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ABSTRACT: Solid tumors constitute the majority of diagnosed cancers. For effective killing, therapeutic agents should ideally be delivered uniformly and at lethal doses to all cancer cells comprising the tumors, while keeping normal organ toxicities to a minimum. This requirement sets two of the major challenges in drug delivery to solid cancers: uniformity in delivery, and delivery of at least a minimum amount of therapeutics per cancer cell. Herein we review various approaches that aim to improve the penetration and content release of delivered therapeutic agents from nanocarriers of self-assembling nature. Biophysical characteristics of solid tumors are briefly discussed to motivate and rationalize the design of reported nanoparticle structures. This review does not aim to be exhaustive of the various designs and strategies, but to mostly give a flavor of the general current directions aiming to address these challenges.
Current pharmaceutical biotechnology 09/2012; 13(7):1306-16. · 3.40 Impact Factor
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ABSTRACT: pH-triggered lipid-membranes designed from biophysical principles are evaluated in the form of targeted liposomal doxorubicin with the aim to ultimately better control the growth of vascularized tumors. We compare the antitumor efficacy of anti-HER2/neu pH-triggered lipid vesicles encapsulating doxorubicin to the anti-HER2/neu form of an FDA approved liposomal doxorubicin of DSPC/cholesterol-based vesicles. The HER2/neu receptor is chosen due to its abundance in human breast cancers and its connection to low prognosis. On a subcutaneous murine BT474 xenograft model, superior control of tumor growth is demonstrated by targeted pH-triggered vesicles relative to targeted DSPC/cholesterol-based vesicles (35% vs. 19% decrease in tumor volume after 32 days upon initiation of treatment). Superior tumor control is also confirmed on SKBR3 subcutaneous xenografts of lower HER2/neu expression. The non-targeted form of pH-triggered vesicles encapsulating doxorubicin results also in better tumor control relative to the non-targeted DSPC/cholesterol-based vesicles (34% vs. 41% increase in tumor volume). Studies in BT474 multicellular spheroids suggest that the observed efficacy could be attributed to release of doxorubicin directly into the acidic tumor interstitium from pH-triggered vesicles extravasated into the tumor but not internalized by cancer cells. pH-triggered liposome carriers engineered from gel-phase bilayers that reversibly phase-separate with lowering pH, form transiently defective interfacial boundaries resulting in fast release of encapsulated doxorubicin. Our studies show that pH-triggered liposomes release encapsulated doxorubicin intracellularly and intratumorally, and may improve tumor control at the same or even lower administered doses relative to FDA approved liposomal chemotherapy.
Biomaterials 03/2012; 33(17):4345-52. · 7.40 Impact Factor
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ABSTRACT: Lateral lipid phase separation of titratable PS or PA lipids and their assembly in domains induced by changes in pH are significant in liposome-based drug delivery: environmentally responsive lipid heterogeneities can be tuned to alter collective membrane properties such as permeability (altering drug release) and surface topography (altering drug carrier reactivity) impacting, therefore, the therapeutic outcomes. At the micrometer scale fluorescence microscopy on giant unilamellar fluid vesicles (GUVs) shows that lowering pH (from 7.0 to 5.0) promotes condensation of titratable PS or PA lipids into beautiful floret-shaped domains in which lipids are tightly packed via hydrogen-bonding and van der Waals interactions. The order of lipid packing within domains increases radially toward the domain center. Lowering pH enhances the lipid packing order, and at pH 5.0 domains appear to be entirely in the solid (gel) phase. Domains phenomenologically comprise a circular "core" cap beyond which interfacial instabilities emerge resembling leaf-like stripes. At pH 5.0 stripes are of almost vanishing Gaussian curvature independent of GUVs' preparation path and in agreement with a general condensation mechanism. Increasing incompressibility of domains is strongly correlated with a larger number of thinner stripes per domain and increasing relative rigidity of domains with smaller core cap areas. Line tension drives domain ripening; however, the final domain shape is a result of enhanced incompressibility and rigidity maximized by domain coupling across the bilayer. Introduction of a transmembrane osmotic gradient (hyperosmotic on the outer lipid leaflet) allows the domain condensation process to reach its maximum extent which, however, is limited by the minimal expansivity of the continuous fluid membrane.
