Application and Safety of Erythrocytes as a Novel Drug Delivery System
Gamaledin I. Harisa,
Mohamed F. Ibrahim,
Fars K. Alanazi
Ibrahim A. Alsarra
Carrier erythrocytes are one of biological drug delivery systems have been investigated in recent years. They are biocompatible, biodegradable as well as they have long life span. This review deals with criterion, requirement and methods of drug loading as well as characterizations, safety and application of erythrocyte as drug delivery system. The drugs with high toxicity, such as cardiotoxic and neurotoxic are selected to be encapsulated in the erythrocytes because of their potential for delivery to the liver, spleen and lung. Physical methods like endocytosis and osmotic based methods are used for drug loading into erythrocytes, as well chemical methods are used for the same purpose. After loading of therapeutic agent on erythrocytes, the carrier cells are exposed to physical, cellular as well as biological characterizations. The resealed erythrocytes were used either for sustained release or targeted release of therapeutic agents. The targeted resealed erythrocytes are structurally modified either by glutaraldehyde, oxidizing agents or by drug itself. The modified carrier erythrocytes have tendency to be phagocytosed, therefore, they removed from the circulation and targeted to the organs of reticuloendothelial system.
Received: January 09, 2011;
Accepted: March 16, 2011;
Published: June 18, 2011
The drug delivery systems currently available enlist carriers that are simple
soluble macromolecules such as monoclonal antibodies, soluble synthetic polymers,
polysaccharides in addition to biodegradable polymers. Moreover they include
complex multi-component structures like microcapsules, microparticles, lipoproteins,
liposomes, ghost cells and cells (Pierige et al.,
The cellular carriers have been a useful device as drug delivery system, these
carriers, including, leukocytes, platelets, hepatocytes, fibroblasts and erythrocytes
(Hamidi and Tajerzadeh, 2003; Rossi
et al., 2005). Erythrocytes have many advantages over the other cellular
carriers in its selectivity to deliver the bioactive agents to any organs. The
destruction of modified erythrocytes occurs in liver, spleen and lymph nodes;
therefore, they are used as carriers to deliver the drugs to Reticulo Endothelial
System (RES) (Gopal et al., 2007). In addition,
the possibility of targeting carrier erythrocytes to non-RES organs has been
exploited. Also these cells are non-immunogenic and biodegradable; they freely
circulate throughout the body and offer ease of preparation. Furthermore, they
have the capacity to carry large amounts of drug Hamidi
and Tajerzadeh (2003). Carrier erythrocytes can be used as circulating drug
reservoirs within the circulation (Jain and Jain, 1997,
The normal erythrocyte is a biconcave, ellipsoidal disc with depressions located
in the center on both sides. The average erythrocyte is 8.6 μm in diameter
and 1.9 μm in thickness and has a mean volume and surface area of 86 and
145 μm2, respectively (Patel, 2009).
The blood volume of a normal adult human male is about 7% of body weight and
about 6.5% in a female. For the blood volume of 5 L in a 70 g man, erythrocytes
make up about 40 to 50% of this volume and there are about 5x1012
erythrocytes per liter of the blood (Magnani et al.,
The red blood cell membrane is dynamic, semi-permeable components of the cell,
associated with energy metabolism in the maintenance of the permeability characteristic
of the cell of various cations (Na+, K+) and anions (Cl¯,
HCO3¯) (Patel, 2009).
Each RBC contains about 280 million hemoglobin molecules. A hemoglobin molecules
consists of a protein called globin, composed of four polypeptide chains; a
ring like non-protein pigment called a heme, is bound to each of the four chains.
At the center of the heme ring combine reversibly with one oxygen molecule,
allowing each hemoglobin molecule to bind four oxygen molecules. RBCs include
water (63%), lipids (0.5), glucose (0.8%), mineral (0.7%), non-hemoglobin protein
(0.9%), methehemoglobin (0.5%) and hemoglobin (33.67%) (Gupta
et al., 2010).
CRITERION FOR USE OF ERYTHROCYTES AS DRUG DELIVERY SYSTEM
The normal physiology of erythrocytes gives opportunity to use them as drug
delivery system. The main function of erythrocytes is the transport of O2
from the lungs to tissues and the CO2 produced in tissues back to
lungs. Thus, erythrocytes are a highly specialized carrier systems in the body
due to the following criterion (Gopal et al., 2007):
||The elastic, biconcave shape enables erythrocytes to squeeze
through narrow capillaries
||Erythrocytes make up about 40 to 50% of blood volume therefore; a large
amount of substance can be encapsulated in erythrocytes
||Mature erythrocytes are simple in structure; they have neither nucleus
nor other organelles, for that reason some of the intracellular space exists
for drug transport
||Life-span of RBCs is 100-120 days in the circulation before removing so
can be used for sustained delivery of therapeutic agents
||Erythrocytes are selectively removed from circulation by the macrophages
in the Reticulo Endothelial System (RES), hence be able to used in targeting
of drugs to RES
||The breakdown products are recycled; hemoglobin is break down into globin
and hem. Globin degraded to amino acids for amino acid pools in the body,
while iron reused in hemoglobin synthesis
DISADVANTAGES OF ERYTHROCYTES AS DRUG DELIVERY SYSTEMS
The modifications that occurred during loading procedure of the drugs into
the erythrocytes accelerate their removal by the RES in vivo (Papadatou
et al., 2009). Also certain encapsulated substances may be leaked
from the loaded erythrocytes (Gupta et al., 2010).
