ΑΡΙΣΤΕΙΔΗΣ ΓΕΩΡΓΟΠΟΥΛΟΣ - Academia.edu (original) (raw)

Papers by ΑΡΙΣΤΕΙΔΗΣ ΓΕΩΡΓΟΠΟΥΛΟΣ

Research paper thumbnail of Dendrimers as Biopharmaceuticals: Synthesis and Properties

Current Topics in Medicinal Chemistry, Sep 1, 2008

Two general aspects which need to be considered for the successful application of dendrimers for ... more Two general aspects which need to be considered for the successful application of dendrimers for biomedical purposes are their availability at an acceptable cost and their suitability as regards their pharmacodynamic and pharmacokinetic properties. These two aspects are covered in this review. In the first part, synthetic strategies for the preparation of dendrimers are outlined and emphasis is given to recent work on methodologies whose aim is the development of more efficient routes to dendrimers in terms of the materials used for their synthesis as well as in terms of the procedures required for their purification. These include procedures involving double-stage and double exponential synthesis, orthogonal coupling strategies, self-assembly and solid-phase approaches, as well as particularly useful synthetic protocols such as those used in "click chemistry". The second part of the review deals with the way in which the size, chemical constitution and physicochemical properties of dendrimers used for drug delivery may affect pharmacodynamic and pharmacokinetic parameters which are important considerations for drug bioavailability. This is illustrated by an overview of examples from recent work involving non-steroidal anti-inflammatory drugs, anticancer drugs and antibacterials.

Research paper thumbnail of Cytotoxic and antitumor activity of liposome-incorporated sclareol against cancer cell lines and human colon cancer xenografts

Pharmacological Research, 2006

The aim of this study was to design and prepare liposome-incorporated sclareol-a highly lipophili... more The aim of this study was to design and prepare liposome-incorporated sclareol-a highly lipophilic natural product-to overcome its water insolubility and develop suitable formulations for in vivo administration. The bioactive labdane-type diterpene sclareol was incorporated into liposomes composed of egg phosphatidylcholine and dipalmitoylphosphatidylglycerol prepared by the thin-film hydration method followed by sonication. A formulation of egg phosphatidylcholine/dipalmitoylphosphatidylglycerol/sclareol (9:0.1:5 molar ratio) was developed and characterized. The lipid recovery and the sclareol to lipid molar ratio were measured using high-performance thin-layer chromatography/flame ionization detection. In vitro drug release was measured in supplemented RPMI-1640 at 37 • C. The liposomal and the free sclareol were initially tested in vitro for their activity against human cancer cell lines using the sulphorhodamine B assay. Liposomes incorporating sclareol at a drug to lipid molar ratio of 0:43, suggesting an incorporation efficiency of almost 80%, showed reduced growth rate of human colon cancer tumors (HCT116) developed in SCID mice, without any significant side effects.

Research paper thumbnail of Lipid analysis of Greek broad bean oil: Preparation of liposomes and physicochemical characterization

European Journal of Lipid Science and Technology, Nov 1, 2005

ABSTRACT Liposomes were prepared from the isolated phospholipids of mature broad bean [Vicia faba... more ABSTRACT Liposomes were prepared from the isolated phospholipids of mature broad bean [Vicia faba L. (syn. Fabae calabaricae)] oil and their physical properties were studied. The method of preparation was the hydration of the thin lipid film, while the probe sonication methodology was used for reducing the size of the vesicles. The seeds of the broad bean were collected in two different periods of maturity and extracted by the Bligh-Dyer method, and the lipid classes were studied by HPTLC/FID. The oils were found to be rich in polar lipids (63.1% and 60.2% of total lipids) and low in neutral lipids (36.9% and 39.8% of total lipids) for the immature and mature seed oils, respectively. The neutral lipid fraction consisted mainly of triacylglycerides (34.2% and 32.3%) whereas the polar lipids mainly consisted of phospholipids (60.2% and 54.2%) for the mature and immature seed oils, respectively. Sphingolipids (8.9%) were identified only in the immature seed oil. The overall goal of this study was the preparation of a new liposomal formulation with physicochemical properties such as unique lipid composition, size and ζ-potential, which are important factors influencing drug delivery to the target tissues.

Research paper thumbnail of Liposomes modify the subcellular distribution of sclareol uptake by HCT-116 cancer cell lines

Biomedicine & Pharmacotherapy, Feb 1, 2007

The uptake of free and liposome-incorporated sclareol and its effect on the growth of human cance... more The uptake of free and liposome-incorporated sclareol and its effect on the growth of human cancer cell line HCT-116 was investigated. Recovery of free and liposomal sclareol in cytosol, nuclei and crude membranes was monitored over time. HCT-116 cells were incubated with 100 microM of free or liposomal sclareol up to 96 h. Intact cells were subjected to subcellular fractionation in order to obtain highly purified fractions of nuclei, cytosol, and crude membranes. Sclareol was extracted from intact cells and from the subcellular fractions using the Bligh-Dyer method and was measured by HPTLC/FID. The effect of sclareol on cell growth was found time dependent. Free sclareol exhibited high toxicity, while the liposomal sclareol showed reduced cytotoxicity but retained the ability to reduce the cell growth rate. The uptake of sclareol by the cells was faster and higher compared to that of its liposomal form. The concentration of sclareol in the three subcellular fractions showed that liposomal sclareol is incorporated in crude membranes and from there it is released in cytosol and nuclei in a time dependent manner, while free sclareol passes directly in the cytosol. These results suggest that liposomal sclareol retains its growth inhibiting activity while its cytotoxic action is diminished. These findings could be due to the sustained delivery of sclareol to the different subcellular sites.

