European Journal of Pharmaceutical Sciences 38 (2009) 185–196

Contents lists available at ScienceDirect

European Journal of Pharmaceutical Sciences

journal homepage: www.elsevier.com/locate/ejps

Review

Dendrimers: Emerging polymers for drug-delivery systems

Basavaraj K. Nanjwade a,∗ , Hiren M. Bechra a,1 , Ganesh K. Derkar a,2 ,
F.V. Manvi a,3 , Veerendra K. Nanjwade b,4
a
Department of Pharmaceutics, JN Medical College, KLE University, Belgaum – 590010, India
b
Department of Pharmaceutics, KLE College of Pharmacy, KLE University, Bangalore – 560010, India

a r t i c l e i n f o a b s t r a c t

Article history: Dendrimers are new class of polymeric materials. It is generally described as a macromolecule, which is
Received 3 December 2008 characterized by its extensively branched 3D structure that provides a high degree of surface functionality
Received in revised form 26 June 2009 and versatility. The unique properties associated with these dendrimers such as uniform size, high degree
Accepted 19 July 2009
of branching, water solubility, multivalency, well-defined molecular weight and available internal cavities
Available online 29 July 2009
make them attractive for biological and drug-delivery applications. Commercialization of dendrimers is
now forthcoming. The present review briefly describes about dendrimer synthesis strategies, types of
Keywords:
dendrimers with different functionalities, properties which having crucial importance and their potential
Dendrimer
Dendrimer synthesis
applications.
Dendrimer properties © 2009 Elsevier B.V. All rights reserved.
Dendrimer types
Dendrimer applications
PAMAM

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 186
2. Synthesis of dendrimers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 186
2.1. Divergent growth method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 187
2.2. Convergent growth method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 187
2.3. Other approach . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 188
2.3.1. ‘Hypercores’ and ‘Branched Monomers’ growth . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 188
2.3.2. ‘Double Exponential’ growth . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 188
2.3.3. ‘Lego’ chemistry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 188
2.3.4. ‘Click’ chemistry . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 188
3. Properties of dendrimers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 188
3.1. Monodispersity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 188
3.2. Nanoscale size and shape . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 189
3.3. Polyvalency . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 189
3.4. Physicochemical properties of dendrimers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 189
3.5. Biocompatibility of dendrimers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 190

Abbreviations: PAMAM, polyamidoamine; PEG, polyethylene glycol; IgG, immunoglobulin G; PPI, poly(propylene imine); PAMAMOS, poly(amidoamine organosili-
con); MRI, magnetic resonance imaging; Gd, gadolinium; DTPA, diethylenetriaminepentaacetic acid; P-gp, P-glycoproteins; Papp , apparent permeability coefficient; TEER,
transepithelial electrical resistance.
∗ Corresponding author. Tel.: +91 9448716277/9742431000 (Cell); fax: +91 8312472387.
E-mail addresses: bknanjwade@yahoo.co.in (B.K. Nanjwade), hiren.bechra@yahoo.com (H.M. Bechra), ganeshderkar@gmail.com (G.K. Derkar),
fvmanvi05@rediffmail.com (F.V. Manvi), vknanjwade@gmail.com (V.K. Nanjwade).
1
Tel.: +91 9429046096 (Cell).
2
Tel.: +91 9880327989 (Cell).
3
Tel.: +91 9845255552 (Cell).
4
Tel.: +91 9972670590 (Cell).

0928-0987/$ – see front matter © 2009 Elsevier B.V. All rights reserved.
doi:10.1016/j.ejps.2009.07.008
186 B.K. Nanjwade et al. / European Journal of Pharmaceutical Sciences 38 (2009) 185–196

4. Types of dendrimers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 190

4.1. Liquid crystalline dendrimers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 190
4.2. Tecto dendrimers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 190
4.3. Chiral dendrimers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 190
4.4. PAMAMOS dendrimers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 191
4.5. Hybrid dendrimers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 191
4.6. Peptide dendrimers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 191
4.7. Glycodendrimers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 191
4.8. PAMAM dendrimer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 192
5. Applications of dendrimers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 192
5.1. Dendrimers as a carrier for drug delivery . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 192
5.1.1. Dendrimers in transdermal drug delivery . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 192
5.1.2. Dendrimers in oral drug delivery . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 192
5.1.3. Dendrimers in ocular drug delivery . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 192
5.1.4. Dendrimers in pulmonary drug delivery . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 193
5.2. Dendrimers in targeted drug delivery . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 193
5.3. Dendrimers for controlled release drug delivery . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 193
5.4. Dendrimers in gene delivery . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 193
5.5. Dendrimers as imaging agents . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 194
6. Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 194
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 194

