Effect Of Photodynamic Inactivation Research Articles

Boosting porphyrin synthesis and ALA-mediated photoinactivation through near-infrared therapy.

Photodynamic inactivation (PDI) combines the use of photosensitizers with visible light to produce reactive oxygen species that effectively eliminate pathogens. To investigate the impact of near- infrared therapy (NIRT) on heme biosynthesis and permeability of the pro-photosensitizers 5-aminolevulinic acid (ALA) and Hexyl-ALA (H-ALA) through biofilms, we applied sub-lethal conditions for both NIRT and PDI to maintain intact bacterial viability. During NIRT, the temperature remained below 37°C, permitting rapid heating (ΔT = 11°C) without causing thermal damage. NIRT potentiated the bactericidal effects of ALA-PDI by 3 logs in Staphylococcus aureus RN6390 biofilms and 4 logs in ST5-SCCmecI biofilms. With H-ALA-PDI, reductions of 4 and 6 logs, respectively, were observed. In the case of ALA, this enhancement was partly due to increased porphyrin synthesis, a result not replicated by simple heating. For H-ALA, the enhanced effect of PDI was likely due to biofilm or bacterial membrane destabilization caused by NIRT and H-ALA. Unlike biofilms, applying the same ALA-PDI conditions, the treatment was effective in planktonic S. aureus RN6390 cultures, reducing the bacteria by 3 logs, with no additional benefit from NIRT, suggesting that NIRT primarily aids in penetrating the biofilm matrix rather than the bacterial cells. Confocal fluorescence microscopy confirmed that NIRT-treated biofilms exposed to ALA exhibited higher porphyrin fluorescence than untreated biofilms. These findings support the use of NIRT in enhancing the effectiveness of PDI against bacterial biofilms.

Antibacterial mechanism of vitamin K3-mediated photodynamic inactivation against Pseudomonas fluorescens and its influences on blunt bream (Megalobrama amblycephala) quality

Photodynamic inactivation (PDI) has emerged as a novel non-thermal process technology for inactivating microorganisms due to its low cost, safety, and efficiency. This study aimed to investigate the antimicrobial effect of VK3-mediated PDI against Pseudomonas fluorescens (P. fluorescens) and to assess its impact on the quality of the blunt bream contaminated with P. fluorescens. The inhibition efficiency and photosensitization mechanism were investigated under different VK3 concentrations and UV-A light time conditions. The results showed that PDI could significantly inactivate P. fluorescens >7.0 log10 cfu/ml with 1 mM VK3 and 45 min of UV-A irradiation. Type I and type II photoresponses could co-occur in VK3-mediated PDI by quantitatively analyzing the photoresponse product ROS (OH, H2O2, 1O2). It was observed by SEM and TEM that the combined treatment of VK3 and UV-A disrupted the cellular integrity of P. fluorescens, leading to leakage of cell contents. Meanwhile, it degraded intracellular proteins and destroys genomic DNA, ultimately leading to bacterial death. In addition, the PDI system slowed down P. fluorescens’ proliferation during the blunt bream storage at 4 °C. The PDI system effectively killed 59 % of P. fluorescens on the blunt bream fillets under 1.5 mM VK3 and 60 min of UV-A irradiation. The VK3-mediated PDI delayed pH change and TVB-N production, inhibited protein degradation and fat oxidation, and ultimately maintained the quality of fish fillets. This study provides a promising PDI system for controlling food microbial contamination and maintaining the storage quality of aquatic products.

