Papers by Alexander Marin
Journal of Pharmaceutical Sciences, 2002
The intracellular uptake and localization of a fluorescently labeled Pluronic P-105 in HL-60 leuk... more The intracellular uptake and localization of a fluorescently labeled Pluronic P-105 in HL-60 leukemia cells and in A2780 drug-sensitive and A2780/ADR MDR ovarian carcinoma cells were characterized by flow cytometry and fluorescence microscopy. Pluronic P-105 molecules were labeled with a pH-sensitive fluorescent label, 5-(and 6-)carboxy-2′7′-dichlorofluorescein. The fluorescence intensity of labeled Pluronic was about twofold higher at pH 7.4 than at pH 5.5. At Pluronic concentrations exceeding the critical micelle concentration (cmc), flow cytometry histograms manifested bimodal distribution of cell fluorescence for all types of cells. Cell population characterized by higher fluorescence intensity presumably resulted from Pluronic transfer from the acidic environment of cytoplasmic vesicles (endosomes or lysosomes) into the neutral environment of the cytoplasm and cell nuclei, which suggested the permeabilization of the membranes of acidic vesicle by Pluronic molecules. For the MDR cells, the bimodal distribution of cell fluorescence was already observed at very low Pluronic concentrations in the incubation medium (i.e., below the cmc). The data suggest that the membranes of acidic vesicles of MDR cells are more susceptible to the action of polymeric surfactants than those of drug-sensitive cells. Permeabilization of acidic vesicles had a dramatic effect on the intracellular trafficking of drugs: when delivered in PBS, the anthracyclin drug ruboxyl (Rb) sequestered in cytoplasmic vesicles and was excluded from cell nuclei; however, when delivered in Pluronic micelles, drug accumulated in cell nuclei. Drug uptake from/with Pluronic micelles was substantially enhanced by ultrasound. These findings suggest that the nuclear accumulation of drugs internalized via fluid-phase endocytosis can be enhanced by the application of Pluronic micelles and can be further augmented by ultrasonic irradiation. © 2002 Wiley-Liss, Inc. and the American Pharmaceutical Association J Pharm Sci 91:157–170, 2002
Biomacromolecules, 2010
Applications of polyelectrolytes as pharmaceutical excipients or biologically active agents gener... more Applications of polyelectrolytes as pharmaceutical excipients or biologically active agents generated an increased interest in formulations, in which ionic macromolecules share the same milieu with protein drugs or vaccine antigens. Macromolecular interactions, which often take place in such systems, can potentially impact formulation activity and stability. The present article reports that poly[di(carboxylatophenoxy)phosphazene], disodium salt (PCPP), which has been previously shown to be a potent vaccine adjuvant, also displays a strong protein stabilizing effect in aqueous solutions that can be significantly amplified in the presence of nonionic surfactants. The phenomenon is studied in the context of macromolecular interactions in the system and is linked to the formation of PCPP-protein and PCPP-protein-surfactant complexes.
Journal of Applied Polymer Science, 2007
Polyphosphazene polyelectrolytes containing various amounts of hydrophobic fluorinated moieties a... more Polyphosphazene polyelectrolytes containing various amounts of hydrophobic fluorinated moieties and ionic carboxylic acid groups were synthesized. Polymer compositions and molecular weights were characterized by NMR and gel permeation chromatography. Interestingly, poly[(carboxylatophenoxy)(trifluoroethoxy)phosphazene] containing 60 mol % fluorinated groups was found to be soluble in aqueous solutions. The behavior of fluorinated polyelectrolytes in reactions of ionic complexation with multivalent and monovalent salts was studied in aqueous solutions and ethanol–water mixtures. Such reactions led to the formation of ionotropic hydrogels under mild conditions and, thus, are of importance to the development of microencapsulation processes and controlled release formulations. All of the synthesized polymers underwent phase separation in the presence of multivalent ionic crosslinkers, such as spermine and calcium chloride. This included a water-soluble polyelectrolyte containing 40 mol % ionic groups and hydrophobic polymer with only 3 mol % carboxylic acid groups. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 103: 53–58, 2007
Biomacromolecules, 2006
Polyphosphazenes with amino acid ester side groups show potential as hydrolytically degradable ma... more Polyphosphazenes with amino acid ester side groups show potential as hydrolytically degradable materials for biomedical applications. This study focuses on practical aspects of their use as biodegradable materials, such as effects of the hydrolytic environment and sample processing. Poly[di(ethyl glycinato)phosphazene], PEGP, and poly[di(ethyl alaninato)phosphazene], PEAP, were prepared by macromolecular substitution reaction, ensuring the absence of the residual chlorine atoms to avoid their influence on the hydrolysis. The kinetics of polymer degradation was studied by simultaneously measuring polymer mass loss, molecular weight decrease, and the release of phosphates and ammonia. The effect of pH, buffer composition, temperature, casting solvents, and film thickness were investigated.
