Evaluation of the Pro-Inflammatory Potential of
Nanostructured Drug Carriers in Knee-Joints of Rats:
Effect on Nociception, Edema, and Cell Migration
TATIANY JOVITA DE FARIA,1 EDUARDO SOUZA-SILVA,1 DANIEL TEIXEIRA DE OLIVEIRA,1
ELENARA LEMOS SENNA,2 CARLOS ROGÉRIO TONUSSI1
1
Department of Pharmacology, Federal University of Santa Catarina, PO Box 476, Florianópolis, SC 88040-900, Brazil
2
Department of Pharmaceutical Sciences, Federal University of Santa Catarina, PO Box 476, Florianópolis,
SC 88040-900, Brazil
Received 27 October 2008; revised 30 January 2009; accepted 17 March 2009
Published online 19 May 2009 in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/jps.21788
ABSTRACT: Nanocarriers have been developed aiming at drug delivery; however,
the irritating effects of these nanoparticles on naïve or inflamed articular tissues are
not known. Poly(D,L-lactide) (N-PLA), methoxy poly(ethylene glycol)-b-poly(D,L-lactide)
(N-PEG-PLA), and Dynasan 116 (SLN) were used to prepare the nanocarriers. The
average diameter (nm) and zeta potential (mV) of these particles were, respectively, 251
and 33.2, 169 and 22.1, and 105 and 13.0. Naive or carrageenan-primed knee-joints
received 100 mL of nanoparticle suspensions or control solution. Incapacitation and
articular diameter were determined hourly. Synovial leukocytes were counted 6 h after
nanoparticle injection. N-PLA increased the articular diameter and leukocytes, but
did not cause incapacitation. In primed knee-joints, N-PLA caused incapacitation, and
increased the articular diameter and leukocytes. SLN did not produce inflammatory
signals either in naive or primed knees. In primed knee-joints, N-PEG-PLA presented an
intermediate effect characterized by an increase in the articular diameter, and a slight
increase of leukocytes, but not incapacitation. These results suggest that solid lipid
nanoparticles may be safer than polymeric ones, which may be correlated to their
chemical composition and superficial charge. ß 2009 Wiley-Liss, Inc. and the American
Pharmacists Association J Pharm Sci 98:4844 4851, 2009
Keywords: nanocarrier; nanoparticle; arthritis; inflammatory pain; drug delivery
INTRODUCTION among other duties. Due to their submicron size,
nanocarriers can easily penetrate into tissues
Nanostructured carriers can be used either to throughout fine capillaries, across the intercellu-
obtain targeted and sustained delivery of drugs, lar fenestration present in the vascular endothe-
to improve oral bioavailability, to allow the lial lining, and are efficiently taken up by the
administration of hydrophobic drugs by intravas- phagocytes. Generally, nano-encapsulated drugs
cular route, or to protect therapeutic agents from can be delivered into target tissues by using
undergoing chemical or enzymatic degradation, anatomical characteristics, for example, for joints,
muscles, and skin, but they can also be designed
with specific ligands to bind to specific cells,
Correspondence to: Carlos Rogério Tonussi (Telephone:
tissue, or organs in the body.1 3
þ55-48-3721-9491; Fax: þ55-48-3337-5479;
A number of different materials, both synthetic
E-mail: [e-mail])
and natural, have been used to obtain biodegradable
Journal of Pharmaceutical Sciences, Vol. 98, 4844 4851 (2009)
nanoparticles, for example, poly(e-caprolactone)
ß 2009 Wiley-Liss, Inc. and the American Pharmacists Association
4844 JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 98, NO. 12, DECEMBER 2009
NANOSTRUCTURED DRUG CARRIERS IN KNEE-JOINTS OF RATS 4845
(PCL), poly(D,L-lactide) (PLA), poly(D,L-glycolide) ing the cyclo-oxygenase (COX) enzyme. However,
(PGA), and their copolymers. These polymers because of their adverse effects, the use of these
undergo hydrolysis in the body, forming biologi- agents as long-term therapy for chronic inflam-
cally compatible products that are metabolized mation and pain in such diseases as rheumatoid
by the citric acid cycle.4,5 However, many of the arthritis and osteoarthritis is often discouraged. It
problems related to polymer-made nanoparticles is important to note that these adverse effects can
derive from their own cytotoxicity, as well as the be effectively reduced by achieving site-specific/
residues of organic solvents used in the production targeted delivery through new formulation
process. To overcome some of these drawbacks, approaches. These formulations not only restrict
lipidic nanostructures, and more recently solid the drug supply to specific organs but also allow
lipid nanoparticles (SLNs), were introduced as the reduction of the required dose. As a result, the
an alternative to traditional carriers. SLN is a use of new delivery systems such as nanoparticles,
submicron-sized lipid emulsion where the liquid has gained widespread applicability in the man-
lipid (oil) has been substituted by a solid lipid.6 agement of chronic diseases.13 Since the use of
SLN displays several advantages such as good nanomaterials administered into joints has not
physical stability, compatibility with hydrophobic been well studied, the aims of the present
drugs, and lower toxicity when compared to investigation were to evaluate the nociception,
polymeric particles.7 edema and leukocyte chemotaxis caused by
Hydrophobic polymer nanoparticles are rapidly different nanocarriers in naive and previously
taken up by the cells of the mononuclear phago- inflamed rat knee-joints.
