NanoparticlesNanoparticlesPresented by:

Chinchole Pravin Sonu(M.PHARM 2nd SEM)

DEPARTMENT OF PHARMACEUTICS & QUALITY ASSURANCE

R. C. Patel Institute of Pharmaceutical Education and Research, shirpur.

contentscontents

IntroductionIntroductionPreparation techniques of nanoparticlesPreparation techniques of nanoparticlesNovel nanoparticulate systemsNovel nanoparticulate systemsDrug loading & in vitro release processDrug loading & in vitro release processPharmaceutical aspects of nanoparticlesPharmaceutical aspects of nanoparticlesCharacterization of nanoparticlesCharacterization of nanoparticlesIn vivo fate & biodistribution of nanoparticlesIn vivo fate & biodistribution of nanoparticlesTherapeutic applications of nanoparticlesTherapeutic applications of nanoparticles

Nanoparticles are subnanosized colloidal structures composed of Nanoparticles are subnanosized colloidal structures composed of synthetic or semi synthetic polymerssynthetic or semi synthetic polymers

The polymeric nanoparticles can carry drugs or proteinacous substances, The polymeric nanoparticles can carry drugs or proteinacous substances, antigens.antigens.

These bioactives are entrapped in the polymer matrix as particulates These bioactives are entrapped in the polymer matrix as particulates enmesh or solid solution or may be bound to the particle surface by enmesh or solid solution or may be bound to the particle surface by physical adsorption or chemicallyphysical adsorption or chemically

Nanospheres contain drug embedded within the matrix or adsorbed on to Nanospheres contain drug embedded within the matrix or adsorbed on to the surfacethe surface

Nanocapsules are vesicular system in which drug is essentially Nanocapsules are vesicular system in which drug is essentially encapsulated within central volume surrounded by an embryonic encapsulated within central volume surrounded by an embryonic continuous polymeric sheathcontinuous polymeric sheath

Preparation tech. of nanoparticlesPreparation tech. of nanoparticles(Polymers for nanoparticles)(Polymers for nanoparticles)

ProteinsProteinsGelatinGelatinAlbuminAlbuminLectinsLectinsLeguminLeguminVicilinVicilin

polysaccharidespolysaccharidesAlginateAlginateDextranDextranChitosanChitosanAgaroseAgarosePullulanPullulan

Various synthetic polymers used for prepration of Various synthetic polymers used for prepration of nanoparticlesnanoparticles

PrepolymerisedPrepolymerisedPoly(e-caprolactone)Poly(e-caprolactone)Polylactic acidPolylactic acidPoly(lactide-co-glycolide)Poly(lactide-co-glycolide)PolystyrenePolystyrene

Polymerised in processPolymerised in processPoly(isobutylcynoacrylate)Poly(isobutylcynoacrylate)Poly(butylcynoacrylate)Poly(butylcynoacrylate)Poly(hexylcynoacrylate)Poly(hexylcynoacrylate)PolymethylmethacrylatePolymethylmethacrylate

Selection of method for preparing nanoparticles depends on Selection of method for preparing nanoparticles depends on the physicochemical characteristics of polymer & the drug to the physicochemical characteristics of polymer & the drug to be loaded. The preparation tech. determine the inner be loaded. The preparation tech. determine the inner structure, in vitro release profile & biological fate of these structure, in vitro release profile & biological fate of these polymeric delivery system.polymeric delivery system.

The different systems includeThe different systems includeA A matrix typematrix type system consisting of an entanglement system consisting of an entanglement of oligomer or polymer units (nanoparticles of oligomer or polymer units (nanoparticles /nanospheres)/nanospheres)A A reservoir typereservoir type of system consisting of an oily core of system consisting of an oily core surrounded by an embryonic polymeric shell surrounded by an embryonic polymeric shell (nanocapsules)(nanocapsules)The drug can be entrapped or adsorbed on surface The drug can be entrapped or adsorbed on surface of these particulate systemof these particulate system

Method are classified asMethod are classified as

Amphiphilic macromolecule cross linkingAmphiphilic macromolecule cross linking 1.Heat crosslinking1.Heat crosslinking 2.Chemical crosslinking2.Chemical crosslinking

Polymerization based methodsPolymerization based methods1. Polymerization of monomers in situ1. Polymerization of monomers in situ2. Emulsion polymerization2. Emulsion polymerization3. Dispersion polymerization 3. Dispersion polymerization 4. Interfacial condensation polymerization4. Interfacial condensation polymerization5. Interfacial complexation5. Interfacial complexation

Polymer precipitation methodsPolymer precipitation methodsSolvent extraction/evaporationSolvent extraction/evaporationSolvent displacementSolvent displacementSalting outSalting out

Polymers used for preparation of nanoparticles& nanocapsulesPolymers used for preparation of nanoparticles& nanocapsules

Polymer usePolymer use TechniqueTechnique Candidate drugCandidate drugHydrophilicHydrophilic

Albumin, gelatinAlbumin, gelatinHeat denaturation & cross Heat denaturation & cross linking in w/o emulsionlinking in w/o emulsionDesolvation &cross linking in Desolvation &cross linking in aqueous mediumaqueous medium

HydrophilicHydrophilic

Hydrophilic & protein affinityHydrophilic & protein affinity

Alginate,chitosan Alginate,chitosan Cross linking in aq. mediumCross linking in aq. medium Hydrophilic & protein affinityHydrophilic & protein affinity

DextranDextran Polymer precipitation in an Polymer precipitation in an organic solventorganic solvent

HydrophilicHydrophilic

HydrophobicHydrophobicPoly(alkylcyanoacrylates)Poly(alkylcyanoacrylates)