Comparison of the antimicrobial reduction effect of photodynamic inactivation with the addition of chlorophyll and curcumin photosensitizer in Aggregatibacter actinomycetemcomitans and Enterococcus faecalis

Background Aggregatibacter actinomycetemcomitans and Enterococcus faecalis are pathogenic bacteria of the oral cavity that cause various diseases such as periodontitis and endodontics. These bacteria are easily resistant to antibiotics. Photodynamic inactivation (PDI) is a method of inactivating microorganisms that utilizes light to activate a photosensitizer agent (PS) that produces reactive oxygen species causing cell lysis. Methods This study used the PDI method with a 405 nm diode laser at various energy density with the addition PS curcumin or chlorophyll Alfalfa, as much as 1.6 mg/ml on A. actinomycetemcomitans and E. faecalis bacteria. Results The study on E. faecalis bacteria showed that the energy density diode laser irradiation of 1.59 J/cm² gave the percentage of E. faecalis bacteria death 36.7% without PS, 69.30% with the addition of chlorophyll Medicago sativa L and 89.42% with the addition of curcumin. Meanwhile, the bacteria A. actinomycetemcomitans showed that the energy density diode laser irradiation of 1.59 J/cm² gave the percentage of bacterial death 35.81% without PS, 64.39% with the addition of chlorophyll Medicago sativa L and 89.82% with the addition of curcumin. PS was critical to the success of the PDI. Conclusions The addition of PS curcumin increased the effectiveness of reducing bacteria E. faecalis and A. actinomycetemcomitans compared to chlorophyll Medicago sativa L.

Amino acids inhibit the photodynamic inactivation effect by hindering cellular oxidative stress

Photodynamic inactivation (PDI) could utilize light to activate reactive oxygen species (ROS) produced by photosensitizers to kill bacteria for preservation. To delve into the complex effects arising during the post-harvest PDI processing, we conducted experiments using Pseudomonas reinekei, a food spoilage bacteria extracted from rotten Pakchoi. Through analyzing the metabolomics results, we discovered that methionine (Met) and glutamate (Glu) exhibited significant inhibitory effects during the PDI process. The oxidative stress generated by light treatment resulted in a reduction of 30.31% and 36.37% in the levels of Met and Glu, respectively. The data also showed that exogenous Met and Glu reduced intracellular oxidative stress levels, increased peroxidase activity, and prevented the damage of intracellular material and cell membrane deformation. That amino acids could inhibit the effect of PDI by hindering oxidative stress. Therefore, the amino acid content should be considered when applying PDI to treat Met- or Glu-rich foods.

Evaluation of quantification methods to determine photodynamic action on mono- and dual-species bacterial biofilms.

The effect of photodynamic inactivation (PDI) sensitized by 5,10,15,20-tetra(4-N,N,N-trimethylammoniophenyl)porphyrin (TMAP4+) on different components of mono- and dual-species biofilms of Staphylococcus aureus and Escherichia coli was determined by different methods. First, the plate count technique showed that TMAP4+-PDI was more effective on S. aureus than E. coli biofilm. However, crystal violet staining revealed no significant differences between before and after PDI biofilms of both bacteria. On the other hand, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method indicated a reduction in viable cells as the light exposure time increases in both, mono- and dual-species biofilms. Furthermore, it was determined that as the irradiation time increases, the amount of extracellular polymeric substances present in the biofilms decreased. This effect was presented in both strains and in the mixed biofilm, being more evident in S. aureus mono-specie biofilm. Finally, scanning electron microscopy analysis showed a decrease in the number of cells forming the biofilm after photosensitization treatments. This information makes it possible to determine whether the photodynamic action is based on damage to metabolic activity, extracellular matrix and/or biomass, which may be useful in establishing a fully effective PDI protocol for the treatment of microorganisms growing as biofilms.

Comparison of the antimicrobial reduction effect of photodynamic inactivation with the addition of chlorophyll and curcumin photosensitizer in Aggregatibacter actinomycetemcomitans and Enterococcus faecalis

