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ABSTRACT: PurposeTo determine the transscleral permeability of chemotherapeutic drugs vinblastine and doxorubicin for treatment of intraocular
tumors, and to compare the use of doxorubicin encapsulated in PLGA and liposome nanoparticles.
MethodsHuman sclera was isolated and mounted in a Lucite chamber. Fluorescently tagged vinblastine (VIN), innately fluorescent free
doxorubicin (DOX), PLGA doxorubicin (PLGA-DOX), or Doxil (Tibotec Therapeutics) were added to the episcleral donor chamber.
The choroidal side was perfused with Balanced Salt Solution. Perfusate fractions were collected over 24h and measured for
fluorescence. Following the experiment, tissue sections were imaged, underwent a drug wash out procedure, and tissue drug
content was analyzed using an LC–MS/MS method.
ResultsWithin 24h, a total of 68%, 74%, 29%, and 1.9% of the drug dose from VIN, DOX, PLGA-DOX, and Doxil, respectively, diffused
across the sclera. VIN and DOX scleral tissue showed strong fluorescence after 24h. PLGA-DOX displayed scattered fluorescence,
and Doxil indicated minimal fluorescence. LC–MS/MS revealed strong tissue binding of DOX.
ConclusionsThis study suggests both vinblastine and doxorubicin are able to diffuse across human sclera. In addition, PLGA nanoparticles
delivered doxorubicin at a slower rate across the sclera, and the liposome preparation resulted in the slowest delivery of
drug.
Pharmaceutical Research 04/2012; 26(5):1155-1161. · 4.09 Impact Factor
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ABSTRACT: To determine whether long-term expression of intraceptors can be achieved using plasmid albumin nanoparticles and whether nanoparticles can inhibit and cause regression of murine corneal neovascularization induced by mechanical-chemical trauma.
Albumin nanoparticles encapsulating pCMV.Flt23K were developed as a lyophilized product that is easily redispersed in an aqueous medium. Nanoparticles were injected into the corneas of uninjured BALB/c mice and observed for toxicity for 3 weeks. Entry of nanoparticles into corneal cells was demonstrated through transmission electron microscopy and confocal imaging. Naked pCMV.Flt23K, nanoparticles encapsulating pCMV.Flt23K, or empty pCMV nanoparticles were injected into uninjured mouse corneas. These corneas were subjected to mechanical alkali trauma 3 weeks after injection.
Nanoparticles were nontoxic to the cornea and entered into corneal keratocyte cytoplasm. They persisted for at least 4 weeks in the cornea, expressed effective intraceptor levels for at least 5 weeks, and reduced corneal neovascularization by approximately 40% (P = 0.035) at 5 weeks after administration.
Albumin nanoparticles are not toxic to the cornea and can express intraceptors for extended periods that are effective in suppressing injury-induced corneal neovascularization.
Investigative Ophthalmology & Visual Science 06/2007; 48(5):2030-6. · 3.60 Impact Factor
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ABSTRACT: To assess the potential of human serum albumin nanoparticles (HSA NP) as a nonviral vector for ocular delivery of Cu, Zn superoxide dismutase (SOD1) gene.
Cu, Zn superoxide dismutase (SOD1) gene-encapsulated nanoparticles (NP) were developed using human serum albumin (HSA), an endogenous protein, by a desolvation-crosslinking method. The pSOD-loaded HSA NP was evaluated for in vitro release characteristics, stability against DNase I and vitreous humor degradation, cytotoxicity, cellular uptake mechanisms, in vitro transfection efficiency, and in vivo gene expression. In vitro studies employed cultured human retinal pigment epithelial (ARPE-19) cells and in vivo studies employed a mouse model. For cell uptake analysis, fluorescein isothiocyanate (FITC)-labeled human serum albumin (HSA) was used.
Plasmid containing SOD1 gene was encapsulated in HSA by a desolvation-crosslinking method. Gene-loaded HSA NP has a mean size of 120 nm, zeta potential of -44 mV, and plasmid encapsulation efficiency of 84%. At high crosslinking degree, HSA NP sustained the in vitro release of plasmid over 6 days, and stabilized plasmid DNA against DNase I and vitreous humor degradation. No cytotoxicity was observed in ARPE 19 cells treated with blank HSA NP at concentrations up to 5 mg/ml for 96 h. Cellular uptake of HSA NP was via receptor-mediated endocytosis that involves primarily caveolae-pathways. Confocal analysis indicated rapid endo/lysosomal escape of HSA NP. Further, confocal studies indicated that HSA readily enters the cell nucleus. In vitro, pSOD-HSA NP resulted in more than 80% transfection efficiency in ARPE-19 cells, which was 5 fold higher than Lipofectamine. HSA NP-transfected cells exhibited enhanced SOD1 activity that was 5 fold higher than untreated cells, indicating the overexpression of the functional gene. Intravitreal injection of HSA NP to the mouse eye at a dose of 130 ng of plasmid produced detectable level of fusion protein expression at 48 h, compared to non-detectable expression in control animals.
The HSA NP developed in this study offers a very promising approach for nonviral gene delivery to the retina.
Molecular vision 02/2007; 13:746-57. · 2.20 Impact Factor
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ABSTRACT: To potentiate the anticancer activity of paclitaxel-loaded PLGA nanoparticles through surface conjugation with wheat germ agglutinin (WGA).
PLGA nanoparticles loaded with paclitaxel and isopropyl myristate (IPM) as release modifier were prepared by a solvent evaporation method. WGA was conjugated to the nanoparticle surface to give novel WIT-NP of 330+/-3 nm. In vitro cytotoxicity of WIT-NP against malignant (A549 and H1299) and normal (CCL-186) pulmonary cell lines was evaluated alongside control formulations. IC50 doses were determined by the MTT assay, while cellular apoptosis was detected by cell nuclei staining and DeadEndtrade mark Fluorometric TUNEL assay. Cell cycle arrest was confirmed by flow cytometry. Cellular uptake of 3[H]-paclitaxel from the test and control formulations was also quantified. In vivo anticancer efficacy was evaluated in the SCID mice model engrafted with the A549 tumor nodule.
