STARCH-CHITOSAN MODIFIED BLEND AS LONG-TERM CONTROLLED DRUG RELEASE FOR CANCER THERAPY STARCH-CHITOSAN MODIFIED BLEND AS LONG-TERM CONTROLLED DRUG RELEASE FOR CANCER THERAPY

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STARCH-CHITOSAN MODIFIED BLEND AS LONG-TERM CONTROLLED DRUG
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Pak. J. Biotechnol. Vol 15 (4) 947-955 (2018) ISSN Print: 1812-1837 www.pjbt.org ISSN Online: 2312-7791
STARCH-CHITOSAN MODIFIED BLEND AS LONG-TERM CONTROLLED DRUG
RELEASE FOR CANCER THERAPY
Ismaeel Moslam Alwaan1*, Muhannad Ahmed1, Khalida K. Abbas Al-Kelaby2, Zuhair Saleh Mehdi
Allebban3
1Faculty of Engineering, University of Kufa, Najaf, Iraq, 2Faculty of Pharmacy, University of Kufa,
Najaf, Iraq, 3Faculty of Medicine, University of Kufa, Najaf, Iraq.
E.mail: *Ismael.alsallami@uokufa.edu.iq; or ism10alw@yahoo.com
Article received 13.7.2018, Revised 11.12.2018, Accepted 18.12.2018
ABSTRACT
The conventional drug delivery system has serious limitations. These limitations can be overcome by using the
smart materials to produce an effective drug delivery system. Corn starch cross-linked chitosan using potassium
persulfate as an accelerator with different ratios of corn starch/chitosan blends were prepared as smart materials for
drug delivery system. The hydroxyurea drug was loaded in smart materials blends to evaluate the efficacy of new
materials on therapy of Rhabdomyosarcoma cancer cell line (RD). The FT-IR results revealed that the reaction was
occurred between blends materials (starch/chitosan). SEM tests illustrated that the different morphologies were
obtained in blend films surface. The rate of drug release was sensitive to pH and significantly increased at pH 2.2 as
compared to pH 7.4. Furthermore, the percentage of swelling was higher in acidic solution than in neutral solution.
It was concluded that the starch/chitosan smart materials may be suitable for medical applications like drug delivery
system to RD cell line as confirmed by the availability and morphology test of RD cell line.
Keywords: Starch, Chitosan, Drug Release, Rhabdomyosarcoma Cancer, Hydroxyurea
1. INTRODUCTION
Cancer refers to uncontrolled cell division
that can attack healthy tissue (Jemal et al., 2011).
Several different methods using to treatment can-
cer include chemotherapy, radiation therapy, sur-
gical intervention and other techniques. Each met-
hod has its advantages and disadvantages; the
choice of the appropriate method depends on the
type of cancer, the location and the patient's con-
dition. There is a highly targeted method such as
radiotherapy and surgery, it can be removed the
malignant tumor completely. However, when can-
cer spreads in multiple places of the body these
methods become ineffective. Moreover, these met-
hods need good patient health (Helleday, et al.
2008).
Chemotherapy is also widely used throughout
the world to control cancer and its growth by tak-
ing medications. These drugs prevent cell division
in the body, which leads to stopping tumor growth
(Helleday, et al., 2008). However, these drugs not
only kill cancer cells but also healthy cells. There
are many harmful side effects of chemotherapy
drugs such as toxicity, include loss of weight, loss
of hair, the sensitivity of skin, vomiting, and the
impact of immunity. There is another problem in
the chemotherapy drugs that these drugs do not
reach enough to the tumor tissue, which leads to
the increase in the dosage. However, this leads to
increased risk of healthy tissue (Wang et al.,
2012). Therefore, it is necessary to develop che-
motherapy drugs that have the ability to target the
tumor tissue and reduce side effects.
The drug delivery system depends on two
important pathways for unique performance inclu-
ding active and passive pathways. The passive eff-
ect is attributed to enhanced permeation retention
(EPR) effect. Typically, the vessels of the tumor
region contain high leakage to food and oxygen to
maintain rapid growth of cancer cells. However,
this technology remains specific due to there are
certain tumors or certain part of the tumor that do
not have such this effect. To overcome such diffi-
culties can be used active targeting (Peer, et al.,
2007). Compared to passive targeting and free
drugs, active targeting can increase the chance of
tumor uptake, improve therapeutic properties and
reduction undesirable effect to normal cells (Ruo-
slahti, et al., 2012). Moreover, particles can be
activated when they reach their target area, for
example, their high susceptibility to pH (Muller,
et al., 2004).