Background Aggregatibacter actinomycetemcomitans and Enterococcus faecalis are pathogenic bacteria of the oral cavity that cause various diseases such as periodontitis and endodontics. These bacteria are easily resistant to antibiotics. Photodynamic inactivation (PDI) is a method of inactivating microorganisms that utilizes light to activate a photosensitizer agent (PS) that produces reactive oxygen species causing cell lysis. Methods This study used the PDI method with a 405 nm diode laser at various energy density with the addition PS curcumin or chlorophyll Alfalfa, as much as 1.6 mg/ml on A. actinomycetemcomitans and E. faecalis bacteria. Results The study on E. faecalis bacteria showed that the energy density diode laser irradiation of 1.59 J/cm² gave the percentage of E. faecalis bacteria death 36.7% without PS, 69.30% with the addition of chlorophyll Medicago sativa L and 89.42% with the addition of curcumin. Meanwhile, the bacteria A. actinomycetemcomitans showed that the energy density diode laser irradiation of 1.59 J/cm² gave the percentage of bacterial death 35.81% without PS, 64.39% with the addition of chlorophyll Medicago sativa L and 89.82% with the addition of curcumin. PS was critical to the success of the PDI. Conclusions The addition of PS curcumin increased the effectiveness of reducing bacteria E. faecalis and A. actinomycetemcomitans compared to chlorophyll Medicago sativa L.

Synergistic effects of nano curcumin mediated photodynamic inactivation and nano-silver@colistin against Pseudomonas aeruginosa biofilms

BackgroundPatients with burn injuries colonized by multidrug-resistant Pseudomonas aeruginosa face increased mortality risk. The efficacy of colistin, a last-resort treatment, is declining as resistance levels rise. P. aeruginosa's robust biofilm exacerbates antibiotic resistance. Photodynamic Inactivation (PDI) shows promise in fighting biofilm. Materials and methodsNano curcumin (nCur) particles were synthesized, and their chemical characteristics were determined using zeta potential (ZP), dynamic light scattering analysis (DLS), energy-dispersive X-ray (EDX) analysis, and fourier transform infrared (FTIR). We conducted an MTT assay to assess the cytotoxicity of nCur-mediated PDI in combination with nanosilver colistin. The fractional biofilm inhibitory concentration (FBIC) of two P. aeruginosa clinical isolates and P. aeruginosa ATCC 27853 during nCur-mediated PDI@AgNPs@CL was determined using a 3-dimensional (3-D) checkerboard assay. To study the effect of nCur-mediated PDI@AgNPs@CL on lasI, lasR, rhlI, rhlR, pelA, and pslA gene expression, Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was conducted at each isolate's FBIC. The impact of treatments was also investigated using scanning electron microscopy (SEM). ResultsThe ZP and mean DLS values of the nCur were 10.3 mV and 402.6 ± 24.6 nm, respectively. The distinct functional groups of nCur corresponded with the peaks of FTIR absorption. Moreover, the EDX analysis showed the ratios of different metals in nCur. Cell viability percentages of nCur-mediated PDI@AgNPs@CL at FBIC concentrations of clinical isolates Nos. 30, 354, and P. aeruginosa ATCC 27853 were 91.36 %, 83.20 %, and 92.48 %, respectively. nCur-mediated PDI@AgNPs@CL treatment showed synergistic effects in clinical isolates and P. aeruginosa ATCC 27853 in a 3-D checkerboard assay. All six of the investigated genes showed down-regulation after nCur-mediated PDI@AgNPs@CL treatment. The most suppressed gene during nCur-mediated PDI@AgNPs@CL treatment was the rhlR gene (-11.9-fold) of P. aeruginosa ATCC 27853. The SEM micrographs further proved the connecting cement reduction and biofilm mass mitigation following nCur-mediated PDI@AgNPs@CL treatments. ConclusionsThe combined effect of nCur-mediated PDI and AgNPs@CL synergistically reduce the formation of biofilm in P. aeruginosa. This may be attributable to the suppression of the genes responsible for regulating the production of biofilms.

Synergistic Antimicrobial Effect of Photodynamic Inactivation and SWEEPS in Combined Treatment against Enterococcus faecalis in a Root Canal Biofilm Model: An In Vitro Study

