Physical Evaluations of Freeze-Dried Ca-Alginate Microspheres Containing Ovalbumin Using Different Lyoprotectants Repository - UNAIR REPOSITORY
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List of Poster Participants
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List of Research Pitch Participants
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Oral Abstracts
4-28
Poster Abstracts
29-59
Research Pitch Abstracts
60 - 84
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NAME & TITLE
NO
1.
2.
LIST OF PRESENTERS
DATE : 15 & 16 AUGUST 2015
VENUE : DK2 HALL
PAGE
ASSOC. PROF. DR. MOMKLESUR RAHMAN
Preparation, Characterization and Release Profiles of Insulin-Loaded Double-Walled Poly
(lactide-co-glycolide) Microspheres
PROF. DR. HSIN-I CHANG
Influence of Liposomal Compositions on The Balance between Adipogenesis and Osteogenesis in Bone
_
__
Regeneration
7
3.
DR. DEWI MELANI HARIYADI
Physical Evaluations of Freeze-Dried Ca-AIgmate Microspheres Containing Ovalbumin Using Different
Lyoprotectants
9
4.
PROF. DR. ALLAN COOMBES
Evaluation of Tinidazole-Loaded Polycaprolactone Matrices for The Treatment of Trichomoniasis
11
5.
6.
7.
8.
9.
10.
__
DR. MOHD HANIF ZULFAKAR
Probing the influence of fish oil bigel on reducing imiqulmod-induced inflammation and enhancing
inhibition of non-melanoma skin cancer
PROF. DR. SAYYED A. SAJADI TABASSI
Sustained Drug Delivery System for Insulin using Supramolecular Hydrogels Composed of Tri-block
Copolymers PEG-PCL 20000-1000 Complexed with a -Cyclodextrin
DR. LAY LAY 'CONSTANCE' SAW
Targeted Oelivery of Hypericin for Early Bladder Cancer Detection ana Therapy
DR. VICTORIA PIUNOVA
Biodegradable Nanogel Star Polymers: A Platform for Programmable Macromolecular Self-Assembly and
Cargo Delivery
DR. IGOR KHALIN
BDNF Delivered to the Brain Using PLGA Nanoparticles Induces Functional Recovery After Traumatic Brain
Injury In Mice
_
_
_
DR. ABD ALMONEM DOOLAANEA
Differences Between CTAB and Chitosan in Modifying PLGA Nanoparticles for Gene Therapy
13
15
16
17
19
21
11.
ASSISTANT. PROF. DR. PEDRAM EBRAHIMNEJAD
Novel Surface Modified Polymeric Nanoparticles of Irinotecan, In vitro and In vivo Evaluation
23
12.
DR. LIKVOONKIEW
The Renal Targeting Potentials of the Polymeric Drug Carrier Poly-L-Glutamic Acid (PG) in Normal and
Diabetic Rats
25
13.
DR. GEETHA VISWANATHAN
Novel Diblock Copolymer Vesicles for Anti-Cancer Drug Delivery
27
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LIST OF PARTICIPANTS
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VENUE FOYER DK2&DK3
NAME & TITLE
PAGE
MS. AFNAN SHIHAB AHMED & DR. MUHAMMAD TAHER
Formulation of Hydrogel with Synthetic Polymers (PVA and PEG)
30
MS. CHEAH HOAYYAN
2.
3.
4.
Chemical Characterization of Nano-Polymeric-Zinc (II) Phthalocyanlne and Its In Vitro Phototoxicity Profile
in 4T1 Murine Breast Cancer Cells
32
MS. LEANNE SABRINA DE SILVA
Preparation and Characterization of Span 60-Cholesterol-TPGS Niosomes
34
DR. FUJU YEN
Tumour-targeted Niosomes: Improving Therapeutic Efficacy of Tocotrienois in Adjuvant Cancer Therapy
36
5.
MR. KRISADA WUTTIKUL
Influences of Surfactant Mixtures, Oil Types and Cosolvent Amounts on Formation of Nonionic
Microemulsions for Using as Pharmaceutical and Cosmetic Carriers _
38
6.