Langmuir 03/2012; 28(9):4113-22. · 4.19 Impact Factor
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ABSTRACT: The killing efficacy of doxorubicin from liposome-based delivery carriers has been shown to correlate strongly with its intracellular trafficking and, in particular, its fast and extensive release from the delivery carrier. However, previously explored pH-triggered mechanisms that were designed to become activated during liposome endocytosis have also been shown to interfere with the liposome stability in vivo. We have designed pH-triggered gel-phase liposomes with heterogeneous membranes for the delivery of doxorubicin. These liposomes are triggered to form "leaky" interfacial boundaries between gel-gel phase separated domains on the membrane bilayer with lowering pH. The pH-triggered mechanism does not compromise liposome stability in vivo and results in superior in vitro killing efficacy of delivered doxorubicin when liposomes are endocytosed by a clathrin-mediated pathway. In the present work, we evaluate the general applicability of these liposomes when targeted to the folate receptor (FR) of KB cancer cells in vitro and become endocytosed by a less acidic pathway: the caveolae pathway. FR-targeting liposomes exhibit almost 50% decrease in cell association for increase in liposome size from 120 to 280 nm in diameter after relatively short incubation times (up to 4 h). The fraction of internalized vesicles, however, is approximately 60% of the cell associated vesicles independent of their size. Our findings demonstrate that, for the same doxorubicin uptake per cancer cell, the killing effect of doxorubicin delivered by pH-triggered lipid vesicles is greater (IC(50) = 0.032 mM for a 6 h incubation) than when delivered by a conventional non-pH-responsive composition (IC(50) = 0.194 mM). These findings suggest higher bioexposure of cells to the therapeutic agent possibly via faster and more extensive release from the carrier. Animal studies of FR-targeting non-pH-responsive liposomal doxorubicin report stronger therapeutic potential for the targeted approach relative to nontargeted liposomes and to free doxorubicin. The findings of the present study suggest that the targeted pH-triggered liposomes could potentially further enhance the therapeutic outcomes of doxorubicin in vivo.
Molecular Pharmaceutics 09/2011; 8(6):2224-32. · 4.78 Impact Factor
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ABSTRACT: Effective targeting and killing of intraperitoneally disseminated micrometastases remains a challenge.
In this work, we evaluated the potential of antibody-labeled PEGylated large liposomes as vehicles for direct intraperitoneal (i.p.) drug delivery with the aim to enhance the tumor-to-normal organ ratio and to improve the bioexposure of cancer cells to the delivered therapeutics while shifting the toxicities toward the spleen. These targeted liposomes are designed to combine: (1) specific targeting to and internalization by cancer cells mediated by liposome-conjugated tumor-specific antibodies, (2) slow clearance from the peritoneal cavity, and (3) shift of normal organ toxicities from the liver to the spleen due to their relatively large size.
Conjugation of anti-HER2/neu antibodies to the surface of large (approximately 600 nm in diameter) PEGylated liposomes results in fast, specific binding of targeted liposomes to cancer cells in vitro, followed by considerable cellular internalization. In vivo, after i.p. administration, these liposomes exhibit fast, specific binding to i.p. cancerous tumors. Large liposomes are slowly cleared from the peritoneal cavity, and they exhibit increased uptake by the spleen relative to the liver, while targeted large liposomes demonstrate specific tumor uptake at early times. Although tissue and tumor uptake are greater for cationic liposomes, the tumor-to-liver and spleen-to-liver ratios are similar for both membrane compositions, suggesting a primary role for the liposome's size, compared to the liposome's surface charge.
The findings of this study suggest that large targeted liposomes administered i.p. could be a potent drug-delivery strategy for locoregional therapy of i.p. micrometastatic tumors.