Background: Persistent bacterial infections caused by biofilm-associated Enterococcus faecalis are a global public health concern. This study aims to compare the biofilm removal effects of methylene blue (MB) or hydrogen peroxide (HP) mediated photodynamic inactivation (PDI) with a shock wave enhanced emission photoacoustic streaming (SWEEPS) method laser irradiation enhanced bactericidal activity. Methods: A total of fifty extracted single-rooted human teeth were used. Each canal was then inoculated with a bacterial suspension of E. faecalis and left for ten days to induce biofilm formation. The antibacterial effects within treated root canals using MB or HP + diode laser or SWEEPS alone or in combination were assessed and compared with 5.25% sodium hypochlorite (NaOCl) as the positive control group. Data were analyzed using one-way ANOVA and Tukey’s post hoc test. Results: The MB + diode laser + SWEEPS and HP + diode laser + SWEEPS groups displayed significantly lower amounts of bacteria than either the MB + diode laser or SWEEPS and HP + diode laser or SWEEPS groups (p < 0.001). There was no statistically significant difference detected between the diode laser and SWEEPS groups (p = 0.982). Conclusions: SWEEPS can enhance the photosensitizer distribution in the root canal, leading to effective biofilm removal. This technique could thus help promote the future development of endodontic treatments.

Towards Microbial Food Safety of Sprouts: Photodynamic Decontamination of Seeds

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.

Comparison of the antimicrobial reduction effect of photodynamic inactivation with the addition of chlorophyll and curcumin photosensitizer in Aggregatibacter actinomycetemcomitans and Enterococcus faecalis

Background: Aggregatibacter actinomycetemcomitans and Enterococcus faecalis are pathogenic bacteria of the oral cavity that cause various diseases such as periodontitis and endodontics. These bacteria are easily resistant to antibiotics. Photodynamic inactivation (PDI) is a method of inactivating microorganisms that utilizes light to activate a photosensitizer agent (PS) that produces reactive oxygen species causing cell lysis. Methods: This study used the PDI method with a 405 nm diode laser at various energy density with the addition PS curcumin or chlorophyll Alfalfa, as much as 1.6 mg/ml on A. actinomycetemcomitans and E. faecalis bacteria. Results: The study on E. faecalis bacteria showed that the energy density diode laser irradiation of 1.59 J/cm² gave the percentage of E. faecalis bacteria death 36.7% without PS, 69.30% with the addition of chlorophyll Medicago sativa L and 89.42% with the addition of curcumin. Meanwhile, the bacteria A. actinomycetemcomitans showed that the energy density diode laser irradiation of 1.59 J/cm² gave the percentage of bacterial death 35.81% without PS, 64.39% with the addition of chlorophyll Medicago sativa L and 89.82% with the addition of curcumin. PS was critical to the success of the PDI. Conclusions: The addition of PS curcumin increased the effectiveness of reducing bacteria E. faecalis and A. actinomycetemcomitans compared to chlorophyll Medicago sativa L.

Growth models of fractal interfaces in the description of microorganism colony growth: effect of photodynamic inactivation.

Candida albicans is responsible for most of the nosocomial infections that affect immunocompromised individuals. We investigated the application of eosin in photodynamic inactivation as a strategy in the inhibition of the growth of C. albicans and the morphological variation and growth dynamics in light of fractal theory. The damage caused to fungal structures alters the roughness of the colony, and these changes were described by parameters that were defined by mathematical models. Proliferation of the fungi should be inhibited in the center of the colonies and the analysis of the edges gives an indication about the dynamics of growth and cell reproduction.

In vitro study on the effects of photodynamic inactivation using methyl pheophorbide a, PhotoMed, PhotoCure, and 660 nm diode laser on Candida albicans

Background: Candida albicans (C. albicans) is the most common fungal pathogen that causes clinical infections in humans. This study aimed to evaluate the effects of photodynamic inactivation (PDI) using a 660 nm diode laser along with methyl pheophorbide a, PhotoMed, and PhotoCure as photosensitizer for analyzing the viability of in vitro inactivation of C. albicansMethods: In the PDI group, 20 μL of C. albicans suspension and 20 μL of photosensitizer were inoculated in a 90 mm petri dish (63.6 cm2). The samples were placed in an incubator at 37 °C for 30 min, and then they were irradiated with light (660 nm diode laser, 3 J/cm2). After laser irradiation, the cells were stored for 48 h at 37 °C in an incubator with 5% CO2, and the number of colonies was counted.Results: The highest reduction in the number of colony-forming units per milliliter (CFU/mL) after PDI was observed in the presence of methyl pheophorbide a and PhotoMed, followed by PhotoCure. One-way analysis of variance (ANOVA) demonstrated a significant inhibition (F = 384.717; P < 0.05) for each PDI.Conclusions: In this study, we demonstrated that the application of PDI to C. albicans using methyl pheophorbide a and PhotoMed resulted in 100% death rates.PDI could be a treatment method because conventional antifungals have limited effects, and they may not eliminate C. albicans completely.