WIT-NP had superior anti-proliferation activity against the A549 and H1299 cell lines compared with conventional paclitaxel formulations as measured by IC50 doses. This was attributed to a more efficient intracellular accumulation of paclitaxel via WGA-receptor-mediated endocytosis and IPM-facilitated intracellular paclitaxel release. WIT-NP activity was associated with paclitaxel-induced apoptosis and cell arrest in the G2/M phase. A single intratumoral injection of WIT-NP at paclitaxel dose of 10 mg/kg inhibited the growth of A549 tumor nodules without inducing significant weight loss in the SCID mice over a period of 25 days. Tumor doubling time was greater than 25 days, compared with 11 days for nodules treated with conventional paclitaxel formulation.
The formulation of WIT-NP, in which WGA is conjugated to the surface of paclitaxel and IPM-loaded PLGA nanoparticles, significantly potentiates the anticancer activity of paclitaxel.
Journal of Controlled Release 12/2005; 108(2-3):244-62. · 5.73 Impact Factor
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ABSTRACT: The purpose of this study was to develop a novel lectin-conjugated isopropyl myristate (IPM)-incorporated PLGA nanoparticle system (NP) for the local delivery of paclitaxel to the lungs. Wheat germ agglutinin (WGA) was conjugated onto preformed IPM- and paclitaxel-loaded PLGA NPs by a two-step carbodiimide method following comparative uptake studies of Concanavalin A, Ricinus communis-120 and WGA on A549, H1299 and CCL-186 cells. WIT-NP with mean diameter of 331 nm and zeta potential of -4.3 mV were prepared with yield of 66% and paclitaxel encapsulation efficiency of 61%. Particle size was expanded by surface conjugation with WGA, while zeta potential was reduced by the addition of IPM and WGA. In vitro paclitaxel release profile was not affected by WGA but initial drug release was enhanced by adding IPM into the formulation. The WIT-NP showed a burst-release of about 32% of the paclitaxel load within the first 5 h followed by a slow zero-order release of another 7% of the drug load in the next 115 h. Compared with the clinical paclitaxel formulation, paclitaxel-loaded nanoparticles without IPM or WGA, or paclitaxel-loaded nanoparticles with only IPM or WGA, the WIT-NP had superior in vitro cytotoxicity against A549 and H1299 cells. IC50 for WIT-NP after 5 and 24 h incubation with A549 cells were not significantly different (15.5 and 15 microM, respectively) whereas the clinical formulation was not cytotoxic after 5 h but had IC50 of 14 microM after 24 h incubation. WIT-NP exhibited stronger cell-killing effect because of more efficient cellular uptake via WGA-receptor-mediated endocytosis and IPM-facilitated release of paclitaxel from the NPs.
Journal of Controlled Release 10/2005; 107(1):30-42. · 5.73 Impact Factor
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ABSTRACT: The calcium pectinate (CaP) capsule, a novel, colon-specific delivery system, was designed and developed using 5-fluorouracil (5-FU) as a model drug. Technically, CaP capsules were prepared by dipping a glass or stainless steel rod successively into pectin and calcium chloride solutions, followed by subsequent air-drying and coating. In vitro studies showed that the release of 5-FU from CaP capsules markedly increased in the presence of rat cecal contents, and the release characteristic was mainly associated with some capsule parameters such as calcium content, shell thickness, and coat amount. Gamma scintigraphic studies demonstrated that CaP capsules could pass through the stomach and small intestine intact and could release drug in colon. The 5-FU releasing characteristics acquired both from in vitro biomimic dissolution experiments and from healthy volunteers indicated that the newly developed CaP capsule possessed the ideal colon-specific drug delivery characteristic.
Drug Development and Industrial Pharmacy 02/2005; 31(2):127-34. · 1.49 Impact Factor
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ABSTRACT: The purpose of this study was to evaluate the extent and mechanism of uptake of wheat germ agglutinin-conjugated PLGA nanoparticles by A549 cells. PLGA nanoparticles of 150 nm were prepared by a solvent diffusion method and covalently conjugated to FITC-WGA (fWGA) or FITC-bovine serum albumin (fBSA) by a two-step carbodiimide method. Uptake of fWGA-PLGA and fBSA-PLGA nanoparticles by confluent A549 cells was quantified by fluorometry. A549 cellular uptake of fWGA-PLGA nanoparticles at 2 h, 37 degrees C was 5.02-fold that of fBSA-PLGA nanoparticles at a loading concentration of 2.65 mg/mL. The difference in uptake between the two types of nanoparticles was increased to 7.84-fold at a higher loading concentration of 5.3 mg/mL, but was reduced to 2.07-fold by lowering the uptake temperature to 4 degrees C. Coincubation with 5 mg/mL of unlabeled WGA negated the differential uptake of fWGA-PLGA nanoparticles at 4 degrees C, suggesting that the nanoparticles interacted with a specific WGA-binding receptor on the cell membrane. Internalization of the fWGA-PLGA nanoparticles by the A549 cells was confirmed by confocal microscopy. Filipin (1 microg/mL), a known inhibitor of caveolae, reduced the 1-h uptake of the nanoparticles by 75%. Surface modification of PLGA nanoparticles with WGA significantly enhanced its endocytosis by A549 cells by a receptor-mediated, caveola-dependent pathway.
Journal of Pharmaceutical Sciences 02/2004; 93(1):20-8. · 3.06 Impact Factor
