Antimicrobial Photodynamic Therapy against Escherichia coli and Staphylococcus aureus Using Nanoemulsion-Encapsulated Zinc Phthalocyanine.

Multidrug-resistant microorganisms have become a significant public health threat, and traditional antibiotics are becoming ineffective. Photodynamic therapy (PDT) is a promising alternative that utilizes photosensitizers and light to produce Reactive Oxygen Species (ROS) that can kill microorganisms. Zinc phthalocyanine (ZnPc) is a promising photosensitizer due to its strong affinity for encapsulation in nanoemulsions and its antimicrobial properties. In this study, nanoemulsion was prepared using Miglyol 812N, a surfactant, and distilled water to dissolve hydrophobic drugs such as ZnPc. The nanoemulsion was characterized by its particle size, polydispersity index, Transmission Electron Microscope and Zeta potential, and the results showed that it was an efficient nanocarrier system that facilitated the solubilization of hydrophobic drugs in water. The use of ZnPc encapsulated in the nanoemulsion produced through the spontaneous emulsification method resulted in a significant reduction in cell survival percentages of gram-positive Staphylococcus aureus and gram-negative Escherichia coli by 85% and 75%, respectively. This may be attributed to the more complex cell membrane structure of E. coli compared to S. aureus. This demonstrates the potential of nanoemulsion-based PDT as an effective alternative to traditional antibiotics for treating multidrug-resistant microorganisms.

Photothermal versus photodynamic treatment for the inactivation of the bacteria Escherichia coli and Bacillus cereus: An in vitro study.

The widespread occurrence of microbial pathogens, including multidrug-resistant (MDR) bacteria, has ignited research efforts to discover alternative strategies to combat infections in patients. Recently, photodynamic therapy (PDT) and photothermal therapy (PTT) have been proposed for the inactivation of pathogens. Although PDT and PTT are very promising antipathogenic tools, further effort is needed to determine their real impact on pathogens apart from the effects of individual elements involved in the photodynamic/photothermal processes, i.e., light, photosensitizers (PSs), and nanoparticles. Accordingly, in the current study, toluidine blue O (TBO) and gold nanoparticles (GNP) were used as generators of reactive oxygen species (ROS) and hyperthermia in the presence of light, respectively. Escherichia coli (E. coli) and Bacillus cereus (B. cereus) bacteria were chosen as examples of gram-negative and gram-positive bacteria, respectively. Before the bactericidal activity of PDT was assessed, the aggregation of TBO and its effect on the growth of both strains of bacteria were studied. Additionally, E. coli and B. cereus were exposed to a range of doses of 633 nm helium-neon laser light to investigate its effect. In a separate set of experiments, the bactericidal activity of PTT was assessed after the effects of GNP and green light (530 nm) had been assessed. The results showed that PDT and PTT should be considered useful tools for bacterial eradication even when the light, PSs, and nanoparticles are each used at doses safe for bacterial growth. Moreover, different photodynamic responses were observed for E. coli and B. cereus, and light from a 633 nm laser and a 530 nm light-emitting diode (LED) showed disparate responses when applied alone to both bacteria.

Green synthesis and biological activity of silver-curcumin nanoconjugates.

AIM: There is an urgent need to develop alternative antimicrobial agents and, one of which is via the use of nanotechnology. Green synthetic routes are recently being replaced for nanoparticles preparation. Methods results: Silver-curcumin nanoconjugates (Ag-CurNCs) were prepared in an eco-friendly method. The prepared nanomaterials were characterized and the photostability was studied under the influence of UV irradiation. Results showed that, the conjugation between curcumin and silver in the nanoform improve the photostability of curcumin. Cytotoxicity was studied on different skin cell lines, and antibacterial activity was investigated against Escherichia coli. Results revealed the antibacterial activity of the prepared nanoconjugates (Ag-CurNCs) with minimal toxicity to skin cells. CONCLUSION: Silver nanoparticles improve the photostability and antibacterial activity of curcumin, while curcumin helps in preparing biocompatible silver nanoparticles.

Fluorescence-based CdTe nanosensor for sensitive detection of cytochrome C.

