The overproduction of hydrogen peroxide is an inherent feature of some tumour cells and inflamed tissues. We took advantage of this peculiarity to eliminate cells using chemiluminescent peroxyoxalate ...reaction. We designed dispersions containing polyoxalate and tetramethylhematoporhyrin (TMHP) in dimethylphthalate droplets stabilized with Pluronic L64. The porphyrin plays the dual role. On the one hand, it serves as an activator of the peroxyoxalate reaction of polyoxalate with intracellular hydrogen peroxide and experiences excitation as a result of the reaction. The light emitted in the reaction in the model system without cells was used to optimize the dispersion's composition. On the other hand, TMHP acts as a photosensitizer (PS) causing cell damage. The formation of singlet oxygen led to cell elimination if the dispersions were used in combination with inducers of oxidative stress: hydrogen peroxide, paraquat, antitumour drug doxorubicin, or a nutritional additive menadione. The PS-induced cytotoxicity correlated with the level of intracellular ROS. The developed approach targeted to endogenous ROS is orthogonal to the classical chemotherapy and can be applied to increase its efficiency.
Polymeric carriers are extensively used in photodynamic therapy (PDT) for increase of efficacy of photosensitizers. Here, we report the influence of nine Pluronic copolymers on phototoxicity of ...chlorin e6 (Ce6), in particular 5‐ to 7‐fold rise in the phototoxicity caused by hydrophilic Pluronics F127, F108, F68 and F87 and practically no influence on Ce6 of more hydrophobic polymers. The revealed value of 0.2 mg mL−1 of Pluronic F127 concentration sufficient for half‐of‐maximal increase of Ce6 photodynamic activity proved to be close to 0.16 mg mL−1 inherent in well‐documented carrier poly(N‐vinylpyrrolidone) (PVP). The dissociation constants of Ce6 complexes with Pluronic F127 and PVP that were estimated from UV spectra were 0.252 and 0.036 mg mL−1, respectively, indicating higher stability of Ce6 complex with PVP. According to the results of 1H‐NMR studies of Ce6 complexes, the porphyrin interacts not only with hydrophobic regions but also with hydrophilic sides of both polymers.
Non‐covalent binding of Chlorin e6 to hydrophilic polymers results in 5–7‐fold enhancement of its phototoxicity in vitro.
Recently it has been shown that Chlorin e6 (Ce6) when complexed with Pluronics (hydrophilic ethylene and propylene oxide block copolymers) and poly(N-vinylpyrrolidone) (PVP) exhibits considerably ...higher phototoxicity towards tumor cells than free Ce6. The present work aimed to model Ce6 interactions with hydrophilic Pluronic F127 and PVP and find out the nature of intermolecular forces stabilizing these complexes. Modeling included 3 steps: (i) application of molecular dynamics to study polymer folding using AMBER 8 program, (ii) evaluation of partial charges in the Ce6 molecule using different quantum mechanical, semi-empirical and topological approaches and (iii) docking analysis of Ce6 interactions with polymer coils using AUTODOCK 4.2. It was found that the folding in regular polymers does not occur stochastically, but involves the formation of "primary" helical structures, which further combined to form hairpin-like "secondary" structures. The latter in turn associated to form coils with minimal solvent accessible hydrophobic area. The Ce6 ring lies flat on the surface of the polymer coil at the interface between hydrophobic and hydrophilic regions. Calculations showed higher affinity of Ce6 for PVP in comparison to Pluronic and revealed marginal contribution of Coulomb forces to the stabilization of both complexes, which are mainly stabilized by van der Waals and hydrogen interactions.
All polymeric chemosensitizers proposed thus far have a linear poly(ethylene glycol) (PEG) hydrophilic block. To testify whether precisely this chemical structure and architecture of the hydrophilic ...block is a prerequisite for chemosensitization, we tested a series of novel block copolymers containing a hyperbranched polyglycerol segment as a hydrophilic block (PPO-NG copolymers) on multi-drug-resistant (MDR) tumor cells in culture. PPO-NG copolymers inhibited MDR of three cell lines, indicating that the linear PEG can be substituted for a hyperbranched polyglycerol block without loss of the polymers’ chemosensitizing activity. The extent of MDR reversal increased with the polymers affinity toward the cells and the expression level of P-glycoprotein. In contrast with Pluronic L61, which increases viability of tumor cells in the absence of drugs, PPO-NG chemosensitizers are completely devoid of this property undesired in cancer therapy, making them promising candidates for application as novel MDR reversal agents.