1. Introduction
eration’ along with doubling the number of end groups (or active
sites) and approximately double the molecular weight of the previ-
Traditionally polymer technologies were focused mainly on lin-
ous generation, too (Table 1). Recent reviews are mainly describing
ear polymers. Currently highly branched polymers have been found
properties of dendrimers (Klajnert and Bryszewska, 2001; Boas and
in pictures, properties of branched macromolecules are quite dif-
Heegaard, 2004), applications in biomedical (Satija et al., 2007),
ferent from conventional polymers. The unique structural features
for drug-delivery systems (Cheng et al., 2008; Gillies and Fréchet,
of dendritic and hyperbranched macromolecules have number of
2005), and also particularly dendrimer applications for cancer ther-
chains whose ends combined with a high degree of branching
apy (Wolinsky and Grinstaff, 2008; Tomalia et al., 2007). Present
which leads to a variety of new physical properties when com-
review will have main focus on the different synthesis strategies of
pared with traditional linear polymers. Dendrimers offer a plenty
dendrimers, types of dendrimers and recent studies on important
of advantages compared to other architectural forms of polymers
applications of dendrimers.
that have been used in drug-delivery systems. They have nar-
row polydispersity, nanometer size range, which can allow easier
passage across biological barriers (e.g. small enough to undergo 2. Synthesis of dendrimers
extravasations through vascular endothelial tissues). Host–guest
can take place either in the interior (binding groups in the inte- Since 1979, two major strategies have evolved for dendrimer
rior of dendrimers are called endoreceptors) or on the periphery synthesis. The first was introduced by Tomalia, called as “divergent
of the dendrimer (groups involved in complexation chemistry method” in which growth of dendrimers originates from a core site.
on the periphery of the dendrimers are called exoreceptors) This approach involves assembling monomeric modules in a radial,
(Tomalia et al., 1990). Macromolecular architectures offer unique branch-upon-branch motif according to certain dendritic rules
nanoscopic shapes and surfaces which opens the door for either
sub-nanoscopic reagents (i.e. classical organic types) or nanoscopic
reagents (e.g. DNA, IgG antibodies, etc.) to perform reactions. So
the structure of these macromolecules has great impact on their
applications.
The name comes from the Greek word dendron, meaning ‘tree’,
and meros meaning ‘part’. Dendrimer is an internationally accepted
term and other synonymous terms are ‘arborols’ and ‘cascade
molecules’. The first ever dendrimers were described by Vögtle
in 1978 (Bhuleier et al., 1978). Then in early 1980s pioneering
work was done by Denkewalter and co-workers as polylysine den-
drimers at Allied Corporation (Denkewalter et al., 1981), Tomalia
et al. at Dow Chemical (Tomalia and Dewald, 1985; Tomalia et al.,
1985) and Newkome et al. (1985) on dendrimer synthesis. Den-
drimers are large and complex molecules having very well-defined
chemical structures. They possess three distinguishing architec-
tural components, mainly (a) an initiator core, (b) an interior layer
(generations), composed of repeating units, radially attached to the
initiator core and (c) exterior (terminal functionality) attached to
the outermost interior generation (Fig. 1) (Tomalia et al., 1985).
Dendrimers are generally produced in an interative sequence of
reaction steps, in that each additional interation leads to higher
generation dendrimer. Each of the new layer creates a new ‘gen- Fig. 1. Schematic representation of generation 4 dendrimer.
 B.K. Nanjwade et al. / European Journal of Pharmaceutical Sciences 38 (2009) 185–196 187

Table 1 process is repeated until the dendrimer of the described size is

Physical characteristics of PAMAM dendrimers.
obtained (Jain and Khopade, 2001). PAMAM dendrimers are syn-
Generation Number of surface groups Molecular weighta Diameter (nm)b thesized by divergent growth approach. This method involves in
0 4 517 1.5 situ construction in stepwise, interative stages around a desired
1 8 1,430 2.2 core such as ethylenediamine, ammonia or cystamine to produce
2 16 3,256 2.9 well-defined structure. The two step reaction sequences exposed
3 32 6,909 3.6 on a core consists of (a) an exhaustive alkylation of primary amines
4 64 14,215 4.5
(Michael addition) with methyl acrylate, and (b) amidation of
5 128 28,826 5.4
6 256 58,048 6.7 amplified ester groups with a large excess of ethylenediamine
7 512 116,493 8.1 to produce primary amine terminal groups which creates ‘gen-
8 1024 233,383 9.7 eration zero’ (G0) of the dendrimer series. Each amino group
9 2048 467,162 11.4
present in this branched molecule is then subsequently reacted
10 4096 934,720 13.5
with two additional molecules of methylmethacrylate monomer,
a
Molecular weight is based on defect-free, ideal-structure, amine terminated followed by reaction with two more ethylenediamine molecules,
dendrimers.
b
to provide first generation starburst dendrimer (Jain and Khopade,
Molecular dimensions determined by size-exclusion chromatography.
2001; Tomalia, 2005). Most divergent dendrimer synthesis requires
excessive monomer loading and lengthy chromatographic separa-
and principles (Tomalia, 1996). The second method, pioneered tions, particularly at higher generations. This divergent approach
by Hawker and Fréchet follows a “convergent growth process” is currently the preferred commercial strategy used by worldwide
(Hawker and Fréchet, 1990). In which, several dendrons are reacted producers; such as Dendritic Nanotechnology Incorporation, DSM
with a multifunctional core to obtain a product. Over 100 com- and Prestorp.
positionally different dendrimer families have been synthesized
and over 1000 differentiated chemical surface modifications have 2.2. Convergent growth method
been reported, mainly on the basis of these two synthetic strate-
gies (Bosman et al., 1999; Fréchet and Tomalia, 2001; Tomalia and This is an alternative approach to dendrimer synthesis. The diffi-
Majoros, 2000). Often, these reactions have to be performed at culty of many reactions that have to be performed on one molecule
many sites on the same molecule, simultaneously. Clearly, the reac- is overcome by starting at the periphery and ending at the core.
tion must be very ‘clean’ and high yielding for the construction Convergent growth begins at what will end up being the surface
of large targets to be feasible. Tomalia-type PAMAM dendrimers of the dendrimer, and work inwards by gradually linking surface
constitute the first dendrimer family to be commercialized, most units together with more monomers (Fig. 3). When the growing
extensively characterized and the best understood series at this dendrons are large enough, several are attached to a suitable core
time, too. to give a complete dendrimer (Matthews et al., 1998).
In the convergent growth a small number of reactive sites are
2.1. Divergent growth method functionalized in each step, giving a small number of possible side
reactions per steps. The other advantages include the ability to
A schematic representation of divergent growth is shown in precisely control molecular weight and make material having func-
(Fig. 2). In this method, the core is reacted with two or more tionalities in precise positions and numbers. The deactive products
moles of reagent containing at least two protecting branching sites, are easier to separate by purification at each synthesized gener-
followed by removal of the protecting groups. The subsequent lib- ation of dendrimer hence the products are more homogeneous
erated reactive sites lead to the first generation dendrimers. This although purification in the higher generation dendrons becomes

Fig. 2. Divergent growth method.

Fig. 3. Convergent growth method.

