Steffen Hackbarth

Singlet oxygen quantum yields of different photosensitizers in polar solvents and micellar solutions

The singlet oxygen luminescence method and the photochemical methods using 1,3-diphenylisobenzofuran (DPBF) or bilirubin ditaurate (BDT) as chemical quenchers were employed to determine the single oxygen quantum yields (ΦΔ) of different phthalocyanines and tris(2,2″-bipyridyl)ruthenium(II) dichloride in dimethylformamide (DMF) or aqueous micellar solution of 0.1 M CTAC (cetyltrimethylammonium chloride). Additionally, a perylenetetracarboxylic acid diimide derivative was examined in DMF. In a series of tetrasulfonated phthalocyanines (PTS) the following order was found: ZnPTS > GaPTS > AlPTS ≈ H2PTS > CoPTS. In general, the singlet oxygen quantum yields are higher in DMF than in 0.1 M CTAC/H2O. The results obtained with the photochemical systems are comparable with those obtained by the photophysical method. The photochemical DPBF method results in absolute values of ΦΔ. However, in micellar solution, chain reactions occur when DPBF is used as chemical quencher in the photo-oxidative process. This problem ...

Skin Penetration and Cellular Uptake of Amorphous Silica Nanoparticles with Variable Size, Surface Functionalization, and Colloidal Stability

In this study, the skin penetration and cellular uptake of amorphous silica particles with positive and negative surface charge and sizes ranging from 291 ± 9 to 42 ± 3 nm were investigated. Dynamic light scattering measurements and statistical analyses of transmission electron microscopy images were used to estimate the degree of particle aggregation, which was a key aspect to understanding the results of the in vitro cellular uptake experiments. Despite partial particle aggregation occurring after transfer in physiological media, particles were taken up by skin cells in a size-dependent manner. Functionalization of the particle surface with positively charged groups enhanced the in vitro cellular uptake. However, this positive effect was contrasted by the tendency of particles to form aggregates, leading to lower internalization ratios especially by primary skin cells. After topical application of nanoparticles on human skin explants with partially disrupted stratum corneum, only the 42 ± 3 nm particles were found to be associated with epidermal cells and especially dendritic cells, independent of their surface functionalization. Considering the wide use of nanomaterials in industries and the increasing interest for applications in pharmaceutics and cosmetics versus the large number of individuals with local or spread impairment of the skin barrier, e.g., patients with atopic dermatitis and chronic eczema, a careful dissection of nanoparticle-skin surface interactions is of high relevance to assess possible risks and potentials of intended and unintended particle exposure.

Novel photosensitizer-protein nanoparticles for photodynamic therapy: photophysical characterization and in vitro investigations.

In this work two types of pheophorbide-HSA (Pheo-HSA) nanoparticles, PHSA40 and PHSA100, were prepared and their photophysical and photosensitizing properties were investigated. Due to intramolecular interactions the singlet oxygen quantum yield of PHSA40 and PHSA100 is very low (less than 0.1). Intracellular uptake and phototoxicity of pheophorbide a as well as of the Pheo-HSA nanoparticles were studied in Jurkat cells. The HSA nanoparticles do not influence the amount of dye accumulation in cells. After 24h incubation, PHSA40 and PHSA100 showed a higher phototoxicity than Pheo. The reason for this behavior is an efficient nanoparticle decomposition in the cellular lysosomes. The process of drug release during incubation of cells with Pheo-HSA nanoparticles was illustrated by fluorescence lifetime imaging (FLIM) and confocal laser scanning microscopy (CLSM). The final phototoxicity of Pheo-HSA is at the same scale as induced by free Pheo. The drug release ability of HSA nanoparticles shows the possibility to use such formulations as drug carriers in PDT treatment. Therefore, this work constructs a standard for further investigation and optimization of photosensitizer-HSA drug carrier system.

New insights to primary photodynamic effects--Singlet oxygen kinetics in living cells.

The kinetics of chemical singlet oxygen quencher consumption inside living cells during low dose illumination was revealed via time resolved singlet oxygen luminescence detection. Deviations of the measured data from the common theoretical model for (1)O(2) kinetics forced the authors to consider a one-dimensional diffusion model for description of the kinetics of singlet oxygen generated by membrane localized photosensitizers. Our observations reconcile seemingly contradictory reports presenting different values for the efficiency of singlet oxygen interaction with cells.

Photodynamic inactivation of multi-resistant bacteria (PIB) - a new approach to treat superficial infections in the 21st century.

