Aquasome: A Novel Drug Delivery

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International Standard Serial Number (ISSN): 2319-8141

International Journal of Universal Pharmacy and Bio Sciences 6(1): January-February 2017
INTERNATIONAL JOURNAL OF UNIVERSAL

PHARMACY AND BIO SCIENCES
Pharmaceutical
Sciences
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AQUASOME: A NOVEL DRUG DELIVERY

S.B.Gondkar1*,A.B.Darekar2,S.D.Yelmame3, R. B. Saudagar4
1
Department of Pharmaceutics, R. G. Sapkal College of Pharmacy, Anjaneri, Nashik422213, Maharashtra, India.
2
Department of Pharmaceutics, R.G. Sapkal College of Pharmacy, Anjaneri, Nashik422213, Maharashtra, India.
3
Department of Pharmaceutics, R.G. Sapkal College of Pharmacy, Anjaneri, Nashik422213, Maharashtra, India.
4
Department of Pharmaceutical Chemistry, R.G. Sapkal College of Pharmacy, Anjaneri,
Nashik-422213, Maharashtra, India.
ABSTRACT
KEYWORDS:
Aquasomes are the nano biopharmaceutical carrier systems containing
Bioactive molecules,
Nanoparticulate carrier
system, Oligomericfilm,
Self-assembly.
For Correspondence:
S.B.Gondkar *
Address:
Department of
Pharmaceutics, R. G.
Sapkal College of
Pharmacy, Anjaneri,
Nashik-422213,
Maharashtra, India.
particle core composed of nano crystalline calcium phosphate or ceramic

diamond, and is covered by a poly hydroxyl oligomeric film. They
recently developed delivery system for bioactive molecules like peptide,
protein, hormones, antigens and genes to specific sites. Aquasomes are
spherical in shape with 60300 nm particles size. These are nano
particulate carrier systems but instead of being simple nano particles
these are three layered self-assembled structures, comprised of a solid
phase nanocrystalline core coated with oligomeric film to which
biochemically active
molecules are adsorbed with or without
modification. These structures are self-assembled by noncovalent and

ionic bonds. The solid core provides the structural stability, while the
carbohydrate coating protects against dehydration and stabilizes the
biochemically active molecules.
Full Text Available On www.ijupbs.com
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INTRODUCTION:
Aquasomes means water bodies are the combination of biotechnology and nanotechnology.
These drug delivery systems were first discovered by Nirkossovsky. Aquasomes are the nano
biopharmaceutical carrier systems containing the particle core composed of nano crystalline
calcium phosphate or ceramic diamond, and are covered by polyhydroxyl oligomeric film. These
three layered structures are self-assembled by non-covalent bonds. The pharmacologically active
molecule can be incorporated by following methods like copolymerization, diffusion or adsorption
to carbohydrate surface of pre-formed nanoparticles
[1].
Aquasomes are spherical 60-300 nm
particles used for drug and antigen delivery. As these are solid or glossy particle dispersed in
aqueous environment, they exhibit the physical properties of colloids and their mechanism of action
is controlled by their surface chemistry. Aquasomes delivers their contents through a combination
of special targeting molecular shielding and slow &sustained release process. Aquasomes
technology represents a platform system for conformation integrity and biochemical stability of
bioactives. Their intended route of administration is parenteral. Some researchers have extended the
route of administration from parenteral to oral. Properties like protection and preservation of fragile
biological molecules, conformational integrity, and surface exposure made it as a successful carrier
system for bioactive molecules like peptide, protein, hormones, antigens and genes to specific sites
[2].
Three types of core materials are mainly used for producing aquasomes: tin oxide,
nanocrystalline carbon ceramics (diamonds) and brushite (calcium phosphate dihydrate).Calcium

