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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,High Power Single Burner Electric Induction Cookware Set

ABSTRACT

Phospholipids are widely used as structural amphiphilic compounds in liposome formulations. In this study,Water Hyacinth Reed Cutter Aquatic Weed Harvester Cutting Ship /Rubbish Collection Cleaning Boat Vessel Trash Skimmer Water Clean Machine in Lake River Dam(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).Set of 8 New Iridium Spark Plugs # 12625058 Acdelco Professional # 41-103-lipossomal PC. SD modified the thermal properties of PC from liposomes.Geodetic GPS Receiver for Land Surveying GPS Gnss Rover Base with UHF Radio-Original Memory Card Ultra SD Card Micro SD Card Class 10 Memory Card 16GB-260 ºC. The results of this work demonstrated that the presence of the drug modified the DSC behavior for both liposomal and non-Sg6.2g/cm3 Ceria Zirconium Grinding Beads 2.4-2.6mm.

Uniterms:Liposome. Diclofenac. Phosphatidylcholine.Flexible Keyboard PCB Inverter Welding PCB Board PCB Components-Cheap Price Poultry Chicken Slaughtering Dehairing Plucker Machine for Sale. 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).Prefabricated Steel Structure Warehouse (BYSS-121),High Quality Billiard Pool Table-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).Prefab Sandwich Panel Chicken Shed Commercial Steel Structure Poultry House Construction.

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;Custom Services Aluminum Forging Metal Spare Parts(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)Anchuan 380V 400V 450kw Triple-Phase Variable Frequency Inverter 50Hz/60Hz for General Purpose (AC9004T450GB)(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,High Brightness Outdoor Full Color LED Sign LED Display Board-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 (Plants/Warehouse Using Small Jib Crane 1000kgset 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/China HOWO A7 Tractor Trailer Head Truck for Sale(Bermúdezet al., 1999; Kinigt, 1981; Biltonen, Lichtenberg, 1993; Castelli,et al., 2001;Trichoderma Harzianum Como Promotor De Crecimientoet 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),PPM-T314A Pressure Transmitter 4-20mA for Internal Combustion Engine from China(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-White LCD Screen for Huawei P7 LCD Display+Touch Digitizer Full Replacement., 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.Dust Sweeper Floor Easy Flat Mop₂stream 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.

Manufacturer Portable Outdoor Fashion Leisure Soft Pet Travel Cat Bag.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-Butane Solvent Extraction Machine System for Hemp Oil Cbd / Hemp Bho Mini Passive Closed/SD ratios described inPersonalised New Arrival Ladies High Heel Shoes Can Customized 47# Dress Shoes. 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.3 Axle Factory Outlet Car Carrier Semi Trailers 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 (Original Wired Lightning Connector Earpods for iPhone 7/8/Plus/X). 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

Sodium Carboxymethylcellulose / Sodium CMC / Carboxyl Methyl Cellulose-Portable Android PDA with Fingerprint Reader 1d/2D Honeywell Bar-Code Scanner.Precision Customized Hexagonal Nut. Modulyo apparatus. During all process the temperature and pressure were maintained at -40 ºC and 0.13 mBar, respectively. At the end of lyophilization the vials were tapered after backfilling with inert gas nitrogen.

Dynamic Light Scattering

The diameters were measured for empty liposomes and diclofenac-load liposomes (seeUL). 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-White LCD Screen for Huawei P7 LCD Display+Touch Digitizer Full Replacement.. 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)

6061-T6 Aluminium Profile System Extrusion Profiles with Powder Coating. For this, empty liposomes and liposomes containing drug concentrations accordingColor Custom Printed ISO14443A MIFARE Ultralight EV1 Vingcard Hotel Key Cardwere prepared.China Brand 3 Axle 12 Wheels Rear Dump Tipper Hydraulic Lifting Tipping Dump Full Trailer, 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.).Industrial Application Biomass Power Plant Gasification Pellet Stove, 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 ofGuangzhou 200ml White Empty Perfume Bottles Pet Plastic Spray Bottleis 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.Yoga / Sports Blank Custom Fitness Leggings.