Research paper thumbnail of Labdane-type diterpenes: thermal effects on phospholipid bilayers, incorporation into liposomes and biological activity

Chemistry and Physics of Lipids, Dec 1, 2005

Labd-13(E)-ene-8alpha,15-diol (1) and its derivative labd-13(E)-ene-8alpha-ol-15-yl-acetate (2) a... more Labd-13(E)-ene-8alpha,15-diol (1) and its derivative labd-13(E)-ene-8alpha-ol-15-yl-acetate (2) are water insoluble biological active molecules and their structures were elucidated using NMR and X-ray techniques. Differential scanning calorimetry (DSC) was applied to study the thermal effects of 1 and 2 on DPPC bilayers. Liposomes composed of egg phosphatidylcholine/dipalmytoylphosphatidylglycerol (9:0.1 molar ratio) were prepared by the thin-film hydration method and were used for incorporating 1 and 2. Free and liposomal 1 and 2 were tested for their activity against human cancer cell lines using the sulphorhodamine B assay. The effect of 1 and 2 on DPPC bilayers caused abolition of the pre-transition temperature, lowering of the main phase transition and reduction of the transition enthalpy only in the presence of cholesterol. The liposomes that have been designed and developed offer high incorporation efficiency; 62.4% (0.369 drug/lipid molar ratio) and 99.7% (0.661 drug/lipid molar ratio) for 1 and 2, respectively. Liposomal 2 showed growth-inhibiting activity against the majority of the tested cell lines.

Research paper thumbnail of Line optical tweezers: A tool to induce transformations in stained liposomes and to estimate shear modulus

Colloids and Surfaces A: Physicochemical and Engineering Aspects, Oct 1, 2009

ABSTRACT Liposomes have been actively studied as models of cell membranes and are currently used ... more ABSTRACT Liposomes have been actively studied as models of cell membranes and are currently used as drug delivery systems of bioactive molecules. Liposome transformations that mimic cellular processes and are associated with their physicochemical properties have become field of interest the last decade. However, there has been little experimental work on controlled vesicle transformations by non-contact, optical handling methods.In this paper we present the use of line optical tweezers to observe liposome state transitions and transformations. Dynamic shape deformations were induced by line optical tweezers in giant stained liposomes leading to budding transition, fission and pearling creation. Under the controlled effect of line optical tweezers reversible liposome deformations were observed. The shear modulus μ of the membrane was inferred by measuring deformation of stained liposomes induced by the applied optical force. Further laser radiation caused irreversible shape deformations of liposomes, which were transformed from spherical to tubular vesicles. The ability of the selective manipulation of liposomes brings us closer to study their physicochemical properties which play a key role in cellular–liposome interactions, drug encapsulation and delivery efficiency.

Research paper thumbnail of Uptake Studies of Free and Liposomal Sclareol by Mcf-7 and H-460 Human Cancer Cell Lines

Springer eBooks, Sep 18, 2007

Research paper thumbnail of Mitochondria-targeted liposomes improve the apoptotic and cytotoxic action of sclareol

Journal of Liposome Research, Nov 2, 2009

Current efforts toward improving the effectiveness of drug therapy are increasingly relying on dr... more Current efforts toward improving the effectiveness of drug therapy are increasingly relying on drug-targeting strategies to effectively deliver bioactive molecules to their molecular targets. Pharmaceutical nanocarriers represent a major tool toward this aim, and our efforts have been directed toward achieving nanocarrier-mediated subcellular delivery of drug molecules with mitochondria as the primary subcellular target. Meeting the need for specific subcellular delivery is essential to realizing the full potential of many poorly soluble anticancer drugs. In this article, we report that mitochondria-targeted liposomes significantly improve the apoptotic and cytotoxic action of sclareol, a poorly soluble potential anticancer drug. The results support the broad applicability of our nanocarrier-mediated subcellular targeting approach as a means to improve the effectiveness of certain anticancer therapeutics.

Research paper thumbnail of Encapsulation of vinblastine into new liposome formulations prepared from triticum (wheat germ) lipids and its activity against human leukemic cell lines

Anticancer research

Liposomes prepared from lipids isolated from Triticum sp. (wheat germ) were used to investigate t... more Liposomes prepared from lipids isolated from Triticum sp. (wheat germ) were used to investigate the percentage of Vinblastine encapsulation and its retention into liposomes. The wheat germ total lipids (TL) were extracted by the Bligh-Dyer method and the lipid classes have been isolated using chromatographic techniques. The type of lipids and their percentage content have been examined by TLC coupled with an FID (latroscan). Two liposomal formulations, i.e., I and II, with encapsulated vinblastine, and formulation III (empty liposomes) have been prepared by thin film hydration method. The cytotoxic/cytostatic activity of these liposomal formulations have been examined against nine human leukemic cell lines. The results showed that the percentage content of vinblastine into liposomes I and II depended on the lipid composition and it was greater into formulation II (> 90%). The retention of the drug into liposomes was studied and found to be time-dependent at 37 degrees C. For the ...

