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Anionic Polysaccharide Hydrogels with Charges Provided by the Polysaccharide

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Drug Delivery Letters, 2012, 2, 000-000 1

Anionic Polysaccharide Hydrogels with Charges Provided by the Polysac-

charide or the Crosslinking Agent

Zied Souguir, Elie About-Jaudet, Luc Picton and Didier Le Cerf*

Université de Rouen, Laboratoire Polymères, Biopolymères, Surfaces, CNRS-UMR 6270 and FR3038, 76821 Mont
Saint Aignan, France

Abstract: Polysaccharides have attractive advantages for using in the field of pharmaceutics. In most cases, they are non
toxic, biocompatible and plentiful. The presence of hydroxyl groups (always) and carboxylic acid groups (sometimes)
permits crosslinking of the chains and obtaining hydrogels. These hydrogels present interesting swelling and mechanical
properties. They have the capacity to entrap active compounds such as drugs and release them through modifications of
the solvation conditions vias pH or ionic strength changes. This review deals with polysaccharide hydrogels obtained by
the formation of covalent bounds between polymeric chains. We have focused our attention on systems with an anionic
character. These charges are due to the presence of acidic functions on the polysaccharide backbone or are brought by the
crosslinking agent. The anionic character dependents on the ionic strength and pH. We discuss the synthesis of these hy-
drogels. After a short presentation of a few polysaccharides, we describe the different crosslinking agents and their action
mode. Applications in drug delivery are discussed along.
Keywords: Anionic polysaccharide, hydrogel, drug delivery, crosslinking agent.

1. INTRODUCTION polysaccharides are neutral (e.g. pullulan, dextran, galacto-

A hydrogel is an insoluble network of polymer chains
that swell in aqueous media. Due its hydrophilicity the
polymer used can absorb large amounts of water (up to 500 g
of water in 1 g of dry hydrogel). This technology has many
application fields such for as food or cosmetic additives,
biomedical implants, drug delivery.
Two general classes of hydrogels can be defined based
on the crosslinking between the polymer chains. First is the
class of physical hydrogels or pseudogels and are non-
permanent. The chains are connected by interactions of low
energy as hydrophobic interactions, hydrogen bonds, electro-
static forces or chain entanglements. The second class com-
prises chemical hydrogels which are permanent. The
crosslinked structure is obtained by covalent bounds between
the chains.
In terms of ionic charge, hydrogels can be neutral, ani-
onic, cationic, or ampholytic according to the presence of
acid or basic functions. Swelling properties are thus greatly
dependent on pH and ionic strength. For a good characteriza-
tion of hydrogels, it is important to know the mechanical
properties (storage (G’) and loss (G’’) moduli). The storage
modulus and the equilibrium swelling vary in reverse order
because they depend in opposite way on the crosslinking
density.
Polysaccharides are widespread in nature. They can be
found in animals, plants, and microorganisms. Natural
*Address correspondence to this author at the Université de Rouen, Labora-
toire Polymères, Biopolymères, Surfaces, CNRS-UMR 6270 and FR3038,
76821 Mont Saint Aignan, France; Tel: +33-235146543;
Fax: +33-235146702; E-mail: didier.lecerf@univ-rouen.fr
mannan, glucuronan) [1-4] or anionic with the presence of
carboxylate (e.g. pectin, alginate, hyaluronan) or sulphate
groups (e.g. carrageenan, heparin) [5-10]. A chemical modi-
fication brings the anionic character on the initially neutral
polysaccharide. The best known and used anionic industri-
ally polysaccharides are, carboxymethylcellulose [11] (e.g.
Blanose
, Kelcol
, Grindsted
) and carboxymethylstarch
[12] (e.g. Explotab®, Primojel
). The modification permits to
solubilize an polysaccharide which is plentiful in nature yet
insoluble. At a smaller scale, for specific applications, poly-
saccharides can be functionalized. As example we can cite is
the carboxymethylation of pullulan or dextran and the sul-
fation of pullulan or glucuronan [13,14]. In most cases, the
resulting polymers are non-toxic, biocompatible and biode-
gradable.
The great hydrophilicity of polysaccharides and their
ability to chemically react justifies their interesting potential
to form hydrogels. With anionic groups, hydrogels become
pH and ionic strength sensitive, which offers important and
new applications in many fields.
A comprehensive search of the literature was conducted
in June 2012 on ISI Web of Knowledge. The following
search terms were employed: hydrogel* and polysaccharide*
as topic. We have identified 1,120 articles and 20,700 cita-
tions in 13,328 different articles during the period 1992-
2011. The (Figs. 1 and 2) show the exponential increase of
articles and citations.
The aim of this paper is to present the different possibili-
ties for obtaining anionic polysaccharide hydrogels that can
be used industrially, in many instances but not exclusively,
for drug delivery. We discuss the main pathway for synthe-

