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Reasealed erythrocytes

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Information about Reasealed erythrocytes
Science-Technology

Published on March 9, 2014

Author: senthilk

Source: authorstream.com

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PowerPoint Presentation: RESEALED ERYTHROCYTES K.Senthil Kumar , M.Pharm Asst.prof QIS college of Pharmacy, Ongole . PowerPoint Presentation: It is system in which drug is delivered in to specific site in the body by using a colloidal carriers . Liposomes Neosome Nanoparticle Dendrimers Pro niosomes Microsphere Microcapsule Dendrimers Aquasomes Resealed erythrocytes TARGETED DRUG DELIVERY SYSTEM PowerPoint Presentation: blood cells used as a carriers for targeted drug delivery such as leukocytes ( WBC), Platelets ( thrombocytes), erythrocytes( RBC), Erythrocytes are more abundant cells in the human body. Which is used as a carrier for deliver the drug Antineoplasm, Antiviral , Anti bodies , Enzymes, Vitamins , steroids , antibiotics etc.. BLOOD CELL CARRIERS INTRODUCTION: Resealed erythrocyte is prepared by collecting blood sample and separating erythrocyte from plasma entrapping drug in the erythrocyte and resealing the carriers is called resealed erythrocytes. The overall process is carried out under osmotic condition and most of the resealed erythrocytes are quickly uptake by macrophages of RES which is present in liver , lung ,spleen. INTRODUCTION PowerPoint Presentation: The blood contains about 55% of plasma and 45 % - corpuscles and 1%- leukocytes. The plasma contains electrolytes and proteins which helpful in maintain the isotonicity and morphology of the cells . Erythrocytes ( Red Blood Corpuscels or Corpuscles) Erythro = red cytes = cell PowerPoint Presentation: COMPOSITION OF BLOOD PowerPoint Presentation: RBC Discocyte ELECTRON MICROSCOPY OF DRUG LOADED RBC: ELECTRON MICROSCOPY OF DRUG LOADED RBC RBC PowerPoint Presentation: SOURCE AND ISOLATION OF ERYTHROCYTES The resealed erythrocyte is ppd by using red blood cells of mice ,horse ,cattle , pig , monkey, rats, chicken sheep,rabbits , human blood ( o+ ve group ) Large animls - Vein puncture Small animal - Heart cardiac /splenic puncture by using a syringe containg suitable anticoagulant ( heparin or EDTA) Fresh blood is used for loading purpose due to high encapsulation efficiency than the aged blood. CARDIAC VEINPUNCTURE: CARDIAC VEINPUNCTURE RABBIT VENIPUNTURE: RABBIT VENIPUNTURE Inject at Marginal ear vein PowerPoint Presentation: The whole blood is centrifuged at 2500 rpm for 5 min at 4 ±1 O c in a refrigerated centrifuge.the serum and Buffy coat are carefully removed and packed cells washed 3 times with PBS 7.4 ph the washed erythrocytes are diluted with PBS and stored at 4 o c until used. Eppondroff tube 144 mmol of Nacl 5mmol of Mgcl2 10 mmol of glucose PowerPoint Presentation: MORPHOLOGY AND PHYSIOLOGY OF ERYTHROCYTES The normal erythrocyte is a bioconacve disc with depression located in the center on both sides . In healthy male contains 5.4 million cells /mm 3 Female 4.8 million cells / mm 3 Diameter 8.6µm Thickness 1.9 µm Surface area 145 µm Volume capacity 96 fl PowerPoint Presentation: COMPOSITION OF ERYTHROCYTES The RBC is made of SPM which easily permeate the cations and anions. It consist of Water -63% Lipids-0.5% Glucose-0.8% Mineral -0.7% Heme-0.9% Methehemoglobin-0.5% Hemogloblin 33.67% PowerPoint Presentation: The HB consist of protein is called globin and pigment is called heme. Erythrocytes live only 120 days because of wear and tear on their plasma membrane . the process of erythrocyte formation within the body is known as erythropoiesis . Immature erythrocyte is called as reticulocytes . Erythrocytes are produced in red bone marrow under the regulation of haemopoietic hormone is called erythropoietin. PowerPoint Presentation: Biocompatible Biodegradable Large volume of drug can be encapsulated in small volume of cells Non immunogenic system Produce controlled release and site specific action Easily passé through capillaries ( squeeze) Minimize the side effect Increase the bioavailability by reducing fluctuation in s.s Longer life span in circulation ADVANTAGES PowerPoint Presentation: DISADVANTAGE Leakage of encapsulated material Possible to contamination Special precaution require for collection of blood Produce toxic effect due to longer circulation. Mismatching of blood group cause fatal effect Donar must be free from acute or chronic disease. PowerPoint Presentation: CHARACTERIZATION OF ERYTHROCYTES When erythrocytes are suspended in a hypotonic medium they swell two times their normal size and membrane rupture in the formation of pores with diameter of 200-500 A O . Normal RBC drug solution Loaded drug PowerPoint Presentation: while haemolysis the aq solution of drug is entrapped and allow them until equilibrium between intracellular and extracellular solution . The erythrocyte are resealed by suspending the cells are incubated in isotonic solution at 37 o c the pores will closed. PowerPoint Presentation: CRITERIA FOR SELECTION OF DRUG : It should be polar Itshould be