Research paper thumbnail of Development of simple thiol-reactive liposome formulations, one-step analysis and physicochemical characterization

Journal of Pharmacy and Pharmacology, 2005

The aim of this study was to synthesize simple thiol-reactive conjugates from maleimide and lipoa... more The aim of this study was to synthesize simple thiol-reactive conjugates from maleimide and lipoamines (stearylamine or oleylamine) and to develop a simple, fast and low-cost method for the preparation of lyophilized general-purpose thiol-reactive liposomes. A formulation of egg phosphatidylcholine-dipalmitoylphoshatidylglycerol (9:0.1 molar ratio) was developed and characterized. Freeze-drying methodology was established to produce a stock of liposomes and the physicochemical characteristics of the reconstituted liposomes were compared with those of the initial preparation. The physicochemical properties (size and ζ-potential) of the new liposomal formulations were studied. High-performance thin-layer chromatography coupled to a flame ionization detector was applied for one-step analysis of the liposomal components and for determining the maleimide-lipoamine conjugates phospholipid molar ratio. The differences concerning the incorporation efficiency of the synthetic conjugates into...

Research paper thumbnail of In-vitro cytotoxic/cytostatic activity of anionic liposomes containing vinblastine against leukaemic human cell lines

Journal of Pharmacy and Pharmacology, 2002

Liposomes prepared from lipids dipalmitoylphosphatidylcholine (DPPC) and dipalmitoylphosphatidylg... more Liposomes prepared from lipids dipalmitoylphosphatidylcholine (DPPC) and dipalmitoylphosphatidylglycerol (DPPG) with cholesterol were used to investigate the percentage of vinblastine encapsulation and the influence of lipid composition on the retention properties of vinblastine in buffer as well as in cell culture medium. Differential scanning calorimetry (DSC) was applied, to study the effect of cholesterol on the phase transition temperature and on the ΔH of the two liposome formulations. The cytotoxic and cytostatic activity of the liposome-encapsulated vinblastine was also examined against six leukaemic human cell lines. The results showed that encapsulation of vinblastine into liposomes was greater than 98% with a drug-phospholipid molar ratio of 0.13-0.18. The major improvement in vinblastine retention in buffer as well as in culture medium was achieved by employing DPPG. The DSC data showed that vinblastine exerted a more perturbing effect in DPPC-cholesterol bilayers than i...

Research paper thumbnail of Accumulation of vinblastine into transfersomes and liposomes in response to a transmembrane ammonium sulfate gradient and their cytotoxic/cytostatic activity in vitro

Anticancer research

Vinblastine was encapsulated into liposomes composed from lipids dimiristoylphosphatidylcholine (... more Vinblastine was encapsulated into liposomes composed from lipids dimiristoylphosphatidylcholine (DMPC) and dipalmitoylphosphatidylcholine (DPPC), with cholesterol and transfersomes with sodium cholate prepared by the thin film hydration method. The percentage of vinblastine encapsulation, the stability of transfersomes and liposomes and the rate of release of encapsulated vinblastine at 37 degrees C were studied. The results showed that encapsulation of vinblastine into liposomes was higher than 98% at a drug/phospholipid molar ratio from 0.17 to 0.18, while encapsulation of vinblastine into transfersomes varied from 50% to 80% at a drug/phospholipid molar ratio from 0.05 to 0.09. The retention of drug in liposomes and in transfersomes was found to be time/dependent. The retention of drug in transfersomes compared to the liposomes was reduced due to the presence of sodium cholate which caused destabilization and reduced the main phase transition temperature Tm of the PC bilayers. Th...

Research paper thumbnail of Chapter 8 Uptake Studies of Free and Liposomal Sclareol by MCF-7 and H-460 Human Cancer Cell Lines

The aim of this study was to investigate the uptake of free and liposomal sclareol and its effect... more The aim of this study was to investigate the uptake of free and liposomal sclareol and its effect on the growth inhibiting activity against MCF-7 and H-460 human cancer cell lines in vitro. Liposomes composed of EPC/DPPG at molar ratio 9:0.1, used to incorporate sclareol, were prepared by the thin-film hydration method followed by sonication. The final liposomal preparation (EPC/DPPG/Sclareol 9:0.1:5 molar ratio) as well as free sclareol (100μM) were incubated up to 96 hours with both cell lines. Sclareol was extracted from cells using the Bligh-Dyer method and was measured by HPTLC/FID. The results showed that the uptake of free sclareol by both cell lines was faster and higher compared to that of its liposomal form. In both cell lines, free sclareol showed high cytotoxicity, while the liposomal sclareol showed reduced cytotoxicity without affecting its ability to reduce the cell growth rate. These findings suggest that liposomal sclareol may possess chemotherapeutic advantages ove...

Research paper thumbnail of Liposomic Niflumic Acid - New Transdermal Anti-Inflammatory Medicine

Research paper thumbnail of Molecular interactions between dimethoxycurcumin and Pamam dendrimer carriers

International Journal of Pharmaceutics, 2007

Dimethoxycurcumin, a lipophilic analog of curcumin found as a major pigment in the Indian species... more Dimethoxycurcumin, a lipophilic analog of curcumin found as a major pigment in the Indian species turmeric (Curcuma longa Linn.), is known to possess significant activity against various cancer cell lines, but its use as an anticancer drug is hindered by its poor water solubility. The conjugation of dimethoxycurcumin to water-soluble PAMAM dendrimers (generations 3.5 and 4) is demonstrated. The maximum drug-dendrimer incorporation efficiency is 4.3 and 5.0 molar for G3.5 and G4, respectively. The FTIR-ATR investigation of the neat compounds and the drug-dendrimer systems indicate that dimethoxycurcumin is in the enolic form, while its interaction with the integer generation dendrimer involves the major conformational change of the terminal ethylene amine groups.

