CO7-1

Establishment of QA/QC for BNCT Neutron Irradiation Field

Y. Sakurai, T. Fujii1, R. Uchida1 and H. Tanaka
Research Reactor Institute, Kyoto University
1
Graduate School of Engineering, Kyoto University

INTRODUCTION: In May 2010, the operation of
Kyoto University Reactor (KUR) restarted, which had
been suspended during four years for thefuel-low-enrichment. Concurrently with the KUR restart,
clinical irradiation of boron neutron capture therapy(BNCT) at the Heavy Water Neutron Irradiation Facility(HWNIF) also restarted [1]. After the restart, 170 BNCT
irradiations have already been carried out as of May 2013.
In the while, Cyclotron-based BNCT Epi-thermal Neutron Source (C-BENS) was installed in this institute in
the end of 2008 [2]. In November 2012, the BNCT clinical trial using C-BENS started. Thus, this institute became a special institute in the world, in where BNCT is
performed at the two-type neutron sources such as reactor-based one and accelerator-based one. It is one of theimportant subjects that the consistent dose-estimation is
performed between the both neutron sources, and then theequivalence and homogeneity for the deposited dose
during the clinical irradiation are assured. The aim of thisresearch is the establishment of quality assurance andquality control (QA/QC) for BNCT neutron irradiation
field. In 2012, one of the important tools for QA/QC,“Multi Ionization Chamber System (MICS)”, was prepared by way of trial [3], and its characteristics were estimated and its efficacy was confirmed.
METHODS: The prototype of MICS consists of (i) achamber of silicon-nitride wall and nitrogen gas for thethermal neutron component (Si3N4(N2)), (ii) a chamber ofboron-evaporation-coated polyethylene wall and nitrogengas, covered with 6LiF shield, for the epi-thermal neutroncomponent (PolyB(N2)), (iii) a chamber of polyethylene
wall and methane gas for the fast neutron component(Poly(CH4)), and (iv) a chamber of graphite wall andargon gas for the gamma-ray component (G(Ar)). Thesechambers were placed on the bismuth-layer side of thecollimator on the remote patient carrier, as shown inphoto 1. The experiment for the characteristic estimation
was performed for the epi-thermal neutron irradiation
mode.
RESULTS AND DISCUSSIONS: Figure 1 shows thechanges of the separate-estimated values with time forthe four components, such as thermal neutron,
epi-thermal neutron, fast neutron, and gamma ray, obtained using MICS. These values are for the KUR powerof 1 MW. The vertical axis is in flux. The solid lines arefor the experimentally-estimated values and the brokenlines are for the estimated values by simulation. In the
epi-thermal neutron irradiation mode, the thermal neutron
flux was practically zero, because it was lower than thelimit for the separate-estimation. For the epi-thermal andfast neutrons, those experimental values were in good
agreement with the simulation values, in the uncertainty
of 4% and 12%, respectively. For the gamma ray, theexperimental value overestimated the simulation value at
63%. The flux dispersion in the experiment was 2%, 26%
and 27%, respectively for epi-thermal neutron, fast neutron and gamma ray. It was confirmed that the accuracy
in the separate-estimation was better and the response
was more stable for the component with the larger response.
CONCLUSION: The efficacy of the prototype MICS
was confirmed. The possibility of the separate-estimation
for the thermal neutron component is expected, as the
KUR power is 5 MW, during the actual BNCT.

Photo 1.

Placement of MICS on the collimator.

Fig. 1. Changes of the separate-estimated values
with time, obtained using MICS.
REFERENCES:[1] Y. Sakurai et al., Nucl. Instr. Meth. A 453 (2000)
569-596.
[2] H. Tanaka et al., Nucl. Instr. Meth. B 267 (2009)
1970-1977.
[3] T. Fujii et al., Appl. Radiat. Isot. 69 (2011)
1862-1865.