Journal of Liposome Research 12/2010; 20(4):330-40. · 1.71 Impact Factor
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ABSTRACT: To enable selective cell-kill, we designed functionalized lipid vesicles with pH-triggered heterogeneous membranes and encapsulated doxorubicin that exhibit tunable surface topography. These vesicles "hide" (mask) the targeting ligands from their surface during circulation in the blood, and only progressively "expose" these ligands as they gradually penetrate deeper into the tumor interstitium, where after endocytosis they burst release their contents. The stimulus to activate the binding reactivity is the pH gradient between the blood stream (pH 7.4-7.0) and the increasingly acidic pH inside the tumor interstitium (pH 6.7-6.5). Doxorubicin release is activated at the endosomal pH 5.5-5.0. We show that tunable functionalized vesicles exhibit environmentally-dependent (pH-dependent) association with cancer cells resulting in high cell-kill selectivity. When lowering the extracellular pH from 7.4 to 6.5, tunable functionalized vesicles deliver doxorubicin to cancer cells that increases from 41% to 93% of maximum resulting in cancer cell killing that increases from 23 to 71% of maximum, respectively. This proof-of-concept shows the potential of tunable targeted liposomal chemotherapy to selectively kill cancer cells in an environmentally-dependent way.
Biomaterials 02/2010; 31(15):4409-16. · 7.40 Impact Factor
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ABSTRACT: The role of pH-dependent lipid heterogeneities on the fusogenicity of membranes was evaluated on model lipid bilayers in the form of unilamellar vesicles composed of lipid pairs at a fixed equimolar ratio of phosphatidylcholine (PC) and phosphatidic acid (PA) headgroups. The pH and the hydrophobic composition (lipid acyl tails) of membranes were systematically altered, and their effect on vesicle aggregation, membrane fusogenicity, content release, and content mixing was evaluated. Lowering pH increases the extent of protonated PA headgroups forming phase-separated PA-rich heterogeneities and giving rise to molecular packing defects originating at the phase boundaries. Phase boundaries within the membrane's hydrophobic portion are affected by the lipid acyl-tail dynamics (fluidity) and the matching or nonmatching lengths of the acyl tails of lipid pairs comprising the membrane. The aggregates' size increases with lowering pH and is independent of the membrane's hydrophobic composition. Contrary to aggregation, the initial rates of lipid mixing are proportional to pH and also depend on membrane's hydrophobic composition. The apparent lipid-mixing rate constants are greater for membranes containing lipid pairs with mismatched acyl-tail lengths, followed by pairs with matching acyl tails in the gel state and by pairs with matching tails where one lipid is close to its transition temperature. Addition of cholesterol conserves the independence of vesicle aggregation from the membrane's hydrophobic composition. However, it decreases the aggregation rates and inverts the tendency for fusion, among the three types of hydrophobic compositions, suggesting a role of cholesterol's preferential partition in different regions of membrane's heterogeneities. We propose a phenomenological model where the membrane phase boundaries containing molecular packing defects act as "sticking points" on the vesicle exterior via which vesicles aggregate upon contact followed by defect merging via intervesicle lipid exchange and mixing. Such heterogeneous bilayers in the form of drug encapsulating liposomes may potentially improve the therapeutic efficacy by fusing with endosomal membranes, thus increasing drug bioavailability.
Langmuir 10/2009; 26(3):1666-73. · 4.19 Impact Factor
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ABSTRACT: During endocytosis, pH-triggered release of encapsulated therapeutics from delivery carriers may accelerate their intracellular trafficking increasing therapeutic efficacy. To improve the therapeutic potential of targeted immunochemotherapy using anti-HER2/neu liposomal doxorubicin, we exploit the formation of leaky heterogeneities on rigid lipid bilayers to extensively release doxorubicin during endocytosis. We have previously demonstrated that pH-dependent formation of phase-separated lipid heterogeneities on the plane of a bilayer membrane increases the permeability of bilayers when they are composed of lipid pairs with rigid non-matching acyl chain lengths. This was suggested to be due to defective packing among lipids residing at the interfaces of lipid domains. Here we design nanometer-size antiHER2/neu-labeled PEGylated vesicles composed of lipid pairs with longer non-matching acyl chain lengths (n=18 and 21). These vesicles exhibit superior killing efficacy of cancer cells compared to established liposome formulations, and their killing efficacy is similar to the effect of combined free doxorubicin and free antiHER2/neu antibody. Other transport-related properties such as liposome blood circulation times, and specific binding and internalization by cancer cells are unaffected. These results demonstrate the potential of vesicles with pH-triggered leaky heterogeneities to increase the therapeutic potential of targeted immunochemotherapy.
