The dual role of photodynamic therapy to treat cancer and microbial infection.

Photodynamic therapy (PDT) is a minimally invasive treatment showing promise against cancer and microbial infections. PDT targets tumor cells while sparing healthy tissue, reducing side effects. It induces immunogenic cell death, potentially stimulating antitumor immune responses and reducing cancer recurrence. In microbial treatment, PDT effectively combats bacteria, fungi and viruses. Combining PDT with chemotherapy, radiotherapy and immunotherapy enhances its efficacy. However, challenges such as tumor hypoxia, limited tissue penetration and phototoxicity necessitate ongoing research efforts to optimize PDT protocols and overcome limitations. Overall, PDT is versatile and continually advancing with refined protocols to improve its clinical utility against cancer and microbial infections.

Comparative assessment of periodontal status and genotoxicity in orthodontic patients on fixed mechanotherapy with and without adjunct chlorhexidine mouthrinse: A randomized control clinical trial.

Background: Adjunct chlorhexidine mouthrinse is used routinely in orthodontic clinical practice for plaque control. However, chlorhexidine has genotoxic effects on the oral cells. Moreover, orthodontic appliance leach Ni, Cr metals ions into saliva causing toxicity of surrounding mucosa. Hence, the aim of the study was to assess the periodontal status and genotoxicity in orthodontic patients on fixed mechanotherapy with and without adjunct chlorhexidine using micronucleus (MN) test. Materials and Methods: A randomized control clinical trial was conducted in 30 patients who were on fixed mechanotherapy. The patients were randomly assigned into two treatment groups; Group-A (Control Group): Included 15 patients who are on fixed orthodontic therapy with mechanical plaque control measures only., Group-B (Experimental Group:) included 15 patients on fixed orthodontic therapy with mechanical plaque control and adjunct chlorhexidine mouthrinse (0.2%) for 2 weeks. Periodontal status and genotoxicity using MN test were done at following time points; T0: Just before start of the orthodontic treatment., T1: 2 weeks after start of the orthodontic treatment., T2: 6 weeks after start of the orthodontic treatment., T3: 12 weeks after start of the orthodontic treatment. Results: Plaque index (PI) and bleeding on probing (BOP) were significantly decreased in Group B as compared to Group A in the time intervals; T0-T2, T0-T3, T1-T3 (P < 0.05). Probing pocket depth (PPD) and Clinical attachment level (CAL) showed no significant change in both the groups. The genotoxicity assessed by MN test was significantly increased in Group B than Group A at time intervals; T0-T1, T0-T2 and T0-T3. Conclusion: Adjunct chlorhexidine resulted in decreased PI and BOP scores but nonsignificant change in PPD and CAL. However, the genotoxicity increased significantly in both the groups but more with adjunct chlorhexidine.

Broad-Spectrum Inhibitors against Class A, B, and C Type ß-Lactamases to Block the Hydrolysis against Antibiotics: Kinetics and Structural Characterization.

