journal of

controlled
release
ELSEVIER Journal of ControlledRelease 39 (1996) 17-25

Preparation and characterization of chitosan microspheres as drug

carrier for prednisolone sodium phosphate as model for anti-
inflammatory drugs
A. Berthold a, K. Cremer b, j. Kreuter a,.
a Institutfiir Pharmazeutische Technologie, Biozentrum-Niederursel, Johann Wolfgang Goethe-Universitdt, Marie-Curie-Strafle 9, D-60439
Frankfurt am Main, Germany
b LTS Lohmann Therapie-Systeme, Lohmannstrafle 2, D-56626 Andernach, Germany
Received 10 February 1995; accepted 16 August 1995

Abstract

Chitosan microspheres were prepared by a novel precipitation process using sodium sulfate as precipitant. Low, medium, and
high molecular weight chitosan was chosen for the formulation of microspheres. The extent of precipitation was controlled by
the concentration of sodium sulfate and monitored by turbidity measurement. The amount of sodium sulfate required for the
preparation of the microspheres depended on the molecular weight of chitosan. The particle size was determined by photon
correlation spectroscopy (PCS) and centrifugal sedimentation. The morphological characteristics were examined using a
scanning electron microscope (SEM). The surface charge was measured by microelectrophoresis. After preparation the loading
property with various anti-inflammatory drugs was investigated using spectrophotometry. The influence of surface adsorption
on the drug modification was controlled by differential scanning calorimetry (DSC). Drug liberation was tested in vitro using
side-by-side diffusion cells with a dialysis membrane made of cellulose acetate. The highest loading (up to 30.5% relative to
the polymer mass) was achieved with prednisolone sodium phosphate (PSP). The adsorbed drug was present in an amorphous
form. The drug release from the microspheres was dependent on the drug-polymer ratio.

Keywords: Chitosan;Microsphere; Precipitationprocess; Prednisolonesodium phosphate; Sustainedrelease

1. Introduction the inflamed lesions. An interesting natural material for

Colitis ulcerosa is a disease of the colon character-
ized by ulcerative, inflammatory lesions. A number of
authors [ 1 - 3 ] have shown that nanoparticles and
microparticles exhibit a tendency to accumulate in
inflamed areas of the body. For this reason we
attempted to investigate the possibility of use of small
particles for the targeting of corticosteroids that are
routinely used for the treatment of colitis ulcerosa to
* Correspondingauthor.
the preparation of such microparticles is chitosan due
to its excellent biocompatibility [4,5]. Therefore, an
additional irritation of the inflamed areas cannot
expected. Chitosan (CS) is a polysaccharide with a
structure comparable to cellulose. CS and cellulose are
both made by linear /3-(1 ~ 4 ) - l i n k e d monosaccha-
ride. The essential difference to cellulose is that CS is
composed of 2-amino-2-deoxy-/3-D-glucan combined
by glycosidic linkages. The primary amino groups lead
to special properties, that render CS very interesting for