Research paper thumbnail of Cytotoxic and antitumor activity of liposome-incorporated sclareol against cancer cell lines and human colon cancer xenografts

Pharmacological Research, 2006

The aim of this study was to design and prepare liposome-incorporated sclareol-a highly lipophili... more The aim of this study was to design and prepare liposome-incorporated sclareol-a highly lipophilic natural product-to overcome its water insolubility and develop suitable formulations for in vivo administration. The bioactive labdane-type diterpene sclareol was incorporated into liposomes composed of egg phosphatidylcholine and dipalmitoylphosphatidylglycerol prepared by the thin-film hydration method followed by sonication. A formulation of egg phosphatidylcholine/dipalmitoylphosphatidylglycerol/sclareol (9:0.1:5 molar ratio) was developed and characterized. The lipid recovery and the sclareol to lipid molar ratio were measured using high-performance thin-layer chromatography/flame ionization detection. In vitro drug release was measured in supplemented RPMI-1640 at 37 • C. The liposomal and the free sclareol were initially tested in vitro for their activity against human cancer cell lines using the sulphorhodamine B assay. Liposomes incorporating sclareol at a drug to lipid molar ratio of 0:43, suggesting an incorporation efficiency of almost 80%, showed reduced growth rate of human colon cancer tumors (HCT116) developed in SCID mice, without any significant side effects.

Research paper thumbnail of Development and Characterization of Oligonucleotide-Tagged Dye-Encapsulating EPC/DPPG Liposomes

Journal of Nanoscience and Nanotechnology, 2010

Liposomes applications in health care include meanly their ability to carry drugs and genes insid... more Liposomes applications in health care include meanly their ability to carry drugs and genes inside the human body for therapeutic purposes. Nevertheless their applicability can extend far beyond and could be used as analytical tools in order to perform rapid, low-cost, sensitive and specific analyses. Their physical characteristics, such as large internal volume and extended surface area, render them ideal for these applications and specifically for improving the specificity and sensitivity of the analytical assay. The purpose of this study was to develop a simple, stable and low-cost oligonucleotide-tagged liposomal formulation consisting of EggPC and DPPG with a simple to synthesize thiol-reactive conjugate (Mal-SA) incorporated into the lipid bilayer of liposomes. The prepared liposomes, having also the water soluble dye Sulforhodamine B encapsulated in their inner cavity, were characterized in terms of their physicochemical (size, size distribution,-potential, lipid content) and mechanical (morphology, rigidity) properties. The results showed that the final liposomal formulation could be used in the future as analytical tool for detecting pathogen strains of microorganism in biological milieu.

Research paper thumbnail of Mitochondria-targeted liposomes improve the apoptotic and cytotoxic action of sclareol

Journal of Liposome Research, 2009

Current efforts toward improving the effectiveness of drug therapy are increasingly relying on dr... more Current efforts toward improving the effectiveness of drug therapy are increasingly relying on drug-targeting strategies to effectively deliver bioactive molecules to their molecular targets. Pharmaceutical nanocarriers represent a major tool toward this aim, and our efforts have been directed toward achieving nanocarrier-mediated subcellular delivery of drug molecules with mitochondria as the primary subcellular target. Meeting the need for specific subcellular delivery is essential to realizing the full potential of many poorly soluble anticancer drugs. In this article, we report that mitochondria-targeted liposomes significantly improve the apoptotic and cytotoxic action of sclareol, a poorly soluble potential anticancer drug. The results support the broad applicability of our nanocarrier-mediated subcellular targeting approach as a means to improve the effectiveness of certain anticancer therapeutics.

Research paper thumbnail of Lipid analysis of Greek broad bean oil: Preparation of liposomes and physicochemical characterization

European Journal of Lipid Science and Technology, 2005

ABSTRACT Liposomes were prepared from the isolated phospholipids of mature broad bean [Vicia faba... more ABSTRACT Liposomes were prepared from the isolated phospholipids of mature broad bean [Vicia faba L. (syn. Fabae calabaricae)] oil and their physical properties were studied. The method of preparation was the hydration of the thin lipid film, while the probe sonication methodology was used for reducing the size of the vesicles. The seeds of the broad bean were collected in two different periods of maturity and extracted by the Bligh-Dyer method, and the lipid classes were studied by HPTLC/FID. The oils were found to be rich in polar lipids (63.1% and 60.2% of total lipids) and low in neutral lipids (36.9% and 39.8% of total lipids) for the immature and mature seed oils, respectively. The neutral lipid fraction consisted mainly of triacylglycerides (34.2% and 32.3%) whereas the polar lipids mainly consisted of phospholipids (60.2% and 54.2%) for the mature and immature seed oils, respectively. Sphingolipids (8.9%) were identified only in the immature seed oil. The overall goal of this study was the preparation of a new liposomal formulation with physicochemical properties such as unique lipid composition, size and ζ-potential, which are important factors influencing drug delivery to the target tissues.

Research paper thumbnail of Dendrimers as Biopharmaceuticals: Synthesis and Properties

Current Topics in Medicinal Chemistry, 2008

Two general aspects which need to be considered for the successful application of dendrimers for ... more Two general aspects which need to be considered for the successful application of dendrimers for biomedical purposes are their availability at an acceptable cost and their suitability as regards their pharmacodynamic and pharmacokinetic properties. These two aspects are covered in this review. In the first part, synthetic strategies for the preparation of dendrimers are outlined and emphasis is given to recent work on methodologies whose aim is the development of more efficient routes to dendrimers in terms of the materials used for their synthesis as well as in terms of the procedures required for their purification. These include procedures involving double-stage and double exponential synthesis, orthogonal coupling strategies, self-assembly and solid-phase approaches, as well as particularly useful synthetic protocols such as those used in "click chemistry". The second part of the review deals with the way in which the size, chemical constitution and physicochemical properties of dendrimers used for drug delivery may affect pharmacodynamic and pharmacokinetic parameters which are important considerations for drug bioavailability. This is illustrated by an overview of examples from recent work involving non-steroidal anti-inflammatory drugs, anticancer drugs and antibacterials.