Emulsion polymerizationEmulsion polymerizationInterfacial/polymerizationInterfacial/polymerization

HydrophilicHydrophilicHydrophobicHydrophobic

PolyestersPolyestersPoly(lactic acid,poly(lactide-co-Poly(lactic acid,poly(lactide-co-glycolide)poly(glycolide)poly(€€-caprolactone-caprolactone))

Solvent extraction-evaporationSolvent extraction-evaporationSolvent displacementSolvent displacementSalting outSalting out

Hydrophilic ,HydrophobicHydrophilic ,HydrophobicSoluble in polar solventSoluble in polar solventSoluble in polar solventSoluble in polar solvent

Method of preparation for nanoparticlesMethod of preparation for nanoparticles

Amphillic macromolecules that undergo a crosslinking Amphillic macromolecules that undergo a crosslinking reaction during preparation of nanospheresreaction during preparation of nanospheres

Monomers that polymerize during formation of nanospheresMonomers that polymerize during formation of nanospheres

Hydrophobic polymers which are initially dissolved in organic Hydrophobic polymers which are initially dissolved in organic solvents & then precipitated under controlled conditions to solvents & then precipitated under controlled conditions to produce nanospheresproduce nanospheres

Nanoparticles preparation by cross linking of amphiphilic Nanoparticles preparation by cross linking of amphiphilic macromoleculesmacromolecules

It can be prepared from amphiphilic macromolecules, proteins & It can be prepared from amphiphilic macromolecules, proteins & polysaccharides which have affinity for aq.&lipid solvents. polysaccharides which have affinity for aq.&lipid solvents. The tech.of their preparation involves the aggregation of amphiphiles followed The tech.of their preparation involves the aggregation of amphiphiles followed by further stabilization either by heat denaturation or chemical cross linking.by further stabilization either by heat denaturation or chemical cross linking.

Cross linking in w/o emulsionCross linking in w/o emulsion1. The method involves the emulsification of bovine serum albumin/human 1. The method involves the emulsification of bovine serum albumin/human

serum albumin or protein aq. solution in oil using high pressure serum albumin or protein aq. solution in oil using high pressure homogenization or high frequency sonicationhomogenization or high frequency sonication

2. The w/o emulsion so formed is poured into preheated oil. The suspension in 2. The w/o emulsion so formed is poured into preheated oil. The suspension in preheated oil maintained above 100preheated oil maintained above 10000C is held stirred for time in order to C is held stirred for time in order to denature & aggregate the protein contents of aq. pool completely & to denature & aggregate the protein contents of aq. pool completely & to evaporate waterevaporate water

3. The particles are finally washed with organic solvent to remove any oil traces 3. The particles are finally washed with organic solvent to remove any oil traces & collected by centrifugation& collected by centrifugation

4. The main factors are emulsification energy & temperature (used for 4. The main factors are emulsification energy & temperature (used for denaturation & aggregationdenaturation & aggregation

Phase separation in aqueous medium (desolvation)Phase separation in aqueous medium (desolvation)

The protein or polysaccharide from an aq. phase can be desolvated by The protein or polysaccharide from an aq. phase can be desolvated by PH change or temp change or by adding counter ions. Cross linking may PH change or temp change or by adding counter ions. Cross linking may be affected simultaneously or next to the desolvation stepbe affected simultaneously or next to the desolvation step

Steps are protein dissolution, protein aggregation & protein Steps are protein dissolution, protein aggregation & protein deaggregationdeaggregation

Solvent competing agent ,sodium sulphate is mainly used as a Solvent competing agent ,sodium sulphate is mainly used as a desolvating agent while alcohol are added as desolvating or desolvating agent while alcohol are added as desolvating or deaggregating agentdeaggregating agent

Addition can be optimized turbid metrically using Nephelometer.Addition can be optimized turbid metrically using Nephelometer.

Aqueous phase (Protein aq.solution)

Protein aggregates (coacervates)

Desolvation (solvent competing agent)

Protein colloidal dispersion

Nanoparticle suspension (external aq. Phase)

Crosslinking(aldehyde)

Resolvation(alcohol)

Nanoparticle preparation by desolvation in an aq. medium technique

PPH H induced aggregationinduced aggregation

Gelatin & Tween 20 were dissolved in aq. phase & PGelatin & Tween 20 were dissolved in aq. phase & PHH was adjusted to was adjusted to optimum value. Solutions were heated ,then quenching at 4oC for 24h.The optimum value. Solutions were heated ,then quenching at 4oC for 24h.The leads to colloidal dispersion of aggregated gelatin. The aggregates were leads to colloidal dispersion of aggregated gelatin. The aggregates were finally cross linked using glutaraldehyde.The size of nanospheres were of finally cross linked using glutaraldehyde.The size of nanospheres were of 200 nm. The ideal P200 nm. The ideal PH H range is 5.5-6.5.range is 5.5-6.5.

Counter ion induced aggregationCounter ion induced aggregation

Separation of protein phase may occur by the presence of counter ions in Separation of protein phase may occur by the presence of counter ions in the aqueous medium.the aqueous medium.

The aggregation can be propagated by adding counters ions followed by The aggregation can be propagated by adding counters ions followed by rigidization step.rigidization step.

Chitosan nanospheres can be prepared by adding Chitosan nanospheres can be prepared by adding tripolyphosphatetripolyphosphate to the to the medium.medium.

Alginate nanoparticles can be prepared by gelation with Alginate nanoparticles can be prepared by gelation with calcium ionscalcium ions

Nanoparticles prepared by polymerization methodsNanoparticles prepared by polymerization methods

The polymers used for nanospheres preparation include poly The polymers used for nanospheres preparation include poly (methylmethacrylate) poly(acrylamide)poly(butyl cyanoacrylate).(methylmethacrylate) poly(acrylamide)poly(butyl cyanoacrylate).