MR. MANSOUREH NAZARI VISHKAEI
Drug Delivery Enhancement by Lipid-Nano Encapsulation
40
7.
PROF. DR. MOHD CAIRUL IQBAL MOHD AMIN
Doxorubicin-loaded pH-Responsive Triblock CA-PEI-pArg Polymeric Micelles for Drug Delivery
42
8.
DR. NAGIB AU ELMARZUG!
Spectroscopic Study of Polyethylene Glycol and ctDNA Biocomplexes
44
~
9.
10.
11.
n
v.
12.
13.
14.
15.
DR. NG SHIOW FERN
Topical Bilayer Film containing Olive compound Hydroxytyrosol: Characterisation, Acute Dermal Irritancy
and Efficacious study in Freud's Adjuvant induced- Rat
_
45
MS. NURFAEZATIL FARHANA NORAZEMI
Exploiting Gellan Gum, Hydrocarbon and Fatty Acid for Oral Drug Delivery System
48
OR. PRABAKARAN LAKSHMANAN
Cempedak (Artocarpus Integer) Fruit Skin Mucilage as a Biomaterial: Extraction and Physicochemical
Characterization Studies
DR. RAVI SHESHALA
Development and Characterization of Phase-Inversion Based In Situ Forming Implants of Triamcinolone
Acetonide for Ocular Drug Delivery
_
52
MS. SYAHRIAH FADHILAH ABDUL RAZAK
Drug Permeation Study through Rat Skin: Effect of Storage Conditions of Skin
54
ASSOC. PROF. DR. AHMAD FUAD SHAMSUDDIN
Gelatine-based Parenteral Colloidal Formulation from Shank & Toes of Gallus Gallus Domesticus
56
___
__
_
MR. ATTIQUE UR RAHMAN MUFTI
Formulation, Stability Testing and Biochemical Safety Evaluation of Palm Oil-Based Nano Vaccine of
Pasteurella Multocida
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LIST OF PARTICIPANTS
DATE: 16 AUGUST 2015
VENUE : DK2 HALL
NO
NAME & TITLE
PAGE
1.
MR. ADEEL MASOOD BUTT
Folate Conjugated Chltosan Coated Pluronic F127-TPGS Mixed Micelles for the Co-delivery of siRNA and
Doxorubicin
61
2.
MS. KIEW SIAW FUI
Dextrin Coated Graphene Oxide Nanoparticles as a Controlled Release Drug Delivery System
63
3.
MR. KUE CHIN SIANG
TrkC Receptor Targeted Delivery of PDT Agent Inhibits Immune-suppressive Leukocytes and Enhances
Adaptive Immune Response Before and After Photodynamic Therapy (PDT)
65
4.
MS. MARSITA ABDUL RAZAK
Memantine Hydrochloride Loaded Poly N-Vinyl Caprolactum-ltaconic Acid Nanoparticles for Nose to
Brain Delivery
67
r-
S.
MR. MASOUD REZVANIAN
Alginate Based Composite Bio-Polymeric Dressing Loaded with Simvastatin for
70
Accelerated Wound Healing
MR. MUHAMMAD IRFAN SIDDIQUE
6.
7.
8.
V"
9.
Nanoparticles Enhance Targeted Topical Corticosteroids Delivery into the Skin with Improved Safety for
the Treatment of Atopic Dermatitis: In-Vivo Safety of Hydrocortisone-Hydroxytyrosol Loaded Chitosan
Nanoparticles
72
MS. NAJWA MOHAMAO
Characterization, Biocompatibility and In Vivo Evaluation of Bacterial Cellulose/Acrylic Acid Hydrogel
Synthesized Via Electron Beam Irradiation for Wound Dressing Applications
74
_
MR. NG KHEN ENG
Doxorubicin-loaded pH Responsive Triblock CA-PEI-pArg Polymeric Micelles for Drug Delivery
MR. TEO SIEW YONG
Evaluation of Alkyds as Promising Biomaterials for Drug Nanocarrier Systems
76
78
10.
MS. VOON SIEW HUI
Chitosan Decorated Polymeric Nanoparticles Improved Stealth Properties and Tumor
Selectivity in Photodynamic Cancer Therapy In Vitro and In Vivo
80
11.