phosphate is the core of interest, owing to its natural presence in the body. The brushite 6M is
unstable and converts to hydroxyapatite upon prolonged storage. Hydroxyapatite seems, therefore,
a better core for the preparation of aquasomes. It is widely used for the preparation of implants for
drug delivery
[3].
Aquasomes discovery comprises a principle from microbiology, food chemistry,
biophysics and many discoveries including solid phase synthesis, supra molecular chemistry,
molecular shape change and self-assembly which are incapable of forming hydrogen bond,
however, their tendency to repel water helps to organize the moiety to surrounding environment.
The organized water decreases the overall level of disorder/ entropy of the surrounding medium.
Since, organized water is thermodynamically unfavorable, the molecule loose water/dehydrate and
get self-assembled [4]
Principle of self-assembly
Self-assembly implies that the constituent parts of some final product assume spontaneously
prescribed structural orientations in two or three dimensional space. The self-assembly of
macromolecules in the aqueous environment, either for the purpose of creating smart nanostructure
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materials or in the course of naturally occurring biochemistry, is governed basically by three

physicochemical processes: the interactions of charged groups, dehydration effects and structural
stability[1].
1. Interaction between charged groups
The interaction of charged groups, for example, amino, carboxyl, sulphate, phosphate groups
facilitates long range approach of self-assembly sub units. The group plays a role in stabilizing
tertiary structures of folded proteins [1]
2. Hydrogen bonding and dehydration effect
Hydrogen bond helps in base pair matching and stabilization of secondary protein structure such as
alpha helices and beta sheets. Molecules forming hydrogen bonds are hydrophilic and this confers a
significant degree of organization to surrounding water molecules. In case of hydrophobic
molecules, which are incapable of forming hydrogen bond [2].
3. Structural stability
Molecules that carry less charge than formally charged groups exhibit a dipole moment. The forces
associated with dipoles are known as van der waals forces. Structural stability of protein in
biological environment determined by interaction between charged group and hydrogen bonds
largely external to molecule and by van der waals forces largely internal to molecule. The Vander
Waals forces, most often experienced by hydrophobic molecular regions that are shielded from
water play a subtle but critical role in maintaining molecular shape or conformation during selfassembly. The van der waals forces are largely responsible for hardness or softness of molecules.
The van der waals interaction among hydrophobic side chain promotes stability of compact helical
structures which are thermodynamically unfavorable for expanded random coils [1].
Strategies used in chemical synthesis of nanostructure
Aquasomes are self-assembled three layered nanostructures. Therefore the strategies involved in
chemical synthesis of nanostructure need elaboration.
1. Sequential covalent synthesis
This can be used to generate arrays of co-valently linked atoms generated with well defined
composition, connectivity and shape i.e. vitamin B12. It can generate the structures that are far from
the thermodynamic minimum for that collection of atoms [5].
2. Covalent polymerization
This strategy is used for preparing molecules with high molecular weight. Here a relatively simple
low weight substance is allowed to react with itself to produce molecule comprising many
covalently linked monomers. For example: Formation of Polyethylene from ethylene. The
molecular weight of polyethylene can be high (>106 Daltons), and it is easily prepared, but the
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molecular structure is simple and repetitive and the process by which it is formed offers only
limited opportunity for controlled variation in the structure or for control of its three dimensional
shape. Polymerization indirectly provides synthetic routes to stable nano structures e.g. phase
separated polymers [6].
3. Self organizing synthesis
This strategy abandons the covalent bond as required connection between atoms and relies instead
on weaker and less directional bonds such as ionic, hydrogen and van der waals interactions to
organize atoms, ions or molecules into structures. The different type of structures prepared by this
strategy includes molecular crystals, ligand crystals, colloids, micelles, emulsions, phase separated
polymers and self-assembled monolayer. Self-organization is the peculiar feature of these methods.
The molecules or ions adjust their own position to reach thermodynamic minimum. By selforganization, true nano structures can be prepared [7].
4. Molecular self-assembly
It is the spontaneous assembly of molecules into structured, stable, non-covalently joined
aggregates. Molecular self-assembly combines features of each preceding strategies to make large
structurally well-defined assemblies of atoms. Formation of well-defined molecules of intermediate
structural complexity through sequential covalent synthesis. Formation of large, stable structurally
defined aggregates of these molecules through ionic, hydrogen and van der waals interactions or
other non-covalent links. Use of multiple copies of one or several of the constituent molecules or of
a polymer, to simplify the synthetic task. The key to this type of synthesis is to understand and
overcome intrinsically unfavorable entropy together in a single aggregate. For final assembly to be
stable and to have well defined shape, the non-covalent connection between molecules must
bestable. The strength of the individual Vander waals interactions and hydrogen bonds are weak
(0.1 to 5 Kcal/mole) relative to typical covalent bonds (40 to 100 Kcal/mole) and comparable to
thermal energies. Thus to achieve acceptable stability, molecules in self assembled aggregates must
be joined by many of these weak noncovalent interaction or by multiple hydrogen bonds or both[1].
Objective behind development of aquasomes:
The main objective of preparing aquasomes is to protect bio-actives. Various other carrier system
are there like prodrugs and liposomes but they have disadvantage that they are prone to and carrier,
so in that condition in aquasomes can be termed as a important carrier. In aquasomes carbohydrate
coating prevents destructive denaturing interaction between drug and solid carriers. Aquasomes
maintains molecular confirmation and optimum pharmacological activity. An active molecule
possess qualities like unique three-dimensional conformation, a freedom of internal molecular
rearrangement which is induced by molecular interactions, freedom of bulk movement but protein
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undergo irreversible denaturation when desiccated, even unstable in aqueous state. In the aqueous
state pH, temperature, solvents, salts cause denaturation thus bio-activity face many biophysical
constrain and hurdles. In such case, aquasomes with natural stabilizers like various polyhydroxy
sugars act as dehydro protectant aiding in maintaining water like state thereby preserves molecules
in dry solid state[7].
Method of Preparation of Aquasome
The general procedure consists of an inorganic core formation, which will be coated with Lactose
forming the polyhydroxylated core that finally will be loaded by model drug .By using the principle
of self-assembly, the aquasomes are prepared in three steps i.e., preparation of core, coating of core,
and immobilization of drug molecule[9][2].
Fig. 1: Method of Preparation of Aquasomes.