188 B.K. Nanjwade et al. / European Journal of Pharmaceutical Sciences 38 (2009) 185–196

Fig. 4. ‘Hypercores’ and ‘Branched Monomers’ growth.

more cumbersome, because of increasing similarity between reac-
tants and formed products.
On the other hand, convergent synthesis strategies are generally
limited to the construction of only lower generation dendrimers
due to the nanoscale steric issues that are encountered when
attaching the dendrons to the molecular level core (Fréchet and
Tomalia, 2001). The convergent methodology also suffers from low
yields in the synthesis of large structures.
2.3. Other approach
2.3.1. ‘Hypercores’ and ‘Branched Monomers’ growth
After the development of the convergent growth approach by
Fréchet group, their efforts focused on the acceleration of den-
drimer synthesis. The outcome of this research demonstrated in
Fig. 4. This method involved the pre-assembly of oligomeric species
which can be linked together to give dendrimers in fewer steps or
higher yields.
2.3.2. ‘Double Exponential’ growth
It is similar to a rapid growth technique for linear polymer. This
approach allows the preparation of monomers for both divergent
and convergent growth from a single starting material (Fig. 5). Then
resulted two products are reacted to give an orthogonally protected
trimer, which can be used to repeat the growth again. This approach
was studied by Shinkai, Fréchet and Roy. Strength of double expo-
nential growth approach is fast synthesis and can be extended with
either divergent or convergent growth easily.
2.3.3. ‘Lego’ chemistry
In this strategy highly functionalized cores and branched
monomers are applied to prepare phosphorus dendrimers. After
several variations in general synthetic scheme, the developed
scheme allows multiplications of the number of terminal surface
groups from 48 to 250 in one step. This synthesis requires minimum
volume of solvent, allow facile purification and produce environ-
mentally benign byproducts such as water and nitrogen (Svenson
and Tomalia, 2005).
2.3.4. ‘Click’ chemistry
By this approach dendrimers with various surface groups in
high purity and excellent yield can be obtained. All generation 2
and some generation 3 Cu(I)-catalyzed dendrimers were isolated
with only sodium chloride as the major byproduct (Svenson and
Tomalia, 2005).
3. Properties of dendrimers
3.1. Monodispersity
Dendrimers are the class of dendritic polymers that can be
constructed with a well-defined molecular structure, i.e. being

Fig. 5. ‘Double Exponential’ growth.