MS. WAN MAZNAH WAN ISHAK
Design and Development of Coenzyme QlO-loaded Omega Fatty Acids Topical Nanoemulsions
82
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Physical Evaluations of Freeze-Dried Ca-Alginate Microspheres
Containing Ovalbumin Using Different Lyoprotectants
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Dewi Melani Harivadi. Tutiek Purwariti, Destia Wardhani, and Sisilia Enmawahyuningtyas
Pharmaceutics Department, Faculty of Pharmacy, Airlangga University, Surabaya,
Indonesia, 60286
Corresponding authorl: dewi-m-hQff.unair.ac.id: dewiffua96@vahoo.com
ABSTRACT SUMMARY
The main objective of the research was to
investigate physical evaluations of freeze-dried
ovalbumin loaded-Ca alginate microspheres. This
research used alginate, CaCl2 and lyoprotectants
(maltodextrin, lactose or sucrose). The freeze-dried
microspheres were subjected to micromeritics
properties including moisture content, angle of repose
angle of repose, bulk density, tapped density, Carr’s
index, and Hausner's ratio.
Results showed that the prepared microspheres
were spherical with particle size of microspheres
using lyoprotectant maltodextrin, lactose and sucrose
were 2.32, 2.15 and 2.05 pm respectively. Moisture
content of freeze-dried microspheres contains
maltodextrin and sucrose were about 4%, lower than
microspheres using lactose of about 6.66%. Bulk
densities and tapped densities were in the range of
0.2634 to 0.3232 and 0.3160 to 0.4040 respectively.
Carr's index (Cl) and Hausner's ratio (HR) were in the
range from 15 to 23% and 1.1765 to 1.2988
respectively.
Microspheres
produced
using
lyoprotectant sucrose resulted in good flow properties
with Cl of 15% and HR of 1.1765, whereas other
formulas showed fair flow. Almost all freeze-dried
ovalbumin-loaded alginate microspheres resulted in
small spherical microspheres with less water content
and good flow properties.
Keywords: Microspheres, Ovalbumin, Sodium
alginate, Lyoprotectant, Flow properties.
INTRODUCTION
Ovalbumin is an egg white protein that is used as
a model antigen and could stimulate the formation of
antibodies and improve immunity. Administering oral
antigen is the most effective way to induce
immunological tolerance to protein antigens (Mowat,
1985).
Current study applies ionotropic gelation method
to form hydrogel microspheres using polymer and
crosslinking agent. Sodium alginate is a safe and
biodegradable polymer as well as cheap and most
commonly used as polymer in the microparticles
(Maria et al., 2012). Pb2*, Cd2*, Zn2*, Cu2\ Co2*, Ca2*,
Ba2*, ar.d Sr2* are generally used as crosslinking
agents (Gombotz et al., 1998). Ca2* resulted stable
9
microparticles. Lyoprotectant was commonly added
to stabilize microspheres during preparation and
storage. This research was conducted to study the
potential of ovalbumin-Ca-alginate microspheres
using different concentrations of lyoprotectants such
as maltodextrin, lactose and sucrose.
EXPERIMENTAL METHODS
This research used concentrations of alginate of
2.5% w/v, concentrations of CaCte of 1,5M and 5%
lyoprotectants (maltodextrin, lactose or sucrose). The
freeze-dried microspheres were subjected to
micromeritics properties including moisture content,
angle of repose angle of repose, bulk density, tapped
density, Carr's index, and Hausner's ratio.
Preparation of alginate microsphere using
ionotropic gelation method could be explained as
follows: Alginate solution containing ovalbumin was
sprayed into crosslinking agent CaCfe solution and
was stirred continuously for 2 hours at 1000 rpm. The
microspheres were collected by centrifugation at
2500 rpm for 6 minutes and finally freeze dried 20
hours at -80°C. Alginate microspheres using
maltodextrin, lactose and sucrose were then called
F1, F2 and F3 respectively. The particle size of
microspheres was characterized by optic microscope.
Loading of ovalbumin into microspheres and
encapsulation efficiency was analyzed using protein
quantification assay using UV spectrophotometry.
