Carbon Nanomaterials and LED Irradiation as Antibacterial Strategies against Gram-Positive Multidrug-Resistant Pathogens.

BACKGROUND: Due to current antibiotic resistance worldwide, there is an urgent need to find new alternative antibacterial approaches capable of dealing with multidrug-resistant pathogens. Most recent studies have demonstrated the antibacterial activity and non-cytotoxicity of carbon nanomaterials such as graphene oxide (GO) and carbon nanofibers (CNFs). On the other hand, light-emitting diodes (LEDs) have shown great potential in a wide range of biomedical applications. METHODS: We investigated a nanotechnological strategy consisting of GO or CNFs combined with light-emitting diod (LED) irradiation as novel nanoweapons against two clinically relevant Gram-positive multidrug-resistant pathogens: methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant Staphylococcus epidermidis (MRSE). The cytotoxicity of GO and CNFs was studied in the presence of human keratinocyte HaCaT cells. RESULTS: GO or CNFs exhibited no cytotoxicity and high antibacterial activity in direct contact with MRSE and MRSA cells. Furthermore, when GO or CNFs were illuminated with LED light, the MRSE and MRSA cells lost viability. The rate of decrease in colony forming units from 0 to 3 h, measured per mL, increased to 98.5 ± 1.6% and 95.8 ± 1.4% for GO and 99.5 ± 0.6% and 99.7 ± 0.2% for CNFs. CONCLUSIONS: This combined antimicrobial approach opens up many biomedical research opportunities and provides an enhanced strategy for the prevention and treatment of Gram-positive multidrug-resistant infections.

Photodynamic inactivation of bacteria to decolonize meticillin-resistant Staphylococcus aureus from human skin.

BACKGROUND: To prevent infections that arise from the skin surface it is necessary to decolonize human skin prior to any proposed treatment or surgical intervention. Photodynamic inactivation of bacteria (PIB) uses cationic photosensitizers that attach to the surface of bacteria, generate reactive oxygen species on light irradiation and thereby kill bacteria via oxidative mechanisms. OBJECTIVES: To evaluate the potential and the safety of PIB for decolonization of bacteria from skin. METHODS: PIB with the new photosensitizer SAPYR [2-((4-pyridinyl)methyl)-1H-phenalen-1-one chloride] was initially tested against different bacterial species in vitro. Then, ex vivo porcine skin samples were used as a model for decolonization of different bacteria species. The numbers of viable bacteria were quantified and the mitochondrial activity of skin cells was histologically analysed (using nitroblue tetrazolium chloride, NBTC). The same procedure was performed for human skin and meticillin-resistant Staphylococcus aureus (MRSA). RESULTS: The in vitro studies showed a 5 log10 reduction of all tested bacterial species. On ex vivo porcine skin samples, PIB reduced the viability of all tested bacterial species by at least 3 log10 steps. On human skin samples ex vivo, PIB reduced the number of viable MRSA by maximal 4·4 log10 steps (1000 µmol L-1 SAPYR, incubation time 10 min, 60 J cm-2 ). NBTC staining showed normal mitochondrial activity in skin cells after all PIB modalities. CONCLUSIONS: The results of this study show that PIB can effectively and safely kill bacteria like MRSA on the skin surface and might have the potential of skin decolonization in vivo.

Effects of gamma-irradiation on antibiotic resistance and diagnostic molecular markers of methicillin-resistant Staphylococcus aureus in Egyptian cancer patients.