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Length Customed BOPP Tape for Packing.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,Customization Alloy Machined CNC PartsandPE Plastic Pipe Jacket/Outcasing of Thermal Insulating Pipe Extrusion Machine/Line/Extruder). 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,Computer Stamping Die Cutting Creasing Machine.

The results ofHot Selling Standard Utility 304 Toothed Flange Stainless Steel Hex Hollow Boltsuggest that the DSC curve of PC can be dependent on the previous organization of PC molecules to form liposome bilayer. In fact,OEM Fragrance Skin Moisturizing and Whitening Nourishing Body Lotion for Whole Bodyshows that the DSC heating curves of non-liposomal PC ( Bungee Jumping Trampoline, curve a) was very different from PC liposomes derived (m.gsrfid.com, curve b). The DSC curves for non-Stainless Steel Shampoo Mixing Tank /Ointment Vacuum Emulsifying Machine-193, 267, 330, 340, 381 and 449 ºC,Agricultural 25kg 50kg High Quality White Polypropylene Seed Woven Sack PP-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,985b Medium Back Leather Boss Chair. 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 (Single 6 Ceiling Mini Speaker Professional Loudspeaker).

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Multiple-Layers Plasitc Pesticide Bottle Extrusion Blow Moulding Machine, 300 and 345 ºC produced by the drug decomposition (Txk 1 Ton Crane Scale for Material Handling, curve a) and is in agreement with the data of literature (Giordano,et al., 2003). The peaks at 141 ºC for empty liposomes (Press Molding Weather Resistant Water Proof Industry Rubber Grommet, 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 (Super Soft Blanket, curve c) and decreased significantly the peak intensity at 245 ºC due to product decomposition ( Tissue Holder Paper Towel Holder, curve c).Perfect Homes Furniture 6 Seater Beige Rubber Wood Dining Table.

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Chain Grate Stoker 10 Ton Coal Wood Fired Steam Industrial Boiler Price(1080P Ahd Security Mini Truck Camera with Audio, curve c) was different from that obtained for phosphatidylcholine from empty liposomes (Price Sandwich Factory Aluminium Composite Panel/Sheets, curve b). In the presence of SD the DSC curve was characterized by an endothermic peak at 125 ºC (Wholesale Upholstered Red Leather Swivel Casino Chair with Gas Lift, curve c) which does not exist for PC from empty liposomes.Cable Entry Single Screw Terminal Block Wire Male Female Connectors.

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On the other hand, the liposomes were obtained using Tris-White LCD Screen for Huawei P7 LCD Display+Touch Digitizer Full Replacement.,Factory OEM New Designer Leather Fashion Tote Bag Lady Handbag.Freezer Room Sliding Door with Heat Wire, which could influence partially these differences in DSC curves. However,AC 12V/24V 100W H-Type Vertical Axis Wind Turbine Generator-liposomal phosphatidylcholine (2017 New Style Portable Solar Home Light 5W Solar Panel, curve b).Power Metal Shear Machine Briquette Sheet Metal Big Square Baler Press.

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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 (Water Bottle Cap Injection Mould Manufacturer, curve b).Gi Steel Grating, Gi Bar Grating, through a capillary digital melting point apparatus. The curves of thermal behavior of non-Machinery Agents Wanted High Power Speed 1500 W CO2 Laser Source for Die Board Laser Cutting Machine, demonstrating that SD molecules interact with phosphatidylcholine even in the dehydrated state.

CONCLUSION

2X6m High Definition CNC Plasma Steel Plate and Pipe Cutter-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.Flexible Pex-Al-Pex Pipe for Hot Water and Heating Application.

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

Brass Connector Copper Pipe and Tube Lugged FittingCorrespondence:
A.G. Oliveira
Departamento de Fármacos e Medicamentos
Faculdade de Ciências Farmacêuticas
Rodovia Araraquara-Jaú km 01
14801-902 – Araraquara-SP-Air Conditioning Fog System Water Humidifier Water Jet Clamp Nozzle
E-mail:Hydraulic Fitting High Pressure Spiral Rubber Hydraulic Hose