Some requirements for particles that are used
as drug transporter should be provided such as
biodegradation, biocompatibility, mechanical pro-
perties and drug compatibility (Alimohammadi, et
al., 2014). Starch is used as a drug delivery system
because of its unique properties that enable it to
work in this field such as improving drug stability
and solubility, decreasing side effects and drug
toxicity, storage stability and excellent biocompa-
tibility (Najafi, et al., 2016). But the main obstacle
of starch is that it is poor in dimensional stability,
mechanical properties and processability of its end
products. Therefore, decided to use starch in the

Alwaan I.M. et al., Pak. J. Biotechnol.
948
compound material (Bajer and Kaczmarek, 2010,
Bajer and Kaczmarek, 2010) demonstrated the
process of interaction between starch and chitosan
in their blend (Bajer, et al., 2010). Therefore, it is
expected that a polymeric composite material will
be produced delivery system for cancer therapy
and that the release of the drug will be slow and
controlled (Subramanian, et al., 2014).
Chitosan results from nature, for example its
existence in the crabs, lobsters, fungal cell walls,
shrimps and in the insect exoskeletons (Dash, et
al., 2011, Testing 2013). Excellent performance
results from this material (biocompatibility, biode-
gradability, high safety and minimal toxicity with
large numbers of biological activities) which gives
these materials a great chance to develop them to
work in wider and more useful applications (Sub-
ramanian, et al., 2014, Dash, et al., 2011). Chito-
san are the drug vehicle with extensive impro-
vement possibility and have the benefit of contro-
lled and slow drug release, which increases drug
stability and solubility, reduces toxicity and enha-
nces efficacy (Aruna, et al., 2013).
Most kinds of sarcoma that affect the soft tiss-
ues during childhood and adolescents are under
20 years old Rhabdomyosarcoma (RMS) (Ognja-
novic, et al. 2009). Because clinical trials are slow
and difficult to perform, most studies on this type
have been conducted in laboratories for this can-
cer type (Pappo, et al., 1999). There are two main
types of RMS, alveolar and embryonal (Hinson, et
al., 2013). Embryonal RMS contains about 15
types of different cell line and one of them RD
cell line (Kang, et al., 2011).
In the present study, corn starch/chitosan ble-
nds loading hydroxyurea was produced and their
physicochemical and morphological characterizat-
ions were investigated. The swelling and control-
led drug (hydroxyurea) release of blends was stu-
died at different pH value. In vitro anticancer
activity was evaluated against RD cancer cells in
terms of percentage cell viability through MTT
and morphological cell density. It is worth menti-
oning here that previous studies have not explored
these systems for Rhabdomyosarcoma cancer cell
line therapy using hydroxyurea drug and the study
of these systems at different pH media is still
needed. Therefore, this is the first attempt to ass-
ess the suitability of these systems for Rhabdo-
myosarcoma cancer cell line therapy.
2. MATERIALS AND METHODS
2.1. Materials: The supplier of native cornstarch
was Inter-scince (ST. Louis, MO – S 4180) Fra-
nce. The amylase and humidity content were 27%
and 8–9% respectively. The mean particle size of
cornstarch was 12μm. The chitosan (90% deacety-
lated, Mw= 161.16) was supplied by micxy reag-
ent, Beijing, china. Hydroxyurea capsules USP
500mg supplied by Hydrea, india. Potassium per-
sulfate (K2S2O8) was purchased from Merck, New
Jersey, United States. Poly (vinyl alcohol) (PVA)
provided by Sinopharm Chemical Reagent co.,
china. Acetic acid glacial (CH3COOH) was
purchased from Himedia, India.
2.2 Preparation of drug loaded particles: Chit-
osan/ corn starch blends at different concentra-
tions (1/0, 1/3, 1/1, 3/1 and 0/1) were prepared
and the compositions are reported in Table 1.
The solution of chitosan was prepared by disso-
lving specific weight of chitosan in 20ml of aque-
ous acetic acid (2%) solution at room temperature
and stirring for three hours to obtain homogenous
solution (Singh, 2016). The solution of starch was
prepared by dissolving desired amount of starch in
20ml of aqueous acetic acid (2%) solution at room
temperature and stirring for one hour until it bec-
ame homogenous solution (Subramanian et al.,
2014). The two solutions of starch and chitosan
were mixed together and 3ml of potassium per-
sulfate (K2S3O8) solution (6g K2S3O8/100ml disti-
lled water) as a catalyst was added to mixture and
heated at 60oC for 30-45min (Testing, 2013,
Aruna et al., 2013).