In this work, we report on the interaction of polyacrylic acid with phosphatidylcholine bilayers and monolayers in slightly acidic medium. We found that adsorption of polyacrylic acid on liposomes ...composed of egg lecithin at pH 4.2 results in the formation of small pores permeable for low molecular weight solutes. However, the pores were impermeable for trypsin indicating that no solubilization of liposomes occurred. The pores were permeable for both positively charged trypsin substrate N-benzoyl-l-arginine ethyl ester and negatively charged pH-indicator pyranine. Two lines of evidence were obtained confirming the involvement of the membrane dipole potential in the insertion of polyacrylic acid into lipid bilayer. (i) Addition of phloretin, a molecule which is known to decrease dipole potential of lipid bilayer, reduced the rate of a polyacrylic acid induced leakage of pyranine from liposomes. (ii) Direct measurements of air/lipid monolayer/water interface surface potential using Kelvin probe showed that adsorption of polyacrylic acid at pH 4.2 induced a decrease in both boundary and dipole potential by 37 and 62mV for ester lipid dioleoylphosphatidylcholine (DOPC). Replacement of DOPC by ether lipid 1,2-di-O-oleyl-sn-glycero-3-phosphocholine (DiOOPC) which is known to form monolayers and bilayers with only minor dipole component of membrane potential showed that addition of PAA produced similar response in the boundary potential (by 50mV) but negligible response in dipole potential of monolayer. These observations agree with our assumption that dipole potential is an important driving force for the insertion of polyacids into biological membranes.
► Polyarylic acid forms small pores in lipid bilayers permeable for small solutes. ► Insertion of polyacrylic acid decreases the membrane dipole potential. ► Membranes composed of ether lipids are less responsive to polyacids.
•Amino groups in the core of nanogel particles are inaccessible for ion pairing.•Increase in the cross-linking elevates the fraction of inaccessible amino groups.•Elevation of the cross-linking ...favours endosomolytic properties in cell cultures.
The comparison study of interaction of linear poly(2-dimethyl amino)ethyl methacrylate and its cationic nanogels of various cross-linking with both DNA and sodium poly(styrene sulfonate) has been performed. Although all amino groups of the nanogels proved to be susceptible for protonation, their accessibility for ion pairing with the polyanions was controlled and impaired with the cross-linking. The investigation of nanogels complexes with cells in culture that was accomplished by using of calcein pH-sensitive probe revealed a successive increase in the cytoplasmic fluorescence upon the growth in the cross-linking due to calceine leakage from acidic compartments to cytosol. This regularity implies that amino groups which are buried presumably inside the nanogel are protected against the ion-pairing with polyanions of plasma membrane and hence are able to manifest buffer properties while captured into acidic endosomes, i.e. possess lyso/endosomolytic capacity. These findings suggest that network architecture makes an important contribution to proton sponge properties of weak polycations.
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•Nanogels produced in reverse micelles exhibit extremely narrow size distribution.•Nanogels with 2% of the cross-linker produce the most compact complexes with DNA.•The same nanogels ...manifest the highest transfection efficacy.
A series of nanogel particles with the growing cross-linking degree was synthesized in reverse micelles by copolymerization of a cationic monomer N,N-dimethylaminoethylmethacrylate with increasing amounts of N,N′-methylenebisacrylamide (MBA). The growth of MBA content in the studied range of 0.2–15% (mol) resulted in a regular decrease of the particles’ size with a narrowing in their size distribution which occurred at the first of the cross-linker addition. In the second region, where the MBA content was more than 5% (mol), the size distribution was very narrow. The cross-linked polymers prepared at 2–5% of the cross-linker amount demonstrated a minimum size of the polyplexes and enhancement in the transfection with plasmid DNA and small interfering RNA targeted to the luciferase gene. The revealed changes in the properties of nanogels occurring upon the changes in the cross-linking degree appear to be a platform for a controlled manufacturing of efficient cross-linked cationic vectors.
