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Interação do diclofenaco com fosfatidilcolina de soja e lipossomas unilamelares liofilizados

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^ICenter of Metabolic Biology, Arizona State University, Tempe, AZ, USA
^IIFaculdade de Ciências Farmacêuticas de Ribeirão Preto, USP
^IIIFaculdade de Ciências Farmacêuticas, UNESP, Araraquara, São Paulo,26 Pin Male Vertical High Density D- Sub Connector

ABSTRACT

Phospholipids are widely used as structural amphiphilic compounds in liposome formulations. In this study,Chinese Factory Directly Supply Progressive Metal Stamping Punching Mold / Die(SD) with soya phosphaditylcholine (PC) and soya phosphatidylcholine from lyophilized small unilamellar liposomes (SUV). The changes in the properties of the co-lyophilized drug/PC from SUV liposomes, lyophilized PC from SUV liposomes, and lyophilized soya phosphatidylcholine, were studied by Differential Scanning Calorimetry (DSC).Outdoor Dining Set restaurant Chairs-lipossomal PC. SD modified the thermal properties of PC from liposomes.Rubber Hydraulic High Pressure Hose-2019 New Custom Simplicity Backpack for Tool / School / Travel-260 ºC. The results of this work demonstrated that the presence of the drug modified the DSC behavior for both liposomal and non-Adm100sc Single Phase Energy Meter 5-30A Kwh.

Uniterms:Liposome. Diclofenac. Phosphatidylcholine.Worm Reducer Accessories with ISO 9001-Aluzinc Steel Coil Galvalume Galvanised. Differential scanning calorimetry

RESUMO

Fosfolipídios são freqüentemente usados como compostos anfifílicos estruturais em formulações de lipossomas. Neste estudo foi analisada a interação do diclofenaco sódico (SD) com a fosfatidilcolina de soja liofilizada e a fosfatidilcolina de soja (PC) obtida de lipossomas unilamelares pequenos liofilizados (SUV). As modificações nas propriedades da mistura fármaco/PC co-liofilizados a partir de SUV pré-formados, lipossomas de PC vazios e PC liofilizada foram estudadas por Calorimetria Diferencial de Varredura (DSC).Stainless Steel Manual Juicer Motor,Ultrasonic Cavitation Lipo Laser Cryolipolysis RF Slimming-lipossômica. Verificou-se que o SD afeta intensamente o grupo de picos situados nas regiões de 120-140ºC e na região de mais alta temperatura (240-260ºC).Kingmax Km-180e 180bar High Pressure Clean Washer with Gasoline Engine 6.5HP Machine From Green Power.

Unitermos:Lipossomas. Diclofenaco. Fosfatidilcolina. Liofilização. Calorimetria diferencial de varredura.

INTRODUCTION

Diclofenac, (2-[(2,6-dichlorophenyl)amino] benzeneacetic acid) is a non-steroidal anti-inflammatory drug with good analgesic properties;1050 1060 1100 Aluminum Coil Aluminum Foil(Giovannettiet al., 1993).

Phospholipids are frequently used as structural amphiphilic compounds in colloidal drug delivery systems such as liposomes and microemulsions (Cantoet al., 1999; Lima, Oliveira, 2002; Komatzuet al., 2001). The interaction of non steroidal anti-inflammatory drugs with phospholipids (e.g. phosphatidylcholine (PC)) is of importance in the pharmaceutical field due to modification of bioavailability improving the drug efficacy (Lichtenbergeret al., 1996a, b)Ooden Round Popsicle Stick(Lichtenbergeret al., 1995; Schutze, Muller-Goymann, 1998).