Then, the solution was cooled to room tempe-
rature, solid materials were collected and washed
many times to remove the potassium persulfate
catalyst and acetic acid. The hydroxyurea drug
was then added to starch/chitosan solution and
well mixed using magnetic stirrer. Moreover, the
PVA solution was prepared by mixing 60mg PVA
/100ml distilled water at 85-95oC.The 3% of PVA
solution (from the total starch/chitosan blends as
dry weight) was added to starch/chitosan solution,
dispersed the particles using ultrasonic and fina-
lly, the samples were dried at room temperature to
gain the new drug (Pandey et al., 2015).
Table 1: The compositions of starch /chitosan blends.
Materials
Ao
A1
A2
A3
CH
Starch (g)
2
6
4
2
-
Chitosan (g)
-
2
4
6
2
Acetic acid (%)
2
2
2
2
2
K2S2O8((ch.+st.)*0.1)(g)
-
0.8
0.8
0.8
-
PVA (w/v) (%)
3
3
3
3
3
Hydroxyurea (%)
25
25
25
25
25

Vol. 15 (4) 2018 Starch-chitosan modified ………
949
2.3 Fourier Transform Infrared Spectroscopy
(FT-IR): The samples were mixed with KBr pow-
der/sample and tested by pellet method. The con-
ditions were 32 scans and the resolution was 4
cm−1 while its wave number range between 400-
4000cm−1 (Sacithraa, et al., 2013).
2.4. Scanning electron microscopy (SEM): The
shape, surface morphology, size of corn starch
and drug loaded film were examined using a scan
electron microscope (SEM) at 10kV as an accele-
rating voltage. The dry specimens were scattering
on dual twig tape stable on circular copper stubs,
then painted with gold film (∼15nm) to prevent
electrical charge accumulation (Najafi and Bag-
haie, 2016).
2.5 Evaluation of in vitro drug release: The
drug release study through the starch, chitosan
and cross-linked films was carried out in differ-
ent pH solutions at 37ºC under unstirred condi-
tions. A known weight of the film i.e. 0.05g was
put in 50ml phosphate buffer solution at different
pH values (2.2 and 7.4) (A. Hinson, et al., 2013).
The behavior of drug release was studied in the
simulated gastric and intestinal pH conditions
(Virpal, 2014). At predefined intervals of time
the electrical conductivity was measured by con-
ductivity meter (EC214, HANNA Instrument
Inc., Romania) to calculate the percentage of
drug in this solution. The amount of drug relea-
sed through the starch, chitosan and cross-linked
beads was calculated by prepared suitable cali-
bration curve.
2.6 Swelling studies: The swelling behavior of
the cross-linked films was studied at different pH
solutions (7.4 and 2.2) to understand the mech-
anism of transfer of the drug into the cross-linked
films. A specified weight of the dry sample was
placed in the solution at 37ºC. At different periods
of time the film was got out from the solution and
the excess solution on the surface of sample was
removed by tissue paper. The weight of the film
was determined using sensitive balance (± 0.0001
g) (TP-214, Denver Instrument Germany) and the
percentage of swelling was calculated by using
the following equation (Kumari and Kundu,
2008, Singh and Kumari, 2014):
where, Ws is the weight of sample after swelling
and Wd is the dry weight of the sample.
2.7 Cytotoxicity assay: The cytotoxicity of diffe-
rent composites was evaluated by MTT assay.
This assay depends on the change tetrazolium to
formazan during interaction with mitochondrial in
cultured cells, and the formazine amount produ-
ced refers to number of living cells (Sylvester,
2011).
Cells were placed into 96 well plates at 1.0 x
l05 cells/ml concentration. After the incubation at
37°C for 24-48 hrs, a monolayer of RD cells was
formed at 80-100% concentration. Different conc-
entrations of drug composites (1, 10, 100, 500 and
1000µg/ml) were put into wells at a total volume
of 100µl in triplicate for each well except control
cells (Sun, 2015). Before MTT assay and morpho-
logical cell assessment, all samples were incu-
bated with media for 24 hrs to facilitate hydroxy-
urea release into the media (Pandey, et al., 2015).