0168-3659/96/$15.00 © 1996 ElsevierScienceB.V. All rights reserved

SSD10168-3659(95)00129-8
18 A. Berthold et al. / Journal of Controlled Release 39 (1996) 17-25
pharmaceutical applications. For example, in contrast
to most other natural polymers, CS has a positively
charge and is mucoadhesive [6,7]. Commercially
available chitosan has an average molecular weight of
between 3800 and 2 000 000 and is from 66% up to
95% deacetylated. Its excellent biocompatibility [4,5]
and its availability in large quantities render it a very
interesting material for drug delivery systems.
CS has been used for sustained release systems [8-
12], preparation of mucoadhesive formulations
[4,5,13,14], improvement of the dissolution rate of
poorly soluble drugs [ 15-17], drug targeting [ 18,19]
and enhancement of peptide drug absorption
[5,13,20].
So far, few studies on the preparation of CS micro-
spheres have been carried out. Microspheres were pre-
pared by using a cross-linking agent such as glutaral-
dehyde combined with an emulsion technique
[ 19,21,22]. Other approaches employed an emulsion/
solvent evaporation technique [18] and spray drying
[15].
The objective of the present study was to develop a
simple method for the preparation of chitosan micro-
spheres in a size below 5 Izm, that does not require
complex apparatus and special precautions, and which
are loaded with anti-inflammatory drugs. Since none
of the above quoted methods fulfils all the require-
ments, a novel process for the formulation of micros-
pheres was developed by us, and their physicochemical
characterization was carried out. This new method does
not require the use of an organic solvent. Additionally,
fewer purification steps are necessary than in other
described methods. Furthermore, a high loading capac-
ity combined with a sustained drug release were
achieved.
2. Materials and methods
2.1. Reagents and chemicals
Low, medium, and high molecular weight chitosan
(Mr 70 000, 750 000 and 2 000 000) with a deacety-
lation grade of about 87% were provided by Fluka Bio-
Chemika (Buchs, Switzerland). Acetic acid,
phosphoric acid, potassium hydrogen phosphate,
potassium dihydrogen phosphate, and sodium hydrox-
ide were obtained from Merck (Darmstadt, Germany).
Polysorbate 80 was obtained from ICI Specialty Chem-
icals (Essen, Germany). Hydrocortisone, predniso-
lone, and prednisolone sodium phosphate (PSP) were
purchased from Synopharm GmbH (Barsbtittel, Ger-
many). Triamcinolone acetonide and betamethasone
valerate were supplied by Caesar & Lorentz GmbH
(Hilden, Germany). Ethanol was obtained from Aug.
Hedinger (Stuttgart, Germany). All reagents and sol-
vents were of analytical grade. Water was always used
in demineralized form.
2.2. Equipment
For the preparation of microspheres and for the sus-
pension of lyophilized preparations a bath-type ultra-
sonifier (Sonorex Super RK 106, Bandelin electronic,
Berlin, Germany) was used. Transmission measure-
ments and drug determinations were performed with a
spectrophotometer (Hitachi U-3000, Hitachi Ltd.,
Tokyo, Japan). The pH values were determined with a
Metrohm pH-Meter E512 (Herisau, Switzerland).
2.3. Preparation of chitosan microspheres
Chitosan (0.25% w / v ) was dissolved in an aqueous
solution of acetic acid (2% v / v ) containing 1% poly-
sorbate 80. A solution of sodium sulfate (20% w / v )
was added dropwise (5 ml min ~) during stirring with
a blade stirrer RW 18 (IKA-Werk, Staufen, Germany)
at 400 rpm and ultrasonication. The formation of
microspheres was indicated by turbidity, examined by
transmission measurements at 500 nm. After the addi-
tion of sodium sulfate, stirring and sonication were
continued for another 1 h. The microspheres were puri-
fied by centrifugation for 15 rain at 3000 rpm (GPR
Centrifuge, Beckman Instruments, Palo Alto, CA,
USA). The obtained sediment then was suspended in
water. These two purification steps were repeated
twice. All purified microspheres then were lyophilized
(Lyovac GT2, Leybold Heraeus, Htirth, Germany).
2.4. Size determination and morphological
characteristics
Microsphere sizes were measured by photon corre-
lation spectrometry (PCS) using a BI-200 SM Goni-
ometer Ver. 2.0 (Brookhaven Instruments Corp.,
Holtsville, NY, USA) at a scattering angle of 90 ° and
 A. BerthoM et al. / Journal of Controlled Release 39 (1996) 17-25 19

