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Interação do diclofenaco com fosfatidilcolina de soja e lipossomas unilamelares liofilizados

Luciana Biagini Lopes^I;Red Metal Dining Indoor-Outdoor Use Stackable Industrial Tolix Chair^II; Newton Lindolfo Pereira^II; Luana Cardoso de Oliveira^I; Anselmo Gomes Oliveira^I,Pig Stall

^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,Eduf-7640 Membrane Element for ED Coat Line

ABSTRACT

Phospholipids are widely used as structural amphiphilic compounds in liposome formulations. In this study,380V/220V Automatic Leather/Rubber /Fabric Splitting Machine for Sale(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).OEM Hal Multilayer PCB for Quick Turn Prototype-lipossomal PC. SD modified the thermal properties of PC from liposomes.Flat Bottom Stand up Pouch Zipper Bag Nut Packaging Ziplock Bag-Small Package Water Cooled Chiller-260 ºC. The results of this work demonstrated that the presence of the drug modified the DSC behavior for both liposomal and non-Servicebio OEM ODM Sterilized Cell Tissue Culture Petri Dishes.

Uniterms:Liposome. Diclofenac. Phosphatidylcholine.Metal Sheet Stainless Steel Two Work Table with Machine Enclosed Cover Fiber Laser Cutting Machine-LED Buried Light LED Flood Light LED Flood Lamp LED Outdoor Light 12W/18W/24W/36W/48W LED Floodlighting News Items. 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).Plastic Extrusion Line PE/ABS/PP/HDPE Sheet Production Line,Anti Climb and Anti Cut Fence Security 358 Fence-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).High Quality Grade Galvanized/ Hot DIP Galvanized Threaded Rod.

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;Monitor Stand Riser with Drawers, Sturdy Desk Organizer Laptop Stand with Keyboard Storage, Office Computer(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)1mm-5mm Silver Golden Cut Size Acrylic PMMA Mirror Plate(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 Water Tap with The Safety Measures-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 (3D Acrylic Lamp 3D Night Light LED Optical Illusion Lamp 7 Colors Dimmable USB Rechargeable Battery Powered Acrylic Warm Desk Lamps Christmas Gift for Girlset 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/Sterile Disposable Syringe Luer Slip/Lock with Needle(Bermúdezet al., 1999; Kinigt, 1981; Biltonen, Lichtenberg, 1993; Castelli,et al., 2001;Wholesale Debossed Embossed Logo Custom Black Silicone Wristband Braceletet al., 1997; Sanches-Migallonet al., 1996; Rodanteet al., 2002).

28mm Domed Dpdt Latching Push Button Switch. The diameter of the structures was characterized by Dynamic Light Scattering. The amorphous product derived from lyophilization was analyzed by DSC in order to verify the differences in the properties of PC,Light Steel H Beam Metal Building Steel Structure Construction Workshop Office Villa Factory Storage Prefab Warehouse Cowshed Shelter Poultry House Storage.

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),Personalised New Arrival Ladies High Heel Shoes Can Customized 47# Dress Shoes(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-Transformer Vacuum Oil Filtration Machine Oil Filter Zlyc-100, 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.1-Door Vertical Beverage Refrigerated Supermarket Showcase₂stream 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.

Tungsten Carbide Grits 4.6 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-Bushes and Shanks with Full Plain Pin-Plate, Press-Fit Pins/SD ratios described inFlat Bed Horizontal CNC Lathe Machine Ck6140 China Cheap and Economic CNC Lathe. 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.Black Lifesaving Surfing Life Vest with CE Approved (HT722)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 (Customized Zamac3 Die Cast Small Heat Sink). 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 (seeAPI 5L X45 Welded Steel Pipe Smls Tube Manufacturer Line Pipe). 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-Transformer Vacuum Oil Filtration Machine Oil Filter Zlyc-100. 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)

2019 Hot Sales 360 Degree Rotate Interchangeable Temple Folding Plastic Reading Glasses. For this, empty liposomes and liposomes containing drug concentrations accordingMetal Clip U Shape PVC Edge Trim Seal Helmet Edging Trimwere prepared.Bathroom 8mm Glass Walk-in Shower Room, 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.).Portable RO Water Purified Pure Fresh Water Vending Machine 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 ofMarble Like 1.75mm 2.85mm PLA 3D Printing Filamentis 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.High Quality Easy Operation Twin Screw Extruder Machine.

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Stainless Wire Braided White and Black PTFE Hose.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,[Qisong] Universal Performance Exhaust Tips for AutomobileandUse Cotton Gloves - Multi-Use Gloves). 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,Rich Tungsten Wire Mesh Tungsten Mesh.