2210-3031/12 $58.00+.00 © 2012 Bentham Science Publishers

2 Drug Delivery Letters, 2012, Vol. 2, No. 4
Fig. (1). Publications on hydrogels (search using “hydrogel*” and “polysaccharide*”, source: Web of Knowledge).
Fig. (2). Citations on hydrogels (search using “hydrogel*” and “polysaccharide*”, source: Web of Knowledge).
sising a polysaccharide network, with chemical crosslinking
and an anionic character. This review is not a report on the
state of the art of these systems. Many excellent review arti-
cles detailing the swelling processes [15, 16] or the use of
hydrogels for biomedical applications [17] as drug delivery
[18], cell encapsulation [19], regenerative medicine [20] and
agriculture [21] have already been published.
In the first part of this paper we present anionic natural
and modified polysaccharides which we will discuss in terms
of their use as hydrogels.
Hydrogels that have an anionic character brought by the
polysaccharide backbone are presented in the second part.
We show the different possibilities for crosslinking (Fig. 3).
The third part is devoted to crosslinkers that provide the
anionic character.
Souguir et al.
Several examples selected from the literature illustrate
the different possibilities and potential applications in drug
delivery.
2. POLYSACCHARIDES
2.1. Anionic Natural Polysaccharides
2.1.1. Sodium Alginate
As a naturally occurring biopolymer [22], alginate (figure
4) has been used successfully in the food and beverage in-
dustry as a gelling agent and colloidal stabilizer, and holds
strong potential in the area of drug delivery. Extracted from
brown algae, alginate polymers consist of linear, unbranched
polysaccharides with acid residues of 1,4’-linked--D-
mannuronic acid (M) and
-L-gluronic acid (G) residues.
Anionic Polysaccharide Hydrogels with Charges Provided by the Polysaccharide
Fig. (3). Schematic formation of anionic polysaccharide hydrogels.
The residues are arranged in blocks along the chain and vary
in sequence and composition.
2.1.2. Xanthan Gum
Xanthan gum [23] is a high molecular weight extra cellu-
lar polysaccharide produced by the fermentation of the gram-
negative bacterium Xanthomonas campestris (Fig. 4). The
primary structure of this naturally produced cellulose deriva-
tive contains a cellulosic backbone (-D-glucose residues)
and a trisaccharide side chain of -D-mannose--D-
glucuronic acid-
-D-mannose attached with alternate glu-
cose residues of the main chain. The terminal D-mannose
residue may carry a pyruvate function, the distribution of
which is dependent on the bacterial strain and the fermenta-
tion conditions. The non-terminal D-mannose unit in the side
chain contains an acetyl function. The anionic character of
Drug Delivery Letters, 2012, Vol. 2, No. 4 3
this polymer is due to the presence of both glucuronic acid
and pyruvic acid groups in the side chain.
2.1.3. Hyaluronic Acid
Hyaluronic acid (HA) [24] is a naturally-occuring linear
polysaccharide composed of
-1-4-linked D-glucoronic acid
(
(1,3) N-acetyl-D-glucosamine disaccharide units, and it is
the only non-sulfated glycosaminoglycan in the extracellular
matrix of all higher animals (Fig. 4).
2.1.4. Heparin
Heparin [25] is a heterogeneous mixture of unbranched,
acidic, glycosaminoglycans (GAG), which consist of alter-
nating glucosamine and hexuronic acids modified by sul-
fation and acetylation. It is a highly sulfated linear polysac-
charide with overall negative charges. Glycosaminoglycan
4 Drug Delivery Letters, 2012, Vol. 2, No. 4
Fig. (4). Chemical structure of anionic polysaccharides.
(GAG) heparin was discovered in 1916 and has been pre-
pared and used as a clinical anticoagulant since 1939. Hepa-
rin is prepared from animal tissues that are rich in mast cells,
such as porcine mucosa and bovine lung. In clinical practice,
conventional heparin is unfractionated, which means that it
consists of molecules of differing lengths, usually between
10 and 50 saccharides (Fig. 4).
2.2. Anionic Modified Polysaccharides
Several methods of synthesis are used to incorporate
acidic functions into polysaccharides. The acid functions are
generally carboxylic acid functions obtained by car-
boxymethylation or sulfate acid functions obtained by sul-
fation.
2.2.1. Carboxymethylation of Polysaccharides
Carboxymethylation [26] is a well-known etherification