1. Preparation of the core:
The first step of aquasome preparation is the fabrication of the ceramic core. The process of
ceramic core preparation depends on the selection of the materials for core. These ceramic cores
can be fabricated by colloidal precipitation and sonication, inverted rnagnetron sputtering, plasma
condensation and other processes. For the core, ceramic materials were widely used because
ceramics are structurally the most regular materials known. Being crystalline, the high degree of
order in ceramics ensures that any surface modification will have only a limited effect on the nature
of the atoms below the surface layer and thus the bulk properties of the ceramic will be preserved.
The high degree of order also ensures that the surfaces will exhibit high level of surface energy that
will favor the binding of polyhydroxyoligomeric surface film. Two ceramic cores that are most
often used are diamond and calcium phosphate [9][11].
2. Carbohydrate coatings:
The second step involves coating by carbohydrate on the surface of ceramic cores. There are
number of processes to enable the carbohydrate (polyhy-droxy oligomers) coating to adsorb
epitaxially on to the surface of the nano-crystalline ceramic cores. The processes generally entail
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the addition of polyhydroxy oligomer to a dispersion of meticulously cleaned ceramics in ultra pure
water, sonication and then lyophilization to promote the largely irreversible adsorption of
carbohydrate on to the ceramic surfaces. Excess and readily desorbing carbohydrate is removed by
stir cell ultra-filtration. The commonly used coating materials are cellobiose, citrate, pyridoxal-5phosphate, sucrose and trehalose[11].
3. Immobilization of drugs:
The surface modified nano-crystalline cores provide the solid phase for the subsequent non
denaturing self-assembly for broad range of biochemically active molecules. The drug can be
loaded by partial adsorption[12].
Properties:
Aquasomes due to their size and structure stability avoid clearance by reticulo endothelial system or
degradation by other environmental challenges. In normal system, calcium phosphate is
biodegradable. Biodegradation in vivo achieved by monocytes and multicellular cells called
osteoclast. Two types of phagocytosis reported, either crystals taken up alone and then dissolved in
cytoplasm afterdisappearance of phagosome membrane or dissolution after formation of hetero
phagosome. Aquasomes possess large size and active surface hence can be efficiently loaded with
substantial amounts of agents through ionic, non co-valent bonds, van der waals forces and entropic
forces. As solid particles dispersed in aqueous environment, exhibit physical properties of colloids
[9][11]
Characterization of ceramic core:

1. Size distribution
For morphological characterization and size distribution analysis, scanning electron microscopy
(SEM) and transmission electron microscopy (TEM) are generally used. Core, coated core, as well
as drug-loaded aquasomes are analyzed by these techniques. Mean particle size and zeta potential
of the particles can also be determined by using photon correlation spectroscopy[11][24].
2. Structural analysis
FT-IR spectroscopy can be used for structural analysis. Using the potassium bromide sample disk
method, the core as well as the coated core can be analyzed by recording their IR spectra in the
wave number range 4000400 cm1; the characteristic peaks observed are then matched with
reference peaks. Identification of sugar and drug loaded over the ceramic core can also be
confirmed by FT-IR analysis of the sample [1][14].
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3. Crystallinity
The prepared ceramic core can be analyzed for its crystalline or amorphous behavior using X-ray
diffraction. In this technique, the X-ray diffraction pattern of the sample is compared with the
standard diffractogram, based on which the interpretations are made[1][18].
Characterization of coated core:
1. Carbohydrate coating:
Coating of sugar over the ceramic core can be confirmed by concanavalin Ainduced aggregation
method (determines the amount of sugar coated over core) or by anthrone method(determines the
residual sugar unbound or residual sugar remaining after coating). Furthermore, the adsorption of
sugar over the core can also be confirmed by measurement of zeta potential[ 1][14][15].
2. Glass transition temperature:
DSC can be used to analyze the effect of carbohydrate on the drug loaded to aquasomes. DSC
studies have been extensively used to study glass transition temperature of carbohydrates and
proteins. The transition from glass to rubber state can be measured using a DSC analyzer as a
change in temperature upon melting of glass[14][15].
Characterization of drug-loaded aquasomes:
1. Drug payload:
The drug loading can be determined by incubating the basic aquasome formulation (i.e., without
drug) in a known concentration of the drug solution for 24 hours at 4C. The supernatant is then
separated by high-speed centrifugation for 1hour at low temperature in a refrigerated centrifuge.
The drug remaining in the supernatant liquid after loading can be estimated by any suitable method
of analysis [21][22].
2. In vitro drug release studies:
The in vitro release kinetics of the loaded drug is determined to study the release pattern of drug
from the aquasomes by incubating a known quantity of drug-loaded aquasomes in a buffer of
suitable pH at 37C with continuous stirring. Samples are withdrawn periodically and centrifuged at
high speed for certain lengths of time. Equal volumes of medium must be replaced after each
withdrawal. The supernatants are then analyzed for the amount of drug released by any suitable
method [1][24].
3. In-process stability studies:
SDS-PAGE (sodium dodecyl sulphate polyacrylamide gel electrophoresis) can be performed to
determine the stability and integrity of protein during the formulation of the aquasomes [14][15][21][22].
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APPLICATIONS
Oxygen carrier
The deliver hemoglobin using colloidal ceramic carbohydrate composites termed aquasomes. This
study demonstrates that the hemoglobin adsorbed hemoglobin adsorbed aquasomes can carry the
oxygen satisfactorily, and it also establishes the superiority of hemoglobin aquasomal formulation
over the other methods acting as artificial blood substitute[24][14][23]
For immunotherapy
Aquasomes for delivery of model allergen without altering the antigenic and immunogenic
properties of the protein/allergen. His report demonstrates that OVA adsorbed aquasomes are able
to induce a strong T cell specific proliferative response with a cytokine profile suggestive of a Th1
response, prevention of anaphylactic reactions and maintenance of low titers of IgE, without
abrogation of Th2-mediated responses. This suggests that aquasomes could have possible
implications in the future of peptide-based vaccines against allergic disorders [19].
For oral route
Technological innovation for the delivery aquasomes via the peroral route. Piroxicam loaded
aquasomes with their nanometric dimensions, low drug dose, and water like properties were
prepared by using two techniques; namely, coprecipitation by refluxing and coprecipitation by
sonication [1].
For immunopotentiation
Aquasomes that develops immune responses to recombinant or synthetic epitopes which is of