 B.K. Nanjwade et al. / European Journal of Pharmaceutical Sciences 38 (2009) 185–196
Table 2
Some products based on dendrimers.
monodisperse, unlike to linear polymers. The monodispersity of
dendrimers has been extensively verified by high performance liq-
uid chromatography (HPLC), size exclusion chromatography (SEC),
mass spectrometry (MS), gel electrophoresis and transmission
electron microscopy (TEM) (Jackson et al., 1998). As mentioned
above, convergent growth process permits the purification pro-
cess at each step of the synthesis which gives the monodisperse
molecules as determined by mass spectrometry. That is the rea-
son, Tomalia said that convergently produced dendrimers are
probably the most precise synthetic macromolecules that exist
today (Tomalia, 2005). However, mass spectroscopy has shown
that dendrimers produced by divergent method are remarkably
monodisperse for earlier generation (G = 0–5) (Tomalia, 1993).
Monodispersity offers researchers the possibility to work with
a tool for well-defined scalable sizes. This property is useful for
applications such as the synthesis of container molecules, use as
templates or in electronic applications. The two factors affects the
degree of monodispersity includes, (i) dendrimer bridging and (ii)
incomplete removal of ethylenediamine at each of the generation
sequences (Tomalia et al., 1985). This latter factor at any point in
dendrimer growth will function as an initiator core to produce 0.5
generation and subsequent generation dendrimers that leads to
polydispersity. Ethylenediamine can be effectively removed from
the lower generation dendrimers by simple vacuum distillation at
lower temperature and from higher generation dendrimers (i.e.
G > 4) by special techniques like azeotropic (n-butanol) and wiped
film distillation (Maciejewski, 1982).
3.2. Nanoscale size and shape
Based on their dimensional length scaling, narrow size distri-
bution, and other biomimetic properties, dendrimers are often
referred an “artificial proteins” (Svenson and Tomalia, 2005).
Within the PAMAM dendrimer family, the diameter of the genera-
tion 1–10 with ethylenediamine core increases from 1.1 to 12.4 nm
(Cheng et al., 2008). The shape persistence of dendrimers is very
important, as it allows the defined placement of functions not
only on the dendrimer surface but also inside the dendritic scaf-
fold. The PAMAM dendrimers of lower generation (G0–G3) with
ethylenediamine core have ellipsoidal shapes, whereas the PAMAM
dendrimer of higher generation (G4–G10) have roughly spherical
shapes (Cheng et al., 2008). X-ray analysis on dendrimer aggre-
gates have been revealed that the molecular shape of the lower to
higher generations becomes increasingly globular (i.e. more spher-
189
ical compare to linear shaped), in order to spread out the larger
molecular structure with a minimal repulsion between the seg-
ments (Percec et al., 1998). These fundamental properties have in
fact lead to their commercial use for gene therapy, immunodiag-
nostics and variety of other biological applications (Bieniarz, 1998).
3.3. Polyvalency
Polyvalency of dendrimers provides many interactions as they
coordinate to materials. Polyvalency shows the outward presenta-
tion of reactive groups on the dendrimer nanostructure exterior.
This creates more connections between surfaces and bulk materi-
als for applications such as adhesives, surface coatings, or polymer
cross-linking. The product, a topical vaginal microbicide called
VivaGelTM (Table 2), prevents infection by HIV and other sexually
transmitted diseases during intercourse, takes advantage of den-
drimer’s polyvalent properties. Depending upon which functional
groups are attached to the tips of a dendrimer’s branches, these
nanostructures can behave like molecular Velcro. Simultaneously,
these functional groups can participate in multiple interactions
with receptors on biological structures like cell membranes and
viruses (Halford, 2005). In the report by Shaunak et al., the polyva-
lency of a dendrimer glucosamine conjugates is also thought to be
responsible for the ability of these molecule to efficiently inhibit
scar-tissue formation. In preliminary animal studies, the bifunc-
tional dendrimer increased the long-term success rate of glaucoma
surgery from 30% to 80% (Shaunak et al., 2004).
3.4. Physicochemical properties of dendrimers
Already in the history of the dendrimers it was suggested that
nano-sized structure of higher generation dendrimers would serve
as synthetic mimics of proteins. The hyperbranched structure of
the dendrimer is less compact than a protein, i.e. interior is not
packed as efficiently as in typical proteins, and structure of the
dendrimer provides a highly multivalent surface, exposing a much
higher number of functional groups on the surface compared to
proteins of similar molecular size (Farin and Avnir, 1991). The use
of dendrimers as protein mimics has been encouraged scientists to
investigate the physicochemical properties of dendrimers in com-
parison to proteins shows that dendrimers, similar to protein, can
adapt “native” (e.g. tighter) or “denaturated” (e.g. extended) con-
formations dependent on the polarity, ionic strength and pH of the
solvent.
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Amino terminated PPI and PAMAM dendrimers (that is the den-
drimers having basic surface groups as well as a basic interior)
at the low pH region shows the extended conformation due to
electrostatic repulsion between the positively charged ammonium
groups. At neutral pH back folding occurs which may be due to
the hydrogen bonding between the unchanged tertiary amines in
the interior and the positively charged surface amines. At pH ≥10
conformation has a higher degree of back folding so dendrimer
acquires a more spherical (globular) structure, that all because
of the weak “inter-dendron” repulsive forces. In comparison with
carboxylic acid terminated PPI dendrimers, small angle neutron
scattering (SANS) and NMR measurements shows that at pH 2 the
dendrimer core has the most extended conformation due to the
electrostatic repulsion between the positively charged protonated
tertiary amines. At pH 6 slight back folding occurs as a result of
attractive Coulomb interactions between the negatively charged
surface carboxy-groups and the positively charged tertiary amines
in the inner shells of the dendrimer. While at pH 11 the electro-
static repulsion between the negative charged forces the surface
groups apart from which gives a more extended conformation with
a highly expanded surface area (Boas et al., 2006). Polarity of the
solvent greatly influences the structure of the dendrimer. NMR
studies performed on PPI dendrimer revealed that an apolar sol-
vent such as benzene will favour polar intramolecular interactions
(e.g. hydrogen bonding) resulting in back-folding of the dendrimer
arms into the dendrimer interior, whereas the increased acidity
of chloroform, will increase salvation of the dendrimeric structure
via hydrogen bond donation to the interior tertiary amines result-
ing in a more extended conformation of the dendrimer (Chai et al.,
2001). At high ionic strength (high concentration of salts) charged
PPI dendrimers favours a contracted conformation of dendrimers
with a high degree of back-folding. In contrast at low salt concentra-
tion, the repulsive forces between the charged dendrimer segments
result in an extended conformation (Boas et al., 2006).
3.5. Biocompatibility of dendrimers