process for polysaccharides, producing derivatives in which
the hydroxyl groups are modified with carboxymethyl (CM)
groups. The advantages of this modification reaction are the
ease of processing, low cost, and safety of the products.
Carboxymethylation of polysaccharides is a widely stud-
ied conversion since it is simple and leads to products with a
variety of promising properties. In general, the polysaccha-
ride is activated with aqueous alkali hydroxide, most often
sodium hydroxide, and converted with monochloroacetic
acid or its sodium salt according to the Williamson ether
Souguir et al.
synthesis, yielding the carboxymethyl (CM) polysaccharide
derivative. Various polysaccharides from different sources
are used as starting materials (CM-cellulose, CM-starch,
CM-Pullulan, CM-Dextran…).
2.2.2. Sulfation of Polysaccharides
Polysaccharides were first sulfated with the aim of de-
veloping new alternatives to heparin, which suffers from its
heterogeneous and variable structure, and multiple in vivo
effects. Esterification of polysaccharides can be accom-
plished using inorganic and organic acids, anhydrides, and
chlorides of alkyl or aryl carboxylic acids, sulfonyl chlo-
rides, and isocyanates, in the presence of suitable catalysts.
Sulfate is one of the industrially important esterified prod-
ucts [27].
Many methods have been described for the sulfation of
carbohydrates [28]. Various methods involving the combina-
tion of sulfating agents and reaction media have been used.
The common reagents used are sulfuric acid, concentrated
chlorosulfonic acid, chlorosulfonic acid-pyridine mixture,
concentrated sulfuric acid in the presence of an aliphatic
alcohol, and sulfamic acid in an aqueous solution of urea.
3. CHEMICAL CROSSLINKING OF ANIONIC POLY-
SACCHARIDE
Several techniques have been reported for the synthesis
of biomedical hydrogels. Chemically cross-linked gels have
ionic or covalent bonds between the polymer chains. Poly-
Anionic Polysaccharide Hydrogels with Charges Provided by the Polysaccharide
saccharide containing functional groups like alcohol or acid
groups are generally soluble in water. The presence of these
functional groups on the polymer chain can be used to pre-
pare hydrogels by forming covalent linkages between the
polymer chains and induce complementary reactivity, such
as amine-carboxylic acid, or by Schiff base formation or by
radiation (UV,
-ray, electron beam…).
3.1. Crosslinking by Amide or Ester Linkage
Most anionic polysaccharides are characterized by the
presence of carboxylic acid functions over the hydroxyl
functions. The chemical properties of acids allow modifica-
tions by several chemical functions (amine, alcohol and
halogen). The literature shows that the use of di-functional
molecules in the presence of anionic polysaccharides allows
the formation of a three-dimensional network hydrogel. In-
deed the use of diamine compounds [29-31], molecules that
H 2N
n
H2N O
OH
H 2N HN
O
H 2N HN
S
O
O Cl
Epichlorohydrin
O P
Cl P Cl
Cl
Phosphorus oxychloride
Fig. (5). Chemical structure of crosslinking agents.
Drug Delivery Letters, 2012, Vol. 2, No. 4 5
are able to react in the presence of a coupling agent such as
carbodiimide with carboxylic acids lead to the formation of
amide bonds.
Among these compounds is dihydrazide, which contains
two amine functions. Several hydrazide derivatives [32, 33]
(Fig. 5) with suitable physicochemical properties for drug
release have been synthesized. Among these modification we
cite the (succinic, adipic or suberic) acid dihydrazide [34-36]
or as other derivatives modified with groups disulfide or
polyethylene glycol (PEG) or polyhydrazides [37] (Fig. 5)
used for the crosslinking of Hyaluronic acid (HA). The use
of adipic acid dihydrazide (AAD) (Fig. 5) allows the
crosslinking of alginate to obtain a biodegradable hydrogel
[38]. The formation of hydrogels in aqueous medium is an
advantage of using these crosslinking agents. As previously
reported the reaction requires the use of a water-soluble car-
bodiimide 1-Ethyl-3-(3-Dimethylaminopropyl) carbodiimide
NH2 alkyl-diamine
OH
O NH2
n polyethylene glycol-diamine (PEG-diamine)
adipic dihydrazide
NH NH2
O
S disulfide-containing dihydrazide
NH NH2
O OH
O O
HO OH
OH
Citric acid
O O-Na +
O O
+ Na-O
O
P P
O
O O-Na +
sodium trimetaphosphate
6 Drug Delivery Letters, 2012, Vol. 2, No. 4 Souguir et al.

