Materials Science and Engineering C 29 (2009) 22502253

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Materials Science and Engineering C

j o u r n a l h o m e p a g e : w w w. e l s ev i e r. c o m / l o c a t e / m s e c

Preparation of chitosansodium alginate microcapsules containing ZnS nanoparticles

and its effect on the drug release
Zhulai Li a,, Peng Chen b, Xiuzhi Xu a, Xiao Ye a, Jin Wang a
a
b

Faculty of Pharmacy, Fujian Medical University, Fujian 350004, China

The Orthopedic Department, The First Afliated Hospital of Fujian Medical University, Fujian 350004, China

a r t i c l e

i n f o

Article history:
Received 25 January 2009
Received in revised form 1 May 2009
Accepted 18 May 2009
Available online 22 May 2009
Keywords:
Chitosansodium alginate microcapsules
ZnS nanoparticles
Drug release

a b s t r a c t
Chitosansodium alginate microcapsules were prepared in the presence of ZnS nanoparticles via the W/O/W
emulsication solvent-evaporation method. Microscopy showed that the microsphere was about 150 nm and
by the absorption spectra, ZnS nanoparticles incorporated was 4 nm. Aspirin was chosen to investigate the
effect of microcapsules on the drug release. It reveals that comparing with the microsphere without nanoparticles, the release speed of microsphere containing ZnS nanoparticles is signicantly decreased from
complete release at 10 h to 50% release by 50 h. The data of release kinetics for the microcapsules can be well
tted by the classic Higuchi model.
2009 Elsevier B.V. All rights reserved.

1. Introduction
Microencapsulation is dened as a technology of packaging solids,
liquids, or gaseous materials in miniature capsules that can release
their contents at controlled rates under specic conditions [1]. The
choice of materials and the methodology for encapsulation are dependent on the active agent in question and the target application [2].
Over the last few years, absorption of therapeutic microcapsules has
received great attention, for their large surface area suitable for drug
absorption, low thickness epithelial barrier, extensive vascularization
and relatively low proteolytic activity compared to other administration routes [3,4].
Chitosan (CS) is a polysaccharide with well-documented relevant
properties as biocompatibility, low toxicity and biodegradability [5].
Furthermore, it is mucoadhesive and has the capacity of promoting
macromolecules permeation through well-organized epithelia [6].
Obtained from the deacetylation of chitin, CS is formed of d-glucosamine
and N-acetylglucosamine units, whose unions can be destroyed, as previously mentioned, by pulmonary lysozyme [7].
Hydrophilic alginate gels are often reported to be leaky to various
encapsulants, thus would be of interest in controlled release, particularly
when applied to hydrophobic macromolecules which would be released
by restricting diffusion out of the gel matrix [8]. Since alginate gels have
a macroporous structure with pore diameters in the order of 10 mm and
in addition may be unstable [9,10], a membrane coat was applied for

Corresponding author. Tel.: +86 591 83569636; fax: +86 591 22862016.
E-mail address: lizhulai@126.com (Z. Li).
0928-4931/$ see front matter 2009 Elsevier B.V. All rights reserved.
doi:10.1016/j.msec.2009.05.010

stability and to reduce bead permeability. Electrostatic interaction of the

alginate carboxyl groups with a polycationic amine coat, encloses the
encapsulant improving drug retention, or its potential release [9].
The formation of polyelectrolyte complexes of chitosan with polyanions has been frequently used to produce chitosan coatings. For
example, calcium alginate gel beads loaded with hybridoma cells [9],
drugs [11], or haemoglobin [12], have been covered by immersion in
chitosan solution. The resulting calcium alginate/chitosan microcapsules are composed of a calcium alginate gel covered by a membrane
of the alginatechitosan complex, which can be called as interpenetrating polymer network (IPN) hydrogels. The main function of the
calcium alginate gel is to entrap the material to be encapsulated,
rapidly and under mild conditions, giving rise to a spherical particle.
This particle can bind chitosan, forming a strong complex membrane
that stabilizes the ionic gel network and reduces and controls its
permeability. The nal structure of the membrane depends, to a great
extent, on the porosity of the alginate gel structure and its porosity,
the pH, the degree of N-deacetylation of chitosan, and its molecular
weight [13]. Till now, chitosanalginate microspheres have been widely
used for drug release systems [14] and bacteria immobilization [15]
because of their favorable properties, such as mild gelation conditions,
biocompatibility, biodegradability, nontoxicity, and pH dependency [16].
Quantum dots (QDs), such as ZnS and CdSe, have been used widely
as uorescent probes in biological research eld. And many good
results have been achieved in molecular and cell imaging [17], nucleic
acid research [18,19], and drug screening and drug discovery [2022].
QDs have narrow emission bandwidth, symmetrical and tunable proles
according to their size and materials composition, excellent photostability, and broad absorption spectra, making them the best choice as
uorescent probes [23]. The most common method to fabricate stable