Colloids and Surfaces B-biointerfaces, 2002
The effect of the Pluronic P-105 aggregation state on its uptake by HL-60 cells was studied by fl... more The effect of the Pluronic P-105 aggregation state on its uptake by HL-60 cells was studied by flow cytometry, fluorescence spectroscopy, and confocal and fluorescence microscopy using a fluorescently labeled Pluronic P105. In the low concentration region below the critical micelle concentration (CMC), Pluronic uptake was proportional to the concentration in the incubation medium. The proportionality broke sharply above the CMC, revealing a less efficient intracellular uptake of Pluronic micelles than that of unimers. The data suggested that Pluronic micelles were internalized via fluid-phase endocytosis while unimers were internalized via diffusion through plasma membranes. Based on the above findings, the shielding effect of Pluronic micelles on drug intracellular uptake was explained.
Archives of Biochemistry and Biophysics, 2000
To assess the effect of a polymeric surfactant, Pluronic P-105 on the activity of electron transp... more To assess the effect of a polymeric surfactant, Pluronic P-105 on the activity of electron transport chains in the mitochondria of HL-60 cells, the bioreduction rates of two membrane-localized lipophilic spin probes, 16-doxylstearic acid methyl ester (16-DSME) and 5-doxylstearic acid (5-DS), were studied. In addition, the effect of Pluronic on the bioreduction rate of the DNA-intercalating spin-labeled anthracyclin drug, Ruboxyl (Rb) was evaluated. For 16-DSME, the bioreduction kinetics was zero order with regard to the nitroxide concentration, indicating that the rate was controlled by the concentration of the reducing enzyme(s), which depends on the activity of the electron transport chains. The introduction of Pluronic at concentrations higher than 0.01% resulted in the decrease of the 16-DSME bioreduction rate. The data suggested that short-term cell incubation with Pluronic resulted in reduced activity of the electron transport chains in the mitochondria of HL-60 cells. This was corroborated by the results of an MTT assay. For 5-DS, the bioreduction kinetics was first order in the absence of Pluronic, but did not follow any simple kinetic law after a short-term cell incubation with Pluronic. For Rb, the degree of nitroxide bioreduction dropped progressively with increasing Pluronic concentration. Thus, incubating cells with polymeric surfactants modulates the intracellular energy metabolism, which can affect the rates of energy-dependent intracellular processes.
Journal of Controlled Release, 2001
The effect of a continuous wave (CW) and pulsed 20-kHz ultrasound on the Doxorubicin (DOX) uptake... more The effect of a continuous wave (CW) and pulsed 20-kHz ultrasound on the Doxorubicin (DOX) uptake by HL-60 cells from the phosphate buffered saline solution (PBS) and Pluronic micellar solutions was studied. Both CW and pulsed ultrasound enhanced DOX uptake from PBS and Pluronic micelles. The main factor that effected drug uptake was ultrasound power density; however, with increasing power, the enhanced drug uptake was accompanied by the extensive cell sonolysis. For PBS, no significant effect of duration of the ultrasound pulse or inter-pulse interval on the drug uptake was observed. For Pluronic micelles, the uptake increased with increasing pulse duration in the range 0.1-2 s, overall sonication time being the same. For 2-s pulses, the uptake was close to that under CW ultrasound. There was no significant effect of the duration of the inter-pulse interval on the drug uptake from Pluronic micelles. The data on the effect of pulse duration on drug uptake suggest that the characteristic times of drug release from micelles and drug uptake by the cells are comparable. The results point to two independent mechanisms controlling acoustic activation of drug uptake from Pluronic micelles. Both mechanisms work in concert. The first one is related to the acoustically-triggered drug release from micelles that results in higher concentration of the free drug in the incubation medium. The second mechanism is based on the perturbation of cell membranes that results in the increased uptake of the micellar-encapsulated drug. The intracellular uptake of Pluronic micelles was confirmed by fluorescence microscopy.