RESULTS AND DISCUSSION
This technique produced spherical microspheres
with particle size using lyoprotectant maltodextrin,
lactose and sucrose were 2.32; 2.15 and 2.05 pm
respectively. These smaller ovalbumin-loaded Ca
alginate microspheres were suitable for oral
administration (Mishra et al, 2008). Spherical
morphological was caused by ability of lyoprotectant
agent to protect microspheres during freeze drying.
For encapsulation efficiency and loading, all
formulas resulted high encapsulation efficiency of
above 80% and loadings of above 30% (Table 1).
This caused by larger amounts of availability of Ca2*
that crosslinked with carboxylates from guluronic acid
in alginate indicates more ovalbumin was entrapped
within alginate microspheres.
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Table 2: Physical properties of the freeze-dried
microspheres
Table 1: Encapsulation efficiency, protein loading and
moisture content of microspheres
Formula
F1
F2
v
F3
k
Encapsulation
Protem
Efficiency (%)
94.47 ± 2.87
Loading (%)
82.93
* 2.80
81.36 ±8.81
31.33 ±3.94
30.57 ±2.18
Moisture
Content (%)
4.38 ± 0.51
6.66 0.42
37.20 ± 5.52
4.50 ±0.30
*
repose
F1
33.69
F2
30.96
23.96
F3
Moisture content of freeze-dried microcpheres
contains maltodextrin and sucrose were about 4%,
lower than microspheres using lactose of about
6.66% (Table 1). Moreover, the results of angle of
repose were in the range of 33.69°, 30.96° and 23.96°
(Table 2). Bulk densities and tapped densities
respectively were in the range of 0.2634 to 0.3232
and 0.3160 to 0.4040. Carr's index (Cl) and
Hausner's ratio (HR) were in the range from 15% to
23% and 1.1765 to 1.2988 respectively.
Microspheres produced using lyoprotectant sucrose
resulted in good flow properties with Cl of 15% and
HR of 1.1.765, whereas other formulas showed fair
flow.
Angle
Formulatl
on
Code
of
O
Bulk
density
(g/ml)
Tapped
density
(9/ml)
Carr's
index
(Ci) (%)
0.2634
0.3232
0.2686
0.3421
23.00
1.2988
0.4040
0.3160
20.00
15.00
1.2500
1.1765
Hausn
eds
ratio
CONCLUSION
Almost all freeze-dried ovalbumin-loaded alginate
microspheres
resulted
in
small spherical
microspheres with less water content and good flow
properties.
REFERENCES
[1] Gombotz, W.R., Slow Fong Wee, 1998. Protein
Release from Alginate Matrices. Advanced Drug
Delivery Reviews 31, p. 267-285.
[2] Maria, M.S., Scher, Herbert, Jeoh, Tina., 2012.
Microencapsulation of bioactives in cross-linked
alginate matrice. Journal of Microencapsulation, p.
286-295.
[3] Mishra, D. N. and Gilhotra, R. M. 2008. Design
and characterization of bioadhesive in-situ gelling
ocular inserts of gatifloxacin sesquihydrate. DARU
Journal of Pharmaceutical Sciences. Vol. 16.
[41 Mowat, A. M. 1985. The Role of Antigen
Recognition and Suppressor Cells in Mice with
Oral Tolerance to Ovalbumin. Immunology. Vol.
56 p. 25.3
ACKNOWLEDGMENTS
The authors are grateful to DIKTI for providing
the research grant and also thank Faculty of
Pharmacy Airlangga University (UNAIR) for
supporting research facilities.
10
J
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!
UNIVEHSIM
KEAANCSAAN
MALATSIA
lfU!l$SIUMfy
;
I
1
!
m
m
QCDDS 2015
|CRS
CONTROLLED RELEASE &
DREG DELIVERY
SYMPOSIUM 2015
CRS
Controlled Releese Society
c-ES:
iI
I
i j
iLAnETuui KM-ea
:
In conjiuiction witli
The 1st MyCRS Scientific Conference
r
Targeted Delivery : Translating
Ideas Into New Technology
i
l2
£
i
a
::
UKM KUALA LUMPUR CAMPUS,
KUALA LUMPUR, MALAYSIA
o
f
I
r
t
I
ffl
1
m
£
n
*•
::::
I
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m
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1-*’ *
fei
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si
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*i
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1I
j
,
o
15“ÿ& 16th
AUGUST
015
•_
.»