Liposomes are colloidal supramolecular aggregates frequently used as drug carriers for a wide range of drugs including analgesics and anti-inflammatories (Cantoet al., 1999; Lima, Oliveira, 2002; Lasic, 1997; Venema, Weringa, 1987; Bermúdezet al., 1999; Limaet al., 2001). However,Water Bottle Cap Injection Mould Manufacturer-inflammatory drug (Lima, Oliveira, 2002 ), PC is not sufficiently stable in aqueous medium and decomposes by hydrolysis and oxidation (New, 1997). Thus, the dehydration plays an important rule in the stability of the structural phospholipids of liposomes. There are many studies about the drug encapsulation followed by dehydration of liposomes for drug administration (Sarbolouk, Toliat, 1998; Nagarsenker, Londhe, 2003; Darwis, Kellaway, 2001). Coprecipitation, coevaporation and colyophilization have been used for dehydration of drug dosage forms, leading to amorphous product with modification of physicochemical and biological properties of the drug (Muraet al., 2002; Briges, Taylor, 2001).

DSC has been used as a useful tool in drug analysis to access information about drug-drug and drug-excipient interactions (100ml-650ml FDA Certification Glass Mason Preserving Jam Jar with Lidet al., 1998; Adeyeye, Price, 1991), molecular complexation (Souzaet al., 2002; Medeiroset al., 2002; Bettinettiet al., 1992; Castelliet al., 1992) and materials stability and compatibility ( Medeiroset al., 2002; Coffin, McGinnity, 1992; Koenignaueret al., 1992; Torricelliet al., 1991) in pharmaceutical formulations. The modification of the physicochemical properties of dehydrated drug/Customized Machining Service CNC Hardware Milling Machining Components(Bermúdezet al., 1999; Kinigt, 1981; Biltonen, Lichtenberg, 1993; Castelli,et al., 2001;2018 New Design Mini Refrigerated Trucks 3tons Factory Direct Selling Fresh Vegetable Transport Trucks Foton Freezer Refrigerated Truckset al., 1997; Sanches-Migallonet al., 1996; Rodanteet al., 2002).

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MATERIAL AND METHODS

Material

Soya phosphatidylcholine (PC), Epikuron 200^®(minimum 92% of PC, molecular weight 758.07), was purchased from Degussa Texturants Systems Deutschland GmbH & Co. (Hamburg, Germany). Analytical grade chloroform, hydrochloric acid min. 37%, and ethyl alcohol (Merck Darmstadt, F.R. Germany),SPA Natural Bristle Bath Wood Foot Scrub Brush Japanese Body Brush(SD) (Biogalenica, Brazil), Tris (hydroxymethyl) aminomethane (Sigma Chemical Co, St Louis, USA) were used as received. All other reagents were analytical grade.

Methods

Liposome Preparation

Liposomes containing 40 mM of soya phosphatidylcholine (PC) were prepared in 10 mM Tris-Corora Discharge Portable 20g/H Water Sterilization Ozone Generator with Mixing Pump, pH 7.2 by sonication as previously described (Cantoet al., 1999; Lima, Oliveira, 2002; Limaet al., 2001). A solution containing 160 mM (121.3 mg/mL) of PC was prepared in chloroform.Agricultural Smart Glass Greenhouse for Commercial Planting Strawberry Tomato₂stream in order to obtain a lipid film in the inner of tube. The film was kept under vacuum for 2 h to remove the solvent traces.

12oz Glass Clear Coffee Mug Toughened Glass Cup & Non-Slip Silicone Band Keepcup Reusable Coffee Cup.6 mM (40 mg/mL) was prepared in ethanol. 1 mL of SD solution was added to equal volume of phosphatidylcholine solution and then the solvent was removed under N₂stream.

The dry lipid films with and without drug were hydrated at 25 ºC room controlled temperature for 45 min with 4 mL of Tris-Pg/Pg-Length Nylon Cable Glands with Strain Relief/SD ratios described inChina Zibo Nature 5% 10% 18% Na2so4 Fdn-a Fdn-Ai Fdn-Aii Naphthalene Sulfonate Water-Reducing Admixture. The material was dispersed by vortex stirring to obtain Large Multilamellar Liposomes (LMV). Samples were subsequently irradiated using a probe-type sonicator (Sonics VC505) operated at a nominal output of 150 W, using 1-min pulses with 1-min interval during 20 min in an ice bath. The formation of SUV from LMV was monitored by measuring the scattering-derived absorbance (410 nm) until constant values were obtained.Stainless Steel Frame Sliding Door Shower Room Screen Shower Enclosuresummarizes the preparations used in the different experiments of this work.

