Six common water-soluble singlet oxygen ((1)O2) photosensitizers - 5,10,15,20-tetrakis(1-methyl-4-pyridinio) porphine (TMPyP), meso-tetrakis(4-sulfonathophenyl)porphine (TPPS4), Al(III) phthalocyanine chloride tetrasulfonic acid (AlPcS4), eosin Y, rose bengal, and methylene blue - were investigated in terms of their ability to produce delayed fluorescence (DF) in solutions at room temperature. All the photosensitizers dissolved in air-saturated phosphate buffered saline (PBS, pH 7.4) exhibit easily detectable DF, which can be nearly completely quenched by 10 mM NaN3, a specific (1)O2 quencher. The DF kinetics has a biexponential rise-decay character in a microsecond time domain. Therefore, we propose that singlet oxygen-sensitized delayed fluorescence (SOSDF), where the triplet state of a photosensitizer reacts with (1)O2 giving rise to an excited singlet state of the photosensitizer, is the prevailing mechanism. It was confirmed by additional evidence, such as a monoexponential decay of triplet-triplet transient absorption kinetics, dependence of SOSDF kinetics on oxygen concentration, absence of SOSDF in a nitrogen-saturated sample, or the effect of isotopic exchange H2O-D2O. Eosin Y and AlPcS4 show the largest SOSDF quantum yield among the selected photosensitizers, whereas rose bengal possesses the highest ratio of SOSDF intensity to prompt fluorescence intensity. The rate constant for the reaction of triplet state with (1)O2 giving rise to the excited singlet state of photosensitizer was estimated to be ~/>1 × 10(9) M(-1) s(-1). SOSDF kinetics contains information about both triplet and (1)O2 lifetimes and concentrations, which makes it a very useful alternative tool for monitoring photosensitizing and (1)O2 quenching processes, allowing its detection in the visible spectral region, utilizing the photosensitizer itself as a (1)O2 probe. Under our experimental conditions, SOSDF was up to three orders of magnitude more intense than the infrared (1)O2 phosphorescence and by far the most important pathway of DF. SOSDF was also detected in a suspension of 3T3 mouse fibroblast cells, which underlines the importance of SOSDF and its relevance for biological systems.

• MeSH
• buňky 3T3 MeSH
• fluorescenční spektrometrie MeSH
• fotosenzibilizující látky chemie   MeSH
• kinetika MeSH
• koncentrace vodíkových iontů MeSH
• kvantová teorie MeSH
• myši MeSH
• singletový kyslík chemie   MeSH
• teplota MeSH
• voda chemie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The present work provides a proof-of-concept that the singlet oxygen-sensitized delayed fluorescence (SOSDF) can be detected from individual living mammalian cells in a time-resolved microscopy experiment. To this end, 3T3 mouse fibroblasts incubated with 100 μM TPPS4 or TMPyP were used and the microsecond kinetics of the delayed fluorescence (DF) were recorded. The analysis revealed that SOSDF is the major component of the overall DF signal. The microscopy approach enables precise control of experimental conditions - the DF kinetics are clearly influenced by the presence of the (1)O2 quencher (sodium azide), H2O/D2O exchange, and the oxygen concentration. Analysis of SOSDF kinetics, which was reconstructed as a difference DF kinetics between the unquenched and the NaN3-quenched samples, provides a cellular (1)O2 lifetime of τΔ = 1-2 μs and a TPPS4 triplet lifetime of τT = 22 ± 5 μs in agreement with previously published values. The short SOSDF acquisition times, typically in the range of tens of seconds, enable us to study the dynamic cellular processes. It is shown that SOSDF lifetimes increase during PDT-like treatment, which may provide valuable information about changes of the intracellular microenvironment. SOSDF is proposed and evaluated as an alternative tool for (1)O2 detection in biological systems.