Polyphosphazenes for Biomedical Applications, 2009
Skin constitutes a natural barrier against infection and has a high density of dendritic cells, s... more Skin constitutes a natural barrier against infection and has a high density of dendritic cells, such as Langerhans cells, whose function it is to recognize foreign bacteria and viruses and initiate an effective immune response. This makes the immune system of the skin a desirable ...
Biomacromolecules, 2005
Polyphosphazene polyelectrolytes are potent immunostimulants. Their in vivo performance has been ... more Polyphosphazene polyelectrolytes are potent immunostimulants. Their in vivo performance has been demonstrated for various antigens in a number of animal models. To improve understanding of the mechanism of action, we performed a comparative study in a model system: bovine serum albumin, BSA-poly[di(carboxylatophenoxy)phosphazene], PCPP, in vitro and in vivo. Multi-angle laser light scattering (MALLS) and size-exclusion HPLC methods were used to investigate polyphosphazene-protein formulations in an attempt to establish correlations between their physicochemical behavior and immunostimulating activity. These studies revealed the formation of water-soluble noncovalent protein-polymer complexes in the system. It was shown that both the amount of bound protein and the complex conformation could play an important role in the in vivo performance of the polyphosphazene polyelectrolytes.
Pharmaceutical Research, 2011
Macromolecules, 2004
A single-step approach for the synthesis of polyphosphazenes containing sulfonic acid functionali... more A single-step approach for the synthesis of polyphosphazenes containing sulfonic acid functionalities is developed. Polyphosphazene sulfonation is conducted via the direct replacement of chlorine atoms of the macromolecular precursor, poly(dichlorophosphazene), PDCP with a ...
Journal of Pharmaceutical Sciences, 2011
Journal of Controlled Release, 2001
Journal of Controlled Release, 2001
The effect of a continuous wave (CW) and pulsed 20-kHz ultrasound on the Doxorubicin (DOX) uptake... more The effect of a continuous wave (CW) and pulsed 20-kHz ultrasound on the Doxorubicin (DOX) uptake by HL-60 cells from the phosphate buffered saline solution (PBS) and Pluronic micellar solutions was studied. Both CW and pulsed ultrasound enhanced DOX uptake from PBS and Pluronic micelles. The main factor that effected drug uptake was ultrasound power density; however, with increasing power, the enhanced drug uptake was accompanied by the extensive cell sonolysis. For PBS, no significant effect of duration of the ultrasound pulse or inter-pulse interval on the drug uptake was observed. For Pluronic micelles, the uptake increased with increasing pulse duration in the range 0.1-2 s, overall sonication time being the same. For 2-s pulses, the uptake was close to that under CW ultrasound. There was no significant effect of the duration of the inter-pulse interval on the drug uptake from Pluronic micelles. The data on the effect of pulse duration on drug uptake suggest that the characteristic times of drug release from micelles and drug uptake by the cells are comparable. The results point to two independent mechanisms controlling acoustic activation of drug uptake from Pluronic micelles. Both mechanisms work in concert. The first one is related to the acoustically-triggered drug release from micelles that results in higher concentration of the free drug in the incubation medium. The second mechanism is based on the perturbation of cell membranes that results in the increased uptake of the micellar-encapsulated drug. The intracellular uptake of Pluronic micelles was confirmed by fluorescence microscopy.
Journal of Applied Polymer Science, 2011
Journal of Applied Polymer Science, 2007
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Papers by Alexander Marin