Add to this the storage problem that need for conditioning carrier cells (Hirlekar
et al., 2008). Otherwise contamination is possible due to the origin
of the blood, the equipment used and the loading environment. Special precautions
are required for the collection and handling of the erythrocytes (Hamidi
et al., 2007b; Rossi et al., 2006).
NECESSITIES FOR DRUGS ENCAPSULATION INTO ERYTHROCYTES
The molecules should be polar, hydrophilic, resist degradation within erythrocytes,
lack of physical or chemical interaction with erythrocyte membrane and well
defined pharmacokinetic and pharmacodynamic properties (Gupta
et al., 2010; Patel, 2009). Non-polar and
hydrophobic molecules may be entrapped in erythrocyte in their respective salts.
Molecules which interact with the membrane and cause deleterious effects on
membrane structure are not considered to be appropriate for encapsulation in
erythrocyte. Erythrocytes can entrap a wide variety of biologically active substance
(5000-600,000 Daltons in size) (Hamidi et al., 2007b).
APPLICATIONS OF RESEALED ERYTHROCYTES
Resealed erythrocytes have been proposed as delivery systems for a variety of applications in human and animal's medicine. In vivo application of the drugs loaded erythrocytes are used either for prolonged drug released or for drug targeting to RES or non RES.
Targeting of bioactive agents to RES: Damaged erythrocytes are rapidly
cleared from circulation by phagocytic cells in liver and spleen. Targeting
of the drug decreases its side effects and the dose to be administered as well
as drug utilization (Balamuralidhara et al., 2011).
Modifications of erythrocytes membranes accelerate their targeting to the liver
as well as spleen. The treatment of the carrier erythrocytes with certain substances
gives rise to alterations in the properties of the loaded erythrocytes. These
substances include antibodies, gluteraldehyde, sialic acid and sulfhydryl containing
substances (Gupta et al., 2010).
||Glutaraldehyde: The treatment of loaded erythrocytes
with glutaraldehyde enhances their properties as carrier systems. It has
been observed that the erythrocytes treated in this way are more stable
which increases their osmotic resistance, as well as their resistance to
turbulences. It means that the output of the encapsulated substance from
these erythrocytes into the circulatory flow is reduced (Talwar
and Jain, 1992). Similarly, the treatment with glutaraldehyde increases
the selectivity of the erythrocytes towards the RES and specifically, towards
certain organs such as the liver and the spleen (Millan
et al., 2004b)
||Ascorbate and ferrous ions: The chemical alteration of the erythrocyte
membrane with substances as ascorbate/Fe+2, diamide or band 3-cross-linking
reagents can induce increased the uptake of modified red cells by macrophages
(Millan et al., 2004a)
||Biotin: The surface modification of erythrocytes has also been
addressed using phenylhydrazine and N-hydroxysuccinimide ester of biotin
(NHS-biotin) which increase the macrophage uptake of loaded erythrocytes
both in vitro as in vivo (Mishra and Jain,
2002). Moreover, biotinylation of erythrocytes may also be a way of
preparing immuno-erythrocytes attached to biotinylated antibodies that are
stable in circulation and capable of recognizing antigens (Gupta
et al., 2010)
||Antibody: Coating the loaded erythrocytes by anti-Rh or other types
of antibodies is another method that makes the erythrocytes more recognizable
by RES macrophages. In this technique targeting of erythrocytes is either
spleen or liver. If the antibody used as ligand in from of immunoglobin
G, targeting to spleen is preferred; while if used in form of immunoglobin
M type, the liver targeting is dominant (Erchler et
||Other means of modification: Pre-exposing the carrier erythrocytes
to thermal shock increase the up taking of loaded erythrocytes by RES (Ihler
et al., 1973). Also oxidant compounds like azodicarboxilic acid
bis (dimethylamide) increases the uptake of loaded erythrocytes by RES (Arias
et al., 2010), they are reactive toward the sulfhydryl group-containing
proteins of the cell membrane (Ihler et al.,
1973). The enzyme neuraminidase as well as the proteolytic enzymes,
also has been exploited to improve RES targeting of carrier erythrocytes
with some degree of success (Millan et al., 2004b)
Targeting to sites other than RES-rich Organs: Erythrocytes loaded with
drugs have the ability to deliver a drug or enzyme to the macrophage-rich organs,
Also, such cells have been used to target organs outside the RES. Co-encapsulation
of paramagnetic particles, photosensitive agents in erythrocytes along with
the drug to be targeted; application of ultrasound waves as well as site-specific
antibody attachment to erythrocyte membrane (Hamidi et
al., 2007b). The magnetic erythrocytes, resulting from the co-encapsulation
of the drugs with some ferrous fluids such as cobalt-ferrite and magnetite,
have been reported to direct the encapsulated drug predominantly to the desired
sites of the body by means of an external magnetic field. The magnetically guided
erythrocytes have been tested successfully for targeting anti-inflammatory drugs
to inflamed tissues (Markov et al., 2010; Ross,
2009). Photosensitized erythrocytes have been studied as a photo-triggered
carrier and delivery system for methotrexate in cancer treatment. Moreover,
carrier erythrocytes fused to the thermo-responsive liposomes and their localization
using the external thermal source (Hamidi et al.,
Carrier erythrocytes as slow drug release system: Slow release dosage
forms are designed to obtain a prolonged therapeutic effect by continuously
releasing medication over an extended period of time after administration of
single dose (Hossain et al., 2004).
