by Koteswara Rao Yerra, Jennifer M. Soares and Vanderlei S. Bagnato

Int. J. Mol. Sci. 2025, 26(20), 10211; https://doi.org/10.3390/ijms262010211 - 21 Oct 2025

Viewed by 1136

Abstract

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

(This article belongs to the Special Issue New Molecular Insights into Antimicrobial Photo-Treatments)

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13 pages, 1623 KB

Open AccessArticle

The Photodynamic Antibacterial Potential of New Tetracationic Zinc(II) Phthalocyanines Bearing 4-((Diethylmethylammonium)methyl)phenoxy Substituents

by Gennady Meerovich, Dmitry Bunin, Ekaterina Akhlyustina, Igor Romanishkin, Vladimir Levkin, Sergey Kharnas, Maria Stepanova, Alexander Martynov, Victor Loschenov, Yulia Gorbunova and Marina Strakhovskaya

Int. J. Mol. Sci. 2025, 26(19), 9414; https://doi.org/10.3390/ijms26199414 - 26 Sep 2025

Viewed by 761

Abstract

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

(This article belongs to the Special Issue New Molecular Insights into Antimicrobial Photo-Treatments)

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27 pages, 6077 KB

Open AccessArticle

Photodynamic Effectiveness of Copper-Iminopyridine Photosensitizers Coupled to Zinc Oxide Nanoparticles Against Klebsiella pneumoniae and the Bacterial Response to Oxidative Stress

by Dafne Berenice Hormazábal, Ángeles Beatriz Reyes, Matías Fabián Cuevas, Angélica R. Bravo, David Moreno-da Costa, Iván A. González, Daniel Navas, Iván Brito, Paulina Dreyse, Alan R. Cabrera and Christian Erick Palavecino

Int. J. Mol. Sci. 2025, 26(9), 4178; https://doi.org/10.3390/ijms26094178 - 28 Apr 2025

Cited by 1 | Viewed by 1333

Abstract

One of the most urgent threats to public health worldwide is the ongoing rise of multidrug-resistant (MDR) bacterial strains. Among the most critical pathogens are MDR-Klebsiella pneumoniae strains. The lack of new antibiotics has led to an increased need for non-antibiotic antimicrobial therapies. Photodynamic therapy (PDT) has become increasingly significant in treating MDR bacteria. PDT uses photosensitizer compounds (PS) that generate reactive oxygen species (ROS) when activated by light. These ROS produce localized oxidative stress, damaging the bacterial envelope. A downside of PDT is the limited bioavailability of PSs in vivo, which can be enhanced by conjugating them with carriers like nanoparticles (NPs). Zinc nanoparticles possess antibacterial properties, decreasing the adherence and viability of microorganisms on surfaces. The additive or synergistic effect of the combined NP-PS could improve phototherapeutic action. Therefore, this study evaluated the effectiveness of the copper(I)-based PS CuC1 compound in combination with Zinc Oxide NP, ZnONP, to inhibit the growth of both MDR and sensitive K. pneumoniae strains. The reduction in bacterial viability after exposure to a PS/NP mixture activated by 61.2 J/cm² of blue light photodynamic treatment was assessed. The optimal PS/NP ratio was determined at 2 µg/mL of CuC1 combined with 64 µg/mL of ZnONP as the minimum effective concentration (MEC). The bacterial gene response aligned with a mechanism of photooxidative stress induced by the treatment, which damages the bacterial cell envelope. Additionally, we found that the PS/NP mixture is not harmful to mammalian cells, such as Hep-G2 and HEK-293. In conclusion, the CuC1/ZnONP combination could effectively aid in enhancing the antimicrobial treatment of infections caused by MDR bacteria. Full article

(This article belongs to the Special Issue New Molecular Insights into Antimicrobial Photo-Treatments)

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11 pages, 1507 KB

Open AccessCommunication

Insights on the Mechanical Properties of SARS-CoV-2 Particles and the Effects of the Photosensitizer Hypericin

by Matteo Mariangeli, Ana Moreno, Pietro Delcanale, Stefania Abbruzzetti, Alberto Diaspro, Cristiano Viappiani and Paolo Bianchini

Int. J. Mol. Sci. 2024, 25(16), 8724; https://doi.org/10.3390/ijms25168724 - 10 Aug 2024

Cited by 3 | Viewed by 4603

Abstract

SARS-CoV-2 is a highly pathogenic virus responsible for the COVID-19 disease. It belongs to the Coronaviridae family, characterized by a phospholipid envelope, which is crucial for viral entry and replication in host cells. Hypericin, a lipophilic, naturally occurring photosensitizer, was reported to effectively inactivate enveloped viruses, including SARS-CoV-2, upon light irradiation. In addition to its photodynamic activity, Hyp was found to exert an antiviral action also in the dark. This study explores the mechanical properties of heat-inactivated SARS-CoV-2 viral particles using Atomic Force Microscopy (AFM). Results reveal a flexible structure under external stress, potentially contributing to the virus pathogenicity. Although the fixation protocol causes damage to some particles, correlation with fluorescence demonstrates colocalization of partially degraded virions with their genome. The impact of hypericin on the mechanical properties of the virus was assessed and found particularly relevant in dark conditions. These preliminary results suggest that hypericin can affect the mechanical properties of the viral envelope, an effect that warrants further investigation in the context of antiviral therapies. Full article