Cytochrome c (Cyt c) is commonly used as intrinsic biomarker for several characteristics of the cell such as respiration, energy level and apoptosis. In the present study a simple colorimetric sensor should be developed and tested for the real-time detection of Cyt c in living cells. We synthesized cadmium telluride quantum dots (CdTe QDs) capped with thioglycolic acid (TGA) as a fluorometric Cyt c nanosensor. The synthesized TGA/CdTe QDs nanosensor was characterized by Fourier transform infrared spectroscopy, transmission electron microscopy, and absorption as well as fluorescence spectrophotometry. We investigated the developed TGA/CdTe QDs sensor with regard to its applicability in the fluorometric detection of Cyt c. Results showed that the TGA/CdTe QDs could be used as a sensitive fluorescence probe for the quantification of different concentrations of Cyt c ranging from 0.5 - 2.5µM. Increased binding of QDs to Cyt c results in decreasing fluorescence. The fluorescence of the QDs is inversely correlated to the Cyt c concentration. Based on these data, a standard curve up to 2.5µM Cyt c was established. Moreover, the developed nanosensor was applied in different concentrations on primary human dermal fibroblasts. Results showed that TGA/CdTe QDs were taken up by cells and could be visualized by fluorescence microscopy. Quantification of Cyt c within living cells via QDs is, however, influenced by various factors such as cell damage, QD aggregation or the level of reactive oxygen species, which have to be taken into account.

Exploiting biosynthetic gold nanoparticles for improving the aqueous solubility of metal-free phthalocyanine as biocompatible PDT agent.

Increasing the limit of dispersion of metal-free phthalocyanine (H2Pc) in an aqueous medium using biosynthetic gold nanoparticles for photodynamic therapy (PDT) is investigated. Gold nanoparticles (Au NPs) are biosynthesized in one step using Potatoes (Solanum tuberosum) extract and are characterized by UV/VIS spectrophotometry, Fourier transformer infrared spectroscopy (FTIR), and transmission electron microscopy (TEM). The metal-free phthalocyanine is conjugated to the surface of the gold nanoparticles in a side to side regime through the secondary amine groups of H2Pc. The clear violet solution of phthalocyanine-gold (Pc-Au) nanoconjugates is investigated by UV-VIS, FTIR and TEM techniques. Disappearance of the absorption band of the secondary amine in the Pc-Au nanoconjugates compared to that of the parent H2Pc, and detection of the absorption band of H2Pc in the aqueous medium confirmed the dispersion of H2Pc and consequently the loading of H2Pc on the surface of Au NPs. The cytotoxic effect of biosynthetic gold nanoparticles and Pc-Au nanoconjugates compared to chemically synthesized gold nanoparticles on buffalo epithelial cells has been studied in vitro. Interestingly, the results showed that the biosynthetic Au NPs as well as Pc-Au nanoconjugates have no effect on buffalo epithelial cells viability, which indicating their biocompatibility contrary to the chemically synthesized Au NPs. This work will open the door, for the first time, for using H2Pc suspended in water for PDT and other phototherapeutic applications.

A new biocompatible nanocomposite as a promising constituent of sunscreens.

Skin naturally uses antioxidants to protect itself from the damaging effects of sunlight. If this is not sufficient, other measures have to be taken. Like this, hydroxyapatite has the potential to be applied as an active constituent of sunscreens since calcium phosphate absorbs in the ultraviolet region (UV). The objective of the present work was to synthesize a hydroxyapatite-ascorbic acid nanocomposite (HAp/AA-NC) as a new biocompatible constituent of sunscreens and to test its efficiency with skin cell models. The synthesized HAp/AA-NC was characterized by Fourier transform infrared spectroscopy, transmission electron microscopy, absorption spectrophotometry and X-ray diffraction analysis. The protective effect of the construct was tested with respect to viability and intracellular reactive oxygen species (ROS) generation of primary human dermal fibroblasts (SKIN) and human epidermal keratinocytes (HaCaT). Both cell lines were irradiated with UV light, λmax=254 nm with a fluence of 25 mJ cm(-2) to mimic the effect of UV radiation of sunlight on the skin. Results showed that HAp/AA-NC had a stimulating effect on the cell viability of both, HaCaT and SKIN cells, relative to the irradiated control. Intracellular ROS significantly decreased in UV irradiated cells when treated with HAp/AA-NC. We conclude that the synthesized HAp/AA-NC have been validated in vitro as a skin protector against the harmful effect of UV-induced ROS.

Antimicrobial blue light inactivation of Pseudomonas aeruginosa by photo-excitation of endogenous porphyrins: In vitro and in vivo studies.