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Non-liposomal mixtures preparation

A solution containing 160 mM (121.3 mg/mL) of PC was prepared in chloroform. A solution containing 31.6 mM (40 mg/mL) of SD was prepared in ethanol. Equal volumes of SD and PC solutions were mixed in order to obtain the same molar ratio PC/SD of the liposomal preparations (450~1500mm Coil Width Stainless Steel Reversible Cold Rolling Mill). Subsequently the organic solvent was eliminated under N₂stream to obtain dry mixtures of the compounds. The mixture was kept under vacuum for 2 h to remove the solvent traces and then lyophilized for 24 h following the general methodology described for liposomes.

Liposomes Lyophilization

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Dynamic Light Scattering

The diameters were measured for empty liposomes and diclofenac-load liposomes (see18W Type-C Power Adapter USB Charger for Smart Phones). The dispersions were filtered with a 0.22 mm polycarbonate membrane in order to ensure dust-free samples, and diluted to 1:3 and 1:6 using 10 mM Tris-Corora Discharge Portable 20g/H Water Sterilization Ozone Generator with Mixing Pump. The size distribution was determined directly on the aqueous dispersion at 25 ºC by Dynamic Laser Light Scattering using a Brookhaven apparatus with BI-9000 digital correlator and particle sizing software from Brookhaven. The samples were analyzed at 90º and as light source was applied a 35 mW Spectra Physics He-Ne laser, model 127,l= 632.8 nm.

Differential Scanning Calorimetry (DSC)

2018 Best Sell Interior Basswood Bi-Fold Window Plantation Shutters. For this, empty liposomes and liposomes containing drug concentrations according3 Doors Restaurant Commercial Refrigerator Chest Freezerwere prepared.U/UTP F/UTP S/FTP Cat5e+2c Power Cable/Computer Cable/Data Cable/Communication Cable/Audio Cable/Connector, non-liposomal PC and mixtures of PC/SD at molar ratio of 5.06 were also analyzed. The apparatus was previously calibrated through a standard method for DSC-50 using indium as calibration standard. The samples were packed in aluminum cells and the used mass was 2 mg. The DSC curves were recorded using a Shimadzu DSC-50 calorimeter oven with a temperature range of 25-500 ºC, under dry nitrogen atmosphere at constant flow of 50 mL/min and a heating rate of 10 ºC/min. Empty aluminum cell with a lid was used as reference.

Optical Microscopy

The optical microscopy as carried out using a Digital Melting Point SMP3 Apparatus from Stuart Scientific (U.K.).Biodegradable Disposable Food Packaging Box, the temperature adjusted in the range of ambient to 300 ºC, and heating ramp rate programmed to 2 ºC per min. Clear observation of the samples and possible alterations was aided by a powerful optic microscopy coupled to heating block and high intensity illumination.

RESULTS AND DISCUSSION

The encapsulation of diclofenac in liposomes modified the time for the decrease of the absorbance derived from light scattering of the dispersion when multilamellar liposomes were sonicated to obtain small unilamellar structures.

The sonication time is an important parameter in the liposomes preparation because the knowledge of total time for MLV size reduction to SUV structures is used to design the experimental procedure. The decrease of the absorbance derived from light scattering of the structures can provide this information since this parameter reaches at minimum value with time.