ABSTRACT BQOK
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amnmmmuxttmmMmusmnamws
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£
j
Mitt
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List of Oral Presenters
1
List of Poster Participants
2
List of Research Pitch Participants
3
Oral Abstracts
4-28
Poster Abstracts
29-59
Research Pitch Abstracts
60 - 84
cmnouumusuimmiBjmrsmmmiMis
*XqpMMkajtlMMvMaiMiJhv
IM-IHjffmtltUUmlllHmptmlUviiitw+tm+w.Hdwii'
.
£
/
vas
wf
%£3
315
ip
NAME & TITLE
NO
1.
2.
LIST OF PRESENTERS
DATE : 15 & 16 AUGUST 2015
VENUE : DK2 HALL
PAGE
ASSOC. PROF. DR. MOMKLESUR RAHMAN
Preparation, Characterization and Release Profiles of Insulin-Loaded Double-Walled Poly
(lactide-co-glycolide) Microspheres
PROF. DR. HSIN-I CHANG
Influence of Liposomal Compositions on The Balance between Adipogenesis and Osteogenesis in Bone
_
__
Regeneration
7
3.
DR. DEWI MELANI HARIYADI
Physical Evaluations of Freeze-Dried Ca-AIgmate Microspheres Containing Ovalbumin Using Different
Lyoprotectants
9
4.
PROF. DR. ALLAN COOMBES
Evaluation of Tinidazole-Loaded Polycaprolactone Matrices for The Treatment of Trichomoniasis
11
5.
6.
7.
8.
9.
10.
__
DR. MOHD HANIF ZULFAKAR
Probing the influence of fish oil bigel on reducing imiqulmod-induced inflammation and enhancing
inhibition of non-melanoma skin cancer
PROF. DR. SAYYED A. SAJADI TABASSI
Sustained Drug Delivery System for Insulin using Supramolecular Hydrogels Composed of Tri-block
Copolymers PEG-PCL 20000-1000 Complexed with a -Cyclodextrin
DR. LAY LAY 'CONSTANCE' SAW
Targeted Oelivery of Hypericin for Early Bladder Cancer Detection ana Therapy
DR. VICTORIA PIUNOVA
Biodegradable Nanogel Star Polymers: A Platform for Programmable Macromolecular Self-Assembly and
Cargo Delivery
DR. IGOR KHALIN
BDNF Delivered to the Brain Using PLGA Nanoparticles Induces Functional Recovery After Traumatic Brain
Injury In Mice
_
_
_
DR. ABD ALMONEM DOOLAANEA
Differences Between CTAB and Chitosan in Modifying PLGA Nanoparticles for Gene Therapy
13
15
16
17
19
21
11.
ASSISTANT. PROF. DR. PEDRAM EBRAHIMNEJAD
Novel Surface Modified Polymeric Nanoparticles of Irinotecan, In vitro and In vivo Evaluation
23
12.
DR. LIKVOONKIEW
The Renal Targeting Potentials of the Polymeric Drug Carrier Poly-L-Glutamic Acid (PG) in Normal and
Diabetic Rats
25
13.
DR. GEETHA VISWANATHAN
Novel Diblock Copolymer Vesicles for Anti-Cancer Drug Delivery
27
__
1
A
t
5
mmmmmm§9m§amap:smnsmi2iis
k
;
\L
Sir
T
If
LIST OF PARTICIPANTS
DATE 15 & 16 AUGUST 2015
I
: .
.
r,
NO
1.
iwMtilMi+CBSdmdkCmfmma
k<
,
VENUE FOYER DK2&DK3
NAME & TITLE
PAGE
MS. AFNAN SHIHAB AHMED & DR. MUHAMMAD TAHER
Formulation of Hydrogel with Synthetic Polymers (PVA and PEG)
30
MS. CHEAH HOAYYAN
2.
3.
4.