After 24 hr incubation at 37°C in 5% CO2, The
RD cells were washing by PBS solution to remove
drug composites or standard anticancer drugs used
that may be interacts with MTT reagents. Follo-
wed by the addition of 100µl maintenance media
for all wells with 20 µl MTT reagent (Ravikumara
and Madhusudhan, 2011).
The plates were incubated for 4 hrs at 37°C,
under flow of 5% CO2. The formazan particles
were produced during the MTT reaction with
mitochondrial enzymatic for living RD cells.
After this incubation, diluted dimethylsulfoxide
DMSO (1:1) in isopropanol was added that solu-
blized formazan salt particles. By using ELISA
reader, the optical density was calculated at wave-
length 490-630nm (Sun, 2015, Ravikumara and
Madhusudhan, 2011).
3. RESULTS AND DISCUSSION
3.1. Fourier Transform Infrared Spectroscopy (FT
-IR): The results of FTIR for corn starch, chitosan
and cross-linked film are shown in Figure 1. The
results of FT-IR for pure starch revealed that the
peak observed at 3416 is assigned to O–H group
(Nnamonu et al., 2012), peaks at 2928cm-1 was
attributed to C–H stretching bond (Subramanian
et al., 2014), peaks observed at 1649cm-1 was
assigned to the C=O stretching (Singh and
Kumari, 2014), peaks in the range of 1365-1454
cm−1 were due to the H–C–H, C–H and O–H
bending modes (Zeng et al., 2011). There were
several absorbances bands at 1157, 1083, and
1018cm-1 which were referred to C-O stretching
bond (Zeng et al., 2011, Winarti et al., 2014).
The results of FT-IR for pure chitosan rev-
ealed that the main bands in the range from
3750cm-1 to 3000cm-1 were due to O-H groups str-
etching vibrations, which interfered to the N-H
stretching vibration (Marchessault et al., 2006),
the peaks obtained at 2920cm−1 is refer to C–H
stretching (Subramanian , et al., 2014). The peak
at 2875cm-1 represented H-C-H group (Singh and

Alwaan I.M. et al., Pak. J. Biotechnol.
950
Kumari, 2014). The two peaks at 1635cm-1 and
1595cm-1 were noticed and referred to amide I
and amide II, respectively (Bajer and Kaczmarek,
2010; Wang et al., 2007), methyl and methylene
bending vibrations groups were shown at 1421.54
cm−1 and 1379.1cm−1 respectively (Mano et al.,
2003). The peak located at 1155.36cm−1 is related
to asymmetric vibrations of C-O-C which is pro-
duced during chitosan deacetylation (Silva et al.,
2012) and the band at 1083cm−1 was due to the
glycosidic bonds (Bajer and Kaczmarek, 2010).
In the cross-linking film spectra, the significant
shrinkage of the amine peak intensity at 1595cm-1
and it shifts to a lower value of absorption bands
(Bajer and Kaczmarek, 2010). Moreover, the con-
centration of NH2 group was highly decrease as
revealed by decrease its intensity (Subramanian et
al., 2014, Kumari et al., 2016). It may be attribu-
ted to the reaction between NH2 of the chitosan
and OH- of starch was occurred. Moreover, the
small shift of NH2 peak to a lower value indicated
that the interaction of OH- group and NH2 was
occurred (Bajer and Kaczmarek 2010).
Figure1: FT-IR for corn starch, chitosan and cross-linked film.
3.2 Scan Electron Microscope (SEM): SEM
images of corn starch-chitosan films were shown
in Figure2. Figure2a illustrated the microstructure
of (3/1) starch/chitosan film surface and it was
very rough and withered large granules of starch.
Whereas, the addition of 50%w/w of chitosan into
the starch film matrix (Figure 2b) reduced the gra-
nules and improved the surface structure (homog-
eneous and smooth matrix). However, when the
concentration of chitosan was further increased to
(1/3) starch/chitosan, the chitosan was not fully
blended with the starch as seen in Figure 2c and
this is consistent with S. Othman and R. Shapi
2016.
Figure 2: SEM image for cross-linking film for (a)
(3/1) starch/chitosan (b) (1/1) starch/chitosan (c)
(1/3) starch/chitosan.