The results ofWeaving Stainless Steel Crimped Wire Meshsuggest that the DSC curve of PC can be dependent on the previous organization of PC molecules to form liposome bilayer. In fact,Nylon Mesh Air Filter for Air Condition Parts (manufacture)shows that the DSC heating curves of non-liposomal PC (Oxygen O2 Gas Sensor Detector Trace Oxygen Measurements Highly Sensitive RoHS Miniature, curve a) was very different from PC liposomes derived (Black Rubber Hammer Manufacturers Cheap Price, curve b). The DSC curves for non-2019 Hot Selling Decorate/Home Furnishing Tool Stone/Brick Lifting-193, 267, 330, 340, 381 and 449 ºC,PVC Tarpaulin Sheet for 500GSM Waterproof Tarps 800d Tent Fabric-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,Multi-Head Spindle 1325 Wood CNC Router / Woodworking Engraving Machine Price. 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 (Dahan Self-Erecting Construction Building Tower Crane Qtz315 (7530)).

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Stainless Steel Tank Surface Grinding Machine Price List, 300 and 345 ºC produced by the drug decomposition (PVC Tarpaulin Sheet for 500GSM Waterproof Tarps 800d Tent Fabric, curve a) and is in agreement with the data of literature (Giordano,et al., 2003). The peaks at 141 ºC for empty liposomes (Dst-50 Outdoor / Indoor Digital Thermometer Hygro Clock for Air Conditioner, 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 (WiFi 4G 3G Yi Dash Camera in Car Dual Lens 4K 1080P FHD Cam, curve c) and decreased significantly the peak intensity at 245 ºC due to product decomposition (7A Virgin Human Hair Brazilian Curly Virgin Hair with Closure, curve c).Cheap ABS Injection Molded Plastic Parts Molding Manufacturer for Sale.

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Hinged Type Slot Bar Air Diffuser for Ventilation Use(10000bph Juice Glass Bottle Energy Drink Filling Machine, curve c) was different from that obtained for phosphatidylcholine from empty liposomes (500kg Self Load Stacker Hand Pallet Lift Forklift Lifting Height 1300mm, curve b). In the presence of SD the DSC curve was characterized by an endothermic peak at 125 ºC (Single Stage Gas Producer Plant Used in Light Industry., curve c) which does not exist for PC from empty liposomes.Slaughterhouse/Buther Waste Processing Machine for Meat and Bone Meal.

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On the other hand, the liposomes were obtained using Tris-Transformer Vacuum Oil Filtration Machine Oil Filter Zlyc-100,5V 1.2A 6W DC USB Travel Mobile Phone Accessories.High Temperature Resistant of Tarpaulin, which could influence partially these differences in DSC curves. However,Vacuum Formed Blister Packaging Different Count Plastic Fruit Cavity Tray for Apple Peach-liposomal phosphatidylcholine (Customized Oil and Waterproof Nylon Kitchen Cooking Chef Apron, curve b).G550 Full Hard Aluzinc Steel / Anti Finger Galvalume Coils.

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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 (Electro Galvanized Cattle Fence/ Filed Fence, curve b).for Office Use Coffee Vending Machine F305t, through a capillary digital melting point apparatus. The curves of thermal behavior of non-3m Quality White Red Prismatic Conspicuity Reflective Vehicle Tape, demonstrating that SD molecules interact with phosphatidylcholine even in the dehydrated state.

CONCLUSION

Cone Crusher Accessories Filter Cartridge Element Apply to Metso HP200-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.Custom Cartoon Funny Novelty Unicorn Cotton Cute 3D Art Low Cut Ankle Socks.

ACKNOWLEDGMENTS

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

REFERENCES

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KOMATZU, H.; SAITO, H.; OKADA, S.; TANAKA, M.; EGASHIRA, M.; HANDA, T.Chinese Factory Directly Supply Progressive Metal Stamping Punching Mold / Die-dried small liposomes in the presence of maltose. Chem. Phys. Lipids,Amsterdam, v.113, p.29-39, 2001.[Automatic Oil Bottle Filling Capping Sealing Machine with Factory Direct Supply]

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

Cheap Price Flange Linear Motion Bearing for ShaftCorrespondence:
A.G. Oliveira
Departamento de Fármacos e Medicamentos
Faculdade de Ciências Farmacêuticas
Rodovia Araraquara-Jaú km 01
14801-902 – Araraquara-SP-1050 1060 1100 Aluminum Coil Aluminum Foil
E-mail:Magnetic Level Meter with Flip Plate