Chemical Characterization of Nano-Polymeric-Zinc (II) Phthalocyanlne and Its In Vitro Phototoxicity Profile
in 4T1 Murine Breast Cancer Cells
32
MS. LEANNE SABRINA DE SILVA
Preparation and Characterization of Span 60-Cholesterol-TPGS Niosomes
34
DR. FUJU YEN
Tumour-targeted Niosomes: Improving Therapeutic Efficacy of Tocotrienois in Adjuvant Cancer Therapy
36
5.
MR. KRISADA WUTTIKUL
Influences of Surfactant Mixtures, Oil Types and Cosolvent Amounts on Formation of Nonionic
Microemulsions for Using as Pharmaceutical and Cosmetic Carriers _
38
6.
MR. MANSOUREH NAZARI VISHKAEI
Drug Delivery Enhancement by Lipid-Nano Encapsulation
40
7.
PROF. DR. MOHD CAIRUL IQBAL MOHD AMIN
Doxorubicin-loaded pH-Responsive Triblock CA-PEI-pArg Polymeric Micelles for Drug Delivery
42
8.
DR. NAGIB AU ELMARZUG!
Spectroscopic Study of Polyethylene Glycol and ctDNA Biocomplexes
44
~
9.
10.
11.
n
v.
12.
13.
14.
15.
DR. NG SHIOW FERN
Topical Bilayer Film containing Olive compound Hydroxytyrosol: Characterisation, Acute Dermal Irritancy
and Efficacious study in Freud's Adjuvant induced- Rat
_
45
MS. NURFAEZATIL FARHANA NORAZEMI
Exploiting Gellan Gum, Hydrocarbon and Fatty Acid for Oral Drug Delivery System
48
OR. PRABAKARAN LAKSHMANAN
Cempedak (Artocarpus Integer) Fruit Skin Mucilage as a Biomaterial: Extraction and Physicochemical
Characterization Studies
DR. RAVI SHESHALA
Development and Characterization of Phase-Inversion Based In Situ Forming Implants of Triamcinolone
Acetonide for Ocular Drug Delivery
_
52
MS. SYAHRIAH FADHILAH ABDUL RAZAK
Drug Permeation Study through Rat Skin: Effect of Storage Conditions of Skin
54
ASSOC. PROF. DR. AHMAD FUAD SHAMSUDDIN
Gelatine-based Parenteral Colloidal Formulation from Shank & Toes of Gallus Gallus Domesticus
56
___
__
_
MR. ATTIQUE UR RAHMAN MUFTI
Formulation, Stability Testing and Biochemical Safety Evaluation of Palm Oil-Based Nano Vaccine of
Pasteurella Multocida
I
50
2
58
axrmxiamutant* oejnrsrwosim JUS
hi
A
• It he HtJJ/OBMmJ&Cmimam
tk
i
T
J'j
IW
/its P
LIST OF PARTICIPANTS
DATE: 16 AUGUST 2015
VENUE : DK2 HALL
NO
NAME & TITLE
PAGE
1.
MR. ADEEL MASOOD BUTT
Folate Conjugated Chltosan Coated Pluronic F127-TPGS Mixed Micelles for the Co-delivery of siRNA and
Doxorubicin
61
2.
MS. KIEW SIAW FUI
Dextrin Coated Graphene Oxide Nanoparticles as a Controlled Release Drug Delivery System
63
3.
MR. KUE CHIN SIANG
TrkC Receptor Targeted Delivery of PDT Agent Inhibits Immune-suppressive Leukocytes and Enhances
Adaptive Immune Response Before and After Photodynamic Therapy (PDT)
65
4.
MS. MARSITA ABDUL RAZAK
Memantine Hydrochloride Loaded Poly N-Vinyl Caprolactum-ltaconic Acid Nanoparticles for Nose to
Brain Delivery
67
r-
S.
MR. MASOUD REZVANIAN
Alginate Based Composite Bio-Polymeric Dressing Loaded with Simvastatin for
70
Accelerated Wound Healing
MR. MUHAMMAD IRFAN SIDDIQUE
6.
7.
8.
V"
9.
