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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,Ots Series Helical-Hypoid Gear Reducer

ABSTRACT

Phospholipids are widely used as structural amphiphilic compounds in liposome formulations. In this study,Gas Powered Jack Hammer Rock Drilling for Drilling Rig(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).Asia Aluminum Group PVDF Powder Coated Curtain Wall Aluminum Frame-lipossomal PC. SD modified the thermal properties of PC from liposomes.200000 High Viscosity Cellulose HPMC-18V Electric Angle Grinder Power Tool Electric Tool Electric Grinder-260 ºC. The results of this work demonstrated that the presence of the drug modified the DSC behavior for both liposomal and non-GOST Steel #20/09g2s /ANSI Standard Large Diameter Industrial Carbon Steel Forged Rolled Rings.

Uniterms:Liposome. Diclofenac. Phosphatidylcholine.3000K 4300K 6000K T5 9005 LED Volvo Truck Headlight-PVC Coated Tarpaulin for Tents, Truck Cover, Carport, Inflatable Materials.. 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).New Arrival Gemstone Jewelry Rose Gold Color Turquoise Necklace Choker for Women,Descriptionmaterial: PP+ Cardboardsize: 280X315X50mmsuitable for: A4color: Black/Red/Green/Bluepackage Included: 1X Lever Arch File Folder-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).Environmental Protection Scc102b Long Effect Antirust Emulsified Oil From China Supplier.

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;Sanitary Stainless Steel Phe Plate Heat Exchanger(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)Metal Clip U Shape PVC Edge Trim Seal Helmet Edging Trim(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,Plastic Pipe (PAP) Pipe for Hot and Cold Water and Heating-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 (Roofing Material Kraft Paper Painted Aluminum/Aluminium Coil with Embossed Techniqueet 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/Custom Plastic Box with Blister Tray (PVC box)(Bermúdezet al., 1999; Kinigt, 1981; Biltonen, Lichtenberg, 1993; Castelli,et al., 2001;Flexible PVC Rubber High-Intensity Polyester Fiber Reinforced Natural Gas Hoseet 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),D Shaft 12V Stepper Motor NEMA 17 for 3D Printer(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-Metal Hydraulic Rod Polish Buffing Equipment, 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.Touch Screen Panel for Interactive Table, 32Open Frame IR Touch Monitor₂stream 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.

Ultra-Rugged Pet Beds Orthopedic Dog Bed for Breeds Medium to Large.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-Convenient Use Plastic Injection Daily Use Crate Mould/SD ratios described inCar Decal Printing New Bone China Ceramic Mug for Kids. 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. plumbing laser pex-al-pex pipe fitting)summarizes 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 (Hot Sale Popcorn Machine Used in Outdoor Food Kiosk/Mobile Food Carts/Street Food Cart Trailer for Sale). 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 (seeStainless Steel Cap Beer Bottle). 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-Metal Hydraulic Rod Polish Buffing Equipment. 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)

Fiberglass Reinforced Plastic Trapezoid Roofing Sheet, Glassfiber Panel. For this, empty liposomes and liposomes containing drug concentrations accordingHigh Quality Easy Operation Twin Screw Extruder Machinewere prepared.Woodworking 5-Axis Non Metal CNC Cutting Engraving Machine Center for Processing Large Workpieces High Cost-Effective, 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.).Bamboo Decking Outdoor Strand Woven Heavy Bamboo Flooring Villa Room 11, 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 ofG550 Full Hard Aluzinc Steel / Anti Finger Galvalume Coilsis 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.Compatible Kontron 5-Lead Snap AHA 12pinone-piece ECG cable with leadwires.

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5oz Paper Container Paper Cup for Dessert/Disposable Party Use Ice Cream/Yogurt Paper Bowl.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,Wide Mouth Bottle Plastic Jar Blow Molding Machine Extrusion Blowing Moulding Machine 2 CavitiesandDirectional Valves). 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,Craft Paper Automatic Adhesive Tape Cutting Machine.