Vol. 15 (4) 2018 Starch-chitosan modified ………
951
3.3 In Vitro Drug Release Studies: Drug release
profiles of Ao, A1, A2, A3, CH blends in acid and
neutral media are shown in Figure 3 and 4. It was
observed that the release characteristics of chitos-
an-starch film depend on both composition of film
and pH of release medium (Virpal, 2014). The rel-
ease behaviors of hydroxyurea were shown simi-
lar initial release behavior. The release of the drug
was fast in the first hour in both acid and neutral
solution, while it was moderate release over 9
hours and finally, a nearly constant release of the hyd
-roxyurea was observed for remaining 25 hours.
The drug release rate was maximum for first hour
and it may attribute to present some of the drug on
the outer surface of the films and it was released
when the films were immersed in the fluid media
(Raizaday et al., 2015).
Figure 3 shows that the quantity of drug relea-
sed at pH 7.4 was decreased with increasing chito-
san amount in the blend. This may be due to the
formation of a dense matrix as a result of increas-
ing chitosan in the blends which led to reduce the
swelling of the films (Kumari and Kundu, 2008).
Therefore, the solvent penetration was reduced
and hence reduced the amount of drug released.
This result is consistent with (Wang et al., 2007).
Therefore, the drug release at pH 7.4 is largely dep-
ending on solvent diffusion through the blends.
The reason may be the protonation of amino gro-
ups in chitosan is lacked at pH 7.4 (Kumari et al.,
2016, Lim et al., 2013).
Figure 3: Drug Release of Ao, A1, A2, A3, CH at pH
=7.4.
In acid media, it was observed that the rate of
drug release was increased proportionally as boo-
sting in the release time. The reason of raise in
drug release with raising in time may be attributed
to the hydration of starch in acid media and the
other reason is protonation of amino groups of
chitosan to give highest swelling of starch/
chitosan blends as compared with neutral media
(Raizaday et al., 2015, Bakain et al., 2015). The
mechanism of the drug release through the com-
posites films was the penetration of media in to
drug vehicle, dissolve the drug and then it is diff-
usion out drug vehicle through the same path.
Thus, the drug release takes place from the comp-
act matrix due to increase in swelling of the beads
and penetration of the solvent over a period of
time (Kumari et al., 2016).
Figure 4: Drug Release of Ao, A1, A2, A3, CH at pH
=7.4.
The results indicated that the release of hydro-
xyurea from chitosan/starch film is pH dependant,
the hydroxyurea released faster in acidic environ-
ment than neutral environment as a consequence
of starch hydrolysis and protonation of amino gro-
ups in acidic environment. This result is consistent
with (Lim et al., 2013, Jaimes et al., 2014). From
our observation the acidic environment effect on
starch/chitosan blends was higher than neutral env-
ironment because the chitosan is very sensitive to
acidic environment and the protonation of amino
groups in chitosan occurred in acidic environ-m
ent.
3.4 Swelling Studies: It is not feasible to record
the swelling data of pure starch. The chitosan/
starch blends have better strength as compared to
pure starch and they are capable to longer time in
swelling media. Therefore, the swelling studies
are performed only on cross-linked chitosan-sta-
rch films and chitosan film (Singh and Kumari,
2014). When the composite films are put in the
swelling media, the solution starts to diffuse ins-
ide the films and consequently try to swell (Kum-
ari and Kundu, 2008).
The results indicated that the percentage of
swelling in pH =7.4 increased with passage of
(Figure 5) time. The rate of swelling of biode-
gradable cross-linked films increases linearly for
first four hours followed by almost a constant
swelling for rest of the studied time period. The
results revealed that the percentage of swelling is

Alwaan I.M. et al., Pak. J. Biotechnol.
952
also influenced by the content of chitosan and
starch in the film. The swelling percentage of the
cross-linked films was lower with increase the
concentration of chitosan in the blend and this
may be due to the formation of a dense matrix in
pH=7.4. Similar results were found by ( Hinson et
al., 2013, Singh, 2016).
Figure 5: Swelling behavior in pH =7.4 for corn
starch/chitosan composite and pure chitosan.
It was observed that the swelling percentage of
film in pH 2.2 solution was higher than pH 7.4
solution (Figure 6). It was concluded that the
films in pH less than 6 have new structure which
the protonation was occurred in acid media for
amino and imine groups in films surface and
therefore, it followed relaxation of the polymeric
chains (Lim et al., 2013). Initially, amino and
imine groups at the film surface were protonized
and it leads to separate the hydrogen bonds of
amino and imine groups and this lead to diffuse
the solution inside the film faster as compared to
pH 7.4 to separate the hydrogen bonding inside
the film network. Further, the percentage of swe-
lling significantly faster at pH 2.2 as compared to
pH 7.4 (Kumari and Kundu, 2008).