Purpose: This in-vitro study aimed to assess in 120 [40 community-acquired (CA-MRSA) & 80 hospital-acquired (HA-MRSA)] isolates from cancer patients whether the transmissible staphylococcal cassette chromosome mec (SCCmec) typing, and the Panton-Valentine leukocidin (PVL) virulence genes detection could be employed as tools for molecular diagnostic purposes to distinguish both methicillin-resistant Staphylococcus aureus (MRSA) categories in radiotherapy treated cancer patients.Materials and methods: SCCmec typing was determined by the combination of the type of the cassette chromosome recombinase genes (ccr) gene complex and the class of the methicillin resistance (mec) gene complex. Besides, a rapid slide latex agglutination test (LAT) and antibiotic resistance spectrum determination before and after irradiation were performed.Results: In the strict sense, with the effect of irradiation; the presence of SCCmec subtypes IVa (22.5% vs. 10.0%), b (47.5% vs. 25.0%), & d (7.5 vs. 2.5%) or type V (15.0% vs. 7.5%) genetic elements and PVL genes (p < .001) were not proved as a signature for CA-MRSA. While, the larger SCCmec types II, and III elements were not detected in 14, and 19 from the 38, and 36 typed HA-MRSA isolates (p < .001), respectively. Remarkable effects on class A & class B mec gene complex and type2, type 3 & type 5 ccr gene complex and an increase in agglutination reaction strength in response to gamma irradiation external stimulus were observed.Conclusions: Different heterogeneous genetic composition with upregulation mecA gene expression was detected after irradiation in the HA- MRSA studied population. CA-MRSA showed remarkable ability to acquire multi-antibiotic resistance after irradiation and propose a novel paradigm for future chemotherapy against the multi-resistant pathogens whose proliferation especially among immunocompromised cancer patients is on the increase.

Photothermal inactivation of methicillin-resistant Staphylococcus aureus: anti-biofilm mediated by a polypyrrole-carbon nanocomposite.

Widespread resistance to antibiotics amongst pathogens has become a tremendous challenge of high morbidity and mortality rates which increases the needs to exploring novel methods of treatment. An efficient antimicrobial procedure to root out pathogenic bacteria is photothermal therapy. In this study, antimicrobial effects of a polypyrrole-carbon nanocomposite (PPy-C) upon laser irradiation in order to destroy the pathogenic gram-positive bacterium, methicillin-resistant Staphylococcus aureus (MRSA) were assessed. The bacterial cells were incubated with 500, 750 and 1000 µg ml-1 concentrations of PPy-C and irradiated with an 808-nm laser at a power density of 1.0 W cm-2. To indicate the biocompatibility and toxic effect of the nanocomposite without and with laser irradiation, the authors counted the number of CFUs and compared it to an untreated sample. Antibacterial mechanisms of PPy-C were assessed through temperature increment, reactive oxygen species production, and protein and DNA leakages. Photothermal heating assay showed that 26°C temperature increases in the presence of 1000 µg ml-1 PPy-C led to >98% killing of MRSA. Furthermore, 20 min radiation of near-infrared light to PPy-C in different concentrations indicated destruction and reduction in the MRSA biofilm formation. Therefore, PPy-C was introduced as a photothermal absorber with a bactericidal effect in MRSA.

Bactericidal effect of photodynamic inactivation against methicillin-resistant and methicillin-susceptible Staphylococcus aureus is strain-dependent.

Staphylococcus aureus is one of the most important etiological factors responsible for nosocomial infections. Some of them may be life-threatening, especially in the case of immuno-compromised patients, causing bacteremia, endocarditis, sepsis or toxic-shock syndrome. Their multiresistance to antibiotics produces many therapeutic problems, and for this reason the development of a method alternative to antibiotic therapy is needed. It seems that photodynamic inactivation (PDI) may be an effective and alternative therapeutic option against both methicillin resistant (MRSA) and methicillin sensitive (MSSA) S. aureus strains. The aim of this study was to analyze the bactericidal effect of the PDI against 40 clinical MRSA and 40 MSSA clinical strains that were isolated from patients hospitalized in the Provincial Hospital in Gdansk. The ATCC strain 25904 has been used as a reference. Photodynamic inactivation by means of protoporphyrin diarginate as a photosensitizer was examined. It was observed that the bactericidal effect of the PDI was strain-dependent and ranged from 0 to 3 log(10)-unit reduction in viable counts. The mechanism underlying such a phenomenon is still not understood. Nevertheless, the correlation between the biofilm production ability and different strains response to PDI has been observed.

Killing of methicillin-resistant Staphylococcus aureus by low-power laser light.

The purpose of this study was to determine whether a methicillin-resistant strain of Staphylococcus aureus (MRSA) could be sensitised by toluidine blue O (TBO) to killing by light from a low-power helium/neon (HeNe) laser. Suspensions containing c. 10(10) cfu of MRSA were irradiated with light from a 35 mW HeNe laser (energy dose: 0.5-2.1 J) in the presence of TBO (1.6-12.5 micrograms/ml) and the survivors were enumerated. The kills attained depended on both the light energy dose and concentration of TBO employed. A 4.47 log10 reduction in the viable count was achieved with a TBO concentration of 12.5 micrograms/ml and a light dose of 2.1 J (energy density 43 J/cm2). MRSA were susceptible to killing by the laser light within 30 s of exposure to the TBO. The results of this study have demonstrated that MRSA can be rapidly sensitised by TBO to killing by HeNe laser light and that killing depends on the light energy dose and sensitiser concentration.