(This article belongs to the Special Issue New Molecular Insights into Antimicrobial Photo-Treatments)

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18 pages, 3253 KB

Open AccessArticle

Concanavalin A Delivers a Photoactive Protein to the Bacterial Wall

by Andrea Mussini, Pietro Delcanale, Melissa Berni, Stefano Pongolini, Mireia Jordà-Redondo, Montserrat Agut, Peter J. Steinbach, Santi Nonell, Stefania Abbruzzetti and Cristiano Viappiani

Int. J. Mol. Sci. 2024, 25(11), 5751; https://doi.org/10.3390/ijms25115751 - 25 May 2024

Cited by 6 | Viewed by 2801

Abstract

Modular supramolecular complexes, where different proteins are assembled to gather targeting capability and photofunctional properties within the same structures, are of special interest for bacterial photodynamic inactivation, given their inherent biocompatibility and flexibility. We have recently proposed one such structure, exploiting the tetrameric bacterial protein streptavidin as the main building block, to target S. aureus protein A. To expand the palette of targets, we have linked biotinylated Concanavalin A, a sugar-binding protein, to a methylene blue-labelled streptavidin. By applying a combination of spectroscopy and microscopy, we demonstrate the binding of Concanavalin A to the walls of Gram-positive S. aureus and Gram-negative E. coli. Photoinactivation is observed for both bacterial strains in the low micromolar range, although the moderate affinity for the molecular targets and the low singlet oxygen yields limit the overall efficiency. Finally, we apply a maximum entropy method to the analysis of autocorrelation traces, which proves particularly useful when interpreting signals measured for diffusing systems heterogeneous in size, such as fluorescent species bound to bacteria. Full article

(This article belongs to the Special Issue New Molecular Insights into Antimicrobial Photo-Treatments)

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Review

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32 pages, 8009 KB

Open AccessReview

Recent Advances in Nanoparticle-Mediated Antibacterial Photodynamic Therapy

by Nivedita, Shashwat Sharma, Dyah Ika Krisnawati, Tsai-Mu Cheng and Tsung-Rong Kuo

Int. J. Mol. Sci. 2025, 26(22), 10949; https://doi.org/10.3390/ijms262210949 - 12 Nov 2025

Viewed by 1331

Abstract

The escalating threat of antibiotic resistance has prompted the search for alternative antibacterial therapies. Antibacterial photodynamic therapy (aPDT), which utilizes light-activated photosensitizers to generate reactive oxygen species (ROS), offers a promising, non-invasive approach. The aim of this review is to analyze recent advances in nanoparticle-mediated aPDT and synthesize crucial design principles necessary to overcome the current translational barriers, thereby establishing a roadmap for future clinically applicable antimicrobial treatments. Emerging nanoparticle platforms, including upconverting nanoparticles (UCNPs), carbon dots (CDs), mesoporous silica nanoparticles (MSNs), liposomes, and metal–organic frameworks (MOFs), have demonstrated improved photosensitizer delivery, enhanced ROS generation, biofilm disruption, and targeted bacterial eradication. Synergistic effects are observed when aPDT is integrated with photothermal, chemodynamic, or immunotherapeutic approaches. The review further examines the mechanisms of action, biocompatibility, and antibacterial performance of these nanoparticle systems, particularly against drug-resistant strains and in challenging environments such as chronic wounds. Overall, nanomaterial-mediated aPDT presents a highly promising and versatile solution to antimicrobial resistance. Future perspectives include the integration of artificial intelligence to personalize aPDT by predicting optimal light dosage and nanoplatform design based on patient-specific data, rigorous clinical validation through trials, and the development of safer, more efficient nanoparticle platforms. Full article

(This article belongs to the Special Issue New Molecular Insights into Antimicrobial Photo-Treatments)

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28 pages, 2883 KB

Open AccessReview

Natural Biomolecules and Light: Antimicrobial Photodynamic Strategies in the Fight Against Antibiotic Resistance

by Greta Amendola, Mariagrazia Di Luca and Antonella Sgarbossa

Int. J. Mol. Sci. 2025, 26(16), 7993; https://doi.org/10.3390/ijms26167993 - 19 Aug 2025

Cited by 8 | Viewed by 2138

Abstract

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

(This article belongs to the Special Issue New Molecular Insights into Antimicrobial Photo-Treatments)

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