In order to utilize dendrimers as biological agents, they have to
fulfill certain biological demands. The dendrimer should be:
• non-toxic;
• non-immunogenic (except for vaccines);
• able to cross biobarriers (biopermeable) such as, the intestines,
the blood–tissue barriers, cell membranes, etc.;
• able to stay in circulation for the time needed to have a clinical
effect;
• able to target to specific structures.
Till today, the cytotoxicity of dendrimers has been primar-
ily studied in vitro, however, a few in vivo studies have been
reported. Duncan and co-workers have investigated the rela-
tionship between structure and biocompatibility of PAMAM,
poly(propyleneimine), poly(ethylene oxide) grafted carbosilane
dendrimers with cationic (NH2 -terminated) and anionic (COONa-
terminated) dendrimers in vitro. They have reported that,
regardless of structure, cationic dendrimers were generally more
haemolytic and cytotoxic effect at even relatively low concen-
tration in comparison to anionic dendrimers. They have also
reported increase in the haemolysis with increase in the gener-
ation (Malik et al., 2000). One more study reported that anionic
PAMAM dendrimers have shown a significantly lower cytotoxic-
ity than cationic dendrimer using Caco-2 cells (Jevprasesphant et
al., 2003a). Hydroxy- or methoxy-terminated polyester dendrimers
have been reported to have low toxicity in both in vitro and in
vivo studies. During in vitro study, at high concentration, these
dendrimers have induced some inhibition of cell growth but no
increase in cell death was observed. Upon injection into mice, no
acute or long-term toxicity problems observed. The results make
these new dendritic motifs promising candidates for drug-delivery
devices (Padilla De Jesu’s et al., 2002).
Immunogenicity is one of the crucial biological property of the
dendrimers. Studies performed on unmodified amino-terminated
PAMAM dendrimers showing no or only weak immunogenicity
of the G3–G7 dendrimers. However, later studies showed some
immunogenicity of these dendrimers and found that modifica-
tion of amino-terminated PAMAM dendrimers with polyethylene
glycol (PEG) chains reduces immunogenicity and gives longer life-
time in the blood stream in comparison to unmodified dendrimers
(Kobayashi et al., 2001).
4. Types of dendrimers
In recent years, dendrimers with different designed function-
alities have become objects of particular academic and practical
interest because of their unique superbranched architectures, high
densities of peripheral functionalities, symmetrical shapes, and
monodispersity. Here, some of the dendrimers having different
functionalities are briefly described along with their area of uti-
lization.
4.1. Liquid crystalline dendrimers
They consist of mesogenic (liq. crystalline) monomers e.g. meso-
gen functionalized carbosilane dendrimers. Functionalization of
end group of carbosilane dendrimers with 36 mesogenic units,
attached through a C-5 spacer, leads to liquid crystalline den-
drimers that form broad smetic A phase in the temperature range
of 17–130 ◦ C (Lorenz et al., 1996). Boiko et al. claims that they
have synthesized first photosensitive liquid crystalline dendrimer
with terminal cinnamoyl groups. They have confirmed the struc-
ture and purity of this LC dendrimer by 1H NMR and GPC methods.
It was shown that such a dendrimer, under UV irradiation, can
undergo E-Z isomerization of the cinnamoyl groups and [2 + 2] pho-
tocycloaddition leading to the formation of a three-dimensional
network (Boiko et al., 2001).
4.2. Tecto dendrimers
Tecto-dendrimers are composed of a core dendrimer, which
may or may not contain the therapeutic agent, surrounded by
dendrimers. The surrounding dendrimers are of several types,
each type designed to perform a function necessary to a smart
therapeutic nanodevice (Betley et al., 2002). Fig. 6 shows the
PAMAM core–shell tecto dendrimer. The Michigan Nanotechnol-
ogy Institute for Medicine and Biological Sciences (M-NIMBS) are
developing a tecto dendrimers which perform the following func-
tions: diseased cell recognition, diagnosis of disease state, drug
delivery, reporting location and reporting outcome of therapy. They
have already made and tested functional dendrimers which per-
form each of these functions. Even after a many questions on the
way they are planning to produce a smart therapeutic nanodevice
for the diseased cell like a cancer cell or a cell infected with a virus.
4.3. Chiral dendrimers
The chirality in these dendrimers is based upon the construc-
tion of constitutionally different but chemically similar branches
to chiral core. Chiral, nonracemic dendrimer with well-defined
stereochemistry is particularly interesting subclass, with potential
 B.K. Nanjwade et al. / European Journal of Pharmaceutical Sciences 38 (2009) 185–196
Fig. 6. Schematic representation of PAMAM core–shell tecto dendrimer.
applications in asymmetric catalysis and chiral molecular recogni-
tion (Ritzén and Frejd, 1999).
4.4. PAMAMOS dendrimers
Radially layered poly(amidoamine-organosilicon) dendrimers
(PAMAMOS) are inverted unimolecular micelles that consist of
hydrophilic, nucleophilic polyamidoamine (PAMAM) interiors and
hydrophobic organosilicon (OS) exteriors. These dendrimers are
exceptionally useful precursors for the preparation of honeycomb-
like networks with nanoscopic PAMAM and OS domains (Dvornic
et al., 2000).
4.5. Hybrid dendrimers
Hybrid dendrimers are combination of dendritic and linear
polymers in hybrid block or graft copolymer forms. The small den-
Fig. 7. Carbohydrate containing glycodendrimers.
191
drimer segment coupled to multiple reactive chain ends provides
an opportunity to use them as surface active agents, compatibi-
lizers or adhesives, e.g. hybrid dendritic linear polymers (Jain and
Khopade, 2001).
4.6. Peptide dendrimers
Dendrimers having peptides on the surface of the traditional
dendrimer framework and dendrimers incorporating amino acids
as branching or core units are both defined as ‘peptide dendrimers’
(Colinger, 2002). Also peptide dendrimers can be defined as macro-
molecules that contain peptide bonds in their structure. Because of
biological and therapeutical relevance of peptide molecules, pep-
tide dendrimers play an important role in diverse areas including
cancer, antimicrobials, antiviral, central nervous system, analgesia,
asthma, allergy and Ca+2 metabolism. On the basis of their ability to
be taken up by cells, making peptides very useful for drug delivery.
One more interesting application of peptide dendrimers is that can
be used as contrast agents for magnetic resonance imaging (MRI),
magnetic resonance angiography (MRA), fluorogenic imaging and
serodiagnosis (Bruckdorfer et al., 2004; Crespo et al., 2005).
4.7. Glycodendrimers
The term ‘glycodendrimers’ is used to describe dendrimers
that incorporate carbohydrates into their structures (Fig. 7). Gly-
codendrimers can be classified as: (i) carbohydrate-coated; (ii)
carbohydrate centered; and (iii) fully carbohydrate-based. Glyco-
dendrimers have been used for a variety of biologically relevant
applications such as, glycodendrimers with surface carbohydrate
units have been used to study the protein–carbohydrate inter-
actions that are in many intercellular recognition events. The
accessibility of the sugars is an important consideration for gly-
codendrimers used effectively to evaluate protein–carbohydrate
interactions. Like this, study of protein–carbohydrate interactions,
incorporation into analytical devices, formulation of gels, target-
