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Nanoerthyrosomes As Novel drug carriers

  1. 1. Srinivas Dinakar’s presentation
  2. 2. NANOERTHYROSOMES as NOVEL DRUG CARRIERS <ul><li>CO-ORDINATOR : Mrs.Pavani ,M.Pharm </li></ul><ul><li>CO-AUTHOR : Nikhil Reddy .Y </li></ul>
  3. 3. <ul><li>DEFINITION </li></ul><ul><li>Nanoerythrosomes are erythrocytes based new drug carrier which is prepared by extrusion of erythrocyte ghosts to produce small vesicles having an average diameter of 100nm </li></ul><ul><li>RESEALED ERYTHROCYTES </li></ul><ul><li>When erythrocytes are suspended in a hypnotic medium they swell about one and half times their original size and membrane ruptures resulting in the formation of pores with diameter of 200-500Å 0 </li></ul><ul><li>If the ionic strength of the medium is </li></ul><ul><li>adjusted to isotonicity and the cells are </li></ul><ul><li>incubated at 37˚and causes </li></ul><ul><li>the erythrocytes to reseal. </li></ul>
  4. 4. <ul><li>ISOLATION OF ERYTHROCYTES </li></ul><ul><li>Blood is collected in to heparinized tubes by vein puncture . </li></ul><ul><li>Blood is with drawn from cardiac or splenic puncture </li></ul><ul><li>(small animals ) and through veins (large animals) in a syringe </li></ul><ul><li>containing a drop of anti coagulant . </li></ul><ul><li>The whole blood is centrifuged at 2500rpm for 5 min at 3-5˚c </li></ul><ul><li>in refrigerated centrifuge. </li></ul><ul><li>Serum and buffy coats are carefully removed and packed cells </li></ul><ul><li>are washed with PBS. </li></ul><ul><li>The washed erythrocytes are diluted with PBS and stored at 4˚c </li></ul><ul><li>until used. </li></ul>
  5. 5. <ul><li>ERYTHROCYTES GHOST </li></ul><ul><li>RBC’s on haemolysis and washing with large volume </li></ul><ul><li>of hypnotic medium, loose nearly all their hemoglobin </li></ul><ul><li>and on releasing the resultant cells appear as pale or </li></ul><ul><li>transparent in appearance and are referred as erythrocytes </li></ul><ul><li>ghost </li></ul><ul><li>Preparation and loading of nanoerythrosomes : </li></ul><ul><li>Methods employed are: </li></ul><ul><li>Extrusion </li></ul><ul><li>Sonication </li></ul><ul><li>Electrical break down method </li></ul>
  6. 6. <ul><li>Normal erythrocytes </li></ul><ul><li>Preparation of erythrocyte ghosts </li></ul><ul><li>extrusion/ Sonication /electrical break down </li></ul><ul><li>method </li></ul><ul><li>Nano vesicles </li></ul><ul><li>staining with 0.1% uranyl acetate and </li></ul><ul><li>microscopical examination </li></ul><ul><li>Nanoerythrosomes </li></ul>
  7. 7. <ul><li>EXTRUSION </li></ul><ul><li>Erythrocytes ghosts suspension was </li></ul><ul><li>extruded through polycarbonate membrane filter </li></ul><ul><li>which was connected to an adapter </li></ul><ul><li>Nanoerythrocytes were obtained by </li></ul><ul><li>8 consecutive extrusions </li></ul><ul><li>Extrusion was done using nitrogen </li></ul><ul><li>pressure </li></ul>
  8. 8. <ul><li>SONICATION </li></ul><ul><li>Erythrocytes ghosts suspension was </li></ul><ul><li>sonicated at an energy level of 50W </li></ul><ul><li>for 5 min using dismembrator. </li></ul><ul><li>ELECTRIC BREAK DOWN </li></ul><ul><li>Erythrocyte ghost suspension was subjected to </li></ul><ul><li>variable electric voltage for 100 µseconds. </li></ul><ul><li>Temperature maintained at 37˚c </li></ul><ul><li>In extrusion and Sonication the final </li></ul><ul><li>preparation was stored at 3-5 ˚c in refrigerator </li></ul>
  9. 9. <ul><li>DRUG NANOERYTHROSOME CONJUGATION </li></ul><ul><li>Methotrexate was conjugated to nanoerythrosomes membrane using glyceraldehydes as cross linker. </li></ul><ul><li>The complex was washed with PBS and stored at 3-5˚c </li></ul><ul><li>Entrapment efficiency </li></ul><ul><li>Amount of drug loaded x 100 </li></ul><ul><li>Amount of drug added </li></ul>
  10. 10. IN VITRO CHARACTERIZATION <ul><li>SCANNING ELECTRON MICROSCOPY OF nEs </li></ul><ul><li>EG’s (2parts) + saline buffer (25 parts) </li></ul><ul><li>Formulation is dehydrated and dried to critical point </li></ul><ul><li>One drop of the diluted mixture is placed on a silver </li></ul><ul><li>coated copper grid ,then coated with gold </li></ul>
  11. 11. TRANSMISSION ELECTRON MICROSCOPY OF nEs <ul><li>EG’s (2parts) + saline buffer(25parts) </li></ul><ul><li>One drop of the diluted mixture is placed on a coated copper </li></ul><ul><li>grid and negatively stained with 1% uranyl acetate </li></ul><ul><li>Then the drop of mixture is seen through a transmission </li></ul><ul><li>electron microscope. </li></ul>
  12. 12. <ul><li>ROUTE OF ADMINISTRATION </li></ul><ul><li>They can be administered intravenously or intra arterially. </li></ul><ul><li>APPLICATIONS </li></ul><ul><li>They can target the drugs with in reticulo endothelial system. </li></ul><ul><li>Carriers for drugs , proteins , enzymes ,macromolecules. </li></ul><ul><li>Treatment of liver tumors, parasitic diseases, enzyme deficiency. </li></ul><ul><li>FUTURE PERSPECTIVES </li></ul><ul><li>A large amount of valuable work is needed so as to utilize the potentials of nanoerythrosomes </li></ul><ul><li>Disease like cancer could surely find its cure with the help of nanoerythrosomes. </li></ul><ul><li>Genetic engineering aspects can be coupled to give newer dimension to the existing drug carrier concept. </li></ul>
  13. 13. <ul><li>CONCLUSION </li></ul><ul><li>‘ GOLDEN EGGS IN NOVEL DRUG DELIVERY SYSTEM’ considering </li></ul><ul><li>their tremendous potential </li></ul><ul><li>REFERENCES </li></ul><ul><li>Advances in controlled and novel drug delivery ,edited by N.K JAIN ,CBS 332-351 </li></ul><ul><li>Anatomy and Physiology </li></ul><ul><li>Ross and Wilson’s Human Anatomy and Physiology </li></ul>