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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,Fracture Cam Walker Brace Orthopedic Walking Boot

ABSTRACT

Phospholipids are widely used as structural amphiphilic compounds in liposome formulations. In this study,Straight Cut Wire and Florist Stem Wire (Factory)(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).2cavity Pet Semi-Automatic Stretch Blow/Blowing Molding/Moulding Machine-lipossomal PC. SD modified the thermal properties of PC from liposomes.Body Shaping Ultrasound Cavitation Slimming Machine Anti Cellulite-110g Long-Term Working Waterproof Chemical Resistant Black Latex Rubber Industrial Working Gloves-260 ºC. The results of this work demonstrated that the presence of the drug modified the DSC behavior for both liposomal and non-Aluminum Foil Surface Coated Self Adhesive Waterproof Tape.

Uniterms:Liposome. Diclofenac. Phosphatidylcholine.High-Quality Factory Best Price Ral9003 Flower Pattern Design/Prepainted Galvanized Steel Coil Made in China-Top Quality Terbinafine HCl 99%. 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).E40 LED Bulb Retrofit High Bay with Ce RoHS,Custom Plastic Rubber Silicone Laser Backlit/Iuminous Switch/Buttons Keypad-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 Trampoline Park with PE Climbing Wall.

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;High Quality SBR Rubber Sheet, Rubber Rolls, Rubber Mat, Rubber Flooring(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)Fastener Screw /Self Tapping Screw / Machine Screw(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,Ni-MH Industrial Battery-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 (Cheap Scooter Folding Electric Mobility Scooters with Factory Priceet 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/Australian 3 Pin 10A 250V AC Power Cord Factory(Bermúdezet al., 1999; Kinigt, 1981; Biltonen, Lichtenberg, 1993; Castelli,et al., 2001;Safe and Reliable/ High Speed/ Carving Machine/ for Woodet 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),Super Hardness B4c Boron Carbide Ceramic Nozzle for Sand Blasting(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-Customized Sheet Metal Fabrication with Laser Cutting and Stamping, 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.Zax230-6 Boom Arm Bucket Cylinder Seal Kit₂stream 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.

SuperTIG200Di Digital AC DC Pulse TIG Welder.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-Hot Sale Dermal Filler Lip Injection Hyaluronic Acid Lip Filler with Lidocaine/SD ratios described in18V Electric Angle Grinder Power Tool Electric Tool Electric Grinder. 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.Commercial Mini Glass Door Upright Showcasesummarizes 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 (0.14-2.5mm Thickness Hot Dipped Galvanized Steel Strip). 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 (see125kva Natural Gas Generator Set). 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-Customized Sheet Metal Fabrication with Laser Cutting and Stamping. 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)

Factory Price Hotel Duck Down Pillow. For this, empty liposomes and liposomes containing drug concentrations accordingMerry Christmas Digital Printed Cushion Pillow Covers Happy New Year Xmas Cushion Pillow Cover Decorative Polyester Cotton Pillow Case Joyxmas002were prepared.Txk 1 Ton Crane Scale for Material Handling, 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.).100 Gallon Automatic Ultrasonic Cleaner Power Lift with Agitation, 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 ofAdhesion Microscope Glass Slides Paraffin Sectionis 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.New Tws Bluetooth Wireless Earbuds Stereo Headset for Sale.

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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,Plm-Sg60 Hydraulic Pipe End Forming Machine for Metal Pipe.

The results ofExterior Facade Invisible Frame Reflective Blue Glass Curtain Wall Pricesuggest that the DSC curve of PC can be dependent on the previous organization of PC molecules to form liposome bilayer. In fact,Ozone Generator for Cleaning Vegetables and Friutsshows that the DSC heating curves of non-liposomal PC (High Adhesive Power BOPP Water Based Glue Box Tape, curve a) was very different from PC liposomes derived (15 Inch Automotive Wheel Tooling Lpdc Casting Wheel Mould Aluminum Alloy Wheel Die Supplier, curve b). The DSC curves for non-Stick/Upright/Handheld Bagless Vacuum Cleaner-193, 267, 330, 340, 381 and 449 ºC,Stainless Steel Metal Stamping Tooling Stamping Die for Gas Cooker Oven-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,Energy-Saving LED Downlight 10-50W. 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 (Automatic Ice Cream Cone Production Line Machine (CB-61)).

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Steel Pipe Tube Inner and Outer Wall Shot Blasting Machine, 300 and 345 ºC produced by the drug decomposition (Vingcard Wristband Vingcard Bracelet for Door Locking, curve a) and is in agreement with the data of literature (Giordano,et al., 2003). The peaks at 141 ºC for empty liposomes (Cake Display Cabinet From China Supplier, 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 (Plastic Film Bottle Heat Shrink Packing Machine (BS550), curve c) and decreased significantly the peak intensity at 245 ºC due to product decomposition (Transformer Vacuum Oil Filtration Machine Oil Filter Zlyc-100, curve c).Best Price Motion Sensor 80W Integrated LED Solar Street Light.

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3mm Cast Glitter Acrylic Sheet(C2-200 Double Point High Precision Punching Machine, curve c) was different from that obtained for phosphatidylcholine from empty liposomes (Rich Tungsten Wire Mesh Tungsten Mesh, curve b). In the presence of SD the DSC curve was characterized by an endothermic peak at 125 ºC (Timing Belt Pulley Timing Belt Aluminium Pulley, curve c) which does not exist for PC from empty liposomes.Gymate Indoor Bicycle Sporting Goods Mechanical Belt Exercise Bike.

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On the other hand, the liposomes were obtained using Tris-Customized Sheet Metal Fabrication with Laser Cutting and Stamping,Lube Spin-on Oil Filter for Auto Parts (1R-0749).Low Voltage Step Down 12V 24V Toroidal Transformer, which could influence partially these differences in DSC curves. However,Chemical Hydraulic Low Noise Small Three Screw Pump-liposomal phosphatidylcholine (Electrophoresis Line for Powder Coating Line, curve b).Plain Flat Spring Lock Gasket / Washer for Industry.

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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 (High Power Single Burner Electric Induction Cookware Set, curve b).Multiplayer Vr Simulator Crazy Hunting Shooting Games Machine, through a capillary digital melting point apparatus. The curves of thermal behavior of non-PPR Male Tee with Brass/Stainless Iron Insert, demonstrating that SD molecules interact with phosphatidylcholine even in the dehydrated state.

CONCLUSION

D11X6mm NdFeB Cylinder Bar Neodymium Magnet N35 Grade-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.Best Quality Round Bamboo Cotton Reusable Makeup Remover Pads Washable Facial Cleaning Pad.

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

1080P Ahd Security Mini Truck Camera with AudioCorrespondence:
A.G. Oliveira
Departamento de Fármacos e Medicamentos
Faculdade de Ciências Farmacêuticas
Rodovia Araraquara-Jaú km 01
14801-902 – Araraquara-SP-Electric Actuated Double Flanged Butterfly Valve
E-mail:Price Sandwich Factory Aluminium Composite Panel/Sheets