The different methods using in situ polymerization tech. areThe different methods using in situ polymerization tech. are

Methods in which the monomer to be polymerised is emulsified in a non Methods in which the monomer to be polymerised is emulsified in a non solvent phase or solvent phase or (emulsion polymerization)(emulsion polymerization)Methods in which the monomer is dissolved in a solvent that is non Methods in which the monomer is dissolved in a solvent that is non solvent for the resulting polymer solvent for the resulting polymer (Dispersion polymerization)(Dispersion polymerization)In EP method the monomer is dissolved in an internal phase while in In EP method the monomer is dissolved in an internal phase while in case of DP it is taken in dispersed phase case of DP it is taken in dispersed phase In both cases after polymerization polymer tends to be insoluble in In both cases after polymerization polymer tends to be insoluble in internal phase & results into suspension of nanospheresinternal phase & results into suspension of nanospheres

Emulsion polymerizationEmulsion polymerization

Micellar polymerizationMicellar polymerization

The mechanism involved are micellar nucleation & polymerization & The mechanism involved are micellar nucleation & polymerization & homogenous nucleation & polymerizationhomogenous nucleation & polymerization

The first includes swollen monomer micelles as the site of nucleation & The first includes swollen monomer micelles as the site of nucleation & polymerization.polymerization.

The monomer is emulsified in the non solvent phase with the help of The monomer is emulsified in the non solvent phase with the help of surfactant molecules.surfactant molecules.

The process leads to formation of monomer swollen micelles which has The process leads to formation of monomer swollen micelles which has size in nanometric range. size in nanometric range.

The polymerization reaction proceeds through nucleation & propogation The polymerization reaction proceeds through nucleation & propogation in the presence of chemical or physical initiatorin the presence of chemical or physical initiator

Homogeneous polymerizationHomogeneous polymerizationMonomer is sufficiently soluble in the continuous outer phaseMonomer is sufficiently soluble in the continuous outer phase

Formation of primary chains (oligomers)Formation of primary chains (oligomers)

At certain length oligomers precipitate & forms primary particles At certain length oligomers precipitate & forms primary particles which are stabilized by surfactantwhich are stabilized by surfactant

Addition of monomer input or fusion of primary particles forms Addition of monomer input or fusion of primary particles forms nanoparticlesnanoparticles

Water soluble drugs may be associated with PACA Water soluble drugs may be associated with PACA nanoparticles either by dissolving the drug in the nanoparticles either by dissolving the drug in the aqueous polymerization medium or by incubating the aqueous polymerization medium or by incubating the blank nanospheres with an aq. solution of drugblank nanospheres with an aq. solution of drug

Drug molecules may be entrapped within the polymer Drug molecules may be entrapped within the polymer matrix & are also adsorbed onto the surface of matrix & are also adsorbed onto the surface of nanoparticlesnanoparticles

Drug molecules may be physically adsorbed on the Drug molecules may be physically adsorbed on the surfacesurface

Dispersion polymerizationDispersion polymerization

In emulsion polymerization monomer is emulsified in an In emulsion polymerization monomer is emulsified in an immiscible phase using surfactant.In case of dispersion immiscible phase using surfactant.In case of dispersion polymerization monomer is dissolved in an aqueous polymerization monomer is dissolved in an aqueous medium which acts as precipitant for polymermedium which acts as precipitant for polymer

The monomer is introduced into the dispersion medium. The monomer is introduced into the dispersion medium. Polymerization is initiated by adding a catalyst & Polymerization is initiated by adding a catalyst & proceeds with nucleation phase followed by growth proceeds with nucleation phase followed by growth phase.phase.

Inverse emulsification polymerization mechanismInverse emulsification polymerization mechanism

Emulsion polymerization processEmulsion polymerization process

The nucleation is directly induced in the aqueous The nucleation is directly induced in the aqueous monomer solution & monomer solution & the presence of stabilizer or the presence of stabilizer or surfactant is not requiredsurfactant is not requiredThe acrylamide or methyl methacrylate monomer is The acrylamide or methyl methacrylate monomer is dissolved in aqueous phase & polymerized by gamma dissolved in aqueous phase & polymerized by gamma irradiationirradiationBy chemical initiation (ammonium or potassium By chemical initiation (ammonium or potassium peroxodisulphate) combined with heating to temp. above peroxodisulphate) combined with heating to temp. above 65 65 00CCPMMA nanoparticles can be prepared by gamma PMMA nanoparticles can be prepared by gamma irradiation in the presence of antigenic materialirradiation in the presence of antigenic material

e.g. influenza virion, influenza sub unit antigen, bovine e.g. influenza virion, influenza sub unit antigen, bovine serum albumin, HIV-1 & HIV-2 antigensserum albumin, HIV-1 & HIV-2 antigens

Interfacial polymerizationInterfacial polymerization

The preformed polymer phase is finally transformed to an The preformed polymer phase is finally transformed to an embryonic sheath.embryonic sheath.

The polymer & drug are dissolved in a volatile solvent. The The polymer & drug are dissolved in a volatile solvent. The solution is is poured into a non solvent for both polymer & core solution is is poured into a non solvent for both polymer & core phase.phase.

The polymer phase is separated as as a coacervate phase at o/w The polymer phase is separated as as a coacervate phase at o/w interface. The mixture turns milky due to formation of interface. The mixture turns milky due to formation of nanocapsules.nanocapsules.