Carrier erythrocytes have long life span in the circulation, so that they can
be used as circulating depots for antitumor, antiparasitics, antibiotics as
well as cardiovascular drugs. This happened only when the drug and the selected
method for the drug loading dont change the morphological and physiological
parameters of erythrocytes (Gupta et al., 2010).
Various bioactive agents encapsulated in erythrocytes are developed for the
sustained release in circulation to allow effective treatment of diseases. Resealed
erythrocytes serve as an ideal carrier for antineoplastic agents, antimicrobial
drugs, vitamins and steroids (Gupta et al., 2010).
Erythrocytes as circulating bioreactors: Erythrocytes have been realized
as carriers for enzymes to serve as circulating bioreactors. Sometimes it is
desirable to decrease the level of circulating metabolites that can enter erythrocytes.
Erythrocytes have also been used as circulating bioreactors for the controlled
delivery of antiviral drugs (Magnani and DeLoach, 1992).
METHODS OF DRUG LOADING INTO ERYTHROCYTES
Erythrocytes can be isolated from blood using a suitable anti coagulant (Hamidi
et al., 2007b). Different sources such as human (Harisa
et al., 2011), rats (Mishra and Jain, 2002),mice
(Kravtzoff et al., 1990; Wang
et al., 2010), rabbits (Hamidi et al.,
2001a), dogs (Tonetti et al., 1991) are used
as source for erythrocytes.
|| Schematic illustration for methods of drug loading into erythrocytes
Freshly collected blood is centrifuged in a refrigerated centrifuge in order
to separate packed erythrocytes.
Then packed erythrocytes are washed with isotonic solution several times and
centrifugation between washes to remove other blood components. The hematocrit
adjusted between 5 and 95%, although the most usual is to work with a hematocrit
of 70% (Rossi et al., 2006). The following methods
are used for entrapment of the therapeutic agent into erythrocytes (Fig.
Osmosis-based methods: Erythrocytes have the ability to undergo reversible
swelling and shape changes in a hypotonic solution or under stress. Erythrocytes
can increase in volume by 25-50% leading to an initial change in the shape from
biconcave to spherical adapt additional volume while keeping the surface area
constant (Agnihotri et al., 2010; Gupta
et al., 2010). This change is due to the absence of superfluous membrane.
Therefore, the cells can maintain their integrity up to a tonicity of 150 mosm
kg-1, above which the membrane ruptures, releasing the cellular contents.
At this point (just before cell lysis), some transient pores of 200-500 Å
are generated on the membrane. Erythrocyte ghost is the remnant after cell lysis
and depletion of cellular contents which can be resealed by restoring isotonic
conditions having the drug inside. Upon incubation, the cells resume their original
biconcave shape and recover original impermeability (Briones
et al., 2009; Gothoskar, 2004).
Hypotonic preswelling: In this technique erythrocytes are incubated
in a hypotonic buffered solution to produce swelling and centrifuged at low
centrifugation values. The supernatant is discarded and the cell fraction is
brought to the lysis point by adding 100-120 μL portions of an aqueous
solution of the drug to be encapsulated and centrifugation between the drug
addition steps. The tonicity of a cell mixture is restored at the lysis point
by adding a calculated amount of hypertonic buffer. Then, the cell suspension
is incubated at 37°C to re-anneal the resealed erythrocytes (Gopal
et al., 2007).