(EDCI) typically coupled to the N-hydroxysuccinimide
(NHS) or sulfo-NHS and adjusted to pH between 4 and 5.
Other molecules, oligomers or polymers can be used as
crosslinking agents of anionic polysaccharides. For instance
compounds with two alkyl chains terminated by a diamine
function [29-31] or PEG derivatives terminated diamine have
been used for the crosslinking of HA or alginate. The poor
water solubility of these molecules requires the use of an
organic solvent (DMF, DMSO) and the use of other coupling
agents such as N,N'-dicyclohexylcarbodiimide (DCC).
Another function that allows for chemical crosslinking is
the ester function. Chemical crosslinking of anionic polysac-
charides can be achieved by several synthesis strategies.
The first method used in the case of, CMC, CM-Dextran
or Hyaluronic acid [39-41] allows the crosslinking of the
macromolecular chains through the formation of esters from
the reaction of the carboxylic acid and hydroxyl functions of
the anionic polysaccharide chain. This functionalization is
obtained in aqueous medium at acidic pH by using EDCI as
coupling agent.
Anionic Polysaccharides such as CMP, pectin, polyga-
lacturonic acid can be cross-linked with halogens alkane
derivatives [32-44] to form ester functions. The hydrogel
properties can be easily tailored by varying the concentration
of the dissolved polysaccharide and the amount of crosslink-
ing agent. The crosslinking reactions are preferably carried
out in organic solvents (DMSO, DMF) as water can also
react with the crosslinking agent. The crosslinking requires
the transformation of the carboxylate function to carboxylic
acid by a passage through an H+ exchange column or by di-
alysis against HCl, followed by neutralization with tetrabuty-
lammonium hydroxide (TBA). Furthermore, since the
crosslinking agents are generally very toxic, the gels have to
be extensively extracted to remove traces of unreacted agents.
In the following section we present some applications of
hydrogels and drug immobilization and release by these hy-
drogels. More systems Drug / hydrogels are summarized in
Table 1.
An antibody, Immunoglobulin G (IgG), releasing system
was developed by covalently attaching the IgG to Hyaluronic
acid (HA) hydrogel via a hydrolytically unstable hydrazone
linkage [45]. Alginate was chemically modified into low
molecular weight oligomers that were then crosslinked with
a biodegradable spacer, such as adipic dihydrazide, to form
biodegradable hydrogels [46]. This system may provide a
potent vehicle for the localized delivery of antineoplastic
agents (methotrexate, doxorubicin, and mitoxantrone) in a
minimally invasive manner.
Four alginate hydrogels are formed by change of the con-
centration of the adipic dihydrazide used as a crosslinking
agent (50, 75, 100 and 150 mM). The release of different
drug is studied for the formed hydrogels. Methotrexate was
released over a 2 days period from hydrogels formed at
50mM adipic dihydrazide, and the release was extended up
to 7 days from hydrogels formed at higher concentration of
adipic dihydrazide. This rapid release is due to the minimal
drug-hydrogel interaction. Doxorubicin was released from
days to weeks depending on the concentration of adipic di-
hydrazide used. This release is due to the formation of cova-
lently linked between doxorubicin and the polymer back-