Biomaterials 09/2009; 30(30):6055-64. · 7.40 Impact Factor
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ABSTRACT: During direct cell-to-cell communication, lipids on the extracellular side of plasma membranes reorganize, and membrane-associated communication-related molecules colocalize. At colocalization sites, sometimes identified as rafts, the local cell surface topography and reactivity are altered. The processes regulating these changes are largely unknown. On model lipid membranes, study of simplified processes that control surface topography and reactivity may potentially contribute to the understanding and control of related cell functions and associated diseases. Integration of these processes on nanometer-sized lipid vesicles used as drug delivery carriers would precisely control their interactions with diseased cells minimizing toxicities. Here we design such basic pH-dependent processes on model functionalized lipid bilayers, and we demonstrate reversible sharp changes in binding reactivity within a narrow pH window. Cholesterol enables tuning of the membrane reorganization to occur at pH values not necessarily close to the reported pK(a)'s of the constituent titratable lipids, and bilayer reorganization over repeated cycles of induced pH changes exhibits hysteresis.
Langmuir 08/2009; 25(14):8144-51. · 4.19 Impact Factor
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ABSTRACT: This paper reports the application of frozen cyclohexane-in-water emulsions as sacrificial templates for the fabrication of hollow microcapsules through layer-by-layer assembly of polyelectrolytes, poly(styrenesulfonate sodium salt), and poly(allylamine hydrochloride). Extraction of the cyclohexane phase from frozen emulsions stabilized with 11 polyelectrolyte layers by compatibilization with 30% v/v ethanol leads to the formation of water-filled microcapsules while preserving the spherical geometry. The majority of microcapsules (>90%) are prepared with intact polyelectrolyte membranes as measured by their deformation induced by osmotic pressure. This work provides a new route for the synthesis of hollow multilayered microcapsules under mild operating conditions.
Langmuir 06/2009; 25(17):9728-33. · 4.19 Impact Factor
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Advanced Drug Delivery Reviews 10/2008; 60(12):1317-8. · 11.50 Impact Factor
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ABSTRACT: Heterogeneous lipid membranes tuned by pH were evaluated at 37 degrees C in the form of PEGylated vesicles composed of lipid pairs with dipalmitoyl ( n = 16) and distearoyl ( n = 18) chain lengths. One lipid type was chosen to have the titratable moiety phosphatidic acid on its headgroup, and the other lipid type was chosen to have a phosphatidylcholine headgroup. The effect of pH on the formation of lipid heterogeneities and on membrane permeability was studied on vesicles composed of lipid pairs with matching and nonmatching chain lengths. The formation of lipid heterogeneities increases with decreasing pH in membranes composed of lipid pairs with either matching or nonmatching chain lengths. Increased permeability with decreasing pH was exhibited only by membranes composed of lipid pairs with nonmatching chain lengths. Permeability rates correlate strongly with the predicted extent of interfacial boundaries of heterogeneities, suggesting defective packing among nonmatching acyl chains of lipids. In heterogeneous mixtures with one lipid type in the fluid state ( n = 12), the dependence of membrane permeability on pH is weaker. In the presence of serum proteins, PEGylated gel-phase vesicles containing lipid pairs with nonmatching chain lengths exhibit faster release rates with decreasing pH compared to measured release rates in phosphate buffer, suggesting a second mechanism of formation of separated phases. PEGylated vesicles composed of lipid pairs with nonmatching chain lengths labeled with internalizing anti-HER2/neu antibodies that target overexpressed antigens on the surface of SKOV3-NMP2 ovarian cancer cells exhibit specific cancer cell targeting, followed by extensive internalization (more than 84% of bound vesicles) and fast release of contents intracellularly. These PEGylated vesicles composed of rigid membranes for long blood circulation times that exhibit pH-dependent release of contents intracellularly could become potent drug delivery carriers for the targeted therapy of solid tumors.