resist degradation within erythrocyte s It should be lack of physical and chemical interaction with SPM It should have define kinetic and dynamic properties of drug. Non polar drug encapsulate with salt form Wide variety compound can encapsulate with 5000-6000 daltons. PowerPoint Presentation: I.V,-Accumulation in RES( Kupffer cells in liver) S.C ,-Accumulation lymphatic nodes haemolysis in injection site I.P- Peritoneal macrophages ROUTE OF ADMINISTRATIONS PowerPoint Presentation: The erythrocyte suspension was filled into vials and lyophilized at -40 o c to 0.01 torr using a laboratory lyophilizer . the dried powder was filled in amber colour glass vials stored at 4 0 c for month.improvement in shelf life of the carrier erythrocyte was achieved by storing cells in dry powder form ready for reconstitution . STORAGE OF LOADED ERYTHROCYTES PowerPoint Presentation: Drug loading Electroporation endocytosis osmosis based method chemical perturbation Hypotonic hypotonic dialysis isotonic osmotic lysis hypotonic preswelling Dilution METHODS OF DRUG ENCAPSULATION PowerPoint Presentation: ELECTROPORATION or electro insertion / electroencapsulation This method is based up on application of electrical shock via electrodes which inserted into chamber containing a erythrocytes suspension. The application electrical shock with threshold level of voltage is 2kv/cm for 20 µsec.which create pores in the cell membrane . The pore formation depends upon the electric field strength, pulse duration , and ionic strength of the suspending medium. Once membrane is perforated ions are rapidly distributed between extra and intra cellular until equilibrium.under this condition drug molecule is added they permeate into red blood cells. PowerPoint Presentation: Eg. Chlorpromazine Hydrocortisone Vitamin A PowerPoint Presentation: 1 volume of washed packed erythrocytes + 9 volume of buffer ( 2.5 Mm ATP,2.5Mm mgcl2,1Mm cacl2) Incubation for 2 mt at room temp. Pores are created add drug solution and resealed by using isotonic solution at 37 0 c for 2 mts. ENDOCYTOSIS Eg. Primaquine , vinblastine etc.. PowerPoint Presentation: OSMOSIS BASED METHOD ( PHYSICAL ) Erythrocyte are ability to undergo reversible swelling and shape changes in hypotonic solution or stress. By in increase volume of RBC leads to changes in shape from biconacave to spherical erythrocyte ( spherocytes) The cells can maintain their integrity up to tonicity pressure is 150 mosm kg- 1. More than this pressure cause haemolysis.after celllysis cellular contents are depleted these cells is called erythrocyte ghost.   HYPOTONIC DILUTION:   HYPOTONIC DILUTION It is a first method for encapsulation of chemicals into erythrocytes. It is a simple and easiest method In tthis method volume of packed erythrocyte is diluted with 2-20 volume of hypotonic Solution and drug solution. Then it is stored by adding a hypertonic buffer . the resultant mixture is then centrifuged the supernatant is discarded and cells are washed with isotonic buffer.   HYPOTONIC DIALYSIS :   HYPOTONIC DIALYSIS This method is introduced by kilbansky in 1959 for encapsulation of enzymes and lipids suspension of erythrocytes+ drug ( hematocrit value of 70–80 %) placed in a conventional dialysis tube. immersed in 10–60 ml volumes of a hypotonic buffer. The medium is agitated slowly for 2 hr. The dialysis tube is restored by hypertonic buffer to the surrounding medium or by replacing the surrounding medium by isotonic buffer - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - ISOTONIC HAMOLYSIS: ISOTONIC HAMOLYSIS In this method involves isotonic haemolysis which is achieved by erythrocytes are incubated in solutions of a substance containing high permeability substance suchas urea solution , PEG solution,ammonium chloride , DMSO.so that solute slowly diffuse into cell. Followed by water intake until a osmotic equilibrium. HYPOTONIC PRE -SWELLING TECHNIQUE: HYPOTONIC PRE -SWELLING TECHNIQUE In this method first the erythrocyte is palced in slightly hypotonic solution without lysis . The swollen cells are recovered by centrifugation at low( g) speed . The supernatant is discarded and cells are leave for lysis by adding hypotonic solution and add aqueous solution of drug and resealed it. Methotrexate Metrindazole PowerPoint Presentation: Method Encapsulation efficiency Dilution method 20-40% Dialysis 30-45% Preswell dilution 30-90% Isotonic haemolysis Better invivo survival PowerPoint Presentation: CHEMICAL PERTURBATION METHOD : Generally antibiotics like amphotericin b which kill microorganism by increasing the permeability of their membrane to metabolite and ions .this property could be used in encapsulation drug . Amphotericin B can be used to load the antileukaemic drug daunomycin . Amphotericin B interacts with cholesterol of plasma membrane of RBC which cause induce the permeability of the membrane . Then add the drug in to the above solution and resealed by using isotonic solution . Evaluation test for resealed erythrocytes: Evaluation test for resealed erythrocytes PowerPoint Presentation: Thank you for your patience

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