Hypotonic dialysis: The suspension of erythrocytes with hematocrit 50-90%
is placed in a dialysis bag facing a hypoosmotic buffer at 4°C. The time
of dialysis may vary between 20 and 180 min. Subsequently, an annealing process
is performed with the loaded erythrocytes in an isoosmotic medium for 10 min
at 37°C. Finally, a resealing of the erythrocytes is performed at 37°C
using a hyperosmotic buffer. The hyperosmotic buffer usually contains adenosine,
glucose and magnesium chloride (Millan et al., 2004a).
||Comparison between percent of drug loading, advantages as
well as disadvantages of different osmosis based systems (Gopal
et al., 2007)
|| Scheme represents the details of hypotonic dialysis method
It is based on the principle that semi permeable dialysis membrane maximizes
the intracellular/ extracellular volume ratio for macromolecules during lysis
and resealing. In this method, the erythrocyte suspension and the drug to be
loaded were placed in the blood compartment and the hypotonic buffer was placed
in a receptor compartment. This led to the concept of continuous flow dialysis
(Gopal et al., 2007).
Isotonic osmotic lysis: Isotonic hemolysis can be achieved by physical
or chemical means. If erythrocytes are incubated in solutions of a substance
with high membrane permeability, the solute will diffuse into the cells because
of the concentration gradient. This process is followed by an influx of water
to maintain osmotic equilibrium. Chemicals such as urea solution, polyethylene
glycol and ammonium chloride have been used for isotonic hemolysis. However,
this method also is not immune to changes in membrane structure composition.
The suspension was diluted with an isotonic-buffered drug solution. After the
cells were separated, they were resealed at 37°C (Jaitely
et al., 1996). Figure 2 represents the details
of hypotonic dialysis method (Millan et al., 2004b).
Hypotonic dilution: Hypotonic dilution was the first method investigated
for the encapsulation of chemicals into erythrocytes and is the simplest and
fastest. In this method, a volume of packed erythrocytes is diluted with 2-20
volumes of aqueous solution of a drug. The solution tonicity is then restored
by adding a hypertonic buffer. The resultant mixture is then centrifuged, the
supernatant is discarded and the pellet is washed with isotonic buffer solution
(Tajerzadeh and Hamidi, 2000). Comparison between percent
of drug loading, advantages as well as disadvantages of different osmosis based
systems is shown in Table 1.
Chemical perturbation of the membrane: The membrane permeability of
erythrocytes is increased when the cells are exposed to certain chemicals like
polyene antibiotic such as amphotericin B, halothane also was used for the same
purpose (Lin et al., 1999). This induce irreversible
destructive changes in the cell membrane (Gupta et al.,
Electroporation: This method is based on using transient electrolysis
leading to generate pores that produce desirable membrane permeability for drug
loading into red blood cells (Gopal et al., 2007).
The components can be entrapped when an electric pulse of greater than a threshold
voltage of 1-10 kV cm-1 is applied for 20-160 μsec in media
and resealed in osmotic medium.
The extent of pore formation depends upon the electric field strength, pulse
duration and ionic strength of the suspending medium. Once the membrane is perforated,
regardless of the size of the pores, ions rapidly distribute between the extra
and intracellular space to attain equilibrium, however the membrane still remain
impermeable to its cytoplasmic macromolecules (Hamidi et
al., 2007b; Patel, 2009).
Entrapment by endocytosis: Endocytosis performed by addition of one
volume of washed erythrocytes to nine volumes of buffer containing 2.5 mM Adenine
Triphosphate (ATP), 2.5 mM MgCl2 and 1 mM CaCl2, followed
by incubation for 2 min at room temperature. The pores created by this method
are resealed by using 154 mM of NaCl and incubation at 37°C for 2 min. The
entrapment of material occurs by endocytosis. The vesicle membrane separates
endocytosed material from cytoplasm thus protecting it from the erythrocytes
and vice-versa (Harisa et al., 2011). The various
candidates entrapped by this method include primaquine (Alanazi,
2010; Alanazi et al., 2011) and related 8-amino-quinolines,
vinblastine, chlorpromazine and related phenothiazines, hydrocortisone, propranolol,
tetracaine (Alvarez et al., 1998; Hamidi
et al., 2007b) and pravastatin (Abdel-Hamid et
CHARACTERIZATION OF LOADED ERYTHROCYTES
After loading of therapeutic agent on erythrocytes, the carrier cells are exposed
to physical, cellular as well as biological evaluations (Table
2) (Patel, 2009).
Cell counting and cell recovery: This involves counting the number of
red blood cells per unit volume of whole blood, usually by using automated machine.
Red cell recovery may be calculated on the basis of the differences in the hematocrit
and the volume of the suspension of erythrocytes before and after loading. The
goal is to minimize the loss during the encapsulation procedure to maximize
cell recovery (Millan et al., 2004a).