Langmuir 07/2008; 24(11):5679-88. · 4.19 Impact Factor
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ABSTRACT: Targeted alpha-particle emitters are promising therapeutics for micrometastatic disease. Actinium-225 has a 10-day half-life and generates a total of four alpha-particles per parent decay rendering (225)Ac an attractive candidate for alpha-therapy. For cancer cells with low surface expression levels of molecular targets, targeting strategies of (225)Ac using radiolabeled carriers of low specific radioactivities (such as antibodies) may not deliver enough alpha-particle emitters at the targeted cancer cells to result in killing. We previously proposed and showed using passive (225)Ac entrapment that liposomes can stably retain encapsulated (225)Ac for long time periods, and that antibody-conjugated liposomes (immunoliposomes) with encapsulated (225)Ac can specifically target and become internalized by cancer cells. However, to enable therapeutic use of (225)Ac-containing liposomes, high activities of (225)Ac need to be stably encapsulated into liposomes. In this study, various conditions for active loading of (225)Ac in preformed liposomes (ionophore-type, encapsulated buffer solution, and loading time) were evaluated, and liposomes with up to 73 +/- 9% of the initial activity of (225)Ac (0.2-200 microCi) were developed. Retention of radioactive contents by liposomes was evaluated at 37 degrees C in phosphate buffer and in serum-supplemented media. The main fraction of released (225)Ac from liposomes occurs within the first two hours of incubation. Beyond this two hour point, the encapsulated radioactivity is released from liposomes slowly with an approximate half-life of the order of several days. In some cases, after 30 days, (225)Ac retention as high as 81 +/- 7% of the initially encapsulated radioactivity was achieved. The (225)Ac loading protocol was also applied to immunoliposome loading without significant loss of targeting efficacy. Liposomes with surface-conjugated antibodies that are loaded with (225)Ac overcome the limitations of low specific activity for molecular carriers and are expected to be therapeutically useful against tumor cells having a low antigen density.
Bioconjugate Chemistry 06/2008; 19(6):1274-82. · 4.93 Impact Factor
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ABSTRACT: BACKGROUND: Targeted liposomes can be broadly defined as liposomes that are engineered to interact with a particular population of cells with the objective of delivering a payload or increasing their retention within the targeted cell population by means of a chemical interaction with cell-surface molecules or other tissue-specific ligands. OBJECTIVE: The authors review recent advances in the field with an emphasis on pre-clinical studies and place them in the context of historical developments. METHODS: The review focuses on immunoliposomes (antibody-mediated targeting) as these constructs are presently the most prevalent. Conclusion: The field has advanced in tandem with advances in liposome design and antibody and protein engineering. Targeted liposomes have been used in diagnosis to deliver magnetic resonance contrast agents and radionuclides for magnetic resonance and nuclear medicine imaging, respectively. They have been used in gene therapy to deliver a variety of gene expression modifiers, including plasmids, anti-sense oligonucleotides and short interfering RNA. Targeted liposomes provide a delivery advantage over untargeted liposomes not because of increased localization to tumor sites but because of increased interaction with the target cell population once localized to the tumor site. The increased interaction can take on the form of fusion with the cellular membrane or internalization by endocytosis. To the extent that the spatial distribution of targeted liposomes within a solid tumor may become more non-uniform than has been found for untargeted liposomes, this may be a drawback. However, systematic comparisons of the spatial distribution in tumors of targeted versus untargeted liposomes have yet to be performed. The majority of reported studies have been in the area of chemotherapy delivery. Their use in radionuclide and chemo- and radiosensitizer delivery is just emerging. Multifunctional liposomes containing 'layered functionalities' could potentially be the future direction in targeted liposome-based therapy.