This method is used for proteins, enzymes, antibodies, & cellsThis method is used for proteins, enzymes, antibodies, & cells

Interfacial polymeric condensation of 2,2-bis-(4hydroxyphenyl) Interfacial polymeric condensation of 2,2-bis-(4hydroxyphenyl) propane & sebacoyl chloridepropane & sebacoyl chlorideThe size of nanocapsules ranges from 30-300nmThe size of nanocapsules ranges from 30-300nm

Nanoparticle preparation using emulsion solvent Nanoparticle preparation using emulsion solvent evaporation methodevaporation method

Hydrophobic polymer & or hydrophobic drug is dissolved in a organic Hydrophobic polymer & or hydrophobic drug is dissolved in a organic solvent followed by its dispersion in a continuous aq. Phase in which the solvent followed by its dispersion in a continuous aq. Phase in which the polymer is insoluble. External phase contains stabilizer.polymer is insoluble. External phase contains stabilizer.

Depending upon solvent miscibility tech.may called as solvent extraction Depending upon solvent miscibility tech.may called as solvent extraction or evaporation methodor evaporation method

The polymer precipitation is done byThe polymer precipitation is done by

Increasing the solubility of organic solvent in the external medium by Increasing the solubility of organic solvent in the external medium by adding an alcohol (isopropanol)adding an alcohol (isopropanol)

By incorporating water into ultraemulsion(to extract solvent)By incorporating water into ultraemulsion(to extract solvent)

By evaporation of solvent at room temp. by using vacuumBy evaporation of solvent at room temp. by using vacuum

Using an organic solvent which is completely soluble in the continuous Using an organic solvent which is completely soluble in the continuous aq.phase(acetone)nanoprecipitationaq.phase(acetone)nanoprecipitation

Solvent extraction methodSolvent extraction method

This method involves the preparation of o/w emulsion This method involves the preparation of o/w emulsion

The subsequent removal of solvent or the addition of water to the The subsequent removal of solvent or the addition of water to the system so as to affect diffusion of solvent to external phase system so as to affect diffusion of solvent to external phase (emulsification diffusion method)(emulsification diffusion method)

The solvent used for polymer is poorly miscible with dispersion phase & The solvent used for polymer is poorly miscible with dispersion phase & thus diffuses & evaporates out slowly on continual stirringthus diffuses & evaporates out slowly on continual stirring

Dispersion medium miscible polymer solvent (alcohol & acetone Dispersion medium miscible polymer solvent (alcohol & acetone instantaneously diffuses into the aq. phase & polymer precipitates as instantaneously diffuses into the aq. phase & polymer precipitates as tiny nanospherestiny nanospheres

Solvent displacement methodSolvent displacement method

It is based on interfacial deposition of a polymer following It is based on interfacial deposition of a polymer following displacement of a semi polar solvent miscible with water displacement of a semi polar solvent miscible with water from a lipophilic solutionfrom a lipophilic solution

The organic solvent diffuses instantaneously to the external The organic solvent diffuses instantaneously to the external aq. Phase inducing immediate polymer precipitation because aq. Phase inducing immediate polymer precipitation because of complete miscibility of both the phasesof complete miscibility of both the phasesIf drug is highly hydrophilic it diffuses out into the external aq. If drug is highly hydrophilic it diffuses out into the external aq. phase while if drug is hydrophobic it precipitates in aq. phase while if drug is hydrophobic it precipitates in aq. medium as nanocrystalsmedium as nanocrystals

Solvent displacement methodSolvent displacement method

Salting outSalting out

The method involves the incorporation of a saturated aq. solution of The method involves the incorporation of a saturated aq. solution of polyvinyl alcohol into an acetone solution of polymer under magnetic polyvinyl alcohol into an acetone solution of polymer under magnetic stirring to form an o/w emulsion.stirring to form an o/w emulsion.

In nanoprecipitation In nanoprecipitation tech.polymeric solution is tech.polymeric solution is completely miscible completely miscible with external aq. mediumwith external aq. medium. . But in this methodBut in this method miscibility of both the miscibility of both the phases is prevented by phases is prevented by saturation of external aq .phase with PVA.saturation of external aq .phase with PVA.

Precipitation of polymer occurs when sufficient amount of water is Precipitation of polymer occurs when sufficient amount of water is added to external phase to allow complete diffusion of acetone from added to external phase to allow complete diffusion of acetone from internal phase to aq.phaseinternal phase to aq.phase

This method is suitable for drugs & polymers that are soluble in polar This method is suitable for drugs & polymers that are soluble in polar solvents such as acetone & ethanolsolvents such as acetone & ethanol

Solid lipid nanoparticlesSolid lipid nanoparticles

The solid lipid nanoparticles are submicron colloidal The solid lipid nanoparticles are submicron colloidal carriers which are composed of physiological carriers which are composed of physiological lipid ,dispersed in water or in an aqueous surfactant lipid ,dispersed in water or in an aqueous surfactant solution solution

It combines the advantages of polymeric nanoparticles, fat It combines the advantages of polymeric nanoparticles, fat emulsions & liposomesemulsions & liposomes

The liquid lipid was replaced by a solid lipid which The liquid lipid was replaced by a solid lipid which transformed into solid lipid nanoparticlestransformed into solid lipid nanoparticles

AdvantagesAdvantagesSmall size & narrow size distribution provides for site specific drug Small size & narrow size distribution provides for site specific drug delivery by SLNsdelivery by SLNs

Controlled release of active drug over a long period can be achievedControlled release of active drug over a long period can be achieved

Protection of incorporated drug against chemical degradationProtection of incorporated drug against chemical degradation

SLNs can be lyophilized & spray driedSLNs can be lyophilized & spray dried

No toxic metabolites are producedNo toxic metabolites are produced

Sterilization can be done by autoclaving or gamma irradiationSterilization can be done by autoclaving or gamma irradiation