To elucidate factors stirring the manifestation of different biological effects of Pluronics, we exposed multidrug resistant NCI/ADR-RES cells to various concentrations of polymers in the absence or ...presence of 5 μg/mL of doxorubicin. This method of analysis showed that each Pluronic exhibits chemosensitizing, cell supporting, and cytotoxic properties depending on its concentration. The effects developed in certain concentration ranges specific to each polymer. The free energies of Pluronics interaction with the lipids of cell membrane determined from the concentration dependencies of MDR suppression were close to the free energies of Pluronics partitioning between water and lipid bilayer of liposomes, indicating that PPO–lipid interaction underlies MDR suppression. Free energies corresponding to the increase in cell viability correlated with Pluronic’s binding to cells. It follows that cell support by Pluronics results from their attachment to the cell surface. The presented data are feasible for predicting the biological properties of PEG-containing block copolymers.
A synthetic polyanion composed of styrene, maleic anhydride, and methacrylic acid (molar ratio 56:37:7) significantly inhibited the respiration of isolated rat liver mitochondria in a time-dependent ...fashion that correlated with 1) collapse of the mitochondrial membrane potential and 2) high amplitude mitochondrial swelling. The process is apparently Ca(2+) dependent. Since it is blocked by cyclosporin A, the process is ascribed to induction of the mitochondrial permeability transition. In mitoplasts, i.e., mitochondria lacking their outer membranes, the polyanion rapidly blocked respiration. After incubation of rat liver mitochondria with the polyanion, cytochrome c was released into the incubation medium. In solution, the polyanion modified by conjugation with fluorescein formed a complex with cytochrome c. Addition of the polyanion to cytochrome c-loaded phosphatidylcholine/cardiolipin liposomes induced the release of the protein from liposomal membrane evidently due to coordinated interplay of Coulomb and hydrophobic interactions of the polymer with cytochrome c. We conclude that binding of the polyanion to cytochrome c renders it inactive in the respiratory chain due to exclusion from its native binding sites. Apparently, the polyanion interacts with cytochrome c in mitochondria and releases it to the medium through breakage of the outer membrane as a result of severe swelling. Similar properties were demonstrated for the natural polyanion, tobacco mosaic virus RNA. An electron microscopy study confirmed that both polyanions caused mitochondrial swelling. Exposure of cerebellar astroglial cells in culture to the synthetic polyanion resulted in cell death, which was associated with nuclear fragmentation.
Migration of Poly-l-lysine through a Lipid Bilayer Menger, Fredric M; Seredyuk, Victor A; Kitaeva, Marina V ...
Journal of the American Chemical Society,
03/2003, Letnik:
125, Številka:
10
Journal Article
Recenzirano
When a giant vesicle, composed of neutral and anionic lipid (90:10 mol %), comes into contact with various poly-l-lysines (MW 500−29 300), ropelike structures form within the vesicle interior. By ...using fluorescence lipids and epi-fluorescence microscopy, we have shown that both neutral and anionic lipids are constituents of the ropes. Evidence that the ropes are also comprised of poly-l-lysine comes from two experiments: (a) direct microinjection of poly(acrylic acid) into rope-containing vesicles causes the ropes to contract into small particles, an observation consistent with a polycation/polyanion interaction; and (b) direct microinjection of fluorescein isothiocyanate (a compound that covalently labels poly-l-lysine with a fluorescent moiety) into rope-containing vesicles leads to fluorescent ropes. The results may be explained by a model in which poly-l-lysine binds to the vesicle exterior, forms a domain, and enters the vesicle through defects or at the domain boundary. The model helps explain the ability of poly-l-lysine to mediate the permeation of a cancer drug, doxorubicine, into the vesicle interior.
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