Research paper thumbnail of Dendrimers as Biopharmaceuticals: Synthesis and Properties

Current Topics in Medicinal Chemistry, Sep 1, 2008

Two general aspects which need to be considered for the successful application of dendrimers for ... more Two general aspects which need to be considered for the successful application of dendrimers for biomedical purposes are their availability at an acceptable cost and their suitability as regards their pharmacodynamic and pharmacokinetic properties. These two aspects are covered in this review. In the first part, synthetic strategies for the preparation of dendrimers are outlined and emphasis is given to recent work on methodologies whose aim is the development of more efficient routes to dendrimers in terms of the materials used for their synthesis as well as in terms of the procedures required for their purification. These include procedures involving double-stage and double exponential synthesis, orthogonal coupling strategies, self-assembly and solid-phase approaches, as well as particularly useful synthetic protocols such as those used in "click chemistry". The second part of the review deals with the way in which the size, chemical constitution and physicochemical properties of dendrimers used for drug delivery may affect pharmacodynamic and pharmacokinetic parameters which are important considerations for drug bioavailability. This is illustrated by an overview of examples from recent work involving non-steroidal anti-inflammatory drugs, anticancer drugs and antibacterials.

Research paper thumbnail of Cytotoxic and antitumor activity of liposome-incorporated sclareol against cancer cell lines and human colon cancer xenografts

Pharmacological Research, 2006

The aim of this study was to design and prepare liposome-incorporated sclareol-a highly lipophili... more The aim of this study was to design and prepare liposome-incorporated sclareol-a highly lipophilic natural product-to overcome its water insolubility and develop suitable formulations for in vivo administration. The bioactive labdane-type diterpene sclareol was incorporated into liposomes composed of egg phosphatidylcholine and dipalmitoylphosphatidylglycerol prepared by the thin-film hydration method followed by sonication. A formulation of egg phosphatidylcholine/dipalmitoylphosphatidylglycerol/sclareol (9:0.1:5 molar ratio) was developed and characterized. The lipid recovery and the sclareol to lipid molar ratio were measured using high-performance thin-layer chromatography/flame ionization detection. In vitro drug release was measured in supplemented RPMI-1640 at 37 • C. The liposomal and the free sclareol were initially tested in vitro for their activity against human cancer cell lines using the sulphorhodamine B assay. Liposomes incorporating sclareol at a drug to lipid molar ratio of 0:43, suggesting an incorporation efficiency of almost 80%, showed reduced growth rate of human colon cancer tumors (HCT116) developed in SCID mice, without any significant side effects.

Research paper thumbnail of Lipid analysis of Greek broad bean oil: Preparation of liposomes and physicochemical characterization

European Journal of Lipid Science and Technology, Nov 1, 2005

ABSTRACT Liposomes were prepared from the isolated phospholipids of mature broad bean [Vicia faba... more ABSTRACT Liposomes were prepared from the isolated phospholipids of mature broad bean [Vicia faba L. (syn. Fabae calabaricae)] oil and their physical properties were studied. The method of preparation was the hydration of the thin lipid film, while the probe sonication methodology was used for reducing the size of the vesicles. The seeds of the broad bean were collected in two different periods of maturity and extracted by the Bligh-Dyer method, and the lipid classes were studied by HPTLC/FID. The oils were found to be rich in polar lipids (63.1% and 60.2% of total lipids) and low in neutral lipids (36.9% and 39.8% of total lipids) for the immature and mature seed oils, respectively. The neutral lipid fraction consisted mainly of triacylglycerides (34.2% and 32.3%) whereas the polar lipids mainly consisted of phospholipids (60.2% and 54.2%) for the mature and immature seed oils, respectively. Sphingolipids (8.9%) were identified only in the immature seed oil. The overall goal of this study was the preparation of a new liposomal formulation with physicochemical properties such as unique lipid composition, size and ζ-potential, which are important factors influencing drug delivery to the target tissues.

Research paper thumbnail of Liposomes modify the subcellular distribution of sclareol uptake by HCT-116 cancer cell lines

Biomedicine & Pharmacotherapy, Feb 1, 2007

The uptake of free and liposome-incorporated sclareol and its effect on the growth of human cance... more The uptake of free and liposome-incorporated sclareol and its effect on the growth of human cancer cell line HCT-116 was investigated. Recovery of free and liposomal sclareol in cytosol, nuclei and crude membranes was monitored over time. HCT-116 cells were incubated with 100 microM of free or liposomal sclareol up to 96 h. Intact cells were subjected to subcellular fractionation in order to obtain highly purified fractions of nuclei, cytosol, and crude membranes. Sclareol was extracted from intact cells and from the subcellular fractions using the Bligh-Dyer method and was measured by HPTLC/FID. The effect of sclareol on cell growth was found time dependent. Free sclareol exhibited high toxicity, while the liposomal sclareol showed reduced cytotoxicity but retained the ability to reduce the cell growth rate. The uptake of sclareol by the cells was faster and higher compared to that of its liposomal form. The concentration of sclareol in the three subcellular fractions showed that liposomal sclareol is incorporated in crude membranes and from there it is released in cytosol and nuclei in a time dependent manner, while free sclareol passes directly in the cytosol. These results suggest that liposomal sclareol retains its growth inhibiting activity while its cytotoxic action is diminished. These findings could be due to the sustained delivery of sclareol to the different subcellular sites.