Surface modification can be easily done Surface modification can be easily done

Preparation of SLNPreparation of SLN

Melting of the lipid

Dissolution of drug in melted lipid

Mixing of preheated dispersion medium & drug lipid melt

Premix using stirrer to form coarse pre-emulsion

High pressure homogenization at a temp. above the lipids melting point

o/w nanoemulsion

Solidification of the nanoemulsion by cooling down to room temp. to form SLN

Hot Homogenization Technique

Preparation of SLNPreparation of SLN

Melting of the lipid

Dissolution /solubilization of drug in melted lipid

Solidification of drug loaded lipid in liquid nitrogen or dry ice

Grinding in a powder mill(50-100µm particles)

Dispersion of lipid in the cold aqueous dispersion medium

Solid lipid nanoparticles

Cold Homogenization Technique

Nanocrystals and nanosuspensionNanocrystals and nanosuspension

Pharmaceutical aspects of nanoparticlesPharmaceutical aspects of nanoparticlesThe important process parameters performed are Purification, Freeze The important process parameters performed are Purification, Freeze drying, Sterilizationdrying, SterilizationPurification of nanoparticlesPurification of nanoparticles:-Toxic impurities includes organic :-Toxic impurities includes organic solvents, residual monomers, polymerization initiators, electrolytes, solvents, residual monomers, polymerization initiators, electrolytes, stabilizers & large polymer aggregates. Most commonly used method is stabilizers & large polymer aggregates. Most commonly used method is gel filtration & ultra filtrationgel filtration & ultra filtration

Freeze drying of nanoparticlesFreeze drying of nanoparticlesIt includes freezing of nanoparticle suspension & sublimation of water to It includes freezing of nanoparticle suspension & sublimation of water to produce free flowing powder.produce free flowing powder.

Advantages areAdvantages arePrevention from degradation & or solubilization of the polymerPrevention from degradation & or solubilization of the polymer

Prevention from drug leakage, drug desorption, drug degradationPrevention from drug leakage, drug desorption, drug degradation

Nanocapsules containing oily core may be processed in the presence of Nanocapsules containing oily core may be processed in the presence of mono or disaccharides (glucose or sucrose)mono or disaccharides (glucose or sucrose)Readily dispersible in water without modifications in their Readily dispersible in water without modifications in their physicochemical propertiesphysicochemical properties

Sterilization of NanoparticlesSterilization of Nanoparticles

Nanoparticles for parenteral use should be sterilized to be pyrogen free Nanoparticles for parenteral use should be sterilized to be pyrogen free before animal or human use.before animal or human use.

Sterilization in nanoparticles is achieved by using aseptic Sterilization in nanoparticles is achieved by using aseptic tech.throughout their preparation & processing & formulation & by tech.throughout their preparation & processing & formulation & by sterilizing treatments like autoclaving or sterilizing treatments like autoclaving or γγ-- irradiation irradiation

Characterization of nanoparticlesCharacterization of nanoparticlesParameterParameter Characterization methodCharacterization method

Particle size & size distributionParticle size & size distribution Photon correlation spectroscopy,Laser Photon correlation spectroscopy,Laser defractometry,Transmission electron defractometry,Transmission electron microscopy,SEM,Atomic force microscopy,SEM,Atomic force microscopy,Mercury porosimetrymicroscopy,Mercury porosimetry

Charge determinationCharge determination Laser doppler anemometry, Zeta Laser doppler anemometry, Zeta potentiometerpotentiometer

Surface hydrophobicitySurface hydrophobicity Water contact angle measurements, rose Water contact angle measurements, rose bengal binding X-ray photoelectron bengal binding X-ray photoelectron spectroscopyspectroscopy

Chemical analysis of surfaceChemical analysis of surface Static secondary ion mass spectrometry, Static secondary ion mass spectrometry, sorptometersorptometer

Carrier-drug interactionCarrier-drug interaction Differential scanning calorimetryDifferential scanning calorimetry

Nanoparticle dispersion stabilityNanoparticle dispersion stability Critical flocculation temp (CFT)Critical flocculation temp (CFT)

Release profileRelease profile In vitro release charac.under physiologic & In vitro release charac.under physiologic & sink conditionssink conditions

Drug stabilityDrug stability Bioassay of drug extracted from Bioassay of drug extracted from nanoparticlesnanoparticlesChemical analysis of drugChemical analysis of drug

Size & morphologySize & morphologyEM(SEM & TEM) are widely used for determining particle size & its EM(SEM & TEM) are widely used for determining particle size & its distribution.distribution.Freeze fracturing of particles allows for morphological determination of Freeze fracturing of particles allows for morphological determination of inner structure of particles.inner structure of particles.TEM permits differentiation among nanocapsules, nanoparticles, & TEM permits differentiation among nanocapsules, nanoparticles, & emulsion dropletsemulsion dropletsAtomic force microscopy (AFM) images can be obtained in an aq.medium Atomic force microscopy (AFM) images can be obtained in an aq.medium hence used for investigation of nanoparticle behavior in biological hence used for investigation of nanoparticle behavior in biological environment.environment.