The emergence of antibiotic resistance has led to a global crisis for the physician to handle infection control issues. All antibiotics, including colistin, have lost efficiency against emerging drug-resistant bacterial strains due to the production of metallo-ß-lactamases (MBLs) and serine-ß-lactamases (SBLs). Therefore, it is of the utmost importance to design inhibitors against these enzymes to block the hydrolytic action against antibiotics being used. Although various novel ß-lactamase inhibitors are being authorized or are under clinical studies, the coverage of their activity spectrum does not include MDR organisms expressing multiple classes of ß-lactamases at a single time. This study reports three novel broad-spectrum inhibitors effective against both SBLs and MBLs. Virtual screening, molecular docking, molecular dynamics simulations, and an in silico pharmacokinetic study were performed to identify the lead molecules with broad-spectrum ability to inhibit the hydrolysis of ß-lactam. The selected compounds were further assessed by in vitro cell assays (MIC, 50% inhibitory concentration [IC50], kinetics, and fluorescence against class A, B, and C type ß-lactamases) to confirm their efficacies. A 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was performed to check the toxicity of screened lead molecules. All three selected inhibitors were found to reduce MIC and showed good affinity against all the SBLs and MBLs produced by class A, B, and C type ß-lactamases. These nontoxic novel non-ß-lactam broad-spectrum inhibitors bind to the active site residues of selected ß-lactamases, which are crucial for ß-lactam antibiotic hydrolysis. These inhibitors may be proposed as a future drug candidate in combination with antibiotics as a single formulation to control infection caused by resistant strains. Hence, this study plays a significant role in the cure of infections caused by antibiotic-resistant bacteria. IMPORTANCE Several inhibitors for usage in conjunction with antibiotics have been developed. However, to date, there is no commercially available broad-spectrum ß-lactamase inhibitor that targets both MBLs and SBLs. Here, we showed three novel broad-spectrum inhibitors with promising results through computational techniques and in vitro studies. These inhibitors are effective against both SBLs and MBLs and hence could be used as future drug candidates to treat infections caused by multidrug-resistant bacteria producing both types of enzymes (SBLs and MBLs).

A nano phototheranostic approach of toluidine blue conjugated gold silver core shells mediated photodynamic therapy to treat diabetic foot ulcer.

Diabetic foot infection caused by multidrug-resistant bacteria, is becoming serious problem. Moreover, polymicrobial biofilms contribute significantly to the persistent infections. In the present study, we investigated the effectiveness of novel toluidine blue conjugated chitosan coated gold-silver core-shell nanoparticles (TBO-chit-Au-AgNPs) mediated photodynamic therapy and demonstrate their use as a nontoxic antibacterial therapy to combat diabetic foot ulcer (DFU) caused by multi-drug resistant strains both in monomicrobial and polymicrobial state of infection. In vitro efficacy of TBO-chit-Au-AgNPs mediated photodynamic therapy (PDT) against polymicrobial biofilms was determined using standard plate count method and compared with that of monomicrobial biofilms of each species. Different anti-biofilm assays and microscopic studies were performed to check the efficacy of TBO-chit-Au-AgNPs mediated PDT, displayed significant decrease in the formation of biofilm. Finally, its therapeutic potential was validated in vivo type-2DFU. Cytokines level was found reduced, using nano-phototheranostic approach, indicating infection control. Expression profile of growth factors confirmed both the pathogenesis and healing of DFU. Hence, we conclude that TBO-chit-Au-AgNPs mediated PDT is a promising anti-bacterial therapeutic approach which leads to a synergistic healing of DFU caused by MDR bacterial strains.

Antimicrobial photodynamic therapy (aPDT) against vancomycin resistant Staphylococcus aureus (VRSA) biofilm disruption: A putative role of phagocytosis in infection control.

Biofilm mediated infections have major clinical impact. Staphylococcus aureus is a pathogen that frequently causes biofilm forming infections, such as those associated with medical devices and persistent wounds. Microorganisms embedded in biofilm are impervious to antibiotics and other antimicrobial agents, thus they are difficult to eliminate. The upsurge of multi-drug resistant strains makes treating such illnesses even more difficult. Therefore, new strategies are required to combat such type of infections. In this work, we have proposed an alternative therapeutic option to eradicate preformed biofilm of vancomycin resistant Staphylococcus aureus (VRSA) and enhanced phagocytosis by neutrophils in fresh human blood using curcumin mediated antimicrobial photodynamic therapy (aPDT).At sub-MIC of curcumin, different anti-biofilm assays and microscopic examinations were performed, followed by 20 J/cm2 of blue laser light irradiation which corresponds to 52 s only. The result showed significant disruption of VRSA biofilm. Moreover, when curcumin-aPDT treated VRSA biofilm was exposed to whole blood from healthy donors, it was nearly completely eradicated. The present study suggests that curcumin-aPDT enhanced phagocytosis may be a useful strategy for inactivating VRSA biofilms adhering to medical implant surfaces.