Research paper thumbnail of Labdane-type diterpenes: thermal effects on phospholipid bilayers, incorporation into liposomes and biological activity

Chemistry and Physics of Lipids, Dec 1, 2005

Labd-13(E)-ene-8alpha,15-diol (1) and its derivative labd-13(E)-ene-8alpha-ol-15-yl-acetate (2) a... more Labd-13(E)-ene-8alpha,15-diol (1) and its derivative labd-13(E)-ene-8alpha-ol-15-yl-acetate (2) are water insoluble biological active molecules and their structures were elucidated using NMR and X-ray techniques. Differential scanning calorimetry (DSC) was applied to study the thermal effects of 1 and 2 on DPPC bilayers. Liposomes composed of egg phosphatidylcholine/dipalmytoylphosphatidylglycerol (9:0.1 molar ratio) were prepared by the thin-film hydration method and were used for incorporating 1 and 2. Free and liposomal 1 and 2 were tested for their activity against human cancer cell lines using the sulphorhodamine B assay. The effect of 1 and 2 on DPPC bilayers caused abolition of the pre-transition temperature, lowering of the main phase transition and reduction of the transition enthalpy only in the presence of cholesterol. The liposomes that have been designed and developed offer high incorporation efficiency; 62.4% (0.369 drug/lipid molar ratio) and 99.7% (0.661 drug/lipid molar ratio) for 1 and 2, respectively. Liposomal 2 showed growth-inhibiting activity against the majority of the tested cell lines.

Research paper thumbnail of Line optical tweezers: A tool to induce transformations in stained liposomes and to estimate shear modulus

Colloids and Surfaces A: Physicochemical and Engineering Aspects, Oct 1, 2009

ABSTRACT Liposomes have been actively studied as models of cell membranes and are currently used ... more ABSTRACT Liposomes have been actively studied as models of cell membranes and are currently used as drug delivery systems of bioactive molecules. Liposome transformations that mimic cellular processes and are associated with their physicochemical properties have become field of interest the last decade. However, there has been little experimental work on controlled vesicle transformations by non-contact, optical handling methods.In this paper we present the use of line optical tweezers to observe liposome state transitions and transformations. Dynamic shape deformations were induced by line optical tweezers in giant stained liposomes leading to budding transition, fission and pearling creation. Under the controlled effect of line optical tweezers reversible liposome deformations were observed. The shear modulus μ of the membrane was inferred by measuring deformation of stained liposomes induced by the applied optical force. Further laser radiation caused irreversible shape deformations of liposomes, which were transformed from spherical to tubular vesicles. The ability of the selective manipulation of liposomes brings us closer to study their physicochemical properties which play a key role in cellular–liposome interactions, drug encapsulation and delivery efficiency.

Research paper thumbnail of Uptake Studies of Free and Liposomal Sclareol by Mcf-7 and H-460 Human Cancer Cell Lines

Springer eBooks, Sep 18, 2007

Research paper thumbnail of Mitochondria-targeted liposomes improve the apoptotic and cytotoxic action of sclareol

Journal of Liposome Research, Nov 2, 2009

Current efforts toward improving the effectiveness of drug therapy are increasingly relying on dr... more Current efforts toward improving the effectiveness of drug therapy are increasingly relying on drug-targeting strategies to effectively deliver bioactive molecules to their molecular targets. Pharmaceutical nanocarriers represent a major tool toward this aim, and our efforts have been directed toward achieving nanocarrier-mediated subcellular delivery of drug molecules with mitochondria as the primary subcellular target. Meeting the need for specific subcellular delivery is essential to realizing the full potential of many poorly soluble anticancer drugs. In this article, we report that mitochondria-targeted liposomes significantly improve the apoptotic and cytotoxic action of sclareol, a poorly soluble potential anticancer drug. The results support the broad applicability of our nanocarrier-mediated subcellular targeting approach as a means to improve the effectiveness of certain anticancer therapeutics.

Research paper thumbnail of Encapsulation of vinblastine into new liposome formulations prepared from triticum (wheat germ) lipids and its activity against human leukemic cell lines

Anticancer research

Liposomes prepared from lipids isolated from Triticum sp. (wheat germ) were used to investigate t... more Liposomes prepared from lipids isolated from Triticum sp. (wheat germ) were used to investigate the percentage of Vinblastine encapsulation and its retention into liposomes. The wheat germ total lipids (TL) were extracted by the Bligh-Dyer method and the lipid classes have been isolated using chromatographic techniques. The type of lipids and their percentage content have been examined by TLC coupled with an FID (latroscan). Two liposomal formulations, i.e., I and II, with encapsulated vinblastine, and formulation III (empty liposomes) have been prepared by thin film hydration method. The cytotoxic/cytostatic activity of these liposomal formulations have been examined against nine human leukemic cell lines. The results showed that the percentage content of vinblastine into liposomes I and II depended on the lipid composition and it was greater into formulation II (> 90%). The retention of the drug into liposomes was studied and found to be time-dependent at 37 degrees C. For the ...

Research paper thumbnail of Development of simple thiol-reactive liposome formulations, one-step analysis and physicochemical characterization

Journal of Pharmacy and Pharmacology, 2005

The aim of this study was to synthesize simple thiol-reactive conjugates from maleimide and lipoa... more The aim of this study was to synthesize simple thiol-reactive conjugates from maleimide and lipoamines (stearylamine or oleylamine) and to develop a simple, fast and low-cost method for the preparation of lyophilized general-purpose thiol-reactive liposomes. A formulation of egg phosphatidylcholine-dipalmitoylphoshatidylglycerol (9:0.1 molar ratio) was developed and characterized. Freeze-drying methodology was established to produce a stock of liposomes and the physicochemical characteristics of the reconstituted liposomes were compared with those of the initial preparation. The physicochemical properties (size and ζ-potential) of the new liposomal formulations were studied. High-performance thin-layer chromatography coupled to a flame ionization detector was applied for one-step analysis of the liposomal components and for determining the maleimide-lipoamine conjugates phospholipid molar ratio. The differences concerning the incorporation efficiency of the synthetic conjugates into...