cyte system after intravenous injection, probably
due to the adsorption of blood components that
MATERIALS AND METHODS
makes nanoparticles readily recognizable by the
immune system. The phagocytic stimuli induces
Materials
the macrophages to secrete a large number of
substances, including eicosanoids, reactive oxy- PLA (16 kDa) was supplied by Boehringer
gen metabolites, and cytokines8 that can lead Ingelheim (Ingelheim, France). Methoxy poly-
to inflammatory response. These inflammatory (ethylene glycol)-b-poly (D,L-lactide) (mPEG-PLA,
mediators are required for the elimination of 49 kDa, 20% PEG, 5 kDa) was kindly provided by
pathogenic organisms, but are unnecessary and Alkermes (St. Louis, MO). Miglyol 812 N and
potentially deleterious when the ingested parti- tripalmitin (Dynasan 116) were supplied by Sasol
cles are intended to be inert and nonpathogenic.9 (Frankfurt, Germany). Sorbitan monooleate
Thus, it is evident that, for any clinical applica- (Span 80) and polyoxyethylene sorbitan mono-
tion, nanoparticle biocompatibility is crucial. oleate (Tween-80) were obtained from Delaware
The clearance kinetics and in vivo biodistribu- (Sćo Paulo, Brazil) and Beraca (Sćo Paulo, Brazil),
tion of nanocarriers depend on physico-chemical respectively. Pluronic F68 was purchased from
factors like size, surface charge, and hydropho- BASF (Frankfurt, Germany). Soybean lecithin
bicity, which can all be manipulated. In this (Lipoid S75) was obtained from STP Pharm (Sćo
context, therapeutic agents may be loaded into Paulo, Brazil). Carrageenan (multiple type kappa/
nanoparticles that are not identified by the lambda) was from BDH Chemicals Ltd. (Poole,
immune system. Sterically stabilized nanoparti- England, UK).
cles, known as stealth nanoparticles, have been
prepared either by simple adsorptive coating of
Polymeric Nanocapsules Preparation
the particles with hydrophilic surfactants or by
the use of a polymer or lipid covalently bound to a Colloidal suspensions of PLA and mPEG-PLA
hydrophilic block such as poly(ethyleneglycol). nanocapsules were prepared by an interfacial
The coating layer provides an uncharged, extre- deposition process, as described by Fessi et al.14
mely hydrophilic surface that reduces adsorption Briefly, 40 mg PLA or mPEG-PLA was dissolved
of blood proteins, together with an effective in 10 mL of acetone containing 125 mL of Miglyol
steric barrier that temporally prevents them from 812 N, and 40 mg of Span 80. This organic solution
undergoing phagocytosis.10 12 was poured into 20 mL of an aqueous phase
The first-line treatment for many inflammatory containing 0.15% (w/v) Tween-80, under magnetic
diseases is based on the use of nonsteroidal anti- stirring. Acetone was then eliminated by evapora-
inflammatory drugs (NSAIDs) that act by inhibit- tion under reduced pressure, and the final volume
DOI 10.1002/jps JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 98, NO. 12, DECEMBER 2009
4846 DE FARIA ET AL.
of the colloidal suspension was adjusted to PET). Normally, control animals display a PET of
10 mL.15 approximately 10 s, whereas algogenic substances
injected into knee-joints increase this value
only in the affected limb. The injection site was
first shaved and treated with a 10% povidone-
Solid Lipid Nanoparticle (SLN) Preparation
iodine(10) antiseptic solution. The animals
The SLN was prepared using the hot homogeniza-
were gently restrained in a supine position by
tion technique.16 Briefly, the melted lipid com-
the experimenter, and intraarticular injections
posed of lecithin and tripalmitin was dispersed
were quickly performed with a 27-gauge needle.