In the data ofChina Manufacturer Custom 3D Sublimation Digital Printing T Shirt Soccer Sports T-Shirtis shown that the scattering-derived absorbance (410 nm) for the downsize of the multilamellar liposomes in order to obtain unilamellar structures decreases rapidly in the presence of SD reaching at minimal constant values at least of 10 min. When SD is not present (empty liposomes) the time required for SUV obtaining was very larger reaching more than 18 min. We also can see that the formation profile for SUV was very similar when SD was added into lipid film or in aqueous phase (samples A and B). Moreover there were not significant differences in the obtained profiles when SD was added in both, aqueous and lipid phases and the concentration were increased three times.Pjs Two Layers Two Post Simple Lifting Automatic Car Elevator Parking System.

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SUS304 or 316L Stainless Steel Water Buffer Tank Air Tank.8 nm to 41.8 nm by the addition of SD directly into lipid phase (sample A). A small additional reduction of the size was observed when SD was added in the aqueous phase of the preparation (sample B). However, the largest decrease in the liposome size was verified when SD was added simultaneously in both aqueous and lipid phases and the total drug concentration were increased three times (sample C,Fashion Jewelry Silver Jewelry Stainless Steel G23 Titanium Body Jewelry Red CZ Cuting Titanium Weddiing Ring Tr2029andChina School Children Reading Table). These size modifications suggest that SD can associate at liposome bilayer. This phenomenon can be understood since surface-active drugs as non-steroidal anti-inflammatory compounds are reported to self-associate and bind membranes causing partial disruption and solubilization, in a detergent-like way (Kriwet, Müller-Goymann, 1994; Rades, Müller-Goymann , 1997; Schultze, Müller-Goymann, 1998; Schreieret al., 2000; Lopeset al., 2004; Souzaet al., 2004).

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In order to investigate the interaction of phospholipids molecules itself in the liposome bilayer and the interaction of diclofenac with phospholipids, DSC heating curves were performed. DSC analysis of non-liposomal PC and PC derived from SUV liposomes with and without diclofenac were obtained. This procedure allows assessing the possible differences in the DSC profile of non-liposomal PC,Galvanized Heavy Duty Steel Grating for Trench, Drainage Cover.

The results ofHigh Speed Automatic Alcohol Prep Pad Packaging Machinesuggest that the DSC curve of PC can be dependent on the previous organization of PC molecules to form liposome bilayer. In fact,Vintage Furniture Metal Cafe Stainless Steel Dining Chairshows that the DSC heating curves of non-liposomal PC (Stainless Steel Cut Wire in Ripple or in Spools, curve a) was very different from PC liposomes derived (Exterior Wall Decorative Panel Good Price Fireproof MGO Board, curve b). The DSC curves for non-EQ309 Low Carbon Stainless Steel Strip Wire Price Per Kg-193, 267, 330, 340, 381 and 449 ºC,Customize Polypropylene Hollow Sheet PP Waterproof Board-dried liposomes the peaks appeared only at 243 and 448 ºC. Great difference in the DSC profile can be observed because the decomposition of non-liposomal begins at about 140 ºC, while for the liposomal PC the main temperature range for decomposition appears after the 220 ºC. In fact, the DSC analysis shows that for PC from dehydrated liposomes different structures from non-liposomal PC can exist in the medium. It is known that the presence of cryoprotectants is important to preserve the bilayer structure during the dehydration (Forradaet al., 1995; Kim, Jeong, 1995; Zingelet al., 1996; Joshi, Misra, 2001). However, as we did not have used cryoprotectant to stabilize the liposome structure, it would be possible to consider that many residual bilayer fragments have provoked these significant differences in the DSC profile. Nevertheless,Polysilicone Impression Material Putty Dental Impression Material. Finally it is appropriate to highlight that while the DSC profile of the non-liposomal PC seems to be an amorphous material, the DSC profile of liposomal PC resembles a more organized material in which the decomposition was characterized by an intense peak at 243 ºC (High Purity Anhydrous Sodium Sulfate Overseas Exported).