The importance of combining methods to assess Candida albicans biofilms following photodynamic inactivation

Background: Methylene blue (MB)-mediated photodynamic inactivation (PDI) has shown good results in killing Candida spp. Although MB solutions are commonly used, new formulations have been designed to improve PDI. However, chemical substances in the formulation may interfere with the PDI outcome. In this sense, different methodologies should be used to evaluate PDI in vitro. Herein, we report different methodologies to evaluate the effects of PDI with an oral formulation (OF) containing 0.005% MB on Candida albicans biofilm. Methods: Biofilms were treated using the MB-OF, with 5 min pre-irradiation time and exposure to a 640 nm LED device (4.7 J/cm2). PDI was evaluated by the XTT reduction test, counting the colony forming units (CFU), a filamentation assay, crystal violet (CV) staining, and scanning electronic microscopy (SEM). Results: PDI was able to reduce around 1.5 log10 CFU/mL, even though no significant differences were noted in metabolic activity in comparison to the control immediately after PDI. A significant decrease in yeast to hyphae transition was observed after PDI, while the biofilm exhibited flattened cells and a reduced number of yeasts in SEM. The CV assay showed increased biomass. Conclusion: MB-OF-mediated PDI was effective in C. albicans biofilms, as it significantly reduced the CFU/mL and the virulence of surviving cells. The CV data were inconclusive, since the OF components interacted with the CV, making the data useless. Taken together, our data suggest that the association of different methods allows complementary responses to assess how PDI mediated by a formulation impacts biofilms.

Bioadhesive Curcumin-Mediated Photodynamic Inactivation and Its Potential to Cause Undesirable Effects on Dental and Restorative Surfaces.

Curcumin-mediated Photodynamic Inactivation (PDI) has shown great potential to disinfect specific sites on tooth enamel but may involve contact with restorative materials. Thus, before use in dentistry, it is necessary to investigate whether the PDI protocol causes undesirable changes in the surfaces of aesthetic restorative materials and dental enamel. This study investigated the effect of PDI mediated by curcumin (CUR) in a liquid crystal precursor system on color stability (ΔE), surface roughness (Ra), and microhardness (kgf) of three different composite resins and bovine dental enamel specimens. The microhardness and roughness readings were performed 60 days after the treatments while the color readings were performed immediately, 24, 48, and 72 h, 7, 14, 21, 30, and 60 days after the treatments. Results showed that CUR mediated-PDI does not seem to have the potential to promote any esthetic or mechanical changes to the surface of tooth enamel and can be applied safely in clinical practice. However, the results on color, roughness, and hardness obtained for composite resins show that some negative effects can be produced, depending on the type of restorative material; more experiments must be performed with different formulations and, perhaps, with lower concentrations of CUR.

Photodynamic Inactivation of Legionella pneumophila Biofilm Formation by Cationic Tetra- and Tripyridylporphyrins in Waters of Different Hardness.