ing of MRI contrast agents, drugs and gene delivery systems are
Fig. 8. PAMAM generation 3 dendrimer.
192 B.K. Nanjwade et al. / European Journal of Pharmaceutical Sciences 38 (2009) 185–196
some of the areas where glycodendrimers are likely to be beneficial
(Colinger, 2002; Turnbull and Stoddart, 2002).
4.8. PAMAM dendrimer
Perhaps the family of dendrimers most investigated for drug
delivery is the polyamidoamine (PAMAM) dendrimer (Fig. 8).
Many surface modified PAMAM dendrimers are non-immunogenic,
water-soluble and possess terminal-modifiable amine functional
groups for binding various targeting or guest molecules. PAMAM
dendrimers generally display concentration-dependent toxicity
and haemolysis. PAMAM dendrimers are hydrolytically degradable
only under harsh conditions because of their amide backbones,
and hydrolysis proceeds slowly at physiological temperatures (Lee
et al., 2005). The internal cavities of PAMAM dendrimers can
host metals or guest molecules because of the unique functional
architecture, which contains tertiary amines and amide linkages.
PAMAM dendrimers are generally prepared by divergent method
and product up to generation 10 (G10) have been obtained. PAMAM
dendrimers are the most extensively reported moiety for almost all
existing applications of dendrimers.
5. Applications of dendrimers
5.1. Dendrimers as a carrier for drug delivery
Dendrimers have narrow polydispersity; nanometer size range
of dendrimers can allow easier passage across biological barri-
ers. All these properties make dendrimers suitable as host either
binding guest molecules in the interior of dendrimers or on the
periphery of the dendrimers.
5.1.1. Dendrimers in transdermal drug delivery
In recent era, dendrimers have found applications in trans-
dermal drug-delivery systems. Generally, bioactive drugs have
hydrophobic moieties in their structure, resulting in low water-
solubility that inhibits efficient delivery into cells. Dendrimers
designed to be highly water-soluble and biocompatible have been
shown to be able to improve drug properties such as solubil-
ity and plasma circulation time via transdermal formulations and
to deliver drugs efficiently. Nonsteroidal anti-inflammatory drugs
(NSAIDs) are very effective in the treatment of acute and chronic
rheumatoid and osteoarthritis, however, clinical use of NSAIDs
is often limited by adverse events such as gastrointestinal side
effects (dyspepsia, gastrointestinal bleeding), renal side effects
when given orally. Transdermal drug delivery overcome these
adverse effects and also maintains therapeutic blood level for
longer period of time. Transdermal delivery suffers poor rates of
transcutaneous delivery due to barrier function of the skin. PAMAM
dendrimer complex with NSAIDs (e.g. Ketoprofen, Diflunisal) could
be improving the drug permeation through the skin as penetration
enhancers (Cheng et al., 2007). The model drugs Ketoprofen and
Diflunisal were conjugated with G5 PAMAM dendrimer and inves-
tigated for different studies. In vitro permeation studies on excised
rat skin showed 3.4 times higher permeation of Ketoprofen from
Ketoprofen–dendrimer complex than that from 2 mg/mL Keto-
profen suspended in normal saline. Similarly, a 3.2 times higher
permeated amount was observed with Diflunisal–dendrimer com-
plex. Anti-nociception effect of drugs was studied on mice, results
showed that Ketoprofen–dendrimer complex reducing writhing
activity during the period of 1–8 h after Transdermal administra-
tion, while pure Ketoprofen suspension at the equivalent dose of
Ketoprofen significantly decreased number of writhing between 4
and 6 h after drug was transdermally given (Cheng et al., 2007).
Chauhan et al. investigated transdermal ability of PAMAM den-
drimers by using indomethacin as the model drug for study.
In vitro permeation studies showed increase in the steady-state
flux as increase in concentration of all three types G4-NH2 ,
G4-OH and G-4.5 PAMAM dendrimers. For the in vivo phar-
macokinetic and pharmacodynamic studies, indomethacin and
dendrimer formulations were applied to the abdominal skin of
the Wistar rats and blood collected from the tail vein at the
scheduled time. The indomethacin concentration was significantly
higher with PAMAM dendrimers when compared to the pure
drug suspension. The results showed that effective concentra-
tion could be maintained for 24 h in the blood with the G4
dendrimer–indomethacin formulation. Therefore, data suggested
that the dendrimer–indomethacin based transdermal delivery sys-
tem was effective and might be a safe and efficacious method for
treating various diseases (Chauhan et al., 2003).
5.1.2. Dendrimers in oral drug delivery
Oral drug-delivery system has been the dominant route for
many years because of its significant advantages. It is by far the
most convenient administration route with good patient com-
pliance, especially in the patient’s opinions. Along with these
benefits, there are also some defects of oral delivery route like
low solubility in aqueous solutions and low penetration across
intestinal membranes (Csaba et al., 2006). D’Emanuele and his
research group (Jevprasesphant et al., 2003b) investigated effect
of dendrimer generation and conjugation on the cytotoxicity, per-
meation and transport mechanism of PAMAM dendrimer and
surface-modified cationic PAMAM dendrimer using monolayers of
the human colon adenocarcinoma cell line, Caco-2. As increase
in the concentration and generation, there was increase in the
cytotoxicity and permeation of dendrimers. While reduction in
cytotoxicity observed by conjugation with lauryl chloride. Modi-
fied dendrimers also reduced transepithelial electrical resistance
(TEER) and significantly increased the apparent permeability coef-
ficient (Papp ). In another study of transepithelial permeability of
naproxen, a low solubility model drug was investigated (Najlah
et al., 2007). The stability of these G0 PAMAM conjugates in 50%
liver homogenate was compared to that in 80% human plasma
showed the lactate ester linker gave prodrug of high stability
in plasma with slow hydrolysis in liver homogenate; such con-
jugates may have potential in controlled release systems, while
using diethylene glycol as a linker gives conjugate that showed
high chemical stability, but readily released drug in plasma and
liver homogenate. So, these conjugations demonstrate potential
as nanocarriers for the enhancement of oral bioavailability. P-
glycoproteins (P-gp), an efflux transporter, are able to reduce
the absorption of orally administrated drugs and decrease their
bioavailability, whereas propranolol is the substrate of the P-gp
efflux transporter with poor water solubility. D’Emanuele et al.
synthesized the propranolol–PAMAM dendrimers conjugate and
investigated transport of the conjugate across Caco-2 cell mono-
layer. The results showed that propranolol–dendrimer conjugate
was able to bypass the P-gp efflux transporter. Investigators have
lastly concluded that dendrimers conjugate with drug could reduce
the effect of intestinal P-gp on drug absorption of propranolol and
many other orally administered drugs (D’Emanuele et al., 2004).
5.1.3. Dendrimers in ocular drug delivery
The topical application of active drugs to the eye is the most
prescribed route of administration for the treatment of various
ocular disorders. It is generally agreed that the intraocular bioavail-
ability of topically applied drugs is extremely poor. These results
mainly due to drainage of the excess fluid via nasolacrimal duct