Morphological aspect: The morphological characterization of erythrocytes
is undertaken by comparison with untreated erythrocytes using either transmission
(TEM) or Scanning Electron Microscopy (SEM) (Pierige et
al., 2008). These techniques are done to detect morphological changes
in the erythrocytes induced by encapsulation methods. Thus, when erythrocytes
are subjected to isotonic solutions (300 mosm kg-1) they reveal the
typical morphology of discocyte (biconcave). This evolves to a morphology of
stomatocyte (uniconcave) when they are subjected to solutions of 200 mos M kg-1,
attaining the spherocytic shape (the most fragile of the three) when the solution
is of 150 mosm kg-1 (Hamidi et al., 2007a;
Magnani and DeLoach, 1992).
Osmotic behavior: This test is done to detect the effect of loading
process on the fragility of red blood cells to check the status of erythrocytes
membrane. Unloaded and loaded erythrocytes are tested by exposure to different
concentration of sodium chloride, making them swell, in order to determine the
relative fragility of the red cells Turbulence shock (Bektas
and Ayik, 2009; Abdelhalim and Moussa, 2010).
|| Summary of characterization parameters and their determination
for carrier erythrocytes
This test is done to evaluate the stability of the loaded erythrocytes against
the turbulence stress exerted by the cells against in vivo circulation
turbulence (Millan et al., 2004a). Packed erythrocytes
are suspended in 10 mL of PBS in polypropylene test tubes and are shaken vigorously
using a multiple test tubes orbital shaker at 2000 rpm for 4 h. To determine
the time course of hemoglobin release, 0.5 mL portions of each suspension were
withdrawn at 0, 0.5, 1, 2 and 4 h elapsed and after centrifuging at 1000xg for
10 min. The absorbances of the supernatants are determined spectrophotometrically
at 540 nm. The percent of hemoglobin release is determined in reference to a
completely lysed cell suspension with the same cell fraction (i.e., 0.5 mL packed
cells added to 10 mL of distilled water). To compare the turbulence fragilities
of the different types of erythrocytes, a turbulence fragility index is defined
as the shaking time producing 20% hemoglobin release from erythrocytes (Hamidi
et al., 2007a):
In vitro drug release: The drug loading may produce sustained
release of the drug that influences the pharmacokinetic behavior in vivo of
the loaded erythrocytes. In vitro leakage of the drug from loaded erythrocytes
is tested using autologus plasma or an isoosmotic buffer at 37°C with a
hematocrit adjusted between 0.5 and 50%. The supernatant is removed at the time
intervals previously programmed and replaced by an equal volume of autologous
plasma or buffer (Magnani and DeLoach, 1992).
Some authors recommend performing in vitro the release studies from loaded
erythrocytes using a dialysis bag (Millan et al.,
2004a). The drug release is controlled by molecular weight and liposolubility
of the drug (Hamidi et al., 2007b). Lipophilic
drugs may be released from the red cells by a mechanism of passive diffusion,
while hydrophilic drugs need cell lysis to be released (Hamidi
et al., 2001b).
Hemoglobin release: The content of hemoglobin of the erythrocytes may
be diminished by the alterations in the permeability of the membrane of the
red cells during the encapsulation procedure (Hamidi et
al., 2001b). Furthermore, the relationship between the rate of hemoglobin
and the rate of drug release contributes to interpreting the mechanisms involved
in the release of the substance encapsulated from the erythrocytes (Hamidi
et al., 2001b; Pierige et al., 2008).
The hemoglobin leakage is tested using a red cell suspension by recording the
absorbance of supernatant at 540 nm on a spectrophotometer (Millan
et al., 2004a).
Biological characterization: Biological characterization of the developed
erythrocytes includes sterility test, pyrogenicity test and toxicity tests (Gopal
et al., 2007).
IN VITRO STORAGE OF CARRIER ERYTHROCYTES
Preparing drug-loaded erythrocytes on a large scale and maintaining their survival
and drug content can be achieved by using suitable storage methods. The most
common storage media include Hanks balanced salt solution and acid-citrate-dextrose
at 4°C. Cells remain viable in terms of their physiologic and carrier characteristics
for at least 2 weeks at this temperature. The addition of calcium-chelating
agents or the purine nucleosides improve circulation survival time of cells
upon reinjection (Gopal et al., 2007; Millan
et al., 2004a).
SAFETY CONSIDERATION IN CARRIER ERYTHROCYTES
The safety of utilization of erythrocytes as carrier has been illustrated in
our previous report (Adams et al., 2003) and
summery of this consideration shows in Table 3. The use of
erythrocytes as a drug carrier in human has the inherited problems of transfusion
of blood from one to another. If two different blood types are mixed together,
the blood cells may begin to clump together in the blood vessels, causing a
potentially fatal situation. Therefore, it is important to identify the blood
type of the acceptor and the type of erythrocyte carrier to minimize mismatching
before the administration of drug-loaded erythrocytes takes place. Another inherited
problem is the risk of transmitting diseases. Therefore, screening of these
carriers for the absence of diseases is important to eliminate any risk of contamination.