Z. Li et al. / Materials Science and Engineering C 29 (2009) 22502253

and water soluble QDs for biocompatible purpose is to chemically attach

a hydrophilic organic surfactant onto the surface of nanocrystals. In
recent studies, functionalization of quantum dots with biomolecules
such as DNA, protein, and small biomolecules have been well established
[24,25].
In order to enhance the encapsulation efciency, nanoparticles
have been added in our microencapsulation experiment. Furthermore,
when nanoparticles (i.e. gold nanoparticles [26]) are encapsulated
together in microcapsules, nanoparticles may act as a diffusion barrier
for drugs to release. Although ZnS nanoparticles have attracted considerable attention due to their potential applications in optoelectronics, electronics, catalysis, molecular recognition systems and other
areas [27,28], no data regarding utilizing ZnS nanoparticles to enhance
the encapsulation efciency and extend the controlled release time
have ever been reported.
Here, we report a process for the microcapsules (IPN typed hydrogels)
using natural polysaccharides including alginate and chitosan. The objective of this study was to characterize the Aspirin-containing microcapsules and determine quantitatively the effect of ZnS nanoparticles
on the average size, the encapsulation efciency, the morphology, the
Aspirin release and the release kinetics.
2. Materials and methods
2.1. Materials
Sodium alginate SG 300 was supplied by Sinopharm Chemical
Reagent Co., Ltd. and the molecular weight of the alginate was 694 kDa.
CaCl2 was obtained from Shantou Guanghua Chemical Factory Co.,
Ltd., Glutaraldehyde from Shanghai SHENBO Chemical CO., LTD. and
chitosan (75, 100 and 300 kDa) was purchased from Golden-Shell
Biochemical Co., Ltd. EP grade Aspirin (acetylsalicylic acid) was supplied
by Shandong XINHUA Pharmaceutical Company.
2.2. Microsphere preparation
The emulsication of internal gelation technique to form chitosan
alginate microspheres has been described previously [29]. Briey,
sodium alginate was dissolved in distilled water at a concentration of
4% (w/v), a pre-calculated quantity of model drug Aspirin was added.
The solution was stirred thoroughly to ensure complete mixing of
the drug. The gelation medium was prepared by dissolving chitosan
(1% w/v) in one percent acetic acid, followed by addition of CaCl2
at the concentration of 4% (w/v). The sodium alginate solution was
added dropwise (about 60 drops/min) into the gelation medium
under stirring with the speed of 1000 r/min. After suspended for half
an hour, the microspheres were rinsed with distilled water, ltered,
and dried in the oven at 60 C.
For the microspheres containing ZnS nanoparticles, 0.1 M Na2S
solution was mixed with the sodium alginate solution, while equimolar
0.1 M Zn(CH3COO)2 solution was added into the chitosan solution.
Other processes for preparing the microspheres are the same with above
description. UVvis absorption spectra and transmission electron microscopy (TEM) were used to check the size and morphology of ZnS nanoparticles, UVvis absorption spectra were recorded using a Shimadzu
UV-365 spectrophotometer in the wavelength range of 250500 nm.
TEM analyses were performed using a JEOL JEM2010 TEM at 200 kV.

2251

with lter paper, rapidly weighed, and reimmersed into the swelling
media. The same operation as above continues until the weights of the
swollen microspheres were constant.
The degree of swelling was determined as follows:
Swelling ratio k =

Ws Wd
100
Wd

where Ws is the weight of swollen microspheres (g) and Wd is the

weight of dry microspheres (g).
To check the release of ZnS particles during the swelling, the real
amount of ZnS before and after swelling was monitored by inductively
coupled plasma (ICP) (Ultima2).
2.4. Release studies
For the dissolution test, 5 g microspheres with and without ZnS
nanoparticles were respectively contained in a steelwool basket and
suspended in a volumetric ask lled with 100 ml PBS PH 7.2. This
ask was put into a 37 C reciprocal shaking bath at 100 rpm. At
scheduled time intervals, 5 ml of the dissolution medium was collected to check the content of drug. Based on the UVvis absorbance at
222 nm, the Aspirin content was then measured, from which the
release rate of Aspirin could be determined. As a comparison, the
parallel experiment of the release of Aspirin bought without any
disposal was done under the same condition.
3. Results and discussion
3.1. The morphology of microcapsules
As seen from the transmission electron micrograph in Fig. 1,
chitosansodium alginate microcapsules containing ZnS nanoparticles were nearly uniform with an average diameter of ca. 150 nm.
Fig. 2a shows the UV absorption spectrum of ZnS nanoparticles, which
was produced by the absorption of semiconductor band gap. The absorbance onset is at about 320 nm and from the absorption spectrum,
the average particle size and particle size distribution can be determined
[30]. The calculated result is presented in the inset of Fig. 2b, which
reveals a narrow size distribution with the average size of 4 nm.
3.2. Swelling studies of microspheres
Porosity and surface area of the polymeric microspheres are the
most important functional characteristics in many applications. Figs. 3
and 4 illustrate the swelling properties of chitosansodium alginate
microcapsules without and with ZnS nanoparticles by weight ratio
method. These curves clearly show that the maximum swelling levels
of microspheres in PBS solution are higher than those in NaCl solution