Research paper thumbnail of In-vitro cytotoxic/cytostatic activity of anionic liposomes containing vinblastine against leukaemic human cell lines

Journal of Pharmacy and Pharmacology, 2002

Liposomes prepared from lipids dipalmitoylphosphatidylcholine (DPPC) and dipalmitoylphosphatidylg... more Liposomes prepared from lipids dipalmitoylphosphatidylcholine (DPPC) and dipalmitoylphosphatidylglycerol (DPPG) with cholesterol were used to investigate the percentage of vinblastine encapsulation and the influence of lipid composition on the retention properties of vinblastine in buffer as well as in cell culture medium. Differential scanning calorimetry (DSC) was applied, to study the effect of cholesterol on the phase transition temperature and on the ΔH of the two liposome formulations. The cytotoxic and cytostatic activity of the liposome-encapsulated vinblastine was also examined against six leukaemic human cell lines. The results showed that encapsulation of vinblastine into liposomes was greater than 98% with a drug-phospholipid molar ratio of 0.13-0.18. The major improvement in vinblastine retention in buffer as well as in culture medium was achieved by employing DPPG. The DSC data showed that vinblastine exerted a more perturbing effect in DPPC-cholesterol bilayers than i...

Research paper thumbnail of Accumulation of vinblastine into transfersomes and liposomes in response to a transmembrane ammonium sulfate gradient and their cytotoxic/cytostatic activity in vitro

Anticancer research

Vinblastine was encapsulated into liposomes composed from lipids dimiristoylphosphatidylcholine (... more Vinblastine was encapsulated into liposomes composed from lipids dimiristoylphosphatidylcholine (DMPC) and dipalmitoylphosphatidylcholine (DPPC), with cholesterol and transfersomes with sodium cholate prepared by the thin film hydration method. The percentage of vinblastine encapsulation, the stability of transfersomes and liposomes and the rate of release of encapsulated vinblastine at 37 degrees C were studied. The results showed that encapsulation of vinblastine into liposomes was higher than 98% at a drug/phospholipid molar ratio from 0.17 to 0.18, while encapsulation of vinblastine into transfersomes varied from 50% to 80% at a drug/phospholipid molar ratio from 0.05 to 0.09. The retention of drug in liposomes and in transfersomes was found to be time/dependent. The retention of drug in transfersomes compared to the liposomes was reduced due to the presence of sodium cholate which caused destabilization and reduced the main phase transition temperature Tm of the PC bilayers. Th...

Research paper thumbnail of Chapter 8 Uptake Studies of Free and Liposomal Sclareol by MCF-7 and H-460 Human Cancer Cell Lines

The aim of this study was to investigate the uptake of free and liposomal sclareol and its effect... more The aim of this study was to investigate the uptake of free and liposomal sclareol and its effect on the growth inhibiting activity against MCF-7 and H-460 human cancer cell lines in vitro. Liposomes composed of EPC/DPPG at molar ratio 9:0.1, used to incorporate sclareol, were prepared by the thin-film hydration method followed by sonication. The final liposomal preparation (EPC/DPPG/Sclareol 9:0.1:5 molar ratio) as well as free sclareol (100μM) were incubated up to 96 hours with both cell lines. Sclareol was extracted from cells using the Bligh-Dyer method and was measured by HPTLC/FID. The results showed that the uptake of free sclareol by both cell lines was faster and higher compared to that of its liposomal form. In both cell lines, free sclareol showed high cytotoxicity, while the liposomal sclareol showed reduced cytotoxicity without affecting its ability to reduce the cell growth rate. These findings suggest that liposomal sclareol may possess chemotherapeutic advantages ove...

Research paper thumbnail of Liposomic Niflumic Acid - New Transdermal Anti-Inflammatory Medicine

Research paper thumbnail of Molecular interactions between dimethoxycurcumin and Pamam dendrimer carriers

International Journal of Pharmaceutics, 2007

Dimethoxycurcumin, a lipophilic analog of curcumin found as a major pigment in the Indian species... more Dimethoxycurcumin, a lipophilic analog of curcumin found as a major pigment in the Indian species turmeric (Curcuma longa Linn.), is known to possess significant activity against various cancer cell lines, but its use as an anticancer drug is hindered by its poor water solubility. The conjugation of dimethoxycurcumin to water-soluble PAMAM dendrimers (generations 3.5 and 4) is demonstrated. The maximum drug-dendrimer incorporation efficiency is 4.3 and 5.0 molar for G3.5 and G4, respectively. The FTIR-ATR investigation of the neat compounds and the drug-dendrimer systems indicate that dimethoxycurcumin is in the enolic form, while its interaction with the integer generation dendrimer involves the major conformational change of the terminal ethylene amine groups.