in a hot aqueous Pluronic F68 solution, which
Nanoparticle suspensions (100 mL) diluted in
was subsequently homogenized by sonication and
sterile saline (1:1) were injected into the right
cooled to room temperature.
naive knee-joint, or 7 days after a carrageenan
(300 mg/ 20 mL) sensitizing stimulus in the same
knee. In order to quantify the inflammatory
Particle Size and Zeta Potential Determination
edema, a micrometer gauge was used to take
the medio-lateral articular diameter (AD) at each
The mean particle and zeta potential of the nano-
hour after nanoparticle injections, just after each
particles were determined, respectively, by photon
incapacitation test. Data are presented as the
correlation spectroscopy (PCS) and laser Doppler
mean difference between the AD values taken
anemometry (LDA) using a Zetasizer 3000HS
hourly after and values taken just before nano-
(Malvern Instruments, Bristol, England, UK). For
particle injections (AD increase in centimeters).
all measurements, each sample was diluted to the
appropriate concentration with filtered distilled
water. Each size analysis lasted 120 s and was
Leukocyte Sampling from Synovial Fluid
performed at 258C with an angle detection of 908.
For measurements of zeta potential, nanoparti-
After the 6-h period of incapacitation and edema
cles samples were placed in the electrophoretic
evaluation, the animals were euthanized for the
cell, where a potential of 150 mV was estab-
leukocyte count in the synovial fluid lavage. The
lished. The j-potential values were calculated
synovial cavity of the knee-joint was opened and
from the mean electrophoretic mobility values
5 mL of synovial fluid was collected for a smear
using Smoluchowski s equation.
slide preparation. The smear was stained by May
Grunwald Giemsa stain and used for differential
(mononuclear and polymorphonuclear (cells/mm3),
In Vivo Studies
MON and PMN, respectively) leukocyte count in
an optical microscope (100 magnification). Next,
Animals
the synovial cavity was washed out with 100 mL
Experiments were performed on 48 female Wistar
of sterile saline containing EDTA anticoagulant
rats (250 350 g), which were housed in tempera-
(5%), and this synovial fluid lavage was diluted in
ture-controlled rooms (19 218C) under a 12 12 h
Turk s solution (1:20) for 5 min. A sample of this
light/dark cycle with free access to water and food.
lavage was pipetted and placed in a Neubauer s
All experiments were conducted according to the
counting chamber for total leukocyte count (TC
ethical guidelines of the International Association
in cells/mm3).
for the Study of Pain,17 and approved by the local
ethical committee for animal experimentation
(CEUA-UFSC).
Statistical Analyses and Data Correlation
Articular Incapacitation and Edema Measurements
All statistical analyses were carried out using
The rat knee-joint incapacitation test is described GraphPad Prism 41. Multiple comparisons for
in detail elsewhere.18 In this test, a computer- unpaired means were made using one-way
assisted device measures the total time that the ANOVA, and for paired means of time-course
rat s hind paw ipsilateral to the injected knee did curves, ANOVA for repeated measures. Signifi-
not contact the surface of a revolving cylinder cance levels were determined by the post hoc
(30 cm diameter; 3 rpm) during 1-min period of Tukey s multiple comparison test. Data were
enforced locomotion (i.e., paw elevation time plotted as mean SE of mean.
JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 98, NO. 12, DECEMBER 2009 DOI 10.1002/jps
NANOSTRUCTURED DRUG CARRIERS IN KNEE-JOINTS OF RATS 4847
RESULTS the N-PLA treated joints, and control group knee-
joints did not present leukocytes.
Preparation and Characterization of Nanoparticles
Averaged diameters, polydispersion index, and Carrageenan-Primed Knee-Joints
zeta potential values of the polymeric and lipid
Figure 2 shows the PET (A), AD (B), and leukocyte
nanocarriers are listed in Table 1. The mean
content (C) recorded before and after injection of
particle size of colloidal suspensions ranged from
nanoparticles into knee-joints primed with carra-
105 to 251 nm. The values for zeta potential varied
geenan 7 days earlier. The control group was
from 33.2 to 13.0 mV. The low polydispersion
injected only with PBS. The N-PLA increased
index values indicated that monodispersed dis-
the PET when compared to the control group
tributions of particles were obtained. N-PLA
time-course curve ( p < 0.001). The maximal PETs
displayed the highest size and zeta potential
registered in the primed condition were: control,
values when compared to other nanocarriers.