considerable importance in vaccine research for immune potentiation. Bovine serum albuminimmobilized aquasomes were around 200 nm in diameter and spherical in shape and had
approximately 2030% BSA-loading efficiency. The formulated aquasomes was compared with
plain bovine serum albumin (BSA) and transport of immunogen to APCs was found to be a
promising target for gene therapy. Thus, the enhanced transport of conformationally stable antigen
leads to better presentation to APCs. APCs contain both MHC-I and MHC-II molecules leading to
processing and presentation of antigen via both endocytic and cytosolic pathways leading to elicit
both humoral and cellular responses. Aquasome formulations could elicit combined T-helper 1
(Th1) and Th2immune response and proved to be slightly better carrier system compared with other
ceramic-based antigen nanocarriers.
Anti thrombic activity
Formulated nanoparticles based on heparin-poly (isobutylcyanoacrylate) copolymers to carry

hemoglobin. His work constitutes the demonstration of hemoglobin loaded on nanoparticle surface,
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rather than being encapsulated. Antithrombic activity of native nanoparticles was evaluated by antiXa factor activity assay using a coagulometer ST1. Binding of nanoparticles to von. Willebrand
factor (vWF) was measured. Results have shown that the heparin on the nano particles surface
preserved most its antithrombic activity and its capacity to recognize thev WF. The bound
hemoglobin also maintained its capacity to bind ligands. One ml of nano particles (with a size of
100 nm)suspension can be loaded with up to 2.1 mg of hemoglobin, which preserves its ligand
binding capacity as well as make suitable tools in the treatment of thrombosis oxygen deprived
pathologies. These nanoparticles maintain the heparin antithrombic properties and inhibit
complement activation [1].
Antigen delivery
For delivery of gene Aquasomes can be studied for the delivery of genes. It illustrates the attractive
delivery system loaded with genetic material. Studies reveal that aquasomes protect and maintain
structural integrity of the gene segment. A five layered composition comprised of the ceramic
nanocrystalline core, the polyhydroxyloligomeric film coating, the non-covalently bound layer of
therapeutic gene segment, an additional carbohydrate film and a targeting layer of conformationally
conserved viral membrane proteins, have been proposed for gene therapy. The aquasome vehicle
would afford all of the potential advantages of viral vectors and simultaneous overwhelming the
risk of irrelevant gene integration[14][15].
Fig. 2: Gene Delivery through Aquasomes.

CONCLUSION
Aquasomes have given a new hope for the pharmaceutical Scientists to deliver bioactive molecules.
Within the large pool of peptide drugs are a considerable number of candidates with the potential
for delivery via these carriers. Still, considerable further study of aquasomes is necessary with
respect to pharmacokinetics, toxicology, and animal studies to confirm their efficiency as well as
safety, so as to establish their clinical usefulness and to launch them commercially.
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International Standard Serial Number (ISSN): 2319-8141

INTERNATIONAL JOURNAL OF UNIVERSAL

IMPACT FACTOR 2.96***