Ultraviolet light C in the treatment of chronic wounds with MRSA: a case study.

The prevalence of antibiotic-resistant bacteria such as methicillin-resistant Staphylococcus aureus is rapidly increasing in healthcare facilities and spreading to the community. Methicillin-resistant S. aureus colonize the skin and open wounds and can interfere with wound healing. Recent studies have shown that ultraviolet light C can kill antibiotic-resistant strains of bacteria such as methicillin-resistant S. aureus in both laboratory cultures and animal tissue. This clinical report describes the effects of ultraviolet light C on wound bioburden and closure in three people with chronic ulcers infected with methicillin-resistant S. aureus. In all three patients, ultraviolet light C treatment reduced wound bioburden and facilitated wound healing. Two patients had complete wound closure following 1 week of ultraviolet light C treatment. This case study suggests that ultraviolet light C is a promising adjunctive therapy for chronic wounds containing antibiotic-resistant bacteria such as methicillin-resistant S. aureus.

Surface plasmon resonance-induced photoactivation of gold nanoparticles as bactericidal agents against methicillin-resistant Staphylococcus aureus.

Systemic infections caused by methicillin-resistant Staphylococcus aureus (MRSA) and other bacteria are responsible for millions of deaths worldwide, and much of this mortality is due to the rise of antibiotic-resistant organisms as a result of natural selection. Gold nanoparticles synthesized using the standard wet chemical procedure were photoexcited using an 808 nm 2 W laser diode and further administered to MRSA bacteria. Flow cytometry, transmission electron microscopy, contrast phase microscopy, and fluorescence microscopy combined with immunochemical staining were used to examine the interaction of the photoexcited gold nano-particles with MRSA bacteria. We show here that phonon-phonon interactions following laser photoexcitation of gold nanoparticles exhibit increased MRSA necrotic rates at low concentrations and short incubation times compared with MRSA treated with gold nanoparticles alone. These unique data may represent a step forward in the study of bactericidal effects of various nanomaterials, with applications in biology and medicine.

Photodynamic inactivation of Staphylococcus aureus and methicillin-resistant Staphylococcus aureus with Ru(II)-based type I/type II photosensitizers.

BACKGROUND: The introduction of new disinfection and sterilization methods, such as antimicrobial photodynamic therapy, is urgently needed for the healthcare industry, in particular to address the pervasive problem of antibiotic resistance. This study evaluated the efficacy and the mechanisms of photodynamic antimicrobial chemotherapy (PACT), also known as photodynamic inactivation (PDI) of microorganisms, induced by novel Ru(II)-based photosensitizers against Staphylococcus aureus and methicillin-resistant S. aureus strains. METHODS: The photodynamic antibacterial effects of a new class of Ru(II)-based photosensitizers (TLD1411 and TLD1433) were evaluated against a strain of S. aureus (ATCC 25923) and a methicillin-resistant strain of S. aureus (MRSA, ATCC 33592). Bacterial samples were dosed with a range of photosensitizer concentrations (0.3-12 µM) and exposed to 530 nm light (90J cm(-2)) in normoxic conditions (ambient atmosphere) and in hypoxic conditions (0.5% O2). RESULTS: Both photosensitizers exerted photodynamic inactivation (PDI) of the microorganisms in normoxia, and this activity was observed in the nanomolar regime. TLD1411 and TLD1433 maintained this PDI potency under hypoxic conditions, with TLD1433 becoming even more active in the low-oxygen environment. CONCLUSION: The observation of activity in hypoxia suggests that there exists an oxygen-independent, Type I photoprocess for this new class of compounds in addition to the typical Type II pathway mediated by singlet oxygen. The intrinsic positive charge of the Ru(II) metal combined with the oxygen independent activity demonstrated by this class of photosensitizers presents a new strategy for eradicating both gram-positive and gram-negative bacteria regardless of oxygenation level.