1. Magnetite nanoparticles encapsulated
NANO KOREA 2006
“2006” International Nanotech
Symposium & Exhibition
In Korea
Magnetite nanoparticles encapsulated
with β-glucan for biomedical application
Sang Gil Ko1, Jun Hee Cho1, Yang Kyu Ahn1*, Eun Hee Kim2,
Ki Chang Song3, Eun Jung Choi4
1Department of Nanochemistry & Biochemistry, Konyang University
2Pharmaceutical Examination Div., The Korean Intellectual Property Office
3Department of Chemical Engineering, Konyang University
4Department of Othalmic Optics, Konyang University
Tel :

2. DEPARTMENT OF NANOCHEMISTRY AND BIOCHEMISTRY KONYANG UNIVERSITY
Abstract
Embolotherapy, the process in which a blood vessel or organ is obstructed by the lodgment of a material mass, has been used in cancer treatment as well as artery bleeding or malformation. However, most embolic materials such as polyvinyl alcohol(PVA) could not be detected by an MRI, so it was difficult to locate injected embolic materials in a body. Superparamagnetic nanoparticles have an advantage of producing an enhanced proton relaxation in MRI and the Superparamagnetic nanoparticlescan be incorporated into embolic materials to enable MRI detection and thus find a practical application in embolotherapy. Therefore, we would like to develop a novel embolic material capable of MRI contrast enhancement.
The Superparamagnetic nanoparticles synthesized by the sonochemistry spherical. Considering their uniform size and shape, these Superparamagnetic nanoparticles were suitable to use for medical applications. These magnetic nanoparticles were encapsulated with beta-glucan which is known as a safe material to an human body, then we obtained spherical microspheres having an average diameter of ㎛.
Also, we could controlled the size distribution of the microspheres as well as selected the microspheres of ㎛ in diameter that is suitable to be injected through the blood vessel via angiographic catheters. These selected microspheres were nearly spherical and maintained their shape in water for more than 30 days. Then, in order to optimize the concentration of magnetic materials in beta-glucan, various concentration of microspheres were prepared and obtained magnetic resonance images by 4.7 T MRI.
DEPARTMENT OF NANOCHEMISTRY AND BIOCHEMISTRY KONYANG UNIVERSITY

3. DEPARTMENT OF NANOCHEMISTRY AND BIOCHEMISTRY KONYANG UNIVERSITY
Applications of magnetite nanoparticles
Magnetic inks
Magnetic memory device
Magnetic fluids
Clinical applications (NT+BT)
- drug delivery
- cell separation
- hyperthermia
- MRI imaging
- embolotherapy
DEPARTMENT OF NANOCHEMISTRY AND BIOCHEMISTRY KONYANG UNIVERSITY

4. DEPARTMENT OF NANOCHEMISTRY AND BIOCHEMISTRY KONYANG UNIVERSITY
Embolotherapy
Embolization
- is a process in which a blood vessel or organ is obstructed by the
lodgment of a material mass (as an embolus): used in cancer treatment
Diseases treated by embolotherapy
- Blood vessel trouble
- Bleeding
- Tumor treatment
Requirements for Embolization
- Biocompatible and non-toxic
- No carcinogenic
- Relatively benign inflammatory response
- No aggregation
- Spherical shape
We would like to apply the SPIO in embolotherapy.
Embolotherapy is a process in which a blood vessel or organ is obstructed by the lodgment of a material mass.
Diseases need for embolotherapy are
-blood vessel trouble,
-bleeding,
-and tumor treatment.
To be an ideal embolic material,
it should be Biocompatible and non-toxic,
it has No carcinogenic,
it has Relatively benign inflammatory response,
it cause No aggregation.
Of course, spherical shape of material is preferred.
Those are angiographic catheters.
DEPARTMENT OF NANOCHEMISTRY AND BIOCHEMISTRY KONYANG UNIVERSITY