The bacterium Legionella pneumophila is still one of the probable causes of waterborne diseases, causing serious respiratory illnesses. In the aquatic systems, L. pneumophila exists inside free-living amoebae or can form biofilms. Currently developed disinfection methods are not sufficient for complete eradication of L. pneumophila biofilms in water systems of interest. Photodynamic inactivation (PDI) is a method that results in an antimicrobial effect by using a combination of light and a photosensitizer (PS). In this work, the effect of PDI in waters of natural origin and of different hardness, as a treatment against L. pneumophila biofilm, was investigated. Three cationic tripyridylporphyrins, which were previously described as efficient agents against L. pneumophila alone, were used as PSs. We studied how differences in water hardness affect the PSs’ stability, the production of singlet oxygen, and the PDI activity on L. pneumophila adhesion and biofilm formation and in biofilm destruction. Amphiphilic porphyrin showed a stronger tendency for aggregation in hard and soft water, but its production of singlet oxygen was higher in comparison to tri- and tetracationic hydrophilic porphyrins that were stable in all water samples. All three studied porphyrins were shown to be effective as PDI agents against the adhesion of the L. pneumophila to polystyrene, against biofilm formation, and in the destruction of the formed biofilm, in their micromolar concentrations. However, a higher number of dissolved ions, i.e., water hardness, generally reduced somewhat the PDI activity of all the porphyrins at all tested biofilm growth stages.

Photoinactivation of mycobacteria to combat infection diseases: current state and perspectives.

The spread of multi-drug-resistant bacterial strains causing serious infectious diseases dictates the development of new approaches to combat these diseases. In addition to drug resistance, the important causative agent of tuberculosis (Mycobacterium tuberculosis (Mtb)) is able to persist asymptomatically in individuals for many years, causing latent forms of tuberculosis. In such a dormant state, Mtb cells are also resistant to known antibiotics. In this regard, photodynamic inactivation (PDI) could be an effective alternative to antibiotics as its action is based on the generation of active forms of oxygen independently on the presence of specific antibiotic targets, thereby inactivating both drug-resistant and dormant bacteria. In this review, we summarise examples of the application of PDI for the elimination of representatives of the genus Mycobacteria, both in vitro and in vivo. According to published results, including photosensitisers in the PDI regime results in a significantly higher lethal effect. Such experiments were mainly performed using chemically synthesised photosensitisers, which need to be transported to the areas of bacterial infections, limiting PDI usage by surface (skin) diseases. In this regard, endogenous photosensitisers (mainly porphyrins) could be used to solve the problem of transportation. In vitro experiments demonstrate the effective application of PDI for mycobacteria, including Mtb, using endogenous porphyrins; the intracellular contents of these substances can be elevated by administration of 5-aminolevulenic acid, a precursor of porphyrin synthesis. Photodynamic inactivation can also be used for dormant mycobacteria, which are characterised by high levels of endogenous porphyrins. Thus, PDI can effectively eliminate drug-resistant mycobacteria. The exploitation of modern light-transmitting techniques opens new possibilities to use PDI in clinical settings.Key points •The potential effects of photodynamic inactivation of mycobacteria are critically reviewed. •Approaches to photoinactivation of mycobacteria using exogenous and endogenous photosensitisers are described. •Prospects for the use of photodynamic inactivation in the treatment of tuberculosis are discussed.

PH interferes in photoinhibitory activity of curcumin nanoencapsulated with pluronic® P123 against Staphylococcus aureus.

Microbial contamination control is a public health concern and challenge for the food industry. Antimicrobial technologies employing natural agents may be useful in the food industry for these purposes. This work aimed to investigate the effect of photodynamic inactivation using curcumin in Pluronic® P123 nanoparticles (Cur/P123) at different pH and blue LED light against Staphylococcus aureus. Bacterial photoinactivation was conducted using different photosensitizer concentrations and exposure times at pH 5.0, 7.2 and 9.0. A mixture design was applied to evaluate the effects of exposure time (dark and light incubation) on the photoinhibitory effect. S. aureus was completely inactivated at pH 5.0 by combining low concentrations of Cur/P123 (7.80-30.25 μmol/L) and light doses (6.50-37.74 J/cm2). According to the mathematical model, dark incubation had low significance in bacterial inactivation at pH 5.0 and 9.0. No effect in bacterial inactivation was observed at pH 7.2. Cur/P123 with blue LED was effective in inactivating S. aureus. The antimicrobial effect of photodynamic inactivation was also pH-dependent.