Antimicrobial and antibiofilm photodynamic therapy against vancomycin resistant Staphylococcus aureus (VRSA) induced infection in vitro and in vivo.

Biofilm mediated infection caused by multi-drug resistant bacteria are difficult to treat since it protects the microorganisms by host defense system, making them resistant to antibiotics and other antimicrobial agents. Combating such type of nosocomial infection, especially in immunocompromised patients, is an urgent need and foremost challenge faced by clinicians. Therefore, antimicrobial photodynamic therapy (aPDT) has been intensely pursued as an alternative therapy for bacterial infections. aPDT leads to the generation of reactive oxygen species (ROS) that destroy bacterial cells in the presence of a photosensitizer, visible light and oxygen. Here, we elucidated a possibility of its clinical application by reducing the treatment time and exposing curcumin to 20 J/cm2 of blue laser light, which corresponds to only 52 s to counteract vancomycin resistant Staphylococcus aureus (VRSA) both in vitro and in vivo. To understand the mechanism of action, the generation of total reactive oxygen species (ROS) was quantified by 2'-7'-dichlorofluorescein diacetate (DCFH-DA) and the type of phototoxicity was confirmed by fluorescence spectroscopic analysis. The data showed more production of singlet oxygen, indicating type-II phototoxicity. Different anti-biofilm assays (crystal violet and congo red assays) and microscopic studies were performed at sub-MIC concentration of curcumin followed by treatment with laser light against preformed biofilm of VRSA. The result showed significant reduction in the preformed biofilm formation. Finally, its therapeutic potential was validated in skin abrasion wistar rat model. The result showed significant inhibition of bacterial growth. Furthermore, immunomodulatory analysis with rat serum was performed. A significant reduction in expression of proinflammatory cytokines TNF-α and IL-6 were observed. Hence, we conclude that curcumin mediated aPDT with 20 J/cm2 of blue laser treatment (for 52 s) could be used against multi-drug resistant bacterial infections and preformed biofilm formation as a potential therapeutic approach.

Inhibition of multi-drug resistant Klebsiella pneumoniae: Nanoparticles induced photoinactivation in presence of efflux pump inhibitor.

In the current scenario, frontline antibiotics are losing effectiveness against multidrug-resistant (MDR) bacteria because of the single mode of action. The accumulation of mutations and spread of antibiotic resistance markers among the bacteria results into the severe threat to community health. Now, there is an urgent requirement for the development of an alternate and as well as multiple-targeted action of drugs to stop the spread of resistance in bacteria. Here, we showed an alternative nanoparticle based photodynamic therapy (PDT) targeting the bacterial efflux pumps and its cell wall. The dextran capped gold nanoparticles (GNPDEX) were localized to the bacterial surface by nanoparticle attached Concanavalin-A (ConA), where GNPDEX attached methylene blue (MB) photosensitizer as an MB@GNPDEX-ConA formulation induced the killing of MDR Klebsiella pneumoniae clinical isolates in no time. The intervention of efflux pump inhibitor (EPI) further improved the MB@GNPDEX-ConA treatment modality and displayed the maximum bactericidal cytoplasmic phototoxicity. The CCCP EPI (carbonyl cyanide m-chlorophenylhydrazone) with the PDT increased the bacterial killing by>3 log10 as compared with or without EPI intervention. Further, the fractionated (two light treatment after long dark phase) PDT treatment modality decreased the bacterial biofilm growth up to ~90%. The microscopic as well as ROS fluorescent probes showed the singlet oxygen mediated cytotoxicity. The mode of interactions and genomic DNA photo-toxicity confirmed that EPI enhanced the killing mediated by singlet oxygen generation. The multi-targeted (Cell wall, DNA and efflux pump) modality of MB@GNPDEX-ConA in presence of EPI is an effective and alternative therapeutic approach against most potent Klebsiella MDR infections.