5. DEPARTMENT OF NANOCHEMISTRY AND BIOCHEMISTRY KONYANG UNIVERSITY
MRI detectable embolus
A problem in embolotherapy
- Most embolic materials can not be detected by an MRI.
Ex) Gel form, PVA (polyvinyl alcohol), coil, micro balloon, silicon bead,
glass bead, dehydrogenated ethanol, NBCA(N-butyl 2-cyano acrylate)
- MRI-detectable embolus can be effectively used for embolotherapy
The objective is to synthesize an MRI-detectable embolus
- narrow size distribution of SPIO nanoparticles
- chemically stable SPIO ferrofluids
- spherical and homogeneous SPIO-chitosan microspheres
- contrast enhanced T2-weighted MR images of the microspheres medical
application
Most embolic materials can not be detected by an MRI, so, it is difficult to locate embolic material in the body.
We thought that embedding the SPIO in an embolic material enables the material to be detected by an MRI.
These MR-detectable embolus can be effectively used for embolotherapy.
Thus, We developed a novel embolic material made of the SPIO and chitosan to enable in situ monitoring of the ferrofluid with an MRI.
In order to synthesize an MRI-detectable embolus, we have synthesized narrow size distribution of SPIO nanoparticles,
and synthesized chemically stable SPIO-chitosan ferrofluids,
and prepared spherical and homogeneous SPIO-chitosan microspheres,
finally we took T2-weighted MR images of the microspheres in vivo.
Microcapsule
Microsphere
magnetite
biopolymer
DEPARTMENT OF NANOCHEMISTRY AND BIOCHEMISTRY KONYANG UNIVERSITY

6. DEPARTMENT OF NANOCHEMISTRY AND BIOCHEMISTRY KONYANG UNIVERSITY
Synthesis of magnetite nanoparticles
FeCl3·6H2O
(0.3M, 30ml)
FeCl2·4H2O
(0.15M, 30ml)
optimum conditions:
 add Precipitator ( NH4OH, TMAOH or NaOH )
180mmol, 72mmol, 51mmol
 irradiation power : 250 W
 irradiation time : 30min
 irradiation temperature : keeping 70 ~ 80℃ +
metal chloride mixture
aqueous solution
add oleic acid
(sodium form) 8~12ml
heating
to 70℃
sieve
washing
by centrifuge
uniform magnetite
nanoparticles
DEPARTMENT OF NANOCHEMISTRY AND BIOCHEMISTRY KONYANG UNIVERSITY

7. DEPARTMENT OF NANOCHEMISTRY AND BIOCHEMISTRY KONYANG UNIVERSITY
Preparation of magnetic biopolymer microspheres
1. 3g chitosan powder + 147ml 1% acetic acid solution
→ 2% chitosan solution
2. 7.5g β-glucan powder ml 1% NaOH solution
→ 5% β-glucan solution
Magnetite colloid & chitosan solution
or β-glucan solution
microspheres of
magnetic nanoparticles –chitosan
or magnetic nanoparticles- β-glucan
Nozzle
Alkali solution
NaOH/Ethanol/Water,
4/30/66, W/V/V
Acid solution
Acetic acid/Ethanol/Water,
N2 gas
10L/min
magnetite nanoparticles(0.5g) + 2% chitosan solution(150ml)
magnetite nanoparticles(0.5g) + 5% β-glucan solution 150ml)
Optimum conditions:
 Irradiation Power: 665 W
 Irradiation Time: 15min
DEPARTMENT OF NANOCHEMISTRY AND BIOCHEMISTRY KONYANG UNIVERSITY

8. DEPARTMENT OF NANOCHEMISTRY AND BIOCHEMISTRY KONYANG UNIVERSITY
Structure and property of chitosan and β-glucan
Chitosan
β-Glucan
Chitin
Deactylation
Extraction from mushroom
Molecular form : (-C6H10O5-)n
Dissolved in alkali solution (NaOH solution)
Gelation in acid solution (acetic acid)
Anticancer effect
Diet food
Chitosan
Molecular form : (-C6H11NO4-)n
Chitosan has an average molecular
weight ranging between 3.8X103 and 2.0X106
Dissolved in acid solution (acetic acid)
Gelation in alkali solution (NaOH solution)
Anticancer effect
These biopolymers are nontoxic,
biocompatible, non carcinogenic
material
DEPARTMENT OF NANOCHEMISTRY AND BIOCHEMISTRY KONYANG UNIVERSITY