a temperature of 25°C. A 30 m W He-Ne laser (Melles Table 1

Griot, Cincinnati, CA, USA) was used. Before count- Solubility and loading efficacyof corticosteroidsadsorbed to chito-
san microspheres
ing the samples were diluted with water, that was fil-
tered through a 0.22 /zm cellulose nitrate filter Drug Solubilitya in Surface-
(Schleicher & Schuell GmbH, Dassel, Germany). water(ppm) associatedbdrug
A second size examination was carried out by cen- (% w/w)
trifugal photosedimentation using a centrifugal auto-
Betamethasone valerate 325 0.7-1.9
matic particle analyzer (CAPA-500, Horiba Ltd., Hydrocortisone 280 1.3-3.1
Kyoto, Japan) at a rotational frequency of 500 rpm. Prednisolone 770 0.6-3.8
Before centrifugal analysis the microspheres were sus- PSP 33 )< l 0 4 0.6-30.5
pended in 200 parts ( w / v ) of water. Triamcinolone 20 0.6-3.9
Additionally, microspheres were examined by scan- acetonide
ning electron microscopy (SEM) (S-500, Hitachi, aMerck Index.
USA) for their size, shape and surface characteristics. bRefers to the final drug content of microspheres. The extent of drug
adsorption onto the surface was dependent on the amounts of
employed drug added initially to the particle suspension. The given
2.5. Surface charge (zeta-potential)
ranges are caused by the initially variable drug concentration.

The surface charge of the microspheres was deter-
mined with a Lazer Zee Meter ® Model 501 (Penkem
Inc., Bedford Hills, NY, USA). The measurements
were carried out in phosphate buffers (0.02 M ) ranging
in pH from 2 to 11. Immediately before the determi-
nations the lyophilized microspheres were suspended
in buffer by ultrasonication for 30 min. The concentra-
tions of the suspensions were always 1% ( w / v ) . The
measured values were corrected to a standard reference
temperature of 20°C.
2.6. Drug loading onto chitosan microspheres
natant was measured spectrophotometrically after
dilution with water.
In the second part an extended experiment with pred-
nisolone sodium phosphate (PSP) was carried out
using low (Mr 70 000), medium (Mr 750 000), and
high (Mr 2 000 000) molecular weight chitosan and
drug concentrations ranging between 0.5 and 11 mg
ml ~. Loading condition, sample processing and ana-
lytical method were the same as in the first experiment.
Addition of ethanol was not necessary because of the
good drug solubility in water.
2. 7. Differential scanning calorimetry

In a first set of experiments the loading behaviour of
microspheres was determined using medium molecular
weights (Mr 750 000) of chitosan. The steroidal anti-
inflammatory drugs listed in Table 1 were adsorbed on
the surface of the microspheres. Briefly, solutions with
different amounts of drug (0.5-10.0 mg ml -~) were
added to a 2% ( w / v ) aqueous microsphere suspension.
The following drugs, betamethasone valerate, hydro-
cortisone, prednisolone, and triamcinolone acetonide
required addition of ethanol (30% v / v ) , because of
their low solubility in water. The adsorption experi-
ments were carried out in micro test tubes (Brand,
Wertheim, Germany) at 20°C that were shaken vigor-
ously every 15 min. The pH of the loading medium
was maintained at 4.0 + 0.2 with acetic acid. After 3 h,
the samples were centrifuged for 30 min (Capsule HF-
120, Tomy Seiko Co. Ltd, Tokyo, Japan) and the super-
Differential scanning calorimetry (DSC) of pred-
nisolone sodium phosphate, blank microspheres, phys-
ical mixture and drug-loaded microspheres (20% w /
w drug/polymer) was performed with a DSC 7 PC-
Series (Perkin-Elmer Corp., Norwalk, CT, USA). The
physical mixture was prepared by simple blending in a
mortar and had the same composition as the loaded
microspheres. Samples ( 1.5-7.5 mg) were scanned in
aluminium pans over the temperature range between
180 and 440°C at a scanning rate of 10.0°C min-~.
Nitrogen was used for purging the sample holders at a
flow rate of 20 ml min ~.
2.8. Drug liberation
Drug release from the chitosan microspheres was
examined using Higuchi type side-by-side diffusion
20 A. Berthold et al. /Journal of Controlled Release 39 (1996) 17 25