Expert Opinion on Drug Delivery 03/2008; 5(2):189-204. · 4.90 Impact Factor
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Stavroula Sofou
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ABSTRACT: This review describes strategies for the delivery of therapeutic radionuclides to tumor sites. Therapeutic approaches are summarized in terms of tumor location in the body, and tumor morphology. These determine the radionuclides of choice for suggested targeting ligands, and the type of delivery carriers. This review is not exhaustive in examples of radionuclide carriers for targeted cancer therapy. Our purpose is two-fold: to give an integrated picture of the general strategies and molecular constructs currently explored for the delivery of therapeutic radionuclides, and to identify challenges that need to be addressed. Internal radiotherapies for targeting of cancer are at a very exciting and creative stage. It is expected that the current emphasis on multidisciplinary approaches for exploring such therapeutic directions should enable internal radiotherapy to reach its full potential.
International Journal of Nanomedicine 02/2008; 3(2):181-99. · 3.13 Impact Factor
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Stavroula Sofou
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ABSTRACT: An overview of liposome-based drug-delivery carriers to cancer cells is presented. Properties related to interfacial interactions between liposomes and the biological milieu that determine the fate of liposomes in vivo are discussed. Original approaches to improve specificity for the target and to control the structural responsiveness of liposomes, depending on their immediate environment, with the aim of enhancing the delivered therapeutic doses, are summarized. This review is not exhaustive on research examples of liposomes as carriers for cancer therapy but, rather, aims to describe major directions of designs and strategies over recent years. The current therapeutic trends that exhibit increasingly higher complexity in structures and responses are also discussed.
Nanomedicine 11/2007; 2(5):711-24. · 5.05 Impact Factor
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ABSTRACT: Disseminated, metastatic cancer is frequently incurable. Targeted alpha-particle emitters hold great promise as therapeutic agents for disseminated disease. (225)Ac is a radionuclide generator that has a 10-d half-life and results in alpha-emitting daughter elements ((221)Fr, (217)At, (213)Bi) that lead to the emission of a total of 4 alpha-particles. The aim of this study was to develop approaches for stable and controlled targeting of (225)Ac to sites of disseminated tumor metastases. Liposomes with encapsulated (225)Ac were developed to retain the potentially toxic daughters at the tumor site.
(225)Ac was passively entrapped in liposomes. To experimentally test the retention of actinium and its daughters by the liposomes, the gamma-emissions of (213)Bi were measured in liposome fractions, which were separated from the parent liposome population and the free radionuclides, at different times. Under equilibrium conditions the decay rate of (213)Bi was used to determine the concentration of (225)Ac. Measurements of the kinetics of (213)Bi activity were performed to estimate the entrapment of (213)Bi, the last alpha-emitting daughter in the decay chain.
Stable pegylated phosphatidylcholine-cholesterol liposomes of different sizes and charge were prepared. Multiple (more than 2) (225)Ac atoms were successfully entrapped per liposome. (225)Ac retention by zwitterionic liposomes was more than 88% over 30 d. Retention by cationic liposomes was lower. A theoretical calculation showed that for satisfactory (213)Bi retention (>50%), liposomes of relatively large sizes (>650 nm in diameter) are required. (213)Bi retention was experimentally verified to be liposome-size dependent. For large liposomes, the measured (213)Bi retention was lower than theoretically predicted (less than 10%).
This work supports the hypothesis that it may be possible to develop (225)Ac-based therapies by delivering multiple (225)Ac atoms in liposomes. Improvements in the retention of (225)Ac daughters will likely be necessary to fulfill this potential. Because of the size of the liposomal structures required to contain the daughters, the approach is ideally suited for locoregional therapy (e.g., intraperitoneal, intrahepatic artery, or intrathecal).