9. DEPARTMENT OF NANOCHEMISTRY AND BIOCHEMISTRY KONYANG UNIVERSITY
XRD results of synthesized nanoparticles
220
311
400
422
511`
440
TMAOH
[Water] / [Oleic acid]` `
Size
Sample 1
72 mmol
104.16
6.1 nm
Sample 2
72 mmol
133.32
7.5 nm
DEPARTMENT OF NANOCHEMISTRY AND BIOCHEMISTRY KONYANG UNIVERSITY

10. DEPARTMENT OF NANOCHEMISTRY AND BIOCHEMISTRY KONYANG UNIVERSITY
Size distributions of magnetite nanoparticles
(a)
(b)
(d)
(c)
[Water]/
[Oleic acid]
95.23
104.16
114.83
133.32`
Particle size
(nm)
2.0
3.7
5.4
7.4
S.Dev
(nm/%)
0.3/13.3
0.4/11.2
0.5/9.8
0.8/10.8
( a )
( b )
( c )
( d )
DEPARTMENT OF NANOCHEMISTRY AND BIOCHEMISTRY KONYANG UNIVERSITY

11. DEPARTMENT OF NANOCHEMISTRY AND BIOCHEMISTRY KONYANG UNIVERSITY
AFM images of magnetite nanoparticles
Magnetite 1
Magnetite 2
6.4 nm
133
72mmol
Magnetite 2
3.5 nm 95
Magnetite 1
Mean sizes
[Water] / [Oleic acid]
TMAOH `
DEPARTMENT OF NANOCHEMISTRY AND BIOCHEMISTRY KONYANG UNIVERSITY

12. Optical microscope images of magnetic biopolymer microspheres
Magnetic chitosan microspheres
In water system
Magnetic β-glucan microspheres
In water system
DEPARTMENT OF NANOCHEMISTRY AND BIOCHEMISTRY KONYANG UNIVERSITY

13. SEM images of magnetic biopolymer microspheres
prepared by spray method
Magnetic chitosan microspheres
Magnetic β-glucan microspheres
DEPARTMENT OF NANOCHEMISTRY AND BIOCHEMISTRY KONYANG UNIVERSITY

14. Magnetic propertion of magnetite nanoparticles and
magnetic biopolymer microspheres
( a )
( b )
( c )
Magnetite nanoparticle
Magnetic-chitosan microsphere
Magnetic-β-glucan microsphere
DEPARTMENT OF NANOCHEMISTRY AND BIOCHEMISTRY KONYANG UNIVERSITY

15. DEPARTMENT OF NANOCHEMISTRY AND BIOCHEMISTRY KONYANG UNIVERSITY
MR Imaging results of magnetic biopolymer microspheres
chitosan microspheres
Fe concentration M M M M 1 2 3 4
T1(left)- and T2(right) weighted MR Images
β-glucan microspheres
DEPARTMENT OF NANOCHEMISTRY AND BIOCHEMISTRY KONYANG UNIVERSITY

16. DEPARTMENT OF NANOCHEMISTRY AND BIOCHEMISTRY KONYANG UNIVERSITY
Conclusion
(1) We synthesize magnetite nanoparticles by sonochemical method
(2) The size of particles can be controlled by the ratio of oleic acid vs
water
(3) The synthesited particles show narrow size distribution under S.D 5%
(4) 2% chitosan, 5% β-glucan solution are excellent viscosity, and could know
that form of sphere is made because viscosity does not decrease much
even if irradiate ultrasonic
(5) Fe concentration of 2mM, the MRI contrast was much higher than that of
pure water. Therefore, the ferrofluid may potentially be useful in developing
an MR detectable embolic material.
DEPARTMENT OF NANOCHEMISTRY AND BIOCHEMISTRY KONYANG UNIVERSITY