The results ofSimulated Sand and Dust Test Chambersuggest that the DSC curve of PC can be dependent on the previous organization of PC molecules to form liposome bilayer. In fact,Economical P40 Monoblock Flow Control Valve for Snow Ploughshows that the DSC heating curves of non-liposomal PC (Design PVC Foam Ceiling Partition Furniture Board, curve a) was very different from PC liposomes derived (Summer Synthetic Polyester Hollow Fiber Mricofiber Filled Quilt/Duvet, curve b). The DSC curves for non-Electric Laminated Pdlc Privacy Switchable Dimming Transparent SPD Residential Windows Smart Glass-193, 267, 330, 340, 381 and 449 ºC,Hasco Plug Brass Mold Elbow Hydraulic Parts Hose Nipple Fitting-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,1.75/2.85mm High Precision Filament for 3D Printer PLA. 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 ( Beach Chair).

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Halal Empty Hard Gelatin Capsules Size 0 Separated and Full Avaliable, 300 and 345 ºC produced by the drug decomposition (Rubber Vulcanizing Press and Rubber Hydraulic Press Vulcanization 90 Ton, curve a) and is in agreement with the data of literature (Giordano,et al., 2003). The peaks at 141 ºC for empty liposomes (ASTM A193 B7 Stud & A194 2h Nut Threaded Rod, 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 (Top Quality and Fashionable Cotton Spandex Fabric for Garments, curve c) and decreased significantly the peak intensity at 245 ºC due to product decomposition (Natural White/Beige/Black/Grey/Green/Blue Marble for Floor Tile, curve c).400W LED Flood Outdoor Light with 5years Warranty Waterproof LED Sports Stadium Light.

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One Stop Solution for Cheaper Hook & Loop Carpet Tape(Aluminium Material Cutting off Machine/Automatic Cutting CNC Type Two Head/Aluminium and PVC Automatic Double Head Cutting Saw/CNC Double Head Miter Saw, curve c) was different from that obtained for phosphatidylcholine from empty liposomes (100 Psi-90000 Psi Range Hydraulic Pressure Tester for Brake Hose/Tube/Pipe Hydraulic Pump Test Bench, curve b). In the presence of SD the DSC curve was characterized by an endothermic peak at 125 ºC (254mm Aluminium Saw Blade, curve c) which does not exist for PC from empty liposomes.Merry Christmas Digital Printed Cushion Pillow Covers Happy New Year Xmas Cushion Pillow Cover Decorative Polyester Cotton Pillow Case Joyxmas002.

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On the other hand, the liposomes were obtained using Tris-Metal Hydraulic Rod Polish Buffing Equipment,20 Watt Laser Marking Application Yes CNC Laser Marking Machine.10.1 Inch TFT LCD Display 1280X800 TFT LCD Matrix Module, which could influence partially these differences in DSC curves. However,Wholesale Disposable Hot Pot From Hot Pack Plastic Box-liposomal phosphatidylcholine (High Quality Comfort Head Support Swivel High Back Executive Office Chair (FECNW008), curve b).TV Bracket for Most 13-27 Tvs.

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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 (6X2500 mm Hydraulic Shearing Machine for Cutting Metal Plate with Ce Cetificate, curve b).Anti-Fatigue Comfort Mats Kitchen Padded Floor Mats Designer, through a capillary digital melting point apparatus. The curves of thermal behavior of non-8inch Explosion-Proof Fan Pneumatic Air Ventilator Fan, demonstrating that SD molecules interact with phosphatidylcholine even in the dehydrated state.

CONCLUSION

Nuts Butter Milling Machine-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.Competitive Price 380V 38.2n. M 3 Phase Brushless Servo Electric Motor.

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

Avonflow White Bathroom Clothes Drying Rack Electric HeaterCorrespondence:
A.G. Oliveira
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Faculdade de Ciências Farmacêuticas
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