26.97 4.23 s; N-PLA, 35.84 4.32 s; N-PEG-
PLA, 22.16 2.01 s; and SLN, 22.92 2.04 s.
N-PLA also induced a marked increase in AD
when compared to the control time-course curve
In Vivo Studies
( p < 0.001). The other nanocarriers also produced
Naive Knee-Joints AD increases, although less significant in this
condition. The maximal ADs registered in the
Figure 1 shows the PET (A) and the AD (B)
primed condition were: control, 0.057 0.020 cm;
recorded before intraarticular injection of nano-
N-PLA, 0.170 0.03 cm; N-PEG-PLA, 0.100
carriers into naive knee-joints, and hourly there-
0.003 cm; and SLN, 0.051 0.011 cm. N-PLA
after. No incapacitation was found following
increased the leukocyte content in the synovial
injection of N-PLA, N-PEG-PLA, and SLN. The
fluid (Fig. 1C) when compared to the control group
maximal PETs registered in the naïve condition
( p < 0.01). On average, the leukocyte counts were
were: control, 14.22 1.16 s; N-PLA, 11.27 0.75 s;
as follows: control, 2140 964 cells/mm3; N-PLA,
N-PEG-PLA, 11.79 1.25 s; and SLN, 10.35
9587 1169 cells/mm3; N-PEG-PLA, 6350
1.10 s. However, N-PLA significantly increased
2278 cells/mm3; SLN, 2600 203 cells/mm3.
the AD when compared to control group time-
Although N-PEG-PLA and SLN have also pro-
course curve ( p < 0.001). The maximal ADs
duced higher cell migration than in naive synovia,
registered in the naïve condition were: control,
they cannot be detected as different from the
0.083 0.002 cm; N-PLA, 0.107 0.026 cm; N-
control. As in the naïve condition, the leukocyte
PEG-PLA, 0.017 0.016 cm; SLN, 0.025 0.012
migration presented a higher proportion of PMN
cm. The N-PLA formulation also increased the
over MON cells.
cellular infiltration into synovial fluid after 6 h
(Fig. 1C) when compared to the control group
( p < 0.001). On average, 6875 1350 cells/mm3,
DISCUSSION
2633 1366 cells/mm3, and 1583 685 cells/mm3
inflammatory cells were found 6 h after N-PLA,
In the present study, it was shown that nano-
N-PEG-PLA, and SLN knee-joint administration,
particles prepared using different biodegradable
respectively. PMN leukocytes were prominent in
materials can produce different degrees of inflam-
matory reaction after direct administration in
the articular tissue. In fact, this reaction appeared
Table 1. Values of the Average Diameter (nm),
to be proportional depending on whether this
Polydispersion Index (p.i.), and Zeta Potential (mV) of
Polymeric and Lipid Nanoparticles tissue was naive or previously challenged by an
inflammatory stimulus. Inflammatory articular
nm p.i. Zeta Potential
stimulus promote the influx of PMN and MON
inflammatory cells, and although PMN cell
N-PLA 251 0.170 33.2
presents a short-life period, mononuclear cells
N-PEG-PLA 169 0.120 22.1
SLN 105 0.279 13.0 remain in the tissue several days after. This
previous stimulation dramatically enhances a
N-PLA, poly(D,L-lactide) nanoparticle; N-PEG-PLA, meth-
second challenge with the same or other sub-
oxy poly(ethylene glycol)-b-poly(D,L-lactide) nanoparticle; SLN,
Dynasan 116 nanoparticle. stances, even in a previously ineffective lower
DOI 10.1002/jps JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 98, NO. 12, DECEMBER 2009
4848 DE FARIA ET AL.
Figure 1. Effects of nanoparticles on articular incapacitation, articular diameter, and
leukocyte content in naive knee-joints. N-PLA, N-PEG-PLA, and SLN were injected in a
volume of 100 mL. Articular incapacitation (A) and articular diameter (B) were taken
hourly through 6 h, and the synovial leukocyte content after 6 h (C). The control group
received only physiological saline (100 mL), and indicate statistical difference from
the control group with p < 0.01 and 0.001, respectively (one-way analysis of variance for
repeated measures followed by Tukey s post hoc test). All results were expressed as the
mean SEM of six rats per group.
concentration. Different mechanisms may be producing the increase of inflammatory response.