Nanoparticles Enhance Targeted Topical Corticosteroids Delivery into the Skin with Improved Safety for
the Treatment of Atopic Dermatitis: In-Vivo Safety of Hydrocortisone-Hydroxytyrosol Loaded Chitosan
Nanoparticles
72
MS. NAJWA MOHAMAO
Characterization, Biocompatibility and In Vivo Evaluation of Bacterial Cellulose/Acrylic Acid Hydrogel
Synthesized Via Electron Beam Irradiation for Wound Dressing Applications
74
_
MR. NG KHEN ENG
Doxorubicin-loaded pH Responsive Triblock CA-PEI-pArg Polymeric Micelles for Drug Delivery
MR. TEO SIEW YONG
Evaluation of Alkyds as Promising Biomaterials for Drug Nanocarrier Systems
76
78
10.
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Physical Evaluations of Freeze-Dried Ca-Alginate Microspheres
Containing Ovalbumin Using Different Lyoprotectants
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Dewi Melani Harivadi. Tutiek Purwariti, Destia Wardhani, and Sisilia Enmawahyuningtyas
Pharmaceutics Department, Faculty of Pharmacy, Airlangga University, Surabaya,
Indonesia, 60286
Corresponding authorl: dewi-m-hQff.unair.ac.id: dewiffua96@vahoo.com
ABSTRACT SUMMARY
The main objective of the research was to
investigate physical evaluations of freeze-dried
ovalbumin loaded-Ca alginate microspheres. This
research used alginate, CaCl2 and lyoprotectants
(maltodextrin, lactose or sucrose). The freeze-dried
microspheres were subjected to micromeritics
properties including moisture content, angle of repose
angle of repose, bulk density, tapped density, Carr’s
index, and Hausner's ratio.
Results showed that the prepared microspheres
were spherical with particle size of microspheres
using lyoprotectant maltodextrin, lactose and sucrose
were 2.32, 2.15 and 2.05 pm respectively. Moisture
content of freeze-dried microspheres contains
maltodextrin and sucrose were about 4%, lower than
microspheres using lactose of about 6.66%. Bulk
densities and tapped densities were in the range of
0.2634 to 0.3232 and 0.3160 to 0.4040 respectively.
Carr's index (Cl) and Hausner's ratio (HR) were in the
range from 15 to 23% and 1.1765 to 1.2988
respectively.
Microspheres
produced
using
lyoprotectant sucrose resulted in good flow properties
with Cl of 15% and HR of 1.1765, whereas other
formulas showed fair flow. Almost all freeze-dried
ovalbumin-loaded alginate microspheres resulted in
small spherical microspheres with less water content
and good flow properties.
Keywords: Microspheres, Ovalbumin, Sodium
alginate, Lyoprotectant, Flow properties.
INTRODUCTION
Ovalbumin is an egg white protein that is used as
a model antigen and could stimulate the formation of
antibodies and improve immunity. Administering oral
antigen is the most effective way to induce
immunological tolerance to protein antigens (Mowat,
1985).
Current study applies ionotropic gelation method
to form hydrogel microspheres using polymer and
crosslinking agent. Sodium alginate is a safe and
biodegradable polymer as well as cheap and most
commonly used as polymer in the microparticles
(Maria et al., 2012). Pb2*, Cd2*, Zn2*, Cu2\ Co2*, Ca2*,
Ba2*, ar.d Sr2* are generally used as crosslinking
agents (Gombotz et al., 1998). Ca2* resulted stable
9
microparticles. Lyoprotectant was commonly added
to stabilize microspheres during preparation and
storage. This research was conducted to study the
potential of ovalbumin-Ca-alginate microspheres
using different concentrations of lyoprotectants such
as maltodextrin, lactose and sucrose.
EXPERIMENTAL METHODS
This research used concentrations of alginate of
2.5% w/v, concentrations of CaCte of 1,5M and 5%
lyoprotectants (maltodextrin, lactose or sucrose). The
freeze-dried microspheres were subjected to
micromeritics properties including moisture content,
angle of repose angle of repose, bulk density, tapped
density, Carr's index, and Hausner's ratio.