2.3. Swellabillities of microspheres by weight ratio

The relative weight of dry microspheres (0.5 g) was measured within
a cylindrical glass tube (10 mL), which was covered to avoid liquid
evaporation. Water, 0.9%NaCl solution or Phosphate buffer solution
(PBS) (PH = 7.2) was added into the tube, and were allowed to swell at
room temperature with continuous shaking. The microspheres were
removed at equal intervals from the swelling media, blotted to dryness

Fig. 1. TEM of chitosansodium alginate microcapsules containing ZnS nanoparticles.

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Z. Li et al. / Materials Science and Engineering C 29 (2009) 22502253

Fig. 4. Swelling behavior of chitosanalginate microspheres containing ZnS nanoparticles in water, 0.9% NaCl solution, and PBS PH7.2 solution.

3.3. In vitro release studies

Fig. 2. a) UV absorption spectrum of ZnS nanoparticles in microspheres, and b) the size

distribution.

and water. In three different solutions, the swelling levels of microspheres containing nanoparticles slightly decreased comparing with
those without nanoparticles. The reason can be attributed to ZnS
nanoparticles occupying some space in the microspheres. However,
the time to reach the saturation in the weight for the microsphere
containing nanoparticles was greatly reduced from about 300 min to
60 min. As shown in Fig. 4, for the system with nanoparticles, half of
the saturated weight gain was achieved in the rst 30 min after
immersion, and the saturation weight gain was reached in about
60 min for microspheres in three solutions. After that, swelling was
very slow and nearly stopped after about 300 min. The swelling
was also conrmed from the optical microscopic studies. To further
make sure whether ZnS particles will release during the swelling, we
checked the real amount of ZnS before and after swelling in PBS
solution. Before and after swelling, the amount of ZnS was 14.35%
and 14.05%, respectively, which means that little ZnS particles were
released during the swelling.

The release rate of Aspirin from the microspheres was studied to

investigate the dissolution kinetics. Fig. 5 depicts the release proles of
Aspirin from dry powders, microcapsules without and with nanoparticles in PBS pH 7.2 at 37 C. The results show that Aspirin without
encapsulation release was very rapid and at 4 h, Aspirin was released
completely. When in chitosansodium alginate microcapsules, the
release speed of Aspirin became slow and the drug was fully dissolved
at 10 h. For nanoparticle loaded microspheres, the release rate of Aspirin
was dramatically decreased, the amount of Aspirin being released about
50% until 50 h. The results revealed that ZnS nanoparticles with high
surface have obvious effects to absorb the drugs and that the presence
of nanoparticles occupying pores of hydrogel microcapsules makes the
releasing process difcult. Therefore, microcapsules containing nanoparticles greatly slow down the release speed of drugs. To further verify
this point, we checked the release rate of Aspirin in the microcapsules
with different amount of ZnS nanoparticles. As shown in Fig. 6, the
release rate of Aspirin reduced with the amount of nanoparticles
increasing. In another word, nanoparticles in microcapsules play the
remarkable role in controlling the release of drugs.

Fig. 5. The release proles of Aspirin from dry powders (pure Aspirin), microcapsules
without and with nanoparticles in PBS.

Fig. 3. Swelling behavior of chitosanalginate microspheres in water, 0.9% NaCl solution,

and PBS PH7.2 solution.

Fig. 6. The release proles of Aspirin in microcapsules with different amounts of ZnS
nanoparticles in PBS.

Z. Li et al. / Materials Science and Engineering C 29 (2009) 22502253

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Acknowledgement
The work was supported by the Natural Science Foundation of
Fujian Province (C0610024).
References

Fig. 7. The Aspirin release kinetics tting by Higuchi model.

From the release data in Fig. 5, the lack of linearity of these curves
precludes the possibility of a zero-order kinetics, i.e.
Q
= kt
Q0
where Q is the amount of aspirin that has been released, Q0 is the total
aspirin in microsphere before dissolution, and k is the zero-order
release constant.
Here, we use classic Higuchi model to t our release data [31]:
Q
1=2
= kh t
Q0
where kh is the Higuchi constant. The tting results are shown in
the Fig. 7 and a good linear t occurs, suggesting that Aspirin has
been uniformly distributed over the entire microsphere and the drug
release is a diffusion-controlled process [31].
4. Conclusion
Chitosansodium alginate microcapsules containing ZnS nanoparticles were prepared. The results of drug release reveal that the release
speed of microsphere with ZnS nanoparticles is signicantly decreased,
relative to that of the microsphere without nanoparticles. The data of
release kinetics for the microcapsules can be well tted by the classic
Higuchi model. Our results imply that embedding nanoparticles into
microsphere will potentially be an effective route to control the drug
release.