Specific surfaceSpecific surface:-Is determined with the help of Sorptometer:-Is determined with the help of SorptometerSurface charge & electrophoretic mobility:-The nature & intensity of Surface charge & electrophoretic mobility:-The nature & intensity of surface charge determines their interaction with biological environment as surface charge determines their interaction with biological environment as well as with bioactive compounds.It is determined by measuring the well as with bioactive compounds.It is determined by measuring the particle velocity in an electric field. (Laser doppler anemometry)particle velocity in an electric field. (Laser doppler anemometry)The surface charge of colloidal particles is measured as electrophoretic The surface charge of colloidal particles is measured as electrophoretic mobility which is determined in phosphate saline buffer & human serummobility which is determined in phosphate saline buffer & human serum

Surface hydrophobicitySurface hydrophobicity:-influences in interaction with biological :-influences in interaction with biological environmentenvironment

( Protein particles & cell adhesion). It is determined by two phase ( Protein particles & cell adhesion). It is determined by two phase partition,contact angle measurements, adsorption of hydrophobic partition,contact angle measurements, adsorption of hydrophobic fluorescent or radiolabelled probes.fluorescent or radiolabelled probes.

X-ray photoelectron spectroscopy permits the identification of specific X-ray photoelectron spectroscopy permits the identification of specific chemical groups on the surface o nanoparticles.chemical groups on the surface o nanoparticles.DensityDensity:-The density of nanoparticles is determined with helium or :-The density of nanoparticles is determined with helium or air using a gas Pycnometerair using a gas PycnometerMolecular weight measurement of nanoparticlesMolecular weight measurement of nanoparticles:- :- Molecular weight of Molecular weight of the polymer & its distribution in the matrix can be evaluated by gel the polymer & its distribution in the matrix can be evaluated by gel permeation chromatography using a refractive index detector.permeation chromatography using a refractive index detector.Nanoparticle recovery & drug incorporation efficiency:-Nanoparticle recovery & drug incorporation efficiency:-Nanoparticle Nanoparticle yield can be calculated as yield can be calculated as

nanoparticles recovery(%)=nanoparticles recovery(%)= Conc. of drug in nanoparticles Conc. of drug in nanoparticles *100*100

conc of nanoparticles recoveredconc of nanoparticles recovered

Drug incorporation efficiency or drug contentDrug incorporation efficiency or drug content:-:- Drug content(% w/w) =Drug content(% w/w) = Conc of drug in nanoparticles *100 Conc of drug in nanoparticles *100

conc of nanoparticles recoveredconc of nanoparticles recovered

In vitro releaseIn vitro release:-:-In vitro release profile can be determined using standard In vitro release profile can be determined using standard dialysis, diffusion cell or modified ultrafiltration techniquedialysis, diffusion cell or modified ultrafiltration technique

In vivo fate & biodistribution of nanoparticlesIn vivo fate & biodistribution of nanoparticles

The plasma proteins (opsonins) adsorb on to the surface of colloidal The plasma proteins (opsonins) adsorb on to the surface of colloidal carriers & render particles recognizable to RES.carriers & render particles recognizable to RES.

Phagocytosis of particulates by elements of RES( liver, spleen, Phagocytosis of particulates by elements of RES( liver, spleen, bone marrow), liver( Kupffer cells )is regulated by opsonins & bone marrow), liver( Kupffer cells )is regulated by opsonins & disopsonins (IgA)disopsonins (IgA)

Surface engineering of nanoparticlesSurface engineering of nanoparticles

Steric stabilized (stealth) nanoparticlesSteric stabilized (stealth) nanoparticlesBio-mimetic nanoparticlesBio-mimetic nanoparticlesAntibody coated nanoparticlesAntibody coated nanoparticlesMagnetically guided nanoparticlesMagnetically guided nanoparticlesBioadhesive nanoparticlesBioadhesive nanoparticles

Steric stabilized (stealth) nanoparticles:-Steric stabilized (stealth) nanoparticles:-PEO is most commonly used PEO is most commonly used synthetic material to modify interactions of particles with serum synthetic material to modify interactions of particles with serum components. amphiphilic copolymers rather than homopolymer have components. amphiphilic copolymers rather than homopolymer have been shown to be effective in steric stabilization. Polysorbates 20,40,60 & been shown to be effective in steric stabilization. Polysorbates 20,40,60 & 80 & poloxamers are used for surface modification of PMMA 80 & poloxamers are used for surface modification of PMMA nanoparticlesnanoparticles

Bioadhesive nanoparticles:-covalent coupling of different ligands,Bioadhesive nanoparticles:-covalent coupling of different ligands, tomato tomato lectin, asparagus pea lectinlectin, asparagus pea lectin

Bio-mimetic nanoparticles:-Bio-mimetic nanoparticles:- albumin albumin which is main protein has been which is main protein has been reported as biomimitic ligand. reported as biomimitic ligand. Sialic acidSialic acid also serves the same also serves the same

Magnetically guided nanoparticles:-Nanoparticles are rendered magnetic Magnetically guided nanoparticles:-Nanoparticles are rendered magnetic by incorporating by incorporating FeFe33OO4 4 particles(10-20 nm)particles(10-20 nm) with drug during preparation with drug during preparation stage. Particles are injected through artery of tumour tissue & guided stage. Particles are injected through artery of tumour tissue & guided externally by external magnetic fieldexternally by external magnetic field

Iron oxide nanoparticles are also reportedIron oxide nanoparticles are also reported

Antibody coated nanoparticles:- Selectivity in drug targeting can be Antibody coated nanoparticles:- Selectivity in drug targeting can be achieved by attaching achieved by attaching monoclonal antibodymonoclonal antibody as a site directing device. as a site directing device. Mab can be fixed by direct adsorption or via a Mab can be fixed by direct adsorption or via a spacer moleculespacer molecule or by or by covalent linkagecovalent linkage using carbodiimide, glutaraldehyde reaction. using carbodiimide, glutaraldehyde reaction.