Utilization of erythrocyte as a drug carrier raises another potential concern
due to the changes in their biochemical nature. In some instances such changes
created therapeutic benefits whereas in other cases they yielded unwanted results.
For example, Hamidi et al. (2001b) conducted
a study on erythrocytes loaded with enalaprilat (Hamidi
et al., 2001b). The process produced erythrocytes that were more
rigid, less deformed and more therapeutically efficacious than unloaded erythrocytes.
The modification of erythrocytes with proteins such as streptavidin, however,
elicited some negative results. The attachment of streptavidin to biotinylated
red blood cells caused these cells to be lysed, rapidly cleared from the circulation
thereby reducing their biocompatibility (Muzykantov et
al., 1996). In vivo studies involving humans and animals have
also been conducted on biotinylated red blood cells.
|| Safety consideration in carrier erythrocytes
Extensive biotinylation severely altered the biocompatibility of these cells
causing rapid elimination whereas moderate biotinylation generated stable erythrocytes
that circulated for several hours (Muzykantov et al.,
Encapsulation of drug in erythrocytes alters their pharmacokinetics properties
and changes their metabolic pathway. Thus, in some cases using erythrocytes
as drug carriers resulted in undesirable cytotoxicity. Doxorubicin encapsulated
in erythrocytes treated with glutaraldehyde was more toxic than the parent compound
(Kohane et al., 2002). The drug-encapsulated
erythrocytes may also increase the production of unfavorable metabolites. For
example, doxorubicinol, a toxic metabolite of doxorubicin, was produced in higher
quantity when doxorubicin-encapsulated erythrocytes were administrated.
Carrier erythrocytes are one of biological drug delivery systems have been investigated in recent decades that covered a wide variety of drugs and other bioactive agents. This is generally due to their notable degree of biodegradability, biocompatibility, availability and ease of preparation and use. The controlled and/or targeted release of active agents is among the mostly attractive applications of erythrocyte carriers in drug delivery. In this review, different methods of loading, characterization, applications as well as usage safety have been summarized.
The author gratefully acknowledge the generous financial support from the Deanship of Scientific Research grant No. NPAR3-(2).
Abdelhalim, M.A.K. and S.A. Moussa, 2010.
Biochemical changes of hemoglobin and osmotic fragility of red blood cells in high fat diet rabbits. Pak. J. Biol. Sci., 13: 73-77.CrossRef |
Abdel-Hamid, M.M., M.F. Ibrahim, G.I. Harisa and F.K. Alanazi, 2011.
Ultra performance liquid chromatography as a new validated method for determination of pravastatin sodium in erythrocytes. Asian J. Chem., Vol. 23, (In Press).
Adams, T., F. Alanazi and D.R. Lu, 2003.
Safety and utilization of blood components as therapeutic delivery systems. Curr. Pharm. Biotech., 4: 275-282.CrossRef |
Agnihotri, J., V. Gajbhiye and N.K. Jain, 2010.
Engineered cellular carrier nanoerythrosomes as potential targeting vectors for anti-malarial drug. Asian J. Pharm., 4: 116-120.CrossRef |
Alanazi, F., 2010.
Pravastatin provides antioxidant activity and protection of erythrocytes loaded primaquine. Int. J. Med. Sci., 7: 358-365.PubMed | Direct Link |
Alanazi, F.K., G.I. Harisa, A. Maqboul, M. Abdel-Hamid, S.H. Neau and I.A. Alsarra, 2011.
Bochemically altered human erythrocytes as a carrier for targeted delivery of primaquine: An in vitro
study. Arch. Pharm. Res. (In Press).
Alvarez, F.J., J.A. Jordan, P. Calleja, L.A. Lotero, G. Olmos, J.C. Diez and M.C. Tejedor, 1998.
Cross-linking treatment of loaded erythrocytes increases delivery of encapsulated substance to macrophages. Biotechnol. Appl. Biochem., 27: 139-143.PubMed | Direct Link |
Arias, M., J.C. Quijano, V. Haridas, J.U. Gutterman and V.V. Lemeshko, 2010.
Red blood cell permeabilization by hypotonic treatments, saponin, and anticancer avicins. Biochimica et Biophysica Acta (BBA)-Biomembranes, 1798: 1189-1196.PubMed | Direct Link |
Balamuralidhara, V., T.M. Pramodkumar, N. Srujana, M.P. Venkatesh, N.V. Gupta, K.L. Krishna and H.V. Gangadharappa, 2011.
pH sensitive drug delivery systems: A review. Am. J. Drug Discovery Dev., 1: 24-48.CrossRef | Direct Link |
Bektas, S. and O. Ayik, 2009.