100
cells (Fischer, Frankfurt, Germany). A dialysis
membrane consisting of cellulose acetate (Medicell \
International Ltd, London, UK) with a molecular 80
weight cut-off of 12 0 0 0 - 1 4 000 was used to separate
the chambers. Prior to the experiment the membrane
was washed with water for 1 h. As the release medium
0.2 M phosphate-buffered saline (pH 7.0) was chosen.
The buffer was kept at 37 _+0.5°C and stirred continu-
# i
ously. The employed suspensions were prepared by
enabling adsorption of the drug to the particles for 3 h.
For this purpose lyophilized preparations were sus- 20 ~ t ~\~
'1,
pended in water and drug was added. The amount of
microspheres in these suspensions was varied between
1 and 7% w / v (Table 2, 2nd column), while the o ~ - , , ~ , , T ~

0,0 0,2 0,4 0,6 0,8 1,0

amount of drug ( 1% w / v ) was kept constant. 1 ml of
the thus prepared suspension was added to 5.0 ml buffer
solution in the donor side of the diffusion cells. The
receiver chambers were filled with 6.0 ml pure buffer
solution. After regular periods of time samples of 0.5
ml were taken out of the receiver side and replaced by
fresh phosphate buffer. The samples were diluted with
water and assayed by spectrophotometry at 247 nm.
3. Results and discussion
3.1. P r e p a r a t i o n o f c h i t o s a n m i c r o s p h e r e s
The production process of the present microspheres
is based on the solubility behavior of chitosan ( C S ) ,
which is poorly soluble in water. Addition of an acid
improves the solubility as a result of the protonation of
the amino groups. The solubility is also dependent on
other anions present in the solution. In the presence of
acetate, lactate, or glutamate CS shows good solubility,
Sodium sulfate % (m/v)
Fig. 1. Turbidity monitoring during the precipitation step employed
to produce chitosan microspheres; (O) low molecular weight chi-
tosan; (11) medium molecular weight chitosan; ( • ) high molecular
weight chitosan ( mean 4- SD; n - 3 ).
whereas phosphate, polyphosphates, and sulfate
decrease the solubility. For this reason, sulfate was
taken for microsphere formulation. Sulfate leads to a
poorly soluble chitosan derivative, whereby micro-
sphere formulation become possible.
Chitosan was always employed at a concentration of
0.25% ( w / v ) . Higher concentrations were not practi-
cal, because the viscosity became too high. As a con-
sequence, a homogeneous distribution of the added
sodium sulfate was not possible, which would have led
to the formation of agglomerates. An addition of poly-
sorbate 80 was necessary to stabilize the suspensions.
Without polysorbate 80 the formation of agglomerates
occurred.
The formation of microspheres was monitored by
turbidity. In Fig. 1 the transmission in relation to the

Table 2
Composition and adsorption efficacy of PSP-chitosan microsphere preparations used in the release study

Ratio (PSP:carrier) Amount of microspheres Employed drug (mg) Unassociated drug (mg) Surface associated" Adsorption efficacyb
(mg) drug (% w/w) (%)

Drug solution 0 101.7 101.7 0 0

1:1 101.0 101.7 77.33 24.13 23.96
1: 1.45 144.6 99.7 62.27 25.88 37.54
1:3 202.0 101.7 51.53 24.84 49.33
1:7 697.6 99.7 20.59 l 1.34 79.35

"Drug content of microspheres at equilibrium.

