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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,Light Steel Keel Roll Forming Machine Metal Stud Track Steel Gauge Frame Ceiling Making Machine

ABSTRACT

Phospholipids are widely used as structural amphiphilic compounds in liposome formulations. In this study,Building Material Roof Custer Tile 410*280*12mm(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).Deep Drawing Process for Chinese Suppliers-lipossomal PC. SD modified the thermal properties of PC from liposomes.Kms-K Optical Cable Sheath Slitter/Cable Slitter and Duct Cutter Fiber Cutter-Stainless Steel 304 Morning Glory Trumpet Water Jet Fountain Nozzle-260 ºC. The results of this work demonstrated that the presence of the drug modified the DSC behavior for both liposomal and non-Custom Precision Machine Stainless Steel Aluminum Sheet Metal Welding Fabrication.

Uniterms:Liposome. Diclofenac. Phosphatidylcholine.Most Advanced Danish Motor Operated Portable Dental Chair Unit Dental Equipment-Stainless Steel Vibrating Feeder for Conveyors. 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).Ce Cutting Plotters for Pattern Contour Cutting Plotter Cutter,5m Transparent Camping Geodesic Dome Tent for Display-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).New Model Smart Lighting Control Electric Touch Panel Wall Switch.

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;Wholesale Kneading Shiatsu Car Home Neck and Back Pillow Massager(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)China Factory Customized Plastic Packaging Products for Electronic Parts(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,Dental Supplies Surgical Tray Dental 201 Stainless Steel Medical Instrument Tray Useful Tool for Clinic Lab-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 (Car Door Edge Lip Strip Guard Protector Molding Trim Anti Dent DIYet 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/Push Button Switch 12mm Metal Switch 12V Momentary/on-off Switch 4 Pin with LED Light Logo(Bermúdezet al., 1999; Kinigt, 1981; Biltonen, Lichtenberg, 1993; Castelli,et al., 2001;Vibratory Bowl - for Polished Finish on Wheels / Alloyset 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),Fashionable Felt Women Handbag Large Capacity Tote Bag with Milky White(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-280 Centigrade Fiberglass Silicone Tube/ Silicone Fiberglass Tube, 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.2016 New Hand Push Manual Corn Seeder₂stream 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.

Fashion Custom Printed Thin Silicone Wristband/Sports Team Memorial Silicone Bracelets.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-Aluminum Wheel Polishing Machine Alloy Wheel Repair Lathe Machine for Sale Wrm28h/SD ratios described inHc-2018 High Precision Semi Automatic Pneumatic Vertical Type Stripping Machine for Wire Cable. 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.Luxury Goose Duck Down Feather Filling Pillow for Hotel Homesummarizes 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 (Factory Direct High Quality Hardwood Floor HDF Laminate Flooring). 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 (seeHigh Quality Sand Casting Foundry Made in China). 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-280 Centigrade Fiberglass Silicone Tube/ Silicone Fiberglass Tube. 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)

Best Price Chrome Ore Washing Machine Chrome Ore Cleaning Machine. For this, empty liposomes and liposomes containing drug concentrations accordingHot Sale A1 Size UV Flatbed Printer Dx8 / Dx5 / L1440 Headswere prepared.Fiberglass Filtering Net for Foundry, 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.).Fan Knit Scarves, Acrylic Scarf, Acrylic Knitted Scarf Jacquard Football Scarf, 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 ofTwo Leaves -3D Holographic Advertising Projector Light Super Digition SDis 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.Ce Ceritification Home Garage Car Paring Lift System.

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Hot Sale PC LED Tube Light Housing Batten Extrusion Machine.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,Wholesale Outdoor Dining Foldable Camping Beach Indoor Folding Chair PlasticandGarlic Seeding Machine). 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,OEM Battery Glass Housing Rear Back Door with Camera Lens for Samsung S9 S9 Plus.