Research paper thumbnail of Cytotoxic and antitumor activity of liposome-incorporated sclareol against cancer cell lines and human colon cancer xenografts

Pharmacological Research, 2006

The aim of this study was to design and prepare liposome-incorporated sclareol-a highly lipophili... more The aim of this study was to design and prepare liposome-incorporated sclareol-a highly lipophilic natural product-to overcome its water insolubility and develop suitable formulations for in vivo administration. The bioactive labdane-type diterpene sclareol was incorporated into liposomes composed of egg phosphatidylcholine and dipalmitoylphosphatidylglycerol prepared by the thin-film hydration method followed by sonication. A formulation of egg phosphatidylcholine/dipalmitoylphosphatidylglycerol/sclareol (9:0.1:5 molar ratio) was developed and characterized. The lipid recovery and the sclareol to lipid molar ratio were measured using high-performance thin-layer chromatography/flame ionization detection. In vitro drug release was measured in supplemented RPMI-1640 at 37 • C. The liposomal and the free sclareol were initially tested in vitro for their activity against human cancer cell lines using the sulphorhodamine B assay. Liposomes incorporating sclareol at a drug to lipid molar ratio of 0:43, suggesting an incorporation efficiency of almost 80%, showed reduced growth rate of human colon cancer tumors (HCT116) developed in SCID mice, without any significant side effects.

Research paper thumbnail of Development and Characterization of Oligonucleotide-Tagged Dye-Encapsulating EPC/DPPG Liposomes

Journal of Nanoscience and Nanotechnology, 2010

Liposomes applications in health care include meanly their ability to carry drugs and genes insid... more Liposomes applications in health care include meanly their ability to carry drugs and genes inside the human body for therapeutic purposes. Nevertheless their applicability can extend far beyond and could be used as analytical tools in order to perform rapid, low-cost, sensitive and specific analyses. Their physical characteristics, such as large internal volume and extended surface area, render them ideal for these applications and specifically for improving the specificity and sensitivity of the analytical assay. The purpose of this study was to develop a simple, stable and low-cost oligonucleotide-tagged liposomal formulation consisting of EggPC and DPPG with a simple to synthesize thiol-reactive conjugate (Mal-SA) incorporated into the lipid bilayer of liposomes. The prepared liposomes, having also the water soluble dye Sulforhodamine B encapsulated in their inner cavity, were characterized in terms of their physicochemical (size, size distribution,-potential, lipid content) and mechanical (morphology, rigidity) properties. The results showed that the final liposomal formulation could be used in the future as analytical tool for detecting pathogen strains of microorganism in biological milieu.

Research paper thumbnail of Mitochondria-targeted liposomes improve the apoptotic and cytotoxic action of sclareol

Journal of Liposome Research, 2009

Current efforts toward improving the effectiveness of drug therapy are increasingly relying on dr... more Current efforts toward improving the effectiveness of drug therapy are increasingly relying on drug-targeting strategies to effectively deliver bioactive molecules to their molecular targets. Pharmaceutical nanocarriers represent a major tool toward this aim, and our efforts have been directed toward achieving nanocarrier-mediated subcellular delivery of drug molecules with mitochondria as the primary subcellular target. Meeting the need for specific subcellular delivery is essential to realizing the full potential of many poorly soluble anticancer drugs. In this article, we report that mitochondria-targeted liposomes significantly improve the apoptotic and cytotoxic action of sclareol, a poorly soluble potential anticancer drug. The results support the broad applicability of our nanocarrier-mediated subcellular targeting approach as a means to improve the effectiveness of certain anticancer therapeutics.

Research paper thumbnail of Lipid analysis of Greek broad bean oil: Preparation of liposomes and physicochemical characterization

European Journal of Lipid Science and Technology, 2005

ABSTRACT Liposomes were prepared from the isolated phospholipids of mature broad bean [Vicia faba... more ABSTRACT Liposomes were prepared from the isolated phospholipids of mature broad bean [Vicia faba L. (syn. Fabae calabaricae)] oil and their physical properties were studied. The method of preparation was the hydration of the thin lipid film, while the probe sonication methodology was used for reducing the size of the vesicles. The seeds of the broad bean were collected in two different periods of maturity and extracted by the Bligh-Dyer method, and the lipid classes were studied by HPTLC/FID. The oils were found to be rich in polar lipids (63.1% and 60.2% of total lipids) and low in neutral lipids (36.9% and 39.8% of total lipids) for the immature and mature seed oils, respectively. The neutral lipid fraction consisted mainly of triacylglycerides (34.2% and 32.3%) whereas the polar lipids mainly consisted of phospholipids (60.2% and 54.2%) for the mature and immature seed oils, respectively. Sphingolipids (8.9%) were identified only in the immature seed oil. The overall goal of this study was the preparation of a new liposomal formulation with physicochemical properties such as unique lipid composition, size and ζ-potential, which are important factors influencing drug delivery to the target tissues.

Research paper thumbnail of Dendrimers as Biopharmaceuticals: Synthesis and Properties

Current Topics in Medicinal Chemistry, 2008

Two general aspects which need to be considered for the successful application of dendrimers for ... more Two general aspects which need to be considered for the successful application of dendrimers for biomedical purposes are their availability at an acceptable cost and their suitability as regards their pharmacodynamic and pharmacokinetic properties. These two aspects are covered in this review. In the first part, synthetic strategies for the preparation of dendrimers are outlined and emphasis is given to recent work on methodologies whose aim is the development of more efficient routes to dendrimers in terms of the materials used for their synthesis as well as in terms of the procedures required for their purification. These include procedures involving double-stage and double exponential synthesis, orthogonal coupling strategies, self-assembly and solid-phase approaches, as well as particularly useful synthetic protocols such as those used in "click chemistry". The second part of the review deals with the way in which the size, chemical constitution and physicochemical properties of dendrimers used for drug delivery may affect pharmacodynamic and pharmacokinetic parameters which are important considerations for drug bioavailability. This is illustrated by an overview of examples from recent work involving non-steroidal anti-inflammatory drugs, anticancer drugs and antibacterials.