UPercentageof bound drug.
 A. Berthold et al. / Journal of Controlled Release 39 (1996) 17-25
Fig. 2. Scanning electron micrograph (magnification X 1500) of
chitosan microspheres produced using medium molecular weight
chitosan. The bar represents 5/xm.
added amount of sodium sulfate is shown. The required
sodium sulfate concentration for microsphere forma-
tion increased with increasing molecular weights of
chitosan. A linear correlation between the logarithm of
the molecular weight and the amount of sodium sulfate
40
lO
2 4 6 8 10 12
pH
Fig. 3. Influence ofpH on the zeta-potential of chitosan microspheres;
( • ) low molecular weight chitosan; ( • ) medium molecular weight
chitosan; ( • ) high molecular weight chitosan (mean + SD; n = 5).
21
necessary for the reduction of transmission to 50% was
observed (r = 0.9998; y = 5.79 lx + 3.8283). The rea-
son for this result probably is due to the fact that the
solubility of chitosan is dependent on the number of
positive charges on its surface. A specific amount of
positive charges is necessary for the dissolution of one
chitosan polymer molecule. We assume that high
molecular weight chitosan would require relatively
lower numbers of solubilizing charged groups per mass
of polymer assuming that unprotonated groups could
be concealed in the interior of the rolled-up or folded
macromolecule.
3.2. Size and morphological characteristics
A problem of PCS is that the measurements are less
reliable for large particles with higher standard devia-
tions of the microsphere size [ 23-25 ]. For this reason,
a second technique based on another principle was used
to confirm the results of PCS. Because the results of
both methods were in agreement, PCS was used for
further determinations in the course of the experiments.
The average size was 0.9 ___0.2/~m. No dependence of
the molecular weight on microsphere size was
observed. For resuspension lyophilized microspheres
were mixed with water, and these suspensions were
ultrasonicated for 30 min. No change in average parti-
cle size appeared after lyophilization and resuspension.
SEM studies confirm that the molecular weight of
chitosan has no influence on the size and appearance
of microspheres. A typical scanning electron micro-
graph is shown in Fig. 2. Microspheres are spherical
and regular, with a size between 1.5 and 2.5/xm. The
surface is smooth.
3.3. Surface charge
The zeta-potential has a substantial influence on the
stability of suspensions, the interaction of microspheres
with charged drugs, and also on the adhesion of drug
delivery systems onto biological surfaces. Conse-
quently, investigation of the zeta potential is an impor-
tant part of microsphere characterization.
The chitosan microspheres were charged positively,
although sulfate ions were used as precipitant. This
indicates that only a part of the amino groups are neu-
tralized during microsphere formation. The residual
amino groups would be responsible for the positive zeta
22 A. Berthold et al. /Journal of Controlled Release 39 (1996) 17-25
potential. Moreover, these groups are freely accessible
for interaction with drugs as well. The zeta-potential
was higher at low pH than at high pH (Fig. 3). The pH
dependence was strongest between pH 3 and 6 levelling
off at pH values above 6. This correlated with the dis-
sociation constant of chitosan, which is about 6. The
molecular weight had no effect on the zeta-potential.
This was expected since the deacetylation grade of chi-
tosan was not dependent on the molecular weight.
3.4. Drug loading of chitosan microspheres
In the present study the drug was adsorbed to pre-
viously manufactured empty chitosan microspheres.
Table 1 lists the amounts of surface-associated drug
( w / w ) , calculated as the ratio of the drug content to
the weight of the microspheres. The extent of drug
adsorption onto the surface was dependent on the
amounts of employed drug. The given ranges of drug
adsorption are based on the variable initial drug con-
centration. A higher initial concentration led to a higher
loading efficacy.
Lipophilic steroids were adsorbed in lower amounts
than the hydrophilic derivatives. Several reasons may
account for this. The poor solubility of the lipophilic
substances in water required ethanol addition during
the loading experiment. The ethanol led to two effects,
firstly the microspheres began to swell and secondly
the alcohol shifted the distribution equilibrium of the
drug between the solution and the microsphere surface
towards the solution. Due to these reasons, this method
was not pursued further for the preparation of beta-
methasone valerate, hydrocortisone, prednisolone, and
triamcinolone acetonide microspheres.
The adsorption of PSP was 10 times greater than that
of prednisolone. PSP is a hydrophilic drug, therefore,
ethanol addition during loading was not necessary. In
addition and probably most importantly, PSP has a
negatively charged phosphate group. This phosphate
group presumably led to a powerful interaction of drug
with chitosan and facilitated the formation of an ion
pair.
Fig. 4 shows the adsorption isotherms of PSP onto
microspheres with different molecular weights. During
these adsorption experiments the pH of the medium
was kept at 4.0 + 0.2. At this pH PSP is present in an
anionic form and protonation of the amino groups of
chitosan is still considerable. These conditions lead to
35
30
v
~' /)1
o 20
0,¢ , , , -, . . . . . ~-- F ' i
0 1 2 3 4 5 6
Equilibrium concentration (rag ml 1)
Fig. 4. Adsorption isotherms of PSP onto chitosan microspheres;
( • ) low molecular weight chitosan; ( • ) m e d i u m molecular weight
chitosan; ( • ) high molecular weight chitosan ( m e a n + SD; n = 3).
an optimal interaction between PSP and chitosan. At
lower and at higher pH values the drug loading efficacy
was decreased (data not shown). One problem in these
equilibrium experiments was the flocculation of
microspheres at high drug concentrations. Probably
due to binding of the charged drug the positive charge
of the microspheres was neutralized. This in turn
decreased the repulsive forces between the micros-
pheres. In the experiments displayed in Fig. 4 only
those drug concentrations that did not lead to floccu-
lation were used. The adsorbed amount of drug
depended on the molecular weight of the chitosan.
Adsorption increased with decreasing molecular
weights. Probably due to the lower amount of sodium
sulfate required for microsphere preparation with low
molecular weight chitosan, more amino groups in low
molecular weight chitosan were available for drug
binding.
In Table 3 the correlation coefficients of different
types of adsorption isotherms are shown. The best cor-
relation was obtained with the Freundlich isotherm
[26]. This is an indication that during drug binding
more binding groups on the chitosan microsphere sur-
face may become accessible. Consequently, no simple
flat sorption plateau as with the Langmuir isotherm can
be obtained.
 A. Berthold et al. / Journal of Controlled Release 39 (1996) 17-25 23