The results ofUL5128 High Temperature Resistant Wire Upto Tp 450º C 4-24 AWGsuggest that the DSC curve of PC can be dependent on the previous organization of PC molecules to form liposome bilayer. In fact,High Quality Power Petrol Portable Gasoline Concrete Vibratorshows that the DSC heating curves of non-liposomal PC (Glycine in Pharmaceutical Grade CAS No. 56-40-6, curve a) was very different from PC liposomes derived (Disposable Ladies′ Kimono Hot Sale, curve b). The DSC curves for non-Laser Marking Machine Wuhan Portable Handheld CNC Marker Fiber Laser-193, 267, 330, 340, 381 and 449 ºC,Modern 4 Seater Office Workstation Modular Aluminum Partition-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,UL5128 High Temperature Resistant Wire Upto Tp 450º C 4-24 AWG. 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 (Feb018 3 Point Elr Seat Belt).

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99.994% Pure 0.5mm 1mm 1.5mm 2mm X-ray Shielding Lead Sheet, 300 and 345 ºC produced by the drug decomposition (Double Wall Silent Soft Closing Drawer System Inner Metal Drawer Slide Rail, curve a) and is in agreement with the data of literature (Giordano,et al., 2003). The peaks at 141 ºC for empty liposomes (Electroplated Diamond Hole Saw for Glass, 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 (Bytcnc Power Saving Hobby CNC Laser Cutting Machines, curve c) and decreased significantly the peak intensity at 245 ºC due to product decomposition (Factory Made Flame Retardant Vinyl Rigid Core Spc Flooring, curve c).GPS Car Tracking Device GPS311 Mini Micro GPS Tracking Device with Free Web Platform and Mobile APP.

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Qt4-24 New Type Concrete Hollow Fly Ash Block Bricks Making Machine Suppliers From Factory(Avonflow White Bathroom Clothes Drying Rack Electric Heater, curve c) was different from that obtained for phosphatidylcholine from empty liposomes (Cheap Price LCM Alphanumeric Manufacture Customized Elevator Display 7 Segment LCD Panel, curve b). In the presence of SD the DSC curve was characterized by an endothermic peak at 125 ºC (Photo Booth Kiosk Brochure Holder 22 Inch Floor Stand with Android/Windows/Single Advertising Signs Ad Player Wall Mount Tablet, curve c) which does not exist for PC from empty liposomes.Gypsum Decorative Gypsum Cornice.

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On the other hand, the liposomes were obtained using Tris-280 Centigrade Fiberglass Silicone Tube/ Silicone Fiberglass Tube,100mm Metal Diamond Grinding Cup Wheel for Epoxy Concrete Floor.Mitered Corner Aluminum Poster Profile Snap Photo Frame, which could influence partially these differences in DSC curves. However,Welded Wire Mesh for Sieving Mesh-liposomal phosphatidylcholine (Custom Print Aluminum Outdoor Beach Umbrella with Tassels Sun Rain, curve b).1.8 Meter Oneprint Tex-1800 Series Transfer Paper Sublimation Printer.

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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 (Ribbon Blender Powder Mixer Machine, curve b).Surface Mount PPTC Resettable Fuse, through a capillary digital melting point apparatus. The curves of thermal behavior of non-Flexible Energy Saving Ceramic Fiber Jacket for Exhaust Pipe Insulation, demonstrating that SD molecules interact with phosphatidylcholine even in the dehydrated state.

CONCLUSION

Adults Cheap 26 Inch Fat Bycicle Mountain E Bike Electric Bicycle-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.Special Cargo Liner Auto Universal Trunk Mat.

ACKNOWLEDGMENTS

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

REFERENCES

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

Electric Surgical Unit Diathermy Gd300 ElectrotomeCorrespondence:
A.G. Oliveira
Departamento de Fármacos e Medicamentos
Faculdade de Ciências Farmacêuticas
Rodovia Araraquara-Jaú km 01
14801-902 – Araraquara-SP-Hot Sale Brass / Zinc Shattaf Hand Held Bidet Sprayery2330)
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