[1] A.K. Anal, H. Singh, Trends Food Sci. Technol. 18 (2007) 240.
[2] J. Wan, J.B. Gordon, K. Muirhead, M.W. Hickey, M. Coventry, Lett. Appl. Microbiol.
24 (1997) 153.
[3] J.S. Patton, R.M. Platz, Adv. Drug Deliv. Rev. 8 (1992) 179.
[4] A. Clark, Drug Deliv. Syst. Sci. 2 (2002) 73.
[5] S. Hirano, H. Seino, Y. Akiyama, I. Nonaka, Polym. Mater. Sci. Eng. 59 (1988) 897.
[6] P. Artursson, T. Lindmark, S.S. Davisa, L. Illum, Pharm. Res. 11 (1994) 1358.
[7] R.A.A. Muzzarelli, Cell. Mol. Life Sci. 53 (1997) 131.
[8] V.H.K. Robinson, J.R. Lee, Controlled Drug Delivery, Fundamentals and Applications,
Marcel Dekker, New York, 1987, pp. 361.
[9] S. Overgaard, J.M. Scharer, M. Moo-Young, N.C. Bols, Can. J. Chem. Eng. 69 (1991) 439.
[10] A. Polk, B. Amsden, K. De Yao, T. Peng, M.F. Goosen, J. Pharm. Sci. 83 (1994) 178.
[11] P.R. Hari, T. Chandy, C.P. Sharma, J. Microencapsulation 13 (1996) 319.
[12] M.L. Hugnat, A. Grobeillot, R.J. Neufeld, D. Poncelet, E. Dellacherie, J. Appl. Polym.
Sci. 51 (1994) 1427.
[13] A. Bartkowiak, D. Hunkeler, Chem. Mater. 11 (1999) 2486.
[14] A.K. Anal, W.F. Stevens, Int. J. Pharm. 290 (2005) 45.
[15] K.Y. Lee, T.R. Heo, Appl. Environ. Microbiol. 66 (2000) 869.
[16] M. George, T.E. Abraham, J. Control. Release 114 (2006) 1.
[17] M.Y. Han, X.H. Gao, J.Z. Su, S.M. Nie, Nat. Biotechnol. 19 (2001) 631.
[18] H.X. Xu, M.Y. Sha, E.Y. Wong, J. Uphoff, Y.Z. Xu, J.A. Treadway, A. Truong, E. O'Brien,
S. Asquith, M. Stubbins, N.K. Spurr, E.H. Lai, W. Mahoney, Nucleic Acids Res. 31
(2003) e43 1.
[19] H.C. Yeh, Y.P. Ho, T.H. Wang, Nanomed. Nanotechnol. Biol. Med. 1 (2005) 115.
[20] S.K. Sahoo, V. Labhasetwar, Drug Discov. Today 8 (2003) 1112.
[21] M. Yokoyama, J. Artif. Organs 8 (2005) 77.
[22] M. Ozkan, Drug Discov. Today 9 (2004) 1065.
[23] M. Bruchez, M. Moronne, P. Gin, S. Weiss, A.P. Alivisatos, Science 281 (1998) 2013.
[24] X. Michalet, F.F. Pinaud, L.A. Bentolila, J.M. Tsay, S. Doose, J.J. Li, G. Sundaresan, A.M.
Wu, S.S. Gambhir, S. Weiss, Science 307 (2005) 538.
[25] X. Gao, L. Yang, J.A. Petros, F.F. Marshall, J.W. Simons, S. Nie, Curr. Opin. Biotechnol.
16 (2005) 63.
[26] M.K. Lai, C.Y. Chang, Y.W. Lien, R.C.C. Tsiang, J. Control. Release 111 (2006) 352.
[27] W. Vogel, P.H. Borse, N. Deshmukh, S.K. Kulkarni, Langmuir 16 (2000) 2032.
[28] M. Labrenz, G.K. Druschel, T. Thomsen-Ebert, B. Gilbert, S.A. Welch, K.M. Kemner,
Science 290 (2000) 1744.
[29] D. Poncelet, R. Lencki, C. Beaulieu, J.P. Halle, R.J. Neufeld, A. Fournier, Appl. Microbiol.
Biotechnol. 38 (1992) 39.
[30] N.S. Pesika, K.J. Stebe, P.C. Searson, J. Phys. Chem., B 107 (2003) 10412.
[31] T. Higuchi, J. Pharm. Sci. 52 (1963) 1145.