Figure 6: Swelling behavior in pH 2.2 for native
chitosan and its composite.
3.5 In Vitro Anticancer Evaluations Using RD
Cell Line: The viability assays provide report on
cell killing and metabolic activities. The MTT
assay was applied in this work because it is effect-
tive and quick process for examining mitochon-
drial activity (Sabudin et al., 2012).
After loading hydroxyurea drug onto pure
starch, pure chitosan and starch/chitosan film, the
toxicity of the drug was examined via incubation
with RD cell line. The MTT assay was performed
to show the toxicity of the drug vehicles (pure
corn starch and pure chitosan) (Sun, 2015). Figure
7a showed the cells treated with pure corn starch,
pure chitosan and pure hydroxyurea, which gave a
good idea about the health of cells in pure starch
and pure chitosan as biocompatible materials. The
most-dead cells can be seen at pure hydroxyurea,
which means they can be used as an effective
agent to treat this type of cancer cell (Pandey et
al., 2015), while cells health in the pure starch and
pure chitosan did not affect and remind without
change. This meant that pure corn starch and pure
chitosan have not toxicity onto RD cancer cells.
On the other hand, the results of curing the cancer
cells with drug/starch, drug/chitosan and drug/sta-
rch-chitosan (St:Ch) blend with different percen-
tage of St:Ch revealed that the cancer cells were
effectively killed in a certain concentration range
and its behaviors were similar to behavior the pure
hydroxyurea drug. The cell viability with using
drug/starch, drug/chitosan and drug/starch-chito-
san blend with different percentage of St:Ch can
be inferred by the number of dead cells, as show
in Figure7b. Similar results were found by (Pand-
ey et al., 2015).
(a)
(b)
Figure 7: MTT assay of (a) The effect of pure hydroxy-
urea, pure corn starch and pure chitosan on toxicity of
RD cell line (b) The effect of pure hydroxyurea drug
and its blends with chitosan, starch and crosslinking
film with different percentage of St:Ch on toxicity of
RD cell line

Vol. 15 (4) 2018 Starch-chitosan modified ………
953
The death of RD cells for different vehicles
such as pure starch, drug/starch-chitosan (1/1)
vehicles and free hydroxyurea drug was estim-
ated. The results showed that the slow rate of dead
cell was obtained in drug/starch-chitosan (1/1)
vehicles. Moreover, the cytotoxic effects of films
increased with an increase in concentration of
drug (Figure 8). The toxicity was increased as the
concentration of drug raised from 1 to 1000μg/ml
(Ravikumara and Madhusudhan, 2011).
Figure 8: Morphology of RD cell density (a) without any drug (b) after treatment with pure starch (c) after treatment
with crosslinking film in 100 μg/ml (d) after treatment with free hydroxyurea drug.
4. Conclusions:
The starch/chitosan vehicles were successfully
prepared using potassium persulfate (K2S2O8) as a
catalyst to product biocompatible composite films.
The hydroxyurea was loaded these vehicles and
the products were used as an anticancer drug. FT-
IR spectra results revealed that the amine peak
intensity was significantly decreased, and it has
small shifts to a lower value of absorption bands.
It was concluded that the reaction between the
NH+3 groups of the chitosan and OH- groups of
starch were occurred. The SEM results illustrated
that the roughness of films surface was increased
with an increase percentage of starch. In addition,
the results indicated that the release of hydroxy-
urea drug from chitosan/starch film is pH depend-
ant and the hydroxyurea drug released faster in
acidic environment than neutral environment and
the drug release at pH 7.4 decreased with an incre-
asing in chitosan amount in the blends. Moreover,
the results revealed that the percentage of swelling
is also influenced by the content of chitosan and
starch in the film. The swelling percentage of the
chitosan/starch films was low with increase the
concentration of chitosan at pH7. The pure starch
and pure chitosan have not toxicity effect on RD
cells and remind without change while drug/
starch, drug/chitosan and drug/starch-chitosan (St:
Ch) blend with different percentage of St:Ch have
toxicity effect and they effectively killed cancer
cells in a certain concentration. It was concluded
that the corn starch/chitosan smart materials may
be suitable for medical applications like drug deli-
very system to RD cell line as confirmed by the
availability and morphology of RD cell line.
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