The increasing resistance of bacteria against antibiotics is one of the most important clinical challenges of the 21st century. Within the gram‐positive bacteria the methicillin‐resistant Staphylococcus aureus and Enterococcus faecium represent the major obstacle to successful therapy. Apart from the development of new antibiotics it requires additional differently constituted approaches, like photodynamic inactivation in order to have further effective treatment options against bacteria available. Certain dyes, termed photosensitizers, are able to store the absorbed energy in long‐lived electronic states upon light activation with appropriate wavelengths and thus make these states available for chemical activation of the immediate surroundings. The interaction with molecular oxygen, which leads to different, very reactive and thus cytotoxic oxygen species, is highlighted. In this review the application of the photodynamic inactivation of bacteria will be discussed regarding the possible indications in dermatology, like localized skin and wound infections or the reduction of nosocomial colonization with multi‐resistant bacteria on the skin. The crucial advantage of the local application of photosensitizers followed by irradiation of the area of interest is the fact that independent of the resistance pattern of a bacterium a direct inactivation takes place similarly as with an antiseptic. In this review the physical‐chemical and biological basics of photo‐dynamic inactivation of bacteria (PIB) will be discussed as well as the possible dermatological indications.

Globular dendrimers involving a C60 core and a tetraphenyl porphyrin function

The nanoenvironment provided by the neighboring dendra characteristically influences the redox potentials and the photophysical properties of the tetraphenylporphyrin–zinc (Zn-TPP; denoted as P in Figure) chromophore within the dendrimers 1, which are based on C60 as a structure-determining core. Such functional dendrimers represent a new prototype of model systems for heme proteins (1).

Photoinactivation of Escherichia coli (SURE2) without intracellular uptake of the photosensitizer

This study was performed to investigate the possibility to photodynamically inactivate Gram‐negative bacteria without intracellular uptake of the photosensitizer. The efficiency of the photodynamic growth inhibition of Escherichia coli (SURE2) was proved in a comparative study of a neutral and a cationic photosensitizer.

Photophysical properties of pheophorbide-a-substituted diaminobutane poly-propylene-imine dendrimer

Abstract Dendrimers are very promising molecules as hosts for guest embedding as well as for multiple covalent coupling of dye molecules to their peripheral groups. The use of such dendrimer–dye conjugates as part of a modular carrier system for photodynamic therapy has recently been discussed. Using covalent coupling we accomplished an average loading of 12–13 molecules of pheophorbide a to one diaminobutane poly-propylene-imine dendrimer molecule of the third generation (DAB dendrimer) having a total number of 16 binding sites. This was confirmed with various photophysical measurements and MALDI-mass spectrometry. As long as the dye molecules are covalently bound to the dendrimer, their photosensitized generation of singlet oxygen is reduced dramatically due to interactions between different dye molecules. The main interaction is supposed to be a Forster-energy transfer along the surface of the dendrimer–dye complex. Nevertheless the remaining photosensitized generated singlet oxygen leads to a destruction of the dendrimer backbone under illumination. The photophysical properties of the covalently linked dye–dendrimer complexes and especially the remarkable influence of illumination on these properties are presented in this paper.

Exploiting specific interactions toward next-generation polymeric drug transporters.

A generic method describes advanced tailoring of polymer drug carriers based on polymer-block-peptides. Combinatorial means are used to select suitable peptide segments to specifically complex small-molecule drugs. The resulting specific drug formulation agents render insoluble drugs water-soluble and enable precise adjustment of drug-release profiles beyond established block-copolymer carriers. While proof of principle is shown on chlorin as a partially approved drug for photodynamic cancer therapy, the concept is universal and applies to a broad spectrum of difficult drugs.

Photophysical evaluation of mTHPC-loaded HSA nanoparticles as novel PDT delivery systems

Controlled drug release is one of the main goals of recent developments in drug carrier systems. In this work human serum albumin (HSA) nanoparticles as carriers for 5-, 10-, 15-, 20-Tetrakis (3-hydroxyphenyl)-chlorin (mTHPC) were investigated. The photophysical properties of mTHPC-HSA nanoparticles in dependence of loading ratio and level of HSA cross-linking were determined. Further the drug release after uptake by Jurkat cells and in vitro singlet oxygen kinetics were examined. The loading ratio of the mTHPC-HSA nanoparticles turned out to be of major importance for the PDT relevant electronic parameters in solution. Therefore, only HSA nanoparticles with low mTHPC-loading ratio generate singlet oxygen in D(2)O. However, after cellular uptake all mTHPC-HSA samples generate singlet oxygen in Jurkat cells, but the decomposition rate depends on the level of HSA cross-linking.