The combination of antimicrobial photocatalysis and antimicrobial photodynamic therapy to eradicate the extensively drug-resistant colistin resistant Acinetobacter baumannii.

BACKGROUND: The appearance of bacterial resistance to antibiotics a are major health problems. Antimicrobial photodynamic therapy (aPDT) has emerged as an adjunctive treatment for drug-resistant infections. Titanium dioxide (TiO2) and zinc oxide (ZnO) nanoparticles have recently been proposed as effective carriers for photosensitizers. The present study was designed to evaluate the antibacterial activity of ZnO and TiO2-nanoparticles accompanying aPDT and antimicrobial photocatalysis approaches against extensively drug-resistant colistin resistant Acinetobacter baumannii (XDR-CO-Ab). MATERIALS AND METHODS: A XDR-CO-Ab isolated from burn-wound infection was used in this study. The bacterial suspension was exposed to TBO (Toluidine blue O), TiO2, and ZnO-nanoparticles in different groups. Ultraviolet A (UVA) (320 nm) and blue laser light (405 nm) were used to activate the photosensitizer and nanoparticles. The bacterial viability was then determined by counting the number of colonies forming units (CFUs)/mL. RESULTS: Using both aPDT and antimicrobial photocatalysis methods simultaneously, it had an amazing result; so that 99.32 % of XDR-CO-Ab was killed. According to the results, the bactericidal effects of activated TBO using UVA and laser lights was 5.1 % more than TBO was excited with laser light alone. CONCLUSIONS: It was found that the combination of photocatalyst (TiO2+ZnO-nanoparticles) and TBO as photosensitizer at two-wavelength irradiation with blue laser and UVA lights have been shown to have significant antimicrobial effects.

Photoelimination Potential of Chitosan Nanoparticles-Indocyanine Green Complex Against the Biological Activities of Acinetobacter baumannii Strains: A Preliminary In Vitro Study in Burn Wound Infections.

Introduction: Acinetobacter baumannii strains are important agents causing serious nosocomial infections including soft-tissue and skin infections in patients with burn wounds which have become resistant to several classes of antibiotics. Antimicrobial photodynamic therapy (aPDT) as an alternative antimicrobial procedure is suggested for the treatment of these kinds of infections. The aim of the current study is to evaluate the antibacterial and anti-biofilm efficiency of aPDT by the utilization of an improved form of indocyanine green (ICG) which is encapsulated in chitosan nanoparticles (NCs@ICG). Methods: NCs@ICG were synthesized and confirmed by the scanning electron microscope (SEM). aPDT was performed using NCs@ICG with an 810 nm wavelength of the diode laser at the fluency of 31.2 J/cm2 on 50 A. baumannii strains isolated from burn wounds. The antibacterial and antibiofilm potential of NCs@ICG-aPDT was determined via the colony forming unit (CFU)/mL and crystal violet assays, respectively. In addition, microbial biofilm degradation was evaluated by the SEM. Results: According to the results, NCs@ICG-aPDT showed a significant reduction of 93.2% on the CFU/ mL of planktonic A. baumannii strains compared to the control group (untreated group; P < 0.05). In addition, the biofilm formation of A. baumannii strains was significantly reduced by 55.3% when the bacteria were exposed to NCs@ICG-aPDT (P < 0.05). In contrast, NCs@ICG, ICG, and the diode laser alone were not able to inhibit the CFU/mL and biofilm of A. baumannii strains (P > 0.05). Based on the results of SEM images, NCs@ICG-aPDT disrupted the biofilm structure of A. baumannii strains more than other groups. Conclusion: NCs@ICG-aPDT demonstrates a promising treatment candidate for exploitation in wound infections against both planktonic and biofilm forms of A. baumannii strains.

The effect of antimicrobial photodynamic therapy against virulence genes expression in colistin-resistance Acinetobacter baumannii.