Table 1. Examples of Drugs Incorporated in Anionic Polysaccharide Hydrogels

Polysaccharide Crosslinking Method Drug Reference

HA Dihydrazide Diclofenac sodium, pilocarpine, indomethacin, hydrocortisone, 6
-methyl- [47]

prednisolone, prednisolone, cortisone, corticosterone, dexamethasone, prednisone

Cytokine [36]

Ibuprofen [45]
Water soluble carbodiimide
(EDCI) L-lysine methyl ester [40]

Alginate Diydrazide Methotrexate [38]

Doxorubicin, mitoxantrone [46]

Epichlorohydrin Cefotaxime, Chloramphenicol [52]

CM-Cellulose 1,3-diamino propane Ibuprofen lysine [29]

-radiation

Theophylline [58]

Chondroitin sulfate Epichlorohydrin Paracetamol, Theophylline [53]

Xanthan Epichlorohydrin Theophylline, isosorbite dinitrate, methoxyprogesteron, neomycin and furazolidone [54]

Psyllium polysac-
-radiation Insulin [59]

charide

Amylose Sodium trimeta phosphate Sodium diclophenac [80]

Anionic Polysaccharide Hydrogels with Charges Provided by the Polysaccharide
bone. Doxorubicin was released from these gels over 3.5
days. These release time seems too long due to the presence
of interactions between doxorubicin and alginate. Indeed
doxorubicin is a cationic molecule, the presence of this
charge can cause an interaction with the anionic alginate
macromolecule.
The formation of hydrogels by crosslinking HA, via car-
bodiimide coupling of small hydrazides, PEG diamine, or
PEG dihydrazide was reported [47]. Eventually, more bio-
logically compatible gelation chemistries were investigated and
applied to this system. HA was functionalized with free thiols
via carbodiimide coupling and subsequently crosslinked by
Michael addition with PEG diacrylate. These materials have
been shown to release anti-inflammatory drugs, (diclofenac
sodium, pilocarpine, indomethacin, hydrocortisone, 6
-
methyl-prednisolone, prednisolone, cortisone, corticosterone,
dexamethasone and prednisone). Some of the drugs were
rapidly released from HA hydrogel. These drugs were almost
completely released in 10 mn following first order kinetics.
3.2. Crosslinking by Ether Linkage: Epichlorohydrin
Crosslinking Agent
Epichlorohydrin (ECH) (Fig. 5) has been extensively
used to crosslink polysaccharides [48-50]. In its reaction
with hydroxyl and carboxylic functions of anionic polysac-
charide, mono and diethers or mono and diesters are formed.
The reaction of the polysaccharide with epichlorohydrin
starts with the formation of a polysaccharide alkoxide or
carboxylate by addition of sodium hydroxide (NaOH) or
NaOH and NH4OH [51].
Gelatin and CM-cellulose based hydrogels, crosslinked
with epichlorohydrin, have been developed [52]. The hydro-
gels load and release water soluble antibiotic (chlorampheni-
col-sodium hemisuccinate, ClPh) by diffusion. The loading
and release degrees were correlated with different swelling
degrees, which depend on the crosslinking reaction condi-
tions. The release of the ClPh follows zero-order release ki-
netics. Other natural-based hydrogels [53] such as cellulose
and chondroitin sulfate have been evaluated for the release of
paracetamol and theophylline. The release profiles of the
drugs from cellulose / chondroitin sulfate hydrogels depend
on the chondroitin sulfate content: a decrease for paraceta-
mol and, an increase for theophylline, of the percentage re-
leased with the increase of the chondroitin sulfate content
was observed. The results showed good biocompatibility
with the tested formulations. Other hydrogels based on poly-
saccharide xanthan gums for drug delivery have been devel-
oped. Xanthan gum was crosslinked with epichlorohydrin
and its swelling properties were correlated with crosslinking
agent concentration [54]. These hydrogels were loaded with
theophylline, isosorbite dinitrate or methoxyprogesteron by
diffusion, while neomycin was linked to the xanthan and
furazolidone was incorporated into the neomycin derived