Hematological parameters and erythrocyte osmotic fragility in rainbow trout, Oncorhynchus mykiss
, experimentally infected with Pseudomonas putida
. J. Fish. Aquat. Sci., 4: 246-253.CrossRef | Direct Link |
Briones, E., C.I. Colino, C.G. Millan and J.M. Lanao, 2009.
Increasing the selectivity of amikacin in rat peritoneal macrophages using carrier erythrocytes. Eur. J. Pharm. Sci., 38: 320-324.PubMed | Direct Link |
Chikezie, P.C., 2011.
Comparative in vitro
osmotic stability of three human erythrocyte genotypes in the presence of quinine and chloroquine phosphate. Asian J. Biochem., 6: 55-64.CrossRef | Direct Link |
Erchler, H.G., S. Gasic, K. Bauer, A. Korn and S. Bacher, 1986. In vivo
clearance of antibody-sensitized human drug carrier erythrocytes. Clin. Pharmacol. Ther., 40: 300-303.CrossRef |
Gopal, V.S., A.R. Kumar, N.A. Usha, A. Karthik and N. Udupa, 2007.
Effective drug targeting by erythrocytes as carrier systems. Curr. Trends Biotechnol. Pharm., 1: 18-33.
Gothoskar, A.V., 2004.
Resealed erythrocytes: A review. Pharm. Technol., 1: 140-158.Direct Link |
Gupta, A., A.K. Mishra, P. Bansal, S. Kumar, V. Gupta, R. Singh and G.S.X. Kalyan, 2010.
Cell based drug delivery system through resealed erythrocyte: A review. Int. J. Pharm., 2: 23-30.
Millan, C.G., A.Z. Castaneda, M.L.S. Marinero and J.M. Lanao, 2004.
Factors associated with the performance of carrier erythrocytes obtained by hypotonic dialysis. Blood. Cells. Mol. Dis., 33: 132-140.CrossRef | PubMed |
Hossain, M.B., M. Rashid and A.K.M.M. Hossain, 2004.
Effect of different waxy materials on the release of ibuprofen from polyethylene glycol based suppositories. Pak. J. Biol. Sci., 7: 2082-2085.CrossRef | Direct Link |
Hamidi, M. and H. Tajerzadeh, 2003.
Carrier erythrocytes: An overview. Drug. Deliv., 10: 9-20.PubMed | Direct Link |
Hamidi, M., H. Tajerzadeh, A.R. Dehpour and S. Ejtemaee-Mehr, 2001.
Inhibition of serum angiotensin-converting enzyme in rabbits after intravenous administration of enalaprilat-loaded intact erythrocytes. J. Pharm. Pharmacol., 53: 1281-1286.PubMed | Direct Link |
Hamidi, M., H. Tajerzadeh, A. R. Dehpour, M. R. Rouini and S. Ejtemaee-Mehr, 2001. In vitro
characterization of human intact erythrocytes loaded by enalaprilat. Drug. Deliv., 8: 223-230.PubMed | Direct Link |
Hamidi, M., N. Zarei, A.H. Zarrin and S. Mohammadi-Samani, 2007.
Preparation and in vitro
characterization of carrier erythrocytes for vaccine delivery. Int. J. Pharm., 338: 70-78.CrossRef |
Hamidi, M., A. Zarrin, M. Foroozesh and S. Mohammadi-Samani, 2007.
Applications of carrier erythrocytes in delivery of biopharmaceuticals. J. Control Rel., 118: 145-160.PubMed | Direct Link |
Harisa, G., M.F. Ibrahim and F.K. Al-Anazi, 2011.
Characterization of human erythrocytes as potential carrier for pravastatin: An in vitro
study. Int. J. Med. Sci., 8: 222-230.Direct Link |
Hirlekar, R.S., P.D. Patel, N. Dand and V.J. Kadam, 2008.
Drug loaded erythrocytes: As novel drug delivery system. Curr. Pharm. Des., 14: 63-70.CrossRef |
Ihler, G.M., R.H. Glew and F.W. Schnure, 1973.
Enzyme loading of erythrocytes. Proc. Nat. Acad. Sci. USA., 70: 2663-2666.Direct Link |
Jain, S. and N.K. Jain, 1997.
Engineered erythrocytes as a drug delivery system. Ind. J. Pharm. Sci., 59: 275-281.Direct Link |
Jain, S. and N. K. Jain, 1998.
Preparation, characterization and pharmaceutical potential of engineered erythrocytes. Pharmazie, 53: 5-14.PubMed | Direct Link |
Jaitely, V.P., N. Kanaujia, S.V. Jain and S.P. Vyas, 1996.
Resealed erythrocytes: Drug carrier potentials and biomedical applications. Indian Drugs, 33: 589-594.
Kravtzoff, R., C. Ropars, M. Laguerre, J.P. Muh and M. Chassaigne, 1990.