• MeSH
• analýza jednotlivých buněk přístrojové vybavení   metody   MeSH
• azid sodný chemie   MeSH
• buňky 3T3 MeSH
• design vybavení MeSH
• fibroblasty chemie   MeSH
• fluorescence * MeSH
• kinetika MeSH
• kyslík chemie   MeSH
• mikroskopie přístrojové vybavení   metody   MeSH
• myši MeSH
• oxid deuteria chemie   MeSH
• singletový kyslík chemie   MeSH
• voda chemie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Anion exchange polystyrene nanofiber materials (AE) were prepared by electrospinning followed by two-step functionalization of the nanofiber surface by chlorosulfonic acid and ethylendiamine. The photoactive character of these materials was introduced through adsorption of the tetra-anionic 5,10,15,20-tetrakis-(4-sulfonatophenyl)porphyrin photosensitizer (TPPS-AE) on the nanofiber surface or by encapsulation of the nonpolar 5,10,15,20-tetraphenylporphyrin photosensitizer (AE(TPP)) into the nanofibers. Anion exchange nanofiber materials with porphyrins are characterized by a high ion-exchange capacity, photogeneration of singlet oxygen O2((1)Δg), and singlet oxygen-sensitized delayed fluorescence. Due to the photogeneration of cytotoxic O2((1)Δg), the nanofibers exhibited oxidation of the external substrates in aqueous solution and an efficient antibacterial effect when activated by simulated daylight. Adsorption of both TPPS and I(-) on the surface of AE led to the formation of more efficient I-TPPS-AE materials. Rapid photooxidation of I(-) by O2((1)Δg), and the formation of another cytotoxic species, I3(-), on the surface of the nanofibers were responsible for the increased antibacterial properties of I-TPPS-AE and the prolonged antibacterial effect in the dark.

• MeSH
• adsorpce MeSH
• anionty chemie   MeSH
• Escherichia coli účinky léků   metabolismus   účinky záření   MeSH
• fotosenzibilizující látky chemie   farmakologie   MeSH
• iontová výměna MeSH
• kinetika MeSH
• nanovlákna chemie   MeSH
• oxidace-redukce MeSH
• polystyreny chemie   MeSH
• porfyriny chemie   farmakologie   MeSH
• singletový kyslík chemie   metabolismus   MeSH
• světlo * MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Aqueous dispersions of sulfonated polystyrene nanoparticles (average diameter: 30 ± 14 nm) with encapsulated 5,10,15,20-tetraphenylporphyrin (TPP) are promising candidates for antibacterial treatments due to the photogeneration of cytotoxic singlet oxygen species O2(1Δg) under physiological conditions using visible light. The antibacterial effect on gram-negative Escherichia coli was significantly enhanced after the addition of nontoxic potassium iodide (0.001-0.01 M) because photogenerated O2(1Δg) oxidized iodide to I2/I3-, which is another antibacterial species. The improved antibacterial properties were predicted using luminescence measurements of O2(1Δg), transient absorption of TPP triplets and singlet oxygen-sensitized delayed fluorescence (SODF). In contrast to a solution of free photosensitizers, the aqueous dispersion of photoactive nanoparticles did not exhibit any quenching of the excited states after the addition of iodide or any tendency toward aggregation and/or I3--induced photo-aggregation. We also observed a decrease in the lifetime of O2(1Δg) and a significant increase in SODF intensity at higher temperatures, due to the increased oxygen diffusion coefficient in nanoparticles and aqueous surroundings. This effect corresponds with the significantly stronger antibacterial effect of nanoparticles at physiological temperature (37 °C) in comparison with that at room temperature (25 °C).

• MeSH
• antibakteriální látky farmakologie   MeSH
• časové faktory MeSH
• Escherichia coli účinky léků   MeSH
• fotosenzibilizující látky farmakologie   MeSH
• jodidy farmakologie   MeSH
• kinetika MeSH
• luminiscence MeSH
• mikrobiální testy citlivosti MeSH
• nanočástice chemie   ultrastruktura   MeSH
• nanovlákna chemie   ultrastruktura   MeSH
• teplota * MeSH
• Publikační typ
• časopisecké články MeSH