gel. Zero order release kinetics was observed for the release
of neomycin and furazolidone.
3.3. Radiation Crosslinking
Various natural compounds were used for preparation of
polymer hydrogels. Homogeneous hydrogels with acid poly-
saccharides have been prepared for topical delivery of anti-
Drug Delivery Letters, 2012, Vol. 2, No. 4 7
fungal drugs. Biodegradable and affinity-based hydrogels
were also studied. The potential applications of these sys-
tems are in biosensors as affinity separation support, enzyme
immobilization, cell encapsulation and drug delivery de-
vices. The conventional methods involved for producing
hydrogels use toxic additives to bring about crosslinking and
thus are not suitable for production of biocompatible hydro-
gels. Ionizing radiation possesses the unique ability to initi-
ate crosslinking reactions without the need to add toxic
chemicals. Therefore radiation processing is emerging as an
excellent tool to produce hydrogels for a variety of medical
applications. Carboxymethyl-cellulose (CMC) or car-
boxymethyl-starch (CMS) [55-57] can be crosslinked to
form anionic hydrogels. This offers an opportunity to obtain
non-toxic, additive free, totally biodegradable and biocom-
patible crosslinked hydrogels for many applications.
The crosslinking by radiation is usually accompanied by
a degradation of the macromolecular chain. This degradation
is accentuated in solutions of the polysaccharide and remains
moderate in the solid phase. This requires crosslinking in
solution at high polysaccharide concentration. The hydrogel
of anionic polysaccharide is obtained in solid state or in past-
like condition.
Radiation crosslinking of the anionic polysaccharide as
for medicinal applications and drug delivery are generally
performed in the presence of a synthetic monomer such as
acrylic acid, acrylamide.
For example, a natural polymer based colon specific drug
(theophylline) carrier was prepared from carboxymethylcel-
lulose (CMC) and acrylic acid (AAc) in aqueous solution
employing -radiation induced copolymerization and
crosslinking. The release profile of theophylline was studied
as a function of time and pH [58]. Another publication [59]
deals with the modification of psyllium polysaccharide (ani-
onic polysaccharide) with acrylamide through radiation
crosslinked polymerisation for the design of double potential
hydrogels meant for slow release of insulin. The release ki-
netics of insulin was studied by varying the pH. The release
of drug from the polymer matrix has been observed higher in
pH 7.4 as compared to the pH 2.2. The release of the insulin
from the hydrogels occurred through non-Fickian diffusion
mechanism. This observation shows that the method of syn-
thesis affects the mechanism. Here the double potential is
due to the inherent blood sugar lowering ability of psyllium
and the release of insulin in a controlled manner from the
drug loaded hydrogels.
3.4. Anionic crosslinking agent
This section will be devoted to the crosslinking agents
with an anionic character. We present two examples of these
anionic molecules. The first example is citric acid [60-62]
and the second is phosphate derivatives molecules such as
sodium trimetaphosphate (STMP) and phosphorus oxychlo-
ride (POCl3).
3.4.1. Citric Acid Crosslinking Agent
Citric acid (CA) (Fig. 5) is widely used in food and drug
industry and is an excellent crosslinking agent. CA, wide-
spread in nature (lemon juice), is prepared commercially by
fungal fermentation of glucose. Citric acid was selected taking
8 Drug Delivery Letters, 2012, Vol. 2, No. 4
into account its non toxicity, its use in the food industry as a
safe natural additive, and its high chemical reactivity. Car-
boxylic acids are interesting because they differ in the length
of the spacer and in the type and number of functional
groups on the spacer. This has a significant influence on the