Photodynamic inactivation of Streptococcus mutans by curcumin in combination with EDTA

ObjectiveThis study aimed to test the efficacy of photodynamic inactivation (PDI) mediated by curcumin with EDTA against Streptococcus mutans in planktonic suspension using blue LED light. MethodsAntibacterial activity of curcumin and EDTA was evaluated by determination of their minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC). The fractional inhibitory concentration index (FICI) was used to estimate the synergistic effect of various combination ratios of curcumin and EDTA against S. mutans. Cultures of S. mutans (18 h, 37 °C, 5% C02) were prepared to test the effect of curcumin-mediated PDI (50 μM and 500 μM) with or without 0.4% EDTA and 40 s of light-activation with blue light. EDTA and each concentration of curcumin were also tested individually. Chlorhexidine (0.2%), was used as positive control. Planktonic suspensions were also analyzed by viable colony counts (VCC), confocal laser scanning microscopy (CLSM), transmission electron microscopy (TEM), and polymerase chain reaction (PCR). ResultsThe MIC values of curcumin and EDTA were 5 mM and 0.125% respectively. FICI showed a synergistic interaction between curcumin and EDTA. All the combinations with curcumin and blue LED light resulted in a complete inactivation of the S. mutans and CLSM confirms these results, TEM showed morphological changes produced by the PDI. No damage on DNA structure was detected by PCR. SignificanceCurcumin-mediated PDI with EDTA using a blue light, shows a strong inhibitory effect against S. mutans in planktonic culture. Because of the unspecific target mechanism, it could be a promising technique for disinfection of dental tissues.

Enhanced photodynamic inactivation for Gram-negative bacteria by branched polyethylenimine-containing nanoparticles under visible light irradiation

Antibiotic pollution has been a serious global public health concern in recent years, photodynamic inactivation is one of the most promising and innovative methods for antibacterial applications that avoids antibiotic abuse and minimizes risks of antibiotic resistance. However, limited by the weak interaction between the photosensitizers and Gram-negative bacteria, the effect of photodynamic inactivation cannot be fully exerted. Herein, photosensitizer chlorin e6-loaded polyethyleneimine-based micelle was constructed. The synergy of electrostatic and hydrophobic interactions between the nanoparticles and the bacterial surface promoted the anchoring of nanoparticles onto the bacteria, resulting in enhanced photoinactivation activities on Gram-negative bacteria. As expected, an eminent antibacterial effect was also observed on the Gram-positive bacteria Staphylococcus aureus. The cellular uptake results showed that photosensitizer was firmly anchored to the bacterial cell surface of Escherichia coli or Staphylococcus aureus by the introduction of branched polyethylenimine-containing nanoparticles. The light-triggered generation of reactive oxygen species, mainly singlet oxygen, from the membrane-bound nanoparticles caused irreversible damage to the bacterial outer membrane, achieving enhanced bactericidal efficiency than free photosensitizer. The study would provide an efficient and promising antimicrobial alternative to prevent overuse of antibiotics and have enormous potential for human healthcare and the environment remediation.

Effect of Photodynamic Inactivation against Dormant Forms and Active Growing Cells of Mycobacterium smegmatis

A potential method for the destruction of dormant Mycobacteriumsmegmatis (Msm) mycobacteria via photodynamic inactivation (PDI) using their ability to store endogenous porphyrins during the transition to a dormant state was studied. The dormant Msm cells were obtained under gradual acidification of the growth medium in the stationary phase for 14 days at different concentrations of ferric ions and the presence of 5-aminolevulinic acid in the growth medium. Cells were exposed to light at a wavelength of 532 nm emitted by an LLD10 laser for 5–60 min. It was shown that an increase in the coproporphyrin concentration in M. smegmatis after 6 days of growth correlated with the onset of a decrease in metabolic activity in cells and the formation of ovoid dormant forms. The dormant bacteria were sensitive to PDI and were destroyed after 15–30 min of illumination, in contrast to active cells. In the presence of 5-aminolevulinic acid and an optimal concentration of ferric ions (0.5 mg/L), there was an increase in the production of uroporphyrin in both active and dormant mycobacteria, which was accompanied by an increase in sensitivity to the action of PDI. The results demonstrate the promise of PDI for the destruction of dormant mycobacteria and the fight against latent tuberculosis.