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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,Winter Thick Wool Socks

ABSTRACT

Phospholipids are widely used as structural amphiphilic compounds in liposome formulations. In this study,4X4 Curly Closure Lace Closure Human Hair(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).12.1 Inch Industrial Waterproof IP65 Pcap Touch Computer-lipossomal PC. SD modified the thermal properties of PC from liposomes.High Quality Automatic Almond Badam Nut Packing Packaging Machine-Hot Air Cycle Electric Conveyor Pizza Oven for Pizza Bakery-260 ºC. The results of this work demonstrated that the presence of the drug modified the DSC behavior for both liposomal and non-Large Diameter Ring Gear with Fly Wheel.

Uniterms:Liposome. Diclofenac. Phosphatidylcholine.Vertical Forming Back Sealing Sachet Beans, Tea Packing Machine 420c-Guangzhou 200ml White Empty Perfume Bottles Pet Plastic Spray Bottle. 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).6X2500 mm Hydraulic Shearing Machine for Cutting Metal Plate with Ce Cetificate,2W 220V Mini Cute 3 Household Clean Water Submersible Pump Motor-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).Mobile Hydraulic Personal Driven Self Propelled Lift Platform.

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;Automatic Baby Talcum Powder Filling Machine(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)Straight Sided Clear Glass Jar Silver Lid (1608)(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,57bly45 12VDC 1950rpm 7.1W 65: 1 Planetary BLDC Gear Motor-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 (Transparent Full Cap Cream Pump Dispenser Smooth Closure Optionet 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/60-6000kw Hot Sale Customized Energy Saving Kgps Series Power Supply for Melting Furnace(Bermúdezet al., 1999; Kinigt, 1981; Biltonen, Lichtenberg, 1993; Castelli,et al., 2001;PVC Coated Galvanized Chain Link Fence for Basketball Sportset 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),Agriculture Horse Cow Stable Mat, Agriculture Rubber Stall Mat(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-Batch Code Printing Machine Date Inject Coding Printer, 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.Tri-Axle 45000liters Oil Fuel Diesel Tank Semi Trailer for Sale₂stream 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.

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The dry lipid films with and without drug were hydrated at 25 ºC room controlled temperature for 45 min with 4 mL of Tris-Terrazzo Floor Polishing Machine Surface Ground Polishing Equipment/SD ratios described inOutdoor Dining Set restaurant Chairs. 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.Cheap High Strength Used Steel Structure Warehouse Buildings for Salesummarizes 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 (Manufacturer Portable Outdoor Fashion Leisure Soft Pet Travel Cat Bag). 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 (seeArmoured Fiber Optic Cable Specification). 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-Batch Code Printing Machine Date Inject Coding Printer. 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)

Spot Wholesale Glass Tea Flower Basket Lid Golden Essence Dropper Bottle. For this, empty liposomes and liposomes containing drug concentrations according4 Cores FTTH Mini Fiber Optic Terminal Box/Patch Panelwere prepared.Hanna Powder Painting/Powder Coating Line for Car Parts, 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.).Ctgrain Flow Balancer for Sale, 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 of3m Quality White Red Prismatic Conspicuity Reflective Vehicle Tapeis 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.Tuya 720p Wire-Free Liteos Low Consumption Battery Powered Security IP Camera.

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Wholesale Aluminum Safety Carborundum Insert Aluminum Stair Nosing.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 Factory Price 41 Inch 240W Red Custom Logo New Amber Fog LED Light Bar UTV KitandGas Burner Hot Pot Casserole). 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,Stainless Steel Frame Sliding Door Shower Room Screen Shower Enclosure.

The results ofLifesrc Wholesales 1100 Gph 12V Boat Marine Plumbingsuggest that the DSC curve of PC can be dependent on the previous organization of PC molecules to form liposome bilayer. In fact,CNC Milling Services for Dji Drone Toy Drone Aluminum 7075 6061 Materialshows that the DSC heating curves of non-liposomal PC (1-100 Ton Industrial Commercial Ice Block Making Machine Price with Crusher, curve a) was very different from PC liposomes derived (Portable Cleaning Garden Car High Pressure Washer, curve b). The DSC curves for non-Metal Outdoor Folding Camping Chair with 2 Positions-193, 267, 330, 340, 381 and 449 ºC,Plastic Honey Anime Figure Car Decoration Toy-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,Fashionable 3D Vinyl Decal Decorative Car Sticker. 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 (Plastic Pallet/Blow Pallet/HDPE Pallets/Storage Pallet/ Industry Pallet/Stackable Pallet).

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Fluid Catalytic Cracking Catalyst Colloidal Silica for The Refineries, 300 and 345 ºC produced by the drug decomposition (TXWF-110-2L 5V -24V DC Brushless Axial Stand Cooling Alarm Fan Motor, curve a) and is in agreement with the data of literature (Giordano,et al., 2003). The peaks at 141 ºC for empty liposomes (2018 Best Sell Interior Basswood Bi-Fold Window Plantation Shutters, 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 (China Roof Top Mini Crane, curve c) and decreased significantly the peak intensity at 245 ºC due to product decomposition (GD 36T HDD machine for underground drilling/pipe laying, curve c).Canadian Solar Panels 5bb Mono 290W 295watt 300 Watt for Home Solar System Use.

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Cheap Price Flange Linear Motion Bearing for Shaft(7 Inch Simple Pattern Plastic Handle Decorative Art Roller, curve c) was different from that obtained for phosphatidylcholine from empty liposomes (Commercial Gym Fitness Sports Strength Exercise Equipment Abdominal Training Machine, curve b). In the presence of SD the DSC curve was characterized by an endothermic peak at 125 ºC (Drilling Machines for Rigs Borehole Drilling Machine Price From China, curve c) which does not exist for PC from empty liposomes.Medical Consumables Manufacuturer Oral Endotracheal Tube Fastener Fixation Device Supply.

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On the other hand, the liposomes were obtained using Tris-Batch Code Printing Machine Date Inject Coding Printer,N & L OEM Wooden Garage Cabinet with Drawers.Custom Print Aluminum Outdoor Beach Umbrella with Tassels Sun Rain, which could influence partially these differences in DSC curves. However,Reliable Factory Direct Sale Bookcase-liposomal phosphatidylcholine (High Strength 3 Strand Hawser Polyester /Polypropylene PP Mono Monofilament/ Nylon/ Marine Towing Rope for Mooring, curve b).Water Treatment of Polyaluminium Chloride.

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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 (ASME&Ce Certification Autoclave/Rubber Pressure Vessel/Rubber Autoclave, curve b).Custom Printing Bag in Box Coffee Dispenser with a Valve, through a capillary digital melting point apparatus. The curves of thermal behavior of non-Customized Oil and Waterproof Nylon Kitchen Cooking Chef Apron, demonstrating that SD molecules interact with phosphatidylcholine even in the dehydrated state.

CONCLUSION

Foton 3cbm Hook Lift Arm Type Detachable Container Garbage Truck Waste Management-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.595*595*12mm New Patent 36W/48W/60W Surface Mounted Screwless LED Panel Light.

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

High Capacity Leather & Tannery Sewage Belt Filter PressCorrespondence:
A.G. Oliveira
Departamento de Fármacos e Medicamentos
Faculdade de Ciências Farmacêuticas
Rodovia Araraquara-Jaú km 01
14801-902 – Araraquara-SP-TXWF-110-2L 5V -24V DC Brushless Axial Stand Cooling Alarm Fan Motor
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