Pseudomonas aeruginosa is among the most common pathogens that cause nosocomial infections and is responsible for about 10% of all hospital-acquired infections. In the present study, we investigated the potential development of tolerance of P. aeruginosa to antimicrobial blue light by carrying 10 successive cycles of sublethal blue light inactivation. The high-performance liquid chromatographic (HPLC) analysis was performed to identify endogenous porphyrins in P. aeruginosa cells. In addition, we tested the effectiveness of antimicrobial blue light in a mouse model of nonlethal skin abrasion infection by using a bioluminescent strain of P. aeruginosa. The results demonstrated that no tolerance was developed to antimicrobial blue light in P. aeruginosa after 10 cycles of sub-lethal inactivation. HPLC analysis showed that P. aeruginosa is capable of producing endogenous porphyrins in particularly, coproporphyrin III, which are assumed to be responsible for the photodynamic effects of blue light alone. P. aeruginosa infection was eradicated by antimicrobial blue light alone (48 J/cm(2) ) without any added photosensitizer molecules in the mouse model. In conclusion, endogenous photosensitization using blue light should gain considerable attention as an effective and safe alternative antimicrobial therapy for skin infections. Lasers Surg. Med. 48:562-568, 2016. © 2016 Wiley Periodicals, Inc.

Microplate assay for screening the antibacterial activity of Schiff bases derived from substituted benzopyran-4-one.

Schiff bases (SB(1)-SB(3)) were synthesized from the condensation of 6-formyl-7-hydroxy-5-methoxy-2-methylbenzopyran-4-one with 2-aminopyridine (SB(1)), p-phenylenediamine (SB(2)) and o-phenylenediamine (SB(3)), while Schiff bases (SB(4)-SB(6)) were synthesized by condensation of 5,7-dihydroxy-6-formyl-2-methylbenzopyran-4-one with 2-aminopyridine (SB(4)), p-phenylenediamine (SB(5)) and o-phenylenediamine (SB(6)). Schiff bases were characterized using elemental analysis, IR, UV-Vis, (1)H NMR, (13)C NMR and mass spectroscopy. These compounds were screened for antibacterial activities by micro-plate assay technique. Escherichia coli and Staphylococcus capitis were exposed to different concentrations of the Schiff bases. Results showed that the antibacterial effect of these Schiff bases on Gram-negative bacteria were higher than that on Gram-positive bacteria moreover, the Schiff bases containing substituent OCH(3) on position five have higher antibacterial activity than that containing hydroxy group on the same position.

Evaluation of photodynamic treatment using aluminum phthalocyanine tetrasulfonate chloride as a photosensitizer: new approach.

Photodynamic therapy (PDT) has been the subject of several clinical studies. Evidence to date suggests that direct cell death may involve apoptosis. T(24) cells (bladder cancer cells, ATCC-Nr. HTB-4) were subjected to PDT with aluminum phthalocyanine tetrasulfonate chloride (AlS(4)Pc-Cl) and red laser light at 670 nm. Morphological changes after PDT were visualized under confocal microscopy. Raman microspectroscopy is considered as one of the newly established methods used for the detection of cytochrome c as an apoptotic marker. Results showed that PDT treated T(24) cells seem to undergo apoptosis after irradiation with 3 J cm(-2). Cytochrome c could not be detected from cells incubated with AlS(4)Pc-Cl using Raman spectroscopy whereas AlS(4)Pc-Cl seems to interfere with the Raman spectrum of cytochrome c.

Rapid and sensitive microplate assay for screening the effect of silver and gold nanoparticles on bacteria.

BACKGROUND & AIM: Nanomaterials are the leading requirement of the rapidly developing field of nanomedicine and bionanotechnology, and in this respect, nanotoxicology research is gaining great importance. In the field of infections, nanoparticles are being utilized as therapeutic tools against microbes, thus understanding the properties of nanoparticles and their effect on microbes is essential prior to clinical application. The aim of this study was to evaluate a microplate-based assay for monitoring the toxicity of silver and gold nanoparticles on bacteria. METHOD: Escherichia coli, a Gram-negative bacteria, and Staphylococcus capitis, a Gram-positive bacteria, were exposed to different concentrations of gold and silver nanoparticles. RESULTS: Analysis of bacterial growth showed that the toxicity of silver nanospheres is higher than that of gold nanospheres. The toxicity of silver nanoparticles is dependent on their concentration, whereas in the case of gold nanoparticles, there is no significant toxic effect. Therefore, the described microplate assay could be used as a rapid and sensitive method for detection of bacterial growth inhibition.