Preparation of alginate microsphere using
ionotropic gelation method could be explained as
follows: Alginate solution containing ovalbumin was
sprayed into crosslinking agent CaCfe solution and
was stirred continuously for 2 hours at 1000 rpm. The
microspheres were collected by centrifugation at
2500 rpm for 6 minutes and finally freeze dried 20
hours at -80°C. Alginate microspheres using
maltodextrin, lactose and sucrose were then called
F1, F2 and F3 respectively. The particle size of
microspheres was characterized by optic microscope.
Loading of ovalbumin into microspheres and
encapsulation efficiency was analyzed using protein
quantification assay using UV spectrophotometry.
RESULTS AND DISCUSSION
This technique produced spherical microspheres
with particle size using lyoprotectant maltodextrin,
lactose and sucrose were 2.32; 2.15 and 2.05 pm
respectively. These smaller ovalbumin-loaded Ca
alginate microspheres were suitable for oral
administration (Mishra et al, 2008). Spherical
morphological was caused by ability of lyoprotectant
agent to protect microspheres during freeze drying.
For encapsulation efficiency and loading, all
formulas resulted high encapsulation efficiency of
above 80% and loadings of above 30% (Table 1).
This caused by larger amounts of availability of Ca2*
that crosslinked with carboxylates from guluronic acid
in alginate indicates more ovalbumin was entrapped
within alginate microspheres.
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Table 2: Physical properties of the freeze-dried
microspheres
Table 1: Encapsulation efficiency, protein loading and
moisture content of microspheres
Formula
F1
F2
v
F3
k
Encapsulation
Protem
Efficiency (%)
94.47 ± 2.87
Loading (%)
82.93
* 2.80
81.36 ±8.81
31.33 ±3.94
30.57 ±2.18
Moisture
Content (%)
4.38 ± 0.51
6.66 0.42
37.20 ± 5.52
4.50 ±0.30
*
repose
F1
33.69
F2
30.96
23.96
F3
Moisture content of freeze-dried microcpheres
contains maltodextrin and sucrose were about 4%,
lower than microspheres using lactose of about
6.66% (Table 1). Moreover, the results of angle of
repose were in the range of 33.69°, 30.96° and 23.96°
(Table 2). Bulk densities and tapped densities
respectively were in the range of 0.2634 to 0.3232
and 0.3160 to 0.4040. Carr's index (Cl) and
Hausner's ratio (HR) were in the range from 15% to
23% and 1.1765 to 1.2988 respectively.
Microspheres produced using lyoprotectant sucrose
resulted in good flow properties with Cl of 15% and
HR of 1.1.765, whereas other formulas showed fair
flow.
Angle
Formulatl
on
Code
of
O
Bulk
density
(g/ml)
Tapped
density
(9/ml)
Carr's
index
(Ci) (%)
0.2634
0.3232
0.2686
0.3421
23.00
1.2988
0.4040
0.3160
20.00
15.00
1.2500
1.1765
Hausn
eds
ratio
CONCLUSION
Almost all freeze-dried ovalbumin-loaded alginate
microspheres
resulted
in
small spherical
microspheres with less water content and good flow
properties.
REFERENCES
[1] Gombotz, W.R., Slow Fong Wee, 1998. Protein
Release from Alginate Matrices. Advanced Drug
Delivery Reviews 31, p. 267-285.
[2] Maria, M.S., Scher, Herbert, Jeoh, Tina., 2012.
Microencapsulation of bioactives in cross-linked
alginate matrice. Journal of Microencapsulation, p.
286-295.
[3] Mishra, D. N. and Gilhotra, R. M. 2008. Design
and characterization of bioadhesive in-situ gelling
ocular inserts of gatifloxacin sesquihydrate. DARU
Journal of Pharmaceutical Sciences. Vol. 16.
[41 Mowat, A. M. 1985. The Role of Antigen
Recognition and Suppressor Cells in Mice with
Oral Tolerance to Ovalbumin. Immunology. Vol.
56 p. 25.3
ACKNOWLEDGMENTS
The authors are grateful to DIKTI for providing
the research grant and also thank Faculty of
Pharmacy Airlangga University (UNAIR) for
supporting research facilities.
10
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