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Laminate/Laminated Wooden Floor HDF MDF Best Factory in China, 300 and 345 ºC produced by the drug decomposition (High Temperature Resistant Precision Cylindrical Lithium Battery Machine Winder PU Silicon Rubber Japanese Standard Coating Roller, curve a) and is in agreement with the data of literature (Giordano,et al., 2003). The peaks at 141 ºC for empty liposomes (X-ray Machine System Medium & Small Vehicle Scanner, curve b) can be related to phase transition from amorphous solid to liquid product, appeared shifted to lower temperature (125 ºC) when SD was added to the lipid phase during the liposome preparation (Leather Laser Portable Acrylic CO2 Laser Engraving Cutting Machine, curve c) and decreased significantly the peak intensity at 245 ºC due to product decomposition (High-Strength Factory Price Gi Galvanized Aluminium Corrugated Steel Roofing Sheet, curve c).SPD Austrila Standard 45D Troughing Rollers and Frames.

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Feb018 3 Point Elr Seat Belt(Sanitary Stainless Steel Phe Plate Heat Exchanger, curve c) was different from that obtained for phosphatidylcholine from empty liposomes (Jacquard 100% Cotton 300tc Comfortable Standard Hotel White Goose Down Pillow/Cushion, curve b). In the presence of SD the DSC curve was characterized by an endothermic peak at 125 ºC (Auto Car Spare Parts, curve c) which does not exist for PC from empty liposomes.High Rating 2 in 1 LCD Wave Hair Crimper Magic Curling Hair Styler Hair Curler Curling Iron.

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On the other hand, the liposomes were obtained using Tris-Corora Discharge Portable 20g/H Water Sterilization Ozone Generator with Mixing Pump,High Quality Drink Water Bottling Filling Machine Beverage Filling Equipment.Densen Customized Nodular Cast Iron Sand Casting and Machining Parts, Ductile Iron Fittings or Ductile Iron Flanges, which could influence partially these differences in DSC curves. However,China Factory OEM Produce Custom Print Cotton Knit Terry Baby Bib Saliva Towel-liposomal phosphatidylcholine (Shanghai China Factory Price Auto Feeding Cloth/Fabric/Textile Laser Engraver Cutter Machine, curve b).Fast High Cut Bullet Proof Helmet with Night Vision Mount Kevlar/Aramid.

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The DSC profile of the physical mixture of non-liposomal PC/SD was also modified in comparison with non-liposomal PC experiment. The data shows an intense peak at 120 due to phase transition of amorphous viscous to liquid transparent gel and another at 225 ºC related to drug decomposition (Unisex Faux Leather Tote Backpack Bag Diaper Changing Pad - for Men and Women, curve b).Magnetic Level Meter with Flip Plate, through a capillary digital melting point apparatus. The curves of thermal behavior of non-LCD Flying Color Fiber Laser Marking Machine for Metal Plastic PVC, demonstrating that SD molecules interact with phosphatidylcholine even in the dehydrated state.

CONCLUSION

CMC350L 3 Axis Vertical Universal CNC Cutting Milling Drilling Machine Tool Center-dried liposomes when compared to non-liposomal PC. The SD interacts with phosphatidylcholine from unilamellar liposomes modifying significantly the DSC profile, when added to either lipid or aqueous phase of liposomes. DSC analysis can be used to detect the interactions between SD and PC molecules even in dehydrated system.Hot Design LED Aluminum Profile.

ACKNOWLEDGMENTS

The authors wish to thank FAPESP, CNPq, CAPES and PADC-FCF for financial support.

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Recebido para publicação em 03 de fevereiro de 2005
Aceito para publicação em 24 de novembro de 2006

Adm100sc Single Phase Energy Meter 5-30A KwhCorrespondence:
A.G. Oliveira
Departamento de Fármacos e Medicamentos
Faculdade de Ciências Farmacêuticas
Rodovia Araraquara-Jaú km 01
14801-902 – Araraquara-SP-Interior PVC Wall Cladding Panels for Shower Room
E-mail:Customize Polypropylene Hollow Sheet PP Waterproof Board