involved in this articular sensitization,19 and these Polymeric nanoparticles constituted of hydropho-
remaining mononuclear cells may also be parti- bic materials are fastly cleared from circulation by
cularly important.20 The observation that nano- phagocytosis given that a higher level of opsoniza-
particles produced more intense reaction in the tion occurs on hydrophobic than on hydrophilic
postinflammed condition is particularly relevant surfaces.22 Since the protein surface is hetero-
over tests made in naïve tissues, when the object- geneous and exposes negatively and positively
ive is the therapeutic use of such preparations. charged groups with hydrogen-bonding abilities
The toxicological effects of the nanostructured as well as nonpolar regions, it is conceivable that
systems depend at least on the size, dispersion not only hydrophobic, but also ionic interactions
state, surface charge, shape, chemical composi- are guiding forces for protein adsorption, driving
tion, and surface area. Mostly, the hydrodynamic particle uptake by the immune cells. Concerning
size and surface properties of nanoparticle dis- the nanostructured systems employed in this
persions can exert a dramatic effect on the way an study, N-PLA exhibited the highest zeta potential
organism reacts to it. While the size of nano- ( 33.2 mV) due to the presence of terminal
particles governs its interaction with biological carboxylic groups of the polymer located at the
systems, including absorption, metabolism, and surface of the particle. The reduction of the zeta
excretion, surface characteristics affect agglom- potential observed for N-PEG-PLA can be attrib-
eration, as well as its uptake and translocation uted to the presence of PEG chains on the particle
by the organism.21 Poly D,L-lactide-co-glycolide surface, which cover these PLA carboxylic groups,
(PLGA) and polylactide (PLA) nanoparticles masking the negative charge.23 Due to the chain
activate immune cells (adjuvant activity), also flexibility and electrical neutrality of the PEG
JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 98, NO. 12, DECEMBER 2009 DOI 10.1002/jps
NANOSTRUCTURED DRUG CARRIERS IN KNEE-JOINTS OF RATS 4849
Figure 2. Effect of nanoparticles on articular incapacitation, diameter, and leukocyte
content in primed knee-joints. N-PLA, N-PEG-PLA, and SLN were injected in a volume
of 100 mL, 7 days after carrageenan injection (300 mg/20 mL) into knee-joints. Articular
incapacitation (A) and articular diameter (B) were taken hourly through 6 h, and the
synovial leukocyte content after 6 h (C). The control group received only phosphate-
buffered saline (100 mL), and and indicate statistical difference from the control
group with p < 0.05, 0.01, and 0.001, respectively (one-way analysis of variance for
repeated measures followed by Tukey s post hoc test). All results were expressed as the
mean SEM of six rats per group.
backbone, it is proposed that PEG molecules form polymer nanocarriers. These systems combine
a dynamic molecular   cloud  over the particle the liposome advantages related to the use of
surface, producing a repulsive effect that is biocompatible compounds in their preparation
energetically unfavorable for protein adsorp- with the higher in vivo stability than polymer
tion.11,24 These differences in the surface proper- systems, not undergoing lipid fusion or destabi-
ties may explain why N-PEG-PLA produced less lization after protein adsorption or after lipid
inflammation than N-PLA in the knee-joints. transfer. However, some studies have shown that
Regarding the carriers size, many studies have the use of biocompatible lipid compounds does
shown its influence on the capture by macro- not guarantee a low immune response. In fact, a
phages and consequently the increase of inflam- substantial difference in phagocytosis undergone
matory response. However, in the present study either by PEG-covered SLN (stealth) or naked
the size seems not to be the main factor producing SLN (nonstealth) was found by Bocca et al.25 after
the pro-inflammatory response, since the values incubation of a murine macrophage cell line J774
are not so different among them. An interesting A12 with these nanoparticles.
extension of the present study could evaluate The most frequent lipids used to prepare SLNs
other different sizes for each kind of particle, are triglyceride esters, which do not display
but this variable will never be properly isolated ionizable groups in their structures. Therefore,
since for variate particle sizes their chemical the negative charge of lipid nanoparticles can be
composition will also be modified. attributed to the presence of lecithins used as
Over the last few years, solid lipid nanocarriers hydrophobic stabilizers in their preparation. On
(SLN) have been developed with higher loading the other hand, since the SLN exhibits the lowest
capacities and faster degradation rates than zeta potential value, the stabilization of nano-
DOI 10.1002/jps JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 98, NO. 12, DECEMBER 2009
4850 DE FARIA ET AL.
particles by the hydrophilic, nonionic surfactant 2. Vasir JK, Labhasetwar V. 2005. Targeted drug
delivery in cancer therapy. Technol Cancer Res
poloxamer 188 may also have contributed to
Treat 4:363 374.