Journal of Nuclear Medicine 02/2004; 45(2):253-60. · 6.38 Impact Factor
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ABSTRACT: Phospholipid vesicles are well-studied biomembrane mimics that are of increasing interest in drug delivery, immunoassays, and sensor chips. In a number of biosensor applications it is desirable to be able to adhere vesicles to a surface in a manner which does not result in their rupture or fusion. Such behavior should, in principle, be achievable by controlling the vesicle-surface and vesicle-vesicle interactions. We have varied vesicle composition and charge (phosphatidylcholine, phosphatidylcholine-phosphatidic acid 18 mol%) and solution ionic strength, to study the adhesion of fluorescent vesicles to glass, gold, and gold modified with chemisorbed acetyl-cysteine. The extent of chemisorption was characterized with angle-resolved X-ray photoelectron spectroscopy (ARXPS), and vesicle integrity and behavior was studied using entrapped and lipophilic fluorescent markers, together and in separate measurements. Vesicle fusion (by energy transfer), adhesion of intact vesicles (with entrapped calcein) and diffusion coefficients (by photobleaching recovery) were monitored using confocal fluorescence microscopy. Acetyl-cysteine modified gold surfaces were shown to be appropriate substrates for adhesion of intact vesicles. Finally, as a 'proof of principle' for fluorescence amplification, release of a self-quenching entrapped reporter dye (calcein) by the detergent Triton X-100 was followed in real time.
Biosensors and Bioelectronics 05/2003; 18(4):445-55. · 5.60 Impact Factor
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ABSTRACT: The use of vesicles as amplifiers in biosensors is receiving increasing attention. Because vesicles may entrap thousands of reporter molecules, strong signal amplification can be obtained if a small number of analytes can simply release the entrapped reporters. Surface immobilization of vesicles with sensitivities for different analytes could then provide for simultaneous amplified detection of a number of analytes on a single chip. To achieve this goal, vesicles must first be stably adsorbed to a surface, without rupture. We have varied vesicle composition and charge (phosphatidylcholine, phosphatidylcholine-phosphatidic acid at 4.6 molar ratio) and solution ionic strength, to study the adsorption of fluorescent vesicles to glass, gold, and gold modified with chemisorbed acetylcysteine. Surfaces were characterized with angle-resolved X-ray photoelectron spectroscopy (ARXPS), and vesicle integrity and behavior was studied using entrapped and lipophilic fluorescent markers either together or in separate experiments. Diffusion coefficients (by photobleaching recovery) and vesicle fusion (by energy transfer) were monitored using confocal fluorescence microscopy. Finally, as a “proof of principle”, release of a self-quenching entrapped reporter dye (calcein) by the detergent Triton X-100 was followed in real time.
MRS Proceedings. 12/2000; 705.
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ABSTRACT: Targeted alpha-particle emitters hold great promise as therapeutics for micrometastatic disease. Because of their high energy deposition and short range, tumor targeted alpha-particles can result in high cancer-cell killing with minimal normal-tissue irradiation. Actinium-225 is a potential generator for alpha-particle therapy: it decays with a 10-day half-life and generates three alpha-particle-emitting daughters. Retention of (225)Ac daughters at the target increases efficacy; escape and distribution throughout the body increases toxicity. During circulation, molecular carriers conjugated to (225)Ac cannot retain any of the daughters. We previously proposed liposomal encapsulation of (225)Ac to retain the daughters, whose retention was shown to be liposome-size dependent. However, daughter retention was lower than expected: 22% of theoretical maximum decreasing to 14%, partially due to the binding of (225)Ac to the phospholipid membrane. In this study, Multivesicular liposomes (MUVELs) composed of different phospholipids were developed to increase daughter retention. MUVELs are large liposomes with entrapped smaller lipid-vesicles containing (225)Ac. PEGylated MUVELs stably retained over time 98% of encapsulated (225)Ac. Retention of (213)Bi, the last daughter, was 31% of the theoretical maximum retention of (213)Bi for the liposome sizes studied. MUVELs were conjugated to an anti-HER2/neu antibody (immunolabeled MUVELs) and were evaluated in vitro with SKOV3-NMP2 ovarian cancer cells, exhibiting significant cellular internalization (83%). This work demonstrates that immunolabeled MUVELs might be able to deliver higher fractions of generated alpha-particles per targeted (225)Ac compared to the relative fractions of alpha-particles delivered by (225)Ac-labeled molecular carriers.
Bioconjugate Chemistry 18(6):2061-7. · 4.93 Impact Factor