and elimination of the solution by tear turnover. Several research
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advances have been made in ocular drug-delivery systems by using
specialized delivery systems such as polymers, liposomes, or den-
drimers to overcome some of these disadvantages. Ideal ocular
drug-delivery systems should be nonirritating, sterile, isotonic, bio-
compatible, does not run out from the eye and biodegradable (Tolia
and Choi, 2008). Dendrimers provide unique solutions to com-
plex delivery problems for ocular drug delivery. Recent research
efforts for improving residence time of pilocarpine in the eye was
increased by using PAMAM dendrimers with carboxylic or hydroxyl
surface groups. These surface-modified dendrimers were predicted
to enhance pilocarpine bioavailability (Vandamme and Brobeck,
2005).
5.1.4. Dendrimers in pulmonary drug delivery
Dendrimers have been reported for pulmonary drug delivery
also (Bai et al., 2007). During one study, efficacy of PAMAM den-
drimers in enhancing pulmonary absorption of Enoxaparin was
studied by measuring plasma anti-factor Xa activity, and by observ-
ing prevention efficacy of deep vein thrombosis in a rodent model.
G2 and G3 generation positively charged PAMAM dendrimers
increased the relative bioavailability of Enoxaparin by 40%, while
G2.5 PAMAM, a half generation dendrimers, containing negatively
charged carboxylic groups had no effect. Formulations did not
adversely affect mucociliary transport rate or produce extensive
damage to the lungs. So the positively charged dendrimers are
suitable carrier for Enoxaparin pulmonary delivery (Bai et al., 2007).
5.2. Dendrimers in targeted drug delivery
Nowadays general cancer chemotherapeutics are less effective
in their ability to cure tumors because of the nonselective action
of these highly potent drugs, resulting in dose limiting side effects.
The application of drug carrier systems for targeting tumor cells
has gained credence as an alternative approach for treating cancer
and offers both increased therapeutic index and decreased drug
resistance. An effective targeting drug-delivery system requires a
base that is uniform and able to couple multiple components such
as targeting molecule, drug and cancer imaging agent (Thomas et
al., 2004).
Dendrimers have ideal properties which are useful in targeted
drug-delivery system. One of the most effective cell-specific tar-
geting agents delivered by dendrimers is folic acid. Membrane
associated high-affinity folate receptors are folate-binding proteins
that are over expressed on the surface of different types of cancer
cells (e.g. ovarian). PAMAM dendrimers conjugated with the folic
acid and fluorescein isothiocyanate for targeting the tumor cells
and imaging respectively. Further these two molecules are linked
with complementary oligonucleotides. DNA-assembled nanoclus-
ters were evaluated in vitro which helps in detecting tumor
cell-specific binding and internalization. These DNA-assembled
dendrimer conjugates may allow the combination of different
drugs with different targeting and imaging agents so it is easy
to develop combinatorial therapeutics (Choi et al., 2005). Anti-
bodies specific to CD14 and PSMA (prostate-specific membrane
antigen) were conjugated to a G5 PAMAM dendrimer bearing a
fluorescein imaging tag. Cell binding and internalization of the
antibody-conjugated dendrimers to antigen-expressing cells were
evaluated by flow cytometry and confocal microscopy. It was found
that the conjugates bound specifically to the antigen-expressing
cells in a time- and dose- dependent fashion with an affinity sim-
ilar to that of the free antibody. Confocal microscopy indicated
that cellular internalization of the dendrimer conjugate did occur
(Thomas et al., 2004). During another study, folic acid was con-
jugated to dendrimers as targeting agent and then coupled with
a model drug Methotrexate. These conjugates were injected to
193
immunodeficient mice bearing Human KB tumors and evaluated.
Biodistribution study shows that percentage of the injected dose
particular in tumor cells after one day was around three times
higher with targeted polymer conjugates (with folic acid) com-
pare to nontargeted polymer conjugates (without folic acid). There
was also observed the clear and rapid removal of conjugated den-
drimers from the blood via kidney during the first day postinjection.
Confocal microscopy images obtained of tumors after 15 h of i.v.
injection showed a significant number of fluorescent cells with tar-
geted dye-conjugates compare to the nontargeted dye-conjugates.
Flow cytometry analysis of a single-cell suspension isolated from
the same tumors showed the same results as confocal microscopy
images (Kukowska-Latallo et al., 2005). Patri and co-workers have
investigated that complexing a drug with dendrimer as an inclu-
sion complex improves its solubility in water, a cleavable, while
covalently linked dendrimer conjugate is better for targeted drug
delivery because it does not release the drug prematurely in bio-
logical conditions. They reported less cytotoxic effect with the
covalently linked dendrimer (Patri et al., 2005).
5.3. Dendrimers for controlled release drug delivery
Fréchet and co-workers have prepared polyaryl ether den-
drimers containing dual functionality on the surface. One is used
to attach polyethylene glycol (PEG) units on the surface to improve
water solubility and the other one is utilized to attach hydrophobic
drug molecules. They have also synthesized a series of dendritic
unimolecular micelles with a hydrophobic polyether core sur-
rounded by a hydrophilic PEG shell for drug encapsulation. A
third-generation micelle with indomethacin entrapped as model
drug gives slow and sustained in vitro release, as compared to
cellulose membrane control (Patri et al., 2002; Liu et al., 1999,
2000). PEG-2000 was conjugated to generation G3 PAMAM den-
drimers with varying degree of substitution. Methotrexate drug
was encapsulated (loaded) to the prepared conjugates and inves-
tigated for drug release in a dialysis bag. The results found that
PEG-dendrimers conjugated with encapsulated drug and sustained
release of methotrexate as compare to unencapsulated drug. Con-
trolled release of the Flurbiprofen could be achieved by formation
of complex with amine terminated generation 4 (G4) PAMAM den-
drimers. Prepared dendrimer complexes observed that loaded drug
displayed initial rapid release (more that 40% till 3rd hour) followed
by slow release. Pharmacodynamic study was performed using car-
rageenan induced paw edema model, revealed 75% inhibition at 4th
hour that was maintained above 50% till 8th hour. The dendritic
formulation showed 2-fold and 3-fold increased in mean residence
time and terminal half-life, respectively, as compared to free drugs
(Asthana et al., 2005).
5.4. Dendrimers in gene delivery
Dendrimer-based transfection agents have become routine
tools for many molecular and cell biologists but therapeutic
delivery of nucleic acids remains a challenge. Because of their
immunogenicity, dendrimers are extensively used as non-viral vec-
tor for gene delivery. The use of dendrimers as gene transfection
agents and drug-delivery devices have been extensively reviewed
part (Broeren et al., 2004; Boas and Heegaard, 2004).
The ability of cationic dendrimers to deliver DNA or RNA
has been reviewed previously. Besides of that some research
recently indicated that dendrimer based gene delivery system