BACKGROUND AND AIMS: The emergence of drug-resistant infections is a global problem. Acinetobacter baumannii has attracted much attention over the last few years because of resistance to a wide range of antibiotics. Applying new non-antibiotic methods can save lives of many people around the world. Antimicrobial photodynamic therapy (aPDT) technique can be used as a new method for controlling the infections. In this study we investigated the effect of aPDT on the expression of pathogenic genes in colistin-resistance A. baumannii isolated from a burn patient. MATERIALS AND METHODS: The suspension of colistin-resistance A. baumannii was incubated with 0.01 mg/ml of toluidine blue O (TBO) in the dark; then the light emitting diode device with a wavelength of 630 ± 10 nm and output intensity of 2000-4000 mW /cm2 was irradiated to the suspension at room temperature. Subsequently, after the aPDT, genes expression of ompA and pilZ was investigated by using real-time polymerase chain reaction technique. RESULT: Among the genes studied, the transcript of the ompA gene after aPDT was increased significantly in comparison with control groups (P < 0.05). Whereas, there was no remarkable different in pilZ gene expression (P > 0.05). CONCLUSIONS: It can be concluded from the results that the ompA as an outer membrane of A. baumannii is degraded after exposing aPDT and it will probably be done the penetration of antibiotics into cells of this bacterium easily.

Antimicrobial activity of photodynamic therapy in combination with colistin against a pan-drug resistant Acinetobacter baumannii isolated from burn patient.

Nosocomially-acquired multi-, extensively-, and pandrug resistant (MDR, XDR, and PDR) strains of microorganisms such as Acinetobacter baumannii remain a serious cause of infection and septic mortality in burn patients. Treatment of patients with nosocomial burn wound infections is often complicated by drug-resistant strains of A. baumannii. Today, many researchers are focusing on the investigation of novel non-antibiotic strategies such as photodynamic therapy (PDT). We report a new PDT strategy that suppresses colistin resistance in PDR A. baumannii by interfering with the expression of a pmrA/pmrB two-component system. In the current study, A. baumannii with a PDR feature isolated from a burn patient was used as a test strain. PDT was carried out using toluidine blue O (TBO) and light-emitting diode (LED) as a photosensitizer and radiation source, respectively. The antimicrobial susceptibility profiles were assessed for cells surviving PDT. The effects of sub-lethal PDT (sPDT) on the expression of the pmrA/pmrB two-component signal transduction system were evaluated by real-time quantitative reverse transcription PCR. Results of drug susceptibly testing (DST) in LED and TBO groups separately showed that the bacteria were resistant to all tested antibiotics, while the DST result of the LED+TBO group showed highly declining bacterial growth when compared with the control group. Reduction in the expression of pmrA and pmrB was observed in the treated strains after sPDT. This represents the first conclusive example of a direct role for the PDT in breaking antibiotic resistance by directly modulating two-component system activity.

Modulation of virulence in Acinetobacter baumannii cells surviving photodynamic treatment with toluidine blue.

INTRODUCTION: Widespread resistance to antimicrobial agents has led to a dearth of therapeutic choices in treating Acinetobacter baumannii infections, leading to new strategies for treatment being needed. We evaluated the effects of photodynamic therapy (PDT) as an alternative antimicrobial modality on the virulence features of cell-surviving PDT. MATERIALS AND METHODS: To determine the sublethal PDT (sPDT), a colistin-resistant, extensively drug-resistant A. baumannii (CR-XDR-AB) clinical isolate and A. baumannii and ATCC 19606 strains, photosensitized with toluidine blue O (TBO), were irradiated with light emitting diodes, following bacterial viability measurements. The biofilm formation ability, outer membrane (OM) integrity, and antimicrobial susceptibility profiles were assessed for cell-surviving PDT. The effects of sPDT on the expression of virulent genes were evaluated by real-time quantitative reverse transcription PCR (qRT-PCR). RESULTS: sPDT resulted in the reduction of the biofilm formation capacity, and its metabolic activity in strains. The OM permeability and efflux pump inhibition of the sPDT-treated CR-XDR-AB cells were increased; however, there was no significant change in OM integrity in ATCC 19606 strain after sPDT. sPDT reduced the minimum inhibitory concentrations of the most tested antimicrobials by ≥2-fold in CR-XDR-AB. lpsB, blsA, and dnaK were upregulated after the strains were treated with sPDT; however, a reduction in the expression of csuE, epsA, and abaI was observed in the treated strains after sPDT. CONCLUSION: The susceptibility of CR-XDR-AB to a range of antibiotics was enhanced following sPDT. The virulence of strains is reduced in cells surviving PDT with TBO, and this may have potential implications of PDT for the treatment of A. baumannii infections.