Narrow Results

The spectroscopic and biological properties of the new photosensitizer lutetium texaphyrin (Lu-Tex) were assessed in vitro and in vivo on a C26 colon carcinoma model, in comparison with hematoporphyrin (Hp), photofrin II (PII) and chlorin e₆(Chl). Strong binding of Lu-Tex to lipid bilayer membranes was observed. The results of confocal fluorescence microscopy on C26 cells showed that Lu-Tex was localized in small vesicles in the cytoplasm, possibly in the lysosomes, while Chl and Hp were distributed in larger cytoplasmic vesicles attributed to mitochondria. Scanning electron microscopy and X-ray microanalysis revealed that photodynamic therapy with Lu-Tex induced only slight damage to the cell membrane, leading to a delayed cell response. Chl and Hp caused significant structural damage to the outer cell membrane, resulting in ionic imbalance and fast cell death. The in vitro quantitative assessment of the relative efficiency per absorbed photon of the sensitizers revealed that Lu-Tex was less effective than Chl and Hp. However, the results of our in vivo study showed that at the same light and drug doses the anti-tumor efficiency of the agents was in the following order: Lu-Tex > Chl > PII. The strong in vivo anti-tumor effect of Lu-Tex can be explained by its higher integrated absorption in the long-wavelength range.

The self-assembly of chiral porphyrin molecules HpD (hematoporphyrin IX derivative) has been shown to form helical fibers in low salt aqueous conditions. The spectroscopic (UV and circular dichroism (CD)), thermodynamic (T_m, differential scanning calorimetry (DSC)) and microscopic (light and scanning force microscopy (SFM)) examinations of the HpD properties were performed individually and in the presence of nucleic acid double strands (15–60 °C, 0–50 mM NaCl). The asymmetric HpD molecules themselves at room temperature show sharp positive or negative CD signals, which increase enormously with HpD concentration. The data show strong evidence for the external self-stacking interaction of HpD, pure and in the presence of polynucleotides. At low salt concentration (<40 mM NaCl, pH 7) the spectra change completely by increasing the temperature. At 35 to 40 °C RNA-similar spectra of the pure HpD self-assemblies (without nucleic acids) occur. At higher temperatures the aggregates become unstable and break off. At room temperature the helical structure of the fibers could be visualized by SFM investigations. Molecular modeling analysis offers dynamic arrangements of the self-assemblies from stacks to spiral-like superstructures with increasing temperature. Hydrogen bonding, electron transferring and electrostatic interactions determine the shape of the proposed highly flexible arrangements. Moreover, the interrelation between the HpD stacks and the helix of the polynucleotides was studied. The calculated low transition energies indicate the importance of these structures as a crossing link. All data are discussed in favor of a hypothetical evolutionary matrix role in porphyrin self-assembly for RNA.

In this study we investigated, spectroscopically, the binding of hematoporphyrin (HP) to non-charged lipid vesicles as a function of temperature and the molecular structure of the phospholipid. The temperature dependence of partitioning was employed to evaluate the thermodynamic parameters of the process. We studied the binding of HP to liposomes composed of different phospholipids: natural lecithin and three chemically defined phosphatidylcholines: dimiristoyl-phosphatidylcholine (DMPC), 1-palmitoyl-2-myristoyl-phosphatidylcholine (PMPC) and 1-stearoyl-2-myristoyl-phosphatidylcholine (SMPC), at different temperatures. The last three lipids differ only in the length of the fatty acid on 1 position of the glycerol backbone. Consequently, they have different phase transition temperatures and different order parameters. For SMPC, PMPC and DMPC, we checked the effect of temperatures above and below the phase transition while for lecithin, whose phase transition temperature is well below 0 °C, only temperatures above the phase transition could be tested. A very distinct effect of the phase transition on the binding constant was observed. Below this temperature a dramatic decrease in the binding was observed as the temperature was increased. Above the phase transition, the effect of temperature declined and the changes were minor compared to the changes observed when the bilayers undergo the solid-gel phase transition. Differences in HP binding to the various bilayers were attributed to the differences in the order parameters of DMPC, PMPC, SMPC and lecithin bilayers.