Table 3
Correlation coefficients for different adsorption isotherms

L o w molecular weight chitosan M e d i u m molecular weight chitosan High molecular weight chitosan

Langmuir r = 0.6288 r = 0.7135 r = 0.8690

Freundlich r = 0.9706 r = 0.9905 r = 0.9846
BET" r = 0.9574 r = 0.9543 r = 0.9421

~Brunauer-Emmett-Teller.

3.5. Differential scanning calorimetry
DSC is a possible method to determine the modifi-
cation of a drug [ 27 ]. Fig. 5 shows the DSC thermoan-
alysis of blank microspheres, drug loaded
microspheres, physical mixture, and prednisolone
sodium phosphate (PSP). PSP crystals exhibit an
endothermic peak at 341.5°C due to decomposition of
PSP. Blank microspheres showed two peaks, one at
233.4°C and the other at 271.9°C corresponding to the
a- and/3-form of glucosamine [ 28 ]. The DSC curve
of a physical mixture of the microspheres with PSP
exhibited three peaks at 235.9, 275,4, and 341.9°C,
demostrating no interactions of drug with chitosan. In
contrast, in the case of the microspheres with adsorbed
PSP only two peaks at 235.2 and 263.9°C appeared.
No PSP peak was observable. This leads to the assump-
tion that PSP is present on the microsphere surface in
amorphous form. Furthermore, one peak of chitosan
sulfate was shifted from 271.9 to 263.9°C. These obser-
vations indicate an interaction between the amino
group of chitosan and the phosphate group of the drug
such as the formation of an ion pair.
3.6. Drug liberation
The choice of a suitable drug release model is always
a problem. In addition, for the estimation of drug
release from microspheres the separation of drugs and
spheres is necessary. Conventional separation proce-

/ \ ~-,
"\./ \
'X
/' ./ "'\,
/"////
A . ./J " /
Bj
C~ ,,~x.
\ / \

B
i O.O0 2]000 2~o.oo 2¢o.0o 3c{o.oo ~o. oo 3do.oo a~o.oo 4~o.oo
Temperatur ('C)
Fig. 5. DSC thermoanalysis of blank microspheres ( A ) , drug-loaded microspheres ( B ) , physical mixture ( C ) and PSP ( D ) .
24 A. Berthold et al. / Journal of Controlled Release 39 (1996) 17-25

25

20

-~ /J"
~ 2
==
@ 15 .... E

o.