sturdiness of the crosslinked gel. Actually, CA was used as
crosslinking agent in various polysaccharides derivatives
systems [60-62], and different mechanisms have been pro-
posed in the literature to explain the crosslinking reaction of
polysaccharides with CA. According to [61] the two main
stages of the reaction of polyfunctional carboxylic acids with
polysaccharides are due to the attachment of the polyfunc-
tional carboxylic acids via esterification with a polysaccha-
ride hydroxyl group and further reaction via esterification
with another cellulosic hydroxyl group producing a crosslink
between polysaccharide chains.
3.4.2 Phosphate Links
The anionic character can be brought by phosphate links
with the use of two specific crosslinkers sodium trimeta-
phosphate (STMP) and phosphorus oxychloride (POCl3)
(Fig. 5). Both crosslinkers have the great advantage to be
legal for food use [63]. Hey were used with water-soluble
polysaccharide (pullulan, dextran…) to obtain hydrogels or
microparticles and with granular polysaccharide (e.g. starch)
to minimize granule rupture.
3.4.2.1. Phosphorus Oxychloride
POCl3 is an efficient crosslinker in alkaline medium
(pH>11) in the presence of neutral salt. It disadvantage is to
produce toxic fumes by contact with humid air.
The reactions of POCl3 have been investigated by Hud-
son and Moss (1962) [64] in excess of water. Firstly, one
chloride is substituted very quickly by one hydroxyl to form
dichlorophosphate, which can crosslink two polymeric
chains by a monophosphate [65] if the medium is alkaline
enough (above pH 10) and has sufficient ionic strength. In
this case, grafted monophosphate esters are formed. Increas-
ing the alkalinity increases the crosslinking density, while
increasing salt concentration increases the uptake of alkali by
the polysaccharide as well as the ionic strength of the me-
dium [66].
Dextran hydrogels have been shown to have very high
swelling capacity (up to 7000%) and swelling rate compared
with two other crosslinkers (epichlorohydrin and N,N’-
methylenebisacrylamide) [67, 68]. This behavior is ex-
plained by the presence of the grafted phosphate groups on
the side chains. The authors have studied the penetration of a
copper solution in these hydrogels for waste-water treatment
application.
POCl3 was used to stabilize starch granules from banana
[69] and from waxy maize [70]. On banana starch, micros-
copy, rheology and DSC showed the interest of the
crosslinking. On waxy maize starch, POCl3-treated granules
displayed reduced swelling and higher viscosity with a rigid
external surface area.
3.4.2.2. Sodium Trimetaphosphate
Sodium trimetaphosphate (STMP) is a non toxic
crosslinking agent which is used on several polysaccharides
Souguir et al.
in very alkaline media [71-74]. A reaction mechanism was
proposed as a result of the studies made by high-resolution
NMR spectroscopy. In a first step, sodium tripolyphosphate
is grafted on a polysaccharide. The extremity of tripolyphos-
phate reacts with a second polysaccharide alcoholate, to pro-
duce a crosslinked phosphate network. On the other hand,
tripolyphosphate could lead through hydrolysis to a mono-
phosphate on the chain. [75, 76]
These hydrogels can be used for smooth muscle cell cul-
ture and are promising as scaffolds for vascular engineering
[77].
Microparticles were prepared with carboxymethylpullu-
lan in organic suspension media. Due to the anionic charac-
ter the swelling capacity is much higher than that of the mi-
croparticles obtained using epichlorohydrine [78]. Lysozyme
was immobilized in the particles and the enzymatic activity
was preserved [79]. High amylose cross-linked to different
degrees with sodium trimetaphosphate was evaluated as non-
compacted systems for sodium diclophenac controlled re-
lease [80]. The release profiles were related with classical