Therapeutic applications of nanoparticlesTherapeutic applications of nanoparticles

Biodegradable polymers like gelatin, albumin, casein, Biodegradable polymers like gelatin, albumin, casein, polysaccharide,lectin & synthetic polymers like polycaprolactone, polysaccharide,lectin & synthetic polymers like polycaprolactone, polyesters, polyanhydrides, polycyanoacrylates with various drug polyesters, polyanhydrides, polycyanoacrylates with various drug release charac. have been used to formulate sustained release release charac. have been used to formulate sustained release nanoparticles.nanoparticles.

Intracellular targeting:-Intracellular targeting:-available antibiotics has following limitationsavailable antibiotics has following limitationsPoor intracellular diffusionPoor intracellular diffusion

Most intracellular infections are difficult to irradicate because Most intracellular infections are difficult to irradicate because bacteria inside phagosome are protected from antibioticsbacteria inside phagosome are protected from antibiotics

The need for antibiotics with greater intracellular efficacy led to the The need for antibiotics with greater intracellular efficacy led to the development of endocytosable drug carriers including nanoparticlesdevelopment of endocytosable drug carriers including nanoparticles

Various intracellular bacterial & parasitic infections where Various intracellular bacterial & parasitic infections where Nanoparticle loaded antibiotics are usedNanoparticle loaded antibiotics are usedLeshmaniasisLeshmaniasisSalmonellosisSalmonellosisTrypanosisTrypanosisMalariaMalaria

Prolonged systemic circulationProlonged systemic circulation can be achieved by surface can be achieved by surface modification of nanoparticles with modification of nanoparticles with different block polymers different block polymers (pluronics) or polyethylene glycols(pluronics) or polyethylene glycols

Nanoparticles in chemotherapyNanoparticles in chemotherapy

Antitumour agents that are either entrapped or adsorbed onto Antitumour agents that are either entrapped or adsorbed onto polyalkylcyanoacrylate nanoparticles(doxorubicin)polyalkylcyanoacrylate nanoparticles(doxorubicin)

Chemoembolizatiuon makes use of biodegradable particles Chemoembolizatiuon makes use of biodegradable particles administration to the liver tumours using a cathetor that passes directly administration to the liver tumours using a cathetor that passes directly into an artery of the tumorsinto an artery of the tumorsEnhanced endocytic activity & leaky vasculature of the tumour favours Enhanced endocytic activity & leaky vasculature of the tumour favours accumulation of intravenously administered nanoparticlesaccumulation of intravenously administered nanoparticles

ChemoembolizationChemoembolization

Adjuvant effect for vaccinesAdjuvant effect for vaccinesOn SC injection sustained release of the entrapped antigen induced On SC injection sustained release of the entrapped antigen induced significant antibody response & protected mice against a challenge with significant antibody response & protected mice against a challenge with mouse adapted influenza virus to a greater extentmouse adapted influenza virus to a greater extentA decrease in particle size & an increase in hydrophobicity of the A decrease in particle size & an increase in hydrophobicity of the nanoparticles increased the adjuvant effect.nanoparticles increased the adjuvant effect.

Nanoparticles for peroral administration of proteins & peptides:-Nanoparticles for peroral administration of proteins & peptides:-as as they are susceptible to proteolytic degradation, hence leades to they are susceptible to proteolytic degradation, hence leades to problems of physicochemical & biostability, short biological half life & problems of physicochemical & biostability, short biological half life & inability to pass most of the biological barriers.They are used as peroral inability to pass most of the biological barriers.They are used as peroral carriers with follo.objectivescarriers with follo.objectivesImprovement of bioavailability of drugs with poor absorption Improvement of bioavailability of drugs with poor absorption characteristicscharacteristicsDelivery of vaccine antigen to gut associated lymphoid tissueDelivery of vaccine antigen to gut associated lymphoid tissueReduction of GI irritation due to drugsReduction of GI irritation due to drugsAssurance of stability of drugs in GI tractAssurance of stability of drugs in GI tract

Intra-arterial delivery of nanoparticlesIntra-arterial delivery of nanoparticles

The advantages of nanoparticles includes their subcellular size, targeted The advantages of nanoparticles includes their subcellular size, targeted surface, good suspensibility,& uniform dispersity for catheter based surface, good suspensibility,& uniform dispersity for catheter based therapy & easy penetration into the arterial wall without causing traumatherapy & easy penetration into the arterial wall without causing trauma

Restenosis can be defined as the process of re-obstruction of an artery Restenosis can be defined as the process of re-obstruction of an artery following interventional procedures such as angioplasty, atherectomy,or following interventional procedures such as angioplasty, atherectomy,or stentingstenting

These systems are promised as carriers for genes in restenosis & other These systems are promised as carriers for genes in restenosis & other gene therapy applicationsgene therapy applications

Nanoparticles are carriers for treatment of restenosis as they localize Nanoparticles are carriers for treatment of restenosis as they localize drugs at the site of artery injury rather than systemic administrationdrugs at the site of artery injury rather than systemic administration

Oral delivery of nanoparticlesOral delivery of nanoparticles

Nanoparticles for ocular deliveryNanoparticles for ocular delivery

The short elimination half life of aq.eye drops can be extended from a The short elimination half life of aq.eye drops can be extended from a very short time(1-3 min) to prolonged time(15-20 min) using very short time(1-3 min) to prolonged time(15-20 min) using biodegradable nanoparticlesbiodegradable nanoparticles

The efficacy of pilocarpine loaded nanoparticles is increased by coating The efficacy of pilocarpine loaded nanoparticles is increased by coating them with bioadhesive & viscous polymersthem with bioadhesive & viscous polymers

Methylcellulose, PVA,HPMC for viscosity enhancing propertiesMethylcellulose, PVA,HPMC for viscosity enhancing properties