& 10 ////
u. 1

o_ /~ ////
5 .~/

0 ~ , ~ ' t ' t ' T , I m 0 ~ - 1 I T ~ • I i , T

0 60 120 180 240 300 360 0 60 120 180 240 300 360
Time (min) Time (min)
Fig. 6. Effect of the drug-carrier ratio on the release from micros- Fig. 7. PSP-flux through the cellulose acetate dialysis membrane;
pheres producedby medium molecularweightchitosan. The amount drug solution ( • ) ; suspensions of microspheres containing
of PSP that permeated into the receiver chamber is shown adsorbed PSP in concentrations (drug/polymer) of 1:1 (O), 1:1.45
(mean+SD; n=3); drug solution ( • ) ; Suspensions of micros- (IlL 1:3 ( • ) and 1:7 ( v ) .
pheres containing adsorbed PSP in concentrations (drug/polymer)
of 1:1 (O), 1:1.45 (IlL 1:3 ( • ) and 1:7 ( • ) . relevant decrease in drug release and membrane flux
occurred, indicating that drug liberation from the
dures include ultrafiltration and ultracentrifugation. A microspheres became the rate limiting step.
major problem with these techniques is the time period
between sampling and content determination. During 4. Conclusion
this time period the drug release continues. For this
The linearity of the flux and the potential bioadhe-
reason, dynamic dialysis was chosen to obtain infor-
siveness of these microspheres are important properties
mation about the release rates. Therefore, a two cham-
required for the treatment of cronical gastro-intestinal
ber diffusion cell with an artificial membrane was
inflamations. This bioadhesiveness may even be
employed as an initial drug release model. The sepa-
increased in areas of inflamation [ 1-3,29]. This would
ration of free drug, microspheres and adsorbed drug
of course further improve the efficacy of the above
respectively is guaranteed by the molecular weight cut-
chitosan microspheres. Taking together the good bioac-
off of the dialysis membrane.
ceptibility of chitosan, the positive influence on wound
The release profiles of PSP from the microspheres
and ulcer healing [ 30], its positive charge that seems
made of medium molecular weight chitosan are shown
to be very advantageous for bioadhesion to the nor-
in Fig. 6. The compositions of the various preparations
mally negatively charged biological membranes, the
are shown in Table 2. The release rate decreased with
suitable release profile for PSP, and the potentially
an increase in chitosan/drug ratio. In addition, in Fig. 7
enhanced bioadhesiveness of inflamed areas, the above
the drug flux through the membrane is shown. After a
presented microspheres may represent useful tools for
lag and a following equilibration phase the drug flux
the local gastro-intestinal delivery of corticosteroids to
was approximately constant, and only a slight decrease
inflamed gastro-intestinal areas.
was observable. At a high drug/microsphere ratio no
significant difference in release and flux through the Acknowledgements
membrane appeared between drug solution and micro-
sphere-bound drug. However, at drug/microsphere The authors wish to express their thanks to Mr. M.
ratios of 1:3 and especially of 1:7 a very significant and Ruppel, Botanisches Institut, Universit~it Frankfurt,
 A. Berthold et al. /Journal of Controlled Release 39 (1996) 17-25
Germany, who was responsible for the scanning elec-
tron micrograph. In addition, the financial support of
Lohmann Therapie Systeme (LTS, Andernach, Ger-
many), which made this study possible, is gratefully
acknowledged by the authors.
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