the masking of lecithin-negative charge. These
3. Yih TC, Al-Fandi M. 2006. Engineered nanoparti-
surface properties were probably responsible for
cles as precise drug delivery systems. J Cell Bio-
preventing the SLN uptake by macrophages, and
chem 97:1184 1190.
the knee-joint inflammatory response. In fact,
4. Velez G, Yuan P, Sung C, Tansey G, Reed GF, Chin
SLN injection into naïve or primed knee-joints
CC, Nussenblatt RB, Robinson MR. 2002. Pharma-
did not produce evident signals of inflammatory
cokinetics and toxicity of intravitreal chemotherapy
reaction. The lower inflammatory potential of the
for primary ocular lymphoma. Arch Ophthalmol 45:
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nanoparticles for drug and gene delivery to cells
the nanoparticulate system would be applied in
and tissue. Adv Drug Deliv Rev 55:329 347.
a diseased joint, as in a clinical situation. In
6. Müller RH, Radtke M, Wissing SA. 2002. Nanos-
addition, SLN may present advantages over
tructured lipid matrices for improved microencap-
polymeric nanoparticles when the purpose is
sulation of drugs. Int J Pharm 21:121 128.
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7. Mehnert W, Mäder K. 2001. Solid lipid nanoparti-
systems can also improve dermal penetration.26
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Adv Drug Deliv Rev 47:165 196.
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CONCLUSION
and in vivo interactions of cells with biomaterials.
Biomaterials 9:5 13.
The present study has shown that nanostructured
9. Fernández-Urrusuno R, Fattal E, Porquet D, Féger
systems prepared using different biodegradable
J, Couvreur P. 1995. Influence of surface properties
materials can produce different degrees of inflam- on the inflammatory response to polymeric nano-
particles. Pharm Res 12:1385 1387.
matory reaction in articular tissue. This reaction,
10. Bazile D, Prud homme C, Dassoulet MT, Marlad M,
in fact, seemed to be dependent not only on
Spenlehauer G, Veillard M. 1995. Stealth Me-PEG-
whether this tissue was naive or previously
PLA nanoparticles avoid uptake by the mononuc-
challenged by an inflammatory stimulus, but also
lear phagocytes system. Int J Pharm 84:493 498.
on superficial characteristics affected by chemical
11. Gref R, Luck M, Quellec P, Marchand M, Dellach-
composition of the materials used. In addition,
erie E, Harnisch S, Blunck T, Muller RH. 2000.
single articular injection of poloxamer 188-stabi-
Stealth corona-core nanoparticles surface modified
lized lipid nanocarriers or pegylated nanocarriers
by polyethyleneglycol (PEG): Influences of the cor-
did not cause joint inflammation. This new
ona (PEG chain length and surface density) and
approach may offer important advantages for
core composition on phagocytic uptake and plasma
the improvement of some arthritis treatment protein adsorption. Colloids Surf B Biointerfaces
18:301 313.
modalities, such as pulse therapy, intra-articular
12. Shukla R, Bansal V, Chaudhary M, Basu A, Bhonde
injection, and topical applications of encapsulated
RR, Sastry M. 2005. Biocompatibility of gold nano-
anti-inflammatory drugs.
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cellular compartment: A microscopic overview.
Langmuir 23:10644 10654.
ACKNOWLEDGMENTS
13. Bansal SS, Joshi A, Bansal AK. 2007. New dosage
formulations for targeted delivery of cyclo-oxygenase-2
This work received financial support from Coor-
inhibitors: Focus on use in the elderly. Drugs Aging
denaçćo de Aperfeiçoamento de Pessoal de Nível
24:441 451.
Superior (CAPES Brazil) and Brazillian National
14. Fessi H, Puisieux F, Devissaguet JP, Ammoury N,
Research Council (CNPq). We thank Professor
Benita S. 1989. Nanocapsule formation by interfa-
André Báfica (Federal University of Santa Catar-
cial polymer deposition following solvent displace-
ina) for critical reading of the manuscript.
ment. Int J Pharm 55:R1 R4.
15. Faria TJ, Campos AM, Lemos-Senna E. 2005. Pre-
paration and characterization of poly (D,L-lactide)
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DOI 10.1002/jps JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 98, NO. 12, DECEMBER 2009