Mucin,sodium CMC,hyaluronic acid for bioadhesive propertiesMucin,sodium CMC,hyaluronic acid for bioadhesive properties

Nanoparticles for DNA deliveryNanoparticles for DNA delivery

Nanoparticles have been recently used as a delivery vehicle for the Nanoparticles have been recently used as a delivery vehicle for the transfection of plasmid DNA & to improve their stability in the transfection of plasmid DNA & to improve their stability in the bioenvironmentbioenvironment

Chitosan DNA hybrid colloidal systems either as chitosan DNA complex Chitosan DNA hybrid colloidal systems either as chitosan DNA complex or as a chitosan DNA nanospheres reported better gene expressionor as a chitosan DNA nanospheres reported better gene expression

Biodegradable & biocompatible polymer(DL-lactide-co-glycolide) was Biodegradable & biocompatible polymer(DL-lactide-co-glycolide) was used to encapsulate pDNA in submicron size particles. Nanosize range used to encapsulate pDNA in submicron size particles. Nanosize range pDNA loaded polymer particles with high encapsulation efficiency pDNA loaded polymer particles with high encapsulation efficiency showed sustained release of pDNA over a monthshowed sustained release of pDNA over a month

Nanoparticles for oligonucleotides deliveryNanoparticles for oligonucleotides delivery

Antisense agent bind to DNA or RNA sequences blocking the synthesis Antisense agent bind to DNA or RNA sequences blocking the synthesis of cellular proteins with unparalleledof cellular proteins with unparalleled

Antisense agents are antisense sequenses or nucleotides,ribozymes Antisense agents are antisense sequenses or nucleotides,ribozymes

Antisense sequences hybridize double stranded DNA in the nucleus Antisense sequences hybridize double stranded DNA in the nucleus forming triple helixesforming triple helixes

Riboxymes binds to single targeted mRNARiboxymes binds to single targeted mRNA

ODNs are investigated as HIV infection, Hepatitis B virus infection, ODNs are investigated as HIV infection, Hepatitis B virus infection, Herpes simples virus, cancer, restenosis, Rheumatoid arthritis,& allergic Herpes simples virus, cancer, restenosis, Rheumatoid arthritis,& allergic disordersdisorders

Various therapeutic applications of nanoparticles /nanocapsulesVarious therapeutic applications of nanoparticles /nanocapsules

ApplicationApplication MaterialMaterial PurposePurpose

Cancer therapyCancer therapy Poly(alkylcyanoacrylate)nanoparticlePoly(alkylcyanoacrylate)nanoparticleswith anticancer swith anticancer agents,oligonucleotidesagents,oligonucleotides

Targeting,reduced toxicity, Targeting,reduced toxicity, enhanced uptake of enhanced uptake of antitumour agents, antitumour agents, improved in vivo & in vitro improved in vivo & in vitro stabilitystability

Intracellular Intracellular targetingtargeting

Poly(alkylcyanoacrylate)Poly(alkylcyanoacrylate)Polyester nanoparticles with anti Polyester nanoparticles with anti parasitic or antiviral agentsparasitic or antiviral agents

Target RES for intracellular Target RES for intracellular interactionsinteractions

Prolonged systemic Prolonged systemic circulationcirculation

Polyesters with adsorbed Polyesters with adsorbed polyethylene glycols or pluronicspolyethylene glycols or pluronics

Prolong systemic drug Prolong systemic drug effect,avoid uptake by RESeffect,avoid uptake by RES

Vaccine adjuvantVaccine adjuvant Poly Poly (methylmetghacrylate)nanoparticles (methylmetghacrylate)nanoparticles with vaccines(oral & IM injection)with vaccines(oral & IM injection)

Enhances immune Enhances immune response response

ApplicationApplication MaterialMaterial PurposePurpose

Peroral Peroral absorptionabsorption

Poly(methylmetghacrylate)nanopartPoly(methylmetghacrylate)nanoparticles with proteins & therap.agentsicles with proteins & therap.agents

Enhanced bioavailability Enhanced bioavailability &protection from GI enzymes&protection from GI enzymes

Ocular deliveryOcular delivery Poly(alkylcyanoacrylate)with Poly(alkylcyanoacrylate)with steroids,antiinflammatory steroids,antiinflammatory agents,antibacterial agents for agents,antibacterial agents for glaucomaglaucoma

Improved retention of Improved retention of drugs/reduced wash outdrugs/reduced wash out

DNA deliveryDNA delivery DNA gelatin nanoparticles, DNA DNA gelatin nanoparticles, DNA chitosan nanoparticles,chitosan nanoparticles,

Enhanced delivery & higher Enhanced delivery & higher expression levelsexpression levels

Oligonucleotide Oligonucleotide deliverydelivery

Alginate nanoparticles, Alginate nanoparticles, poly(D,L)lactic acid nanoparticlespoly(D,L)lactic acid nanoparticles

Enhanced delivery of Enhanced delivery of oligonucleotideoligonucleotide

Other Other applicationsapplications

Poly(alkylcyanoacrylate)nanoparticlPoly(alkylcyanoacrylate)nanoparticles with peptideses with peptidesFor transdermal applicationFor transdermal applicationNanooparticles with radioactive or Nanooparticles with radioactive or contrast agentscontrast agentsCopolymerized peptide Copolymerized peptide nanoparticles of n-butyl nanoparticles of n-butyl cyanoacrylate & activated peptides cyanoacrylate & activated peptides

Crosses blood brain barrierCrosses blood brain barrier

Improved absorption & permeationImproved absorption & permeationEnzyme Enzyme immunoassays,Radioimagingimmunoassays,Radioimaging

Oral delivery of peptidesOral delivery of peptides