kinetic mathematical models. An increase in polymer cross-
linking degree resulted in longer release time for both drugs,
although sodium diclophenac generally was released slowly.
Sodium diclophenac release from samples cross-linked at 2%
of sodium hydroxide was driven mainly by Fickian diffu-
sion, while from samples cross-linked at 4% of NaOH fol-
lows anomalous mechanism.
4. CONCLUSION
Polysaccharides in hydrogel shape are interesting candi-
dates for new formulations of drugs. In this review, we have
described only one class of polysaccharide systems, chemi-
cal crosslinked hydrogels with an anionic character. We have
focused on their synthesis and applications. Due to the pres-
ence of anionic charges, the mechanism of entrapment and
release of drugs is based on electrostatic interactions and/or
swelling/deswelling behaviour (for macromolecular species
only). These systems are sensitive to pH and ionic strength.
By appropriate chemical modification, it is possible to mod-
ify the hydrophilic character and to obtain amphiphilic hy-
drogels. These systems form hydrophobic clusters that are
able to entrap hydrophobic drug molecules. Currently, hy-
drophobic modified polysaccharides are obtained by grafting
an alkyl chain on the polymer backbone which reduces the
biocompatibility [81, 82]. Another option in order to main-
tain both biodegradability and biocompatibility properties is
the incorporation of aliphatic polyesters as poly(
-
caprolactone) (PCL) and polylactide (PLA) [83, 84]. We can
try in literature some examples of amphiphilic hydrogels.
Most are obtained by physical crosslinking [83]. Chemical
crosslinking is often used to obtain microparticles [73].
The knowledge of researchers in the field of pharmaceu-
tics allows imagining specific application (drug, temperature,
pH and ionic strength), administration way, kinetics of deliv-
ery… The examples reported here demonstrate the feasibility
to elaborate a polysaccharide hydrogel able to entrap a drug.
The difficulty is the pathway to deliver the drug to a specific
organ target.
Anionic Polysaccharide Hydrogels with Charges Provided by the Polysaccharide
New orientations will be to elaborate hydrogels with
smart behavior. Currently, researchers focus their attention
on thermal sensitive polymers that have Low Critical Solu-
bility Temperature (LCST) behavior. Literature is abundant
on functionalization of polysaccharides with poly(N-
isopropylacrylamide) [85, 86] or polyether as block copoly-
mer of poly(ethylene oxide) and poly(propylene oxide) [87].
All these systems form physical hydrogels by increasing the
temperature. When they are chemically crosslinked, they
present thermoreversible swelling and deswelling properties
and thermosensitive drug release ability [88-91].
Future research will take an interest in other smart re-
sponsive behavior. Two stimuli have particular potentiality
for a better control of the drug delivery in our opinion. The
first one is the use of magnetic fields with “ferrogels”. This
new class of hydrogels is a chemically crosslinked polymer
network containing magnetic nanoparticles [92]. The second
potentially interesting stimulus is electric field with “electro-
gels” in which the polysaccharides are grafted by an organic
conducting polymer such as Poly(aniline) [93, 94].
At any rate, the development of new systems for medical
applications requires cooperation between the academic
world and the industrial world.
CONFLICT OF INTEREST
The author(s) confirm that this article content has no con-
flicts of interest.
ACKNOWLEDGEMENTS
The authors are particularly grateful to Dr. Virginie Du-
long and Dr. Anne-Laurence Dupont for very helpful discus-
sions.
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Received: July 03, 2012 Revised: September 20, 2012 Accepted: September 26, 2012
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