Erythrocytes as carriers for l-asparaginase. Methodological and mouse in-vivo
studies. J. Pharm. Pharmacol., 42: 473-476.PubMed |
Kohane, D.S., N. Plesnila, S.S. Thomas, D. Le, R. Langer and M.A. Moskowitz, 2002.
Lipid-sugar particles for intracranial drug delivery: Safety and biocompatibility. Brain Res., 946: 206-213.CrossRef | PubMed |
Lin, W., D.M. de Freitas, Q. Zhang and K.W. Olsen, 1999.
Nuclear magnetic resonance and oxygen affinity study of cesium binding in human erythrocytes. Arch. Biochem. Biophys., 369: 78-88.CrossRef | PubMed |
Magnani, M. and J.R. DeLoach, 1992.
The Use of Resealed Erythrocytes as Carriers and Bioreactors, Advances in Experimental Medicine and Biology. Vol. 326, Plenum Press, New York, USA., pp: 221-225
Magnani, M., L. Rossi, A. Fraternale, M. Bianchi, A. Antonelli, R. Crinelli and L. Chiarantini, 2002.
Erythrocyte-mediated delivery of drugs, peptides and modified oligonucleotides. Gene. Ther., 9: 749-751.CrossRef |
Markov, D.E., H. Boeve, B. Gleich, J. Borgert, A. Antonelli, C. Sfara and M. Magnani, 2010.
Human erythrocytes as nanoparticle carriers for magnetic particle imaging. Phys. Med. Biol., 55: 6461-6461.CrossRef |
Millan, C.G., M.L.S. Marinero, A.Z. Castaneda and J.M. Lanao, 2004.
Drug, enzyme and peptide delivery using erythrocytes as carriers. J. Control Rel., 95: 27-49.PubMed | Direct Link |
Mishra, P.R. and N.K. Jain, 2002.
Biotinylated methotrexate loaded erythrocytes for enhanced liver uptake. A study on the rat. Int. J. Pharm., 231: 145-153.PubMed | Direct Link |
Muzykantov, V.R., J.C. Murciano, R.P. Taylor, E.N. Atochina and A. Herraez, 1996.
Regulation of the complement-mediated elimination of red blood cells modified withbiotin and streptavidin. Anal. Biochem., 241: 109-119.PubMed | Direct Link |
Papadatou, B., L. Rossi, F. Bracci, D. Knafelz and C. Noto et al
P101 long-term treatment with autologous red blood cells loaded with dexamethasone 21-phosphate in pediatric patients affected by steroid-dependent crohn disease and ulcerative colitis. J. Crohn's Colitis Supplements, 3: 22-22.
Patel, R.P., 2009.
An overview of resealed erythrocyte drug delivery. J. Pharm. Res., 2: 1008-1012.
Pierige, F., S. Serafini, L. Rossi and M. Magnani, 2008.
Cell-based drug delivery. Adv. Drug. Delivery Rev., 60: 286-295.CrossRef | Direct Link |
Ross, R.W., A.L. Zietman, W. Xie, J.J. Coen and D.M. Dahl et al
Lymphotropic nanoparticle-enhanced magnetic resonance imaging (lnmri
) identifies occult lymph node metastases in prostate cancer patients prior to salvage radiation therapy. Clin. Imaging, 33: 301-305.CrossRef |
Rossi, L., S. Serafini, F. Pierige, A. Antonelli and A. Cerasi et al
Erythrocyte-based drug delivery. Expert. Opin. Drug. Deliv., 2: 311-322.CrossRef |
Rossi, L., S. Serafini, F. Pierige, M. Castro and M.I. Ambrosini et al
Erythrocytes as a controlled drug delivery system: Clinical evidences. J. Control Rel., 116: e43-e45.PubMed | Direct Link |
Tajerzadeh, H. and M. Hamidi, 2000.
Evaluation of hypotonic preswelling method for encapsulation of enalaprilat in intact human erythrocytes. Drug. Dev. Ind. Pharm, 26: 1247-1257.PubMed | Direct Link |
Talwar, N. and N.K. Jain, 1992.
Erythrocyte based delivery system of primaquine: In vitro
characterization. J. Microencapsul, 9: 357-364.PubMed | Direct Link |
Tonetti, M., C. Polvani, E. Zocchi, L. Guida and U. Benatti et al
Liver targeting of autologous erythrocytes loaded with doxorubicin. Eur. J. Cancer, 27: 947-948.PubMed | Direct Link |
Wang, G.P., Y.S. Guan, X.R. Jin, S.S. Jiang and Z.J. Lu et al
Development of novel 5-fluorouracil carrier erythrocyte with pharmacokinetics and potent antitumor activity in mice bearing malignant ascites. J. Gastroenterol Hepatol, 25: 985-990.CrossRef |