also have significant potential in clinical trials. Kukowska-Latallo
et al. reported that intravenous administration of G9 PAMAM
dendrimer-complexed pCF1CAT plasmid could result in high
level of gene expression in the lung tissues of rats. It enhances
194 B.K. Nanjwade et al. / European Journal of Pharmaceutical Sciences 38 (2009) 185–196
the transfection efficiency and expression pattern of dendrimer
(Kukowska-Latallo et al., 2000). Amphiphilic dendrimers having
a rigid diphenylethyne core featured a variety of geometries and
substitution patterns, all of which showed high transfection activ-
ity. The hydrophobic parameters influenced the DNA binding and
transport more strongly than anticipated, exhibiting lower toxicity.
In contrast to cationic dendrimers, these dendrimers did not have
any size limitation for transfection (Joester et al., 2003). In another
study, amphiphilic, Tomalia-type PAMAM dendrimers of genera-
tions 1 to 4, were synthesized utilizing di-n-dodecylamine as the
core. It was anticipated that the hydrophobic components would
mimic the membrane transfection ability of natural phospholipids
such as dioleoylphosphatidylethanolamine (DOPE) and enhance
membrane penetration. These constructs forms complexes with
DNA and, in case of the G = 2–4 dendrimers, were able to cross cell
membranes and efficiently deliver DNA (Takahashi et al., 2003).
Polycationic dendrimers also studied with a different cell lines
for ability to transfect DNA and toxicity. In the result, some of
polycation–DNA complexes were less toxic than lipid–DNA sys-
tems (Gebhart and Kabanov, 2001).
Glycoplexes are used to target to the specific cells and/or
to increase gene transfer activity. For example, a galactosylated
polyethyleneimine (PEI) has high transfection efficiency to hep-
atocyte expressing asialoglycoprotein receptor. In addition, a
mannosylated PEI has high transfection efficiency to macrophages
and dendritic cells, which were mediated by the mannose-specific
receptor and DEC-205, respectively. In another approach, Wada
et al. have reported that galactose-␣-cyclodextrines conjugations
with degree of substitution of the galactose moiety was the most
preferential carrier among the prepared series because it provided
good gene transfer activity in vitro with no cytotoxicity. Conse-
quently, the potential use of Gal-␣-CDE conjugate (DSG 4) could be
expected as a non-viral vector to deliver gene and these data may
be useful for design of ␣-cyclodextrins and galactose conjugates
with other non-viral vectors (Wada et al., 2005). Glycosylation of
polymer is one of the effective methods to deliver gene to target
cells and/or to enhance gene transfer.
Cyclodextrins are cyclic (␣-1,4)-linked oligosaccharides of ␣-
D-glucopyranose containing a hydrophobic central cavity and
hydrophilic outer surface, and they are known to function as
novel drug carriers. The most common cyclodextrins are ␣-, ␤-
and ␥-cyclodextrins, which consist of six-, seven- and eight-D-
glucopyranose units, respectively (Wada et al., 2005). PAMAM
dendrimers functionalized with ␣-cyclodextrin showed luciferase
gene expression about 100 times higher than for unfunction-
alized PAMAM or for non-covalent mixtures of PAMAM and
␣-cyclodextrin (Arima et al., 2001). Arima et al. previously reported
the potential use of polyamidoamine (PAMAM) starburst den-
drimer (generation 3, G3) bearing ␣-cyclodextrins with the degree
of substitution (DS) of 2.4 because of the highest transfection effi-
ciency in vitro and in vivo with low cytotoxicity (Wada et al., 2005;
Arima et al., 2001).
Unlike PAMAM dendrimers based gene delivery systems, PPI
dendrimers as gene complexes can lead to liver targeted gene
expression. Dufes et al. indicated those different generations of
PPI dendrimers as transfection agents, gene efficiently expressed in
the liver rather than other organs. Furthermore, they demonstrated
that intravenous administration of a gene medicine and G3 PPI den-
drimer complex could result in intratumoral transgene expression
and regression of the established tumors in all the experimental
animals (Dufes et al., 2005).
For the plant cell gene delivery, Pasupathy and co-workers have
prepared supramolecular complexes via an electrostatic interac-
tions between poly(amidoamine) (PAMAM) dendrimer and green
fluorescence protein (GFP)-encoding plasmid. These complexes
were observed penetrating through the cell walls of turfgrass cal-
lus and expressing foreign genes within the cells. The use of the
delivery system developed here with turfgrass cells as a target can
be easily extended to virtually all plant species having successful
regeneration systems in place. Compared to conventional means
such as viral vectors, biolistic particle bombardment, electropora-
tion or polyethylene glycol attachment, these complexes having a
many advantages such as direct, less invasive, improves transgenic
efficiency and could dramatically enhance the procedures for crop
improvement using transgenic technology (Thomas et al., 2004).
5.5. Dendrimers as imaging agents
The first in vivo diagnostic imaging applications using
dendrimer-based MRI contrast agents were demonstrated in the
early 1990s by Lauterbur and colleagues (Wiener et al., 1994).
In comparison with the commercially available small-molecule
agent (Magnevist, Schering, AG), the dendrimer-based reagents
exhibited blood pool properties and extraordinary relaxivity values
when chelated gadolinium groups (Magnevist® ). These genera-
tion dependent, dramatic enhancements of MRI contrast properties
were some of the first examples of a ‘dendritic effect’ (Tomalia
et al., 2007). Gadolinium is an FDA-approved contrast agent for
MRI which provides greater contrast between normal tissue and
abnormal tissue in the brain and body. It is safer than the iodine
type contrast used in CT scans and also non-radioactive and is
rapidly cleared by kidneys (Patri et al., 2002). Different gener-
ation G1 [n = 4 Gd(III) ions per molecule], G3(n = 16), G5(n = 64)
of Gd(III)DTPA-terminated poly(propylene imine) dendrimers and
Gd(III)DTPA complex [G0 (n = 10)] reference were synthesized and
investigated for relaxivities and concentration detection limits.
Investigator revealed dendrimers G1, G3 and G5 have been shown
an increase in both T1 and T2 relaxivities with increasing molecu-
lar weight. Relaxivity values, measured in blood plasma are in good
agreement with earlier measurements in citrate buffers, it shows
these dendritic contrast agents have very less interactions with
blood plasma proteins. The largest MRI contrast agent G5 with 64
Gd(III) ions gives lowest concentration detection limit would make
G5 potentially the best dendritic MRI contrast agent (Langereis et
al., 2006).
6. Conclusions
Although dendrimer drug delivery is in its infancy, it offers
several attractive features. Dendrimers expect to be a potential
polymer for biomedical, pharmaceutical and biopharmaceutical
fields in the 21st century. Easily controllable features of dendrimers
such as their size, shape, branching length, their surface functional-
ity allow to modify the dendrimers as per the requirements, makes
these compounds ideal carrier in many of the applications. Still
toxicity problems may arise, but they will be resolved by mod-
ifying dendrimer structure. Future work is necessary to find out
cost-effective synthesis strategies and the relationship between
dendrimer and drug molecules for successful commercialization
of this technology.
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