採択課題番号 24009
ＢＮＣＴ用照射場におけるＱＡ／ＱＣの確立
共同通常（京大・原子炉）櫻井良憲、田中浩基、吉永尚生（京大院・工）藤井孝明、武川哲也、内田良平

CO7-2

To Conquer the Clinically Relevant Radioresistant Cell Tumors
Targeting Tumor Endohtelial Cells

Y. Kuwahara, M. Fukumoto, Y. Sakurai1, M. Suzuki1,
K. Ono1 and M. Fukumoto
Department of Pathology, Institute of Development, Aging and Cancer, Tohoku University.
1
Department of Radiation Life Science and Radiation
Medical Science, Research Reactor Institute, Kyoto University
INTRODUCTION: Radiotherapy is one of the major
therapeutic modalities for eradicating malignant tumors.
However, the existence of radioresistant cells remains
one of the most critical obstacles in radiotherapy. To understand the characteristics of radioresistant cells and todevelop more effective radiotherapy, we have establishedclinically relevant radioresistant (CRR) cell lines. Because tumor tissues of CRR cells transplanted into nude
mice were richer in tumor blood vessels compared with
their radiosensitive parental cell lines. So, we performed
boron neutron capture (BNC) method targeting tumor
endothelial cells using PEG-10B. Growth rate of HeLa-R(CRR of HeLa) tumors were not significantly different
from that of non-irradiated control in 4 weeks after irradiation. Tumors of parental cells treated with BNC using
PEG-10B were significantly smaller than those without
radiation. Seven days after BNC CD31 positive bloodvessels were destroyed in BNCT treated HeLa tumors
compared to HeLa tumor without BNCT but were not in
HeLa-R tumors irrespective of BNCT. Moreover, to oursurprise, 4 weeks after BNC the density of CD34, positive blood vessels were almost the same irrespective of
BNCT in both HeLa-R and HeLa tumors.
We need further studies to confirm how the density ofblood vessels contributes to tumor radiotherapy.
EXPERIMENTS: Three days before experiments,
1x106 cells of HeLa and HeLa-R were injected subcutaneously into hind legs of male Balb/c nude mice (4 weeksold). The day of the exposure experiment tumor diameter
was approximately 3-4 mm. For irradiation of tumor endothelial cells by α-particles, PEG-10B were administered.
The compound was suspended in physiological saline at aconcentration of 2500 ppm and was injected via the tailvein. Three hours after the administration, mice were
exposed to neutron radiation at the Research Reactor Institute, Kyoto University (KURRI).
Tumor size was determined by caliper measurements
every three days. Endothelial cells of blood vessels intumor tissues were immunohistochemically stained forCD34 and CD31. Type Ⅳcollagen for pericytes andfunctional blood vessels for injected tomatolectin fromtails. We counted the number of vessels in 10 high power
field (x 400) and calculated the average (n = 3).

RESULTS: We first examined the size of HeLa tumorsand HeLa-R daily after BNCT. Within 24 days after
BNCT, the size of HeLa-R tumors was not different irrespective BNCT. But the size of HeLa tumors was significantly decreased compare to control tumors.
Mice were sacrificed on day 30 after BNCT. Histologicalexamination showed that HeLa-R tumors were richer intumor blood vessels compared with their radiosensitiveparental cell tumors. CD34 positive blood vessels werealso more abundant in SAS-R tumors than in SAS-tumors(Data not shown).
After BNC using PEG-10B, the vessels of HeLa-R
tumors were recovered more rapidly in 7 days. On theother hand, those of parental tumors were destroyed.
However, 4 weeks after BNC, the density of CD34
positive blood vessels was almost the same irrespectiveof BNCT in both HeLa-R tumors and HeLa tumors.
DISCUSSION: In this research, we tried to target tumor endothelial cells of radioresistant HeLa-R tumors,because our preliminary experiment showed that the density of blood vessels in SAS-R tumors was higher thanthat in SAS tumors. We selectively exposed endothelial
cells to α-particles, using BNC with PEG-10B. The tumor
volume of HeLa-R was not significantly different after
exposure to 8 Gy of α-particles for the examination period. But that of HeLa was significantly decreased compare
to control tumors.
7 days after BNC the density of CD31 positive bloodvessels were destroyed in HeLa tumor exposed to
α-particles compared to control HeLa tumor but did not
in HeLa-R tumor irrespective of α-particles exposure.
Moreover, to our surprise, 4 weeks after BNC the densityof CD34, positive blood vessels were almost same irrespective of α-particles exposure in both HeLa-R tumorand HeLa.
Therefore, further studies are needed to confirm how
blood vessel density contributes to tumor radiotherapy.

採択課題番号 24012
血管内皮細胞を標的とする放射線耐性腫瘍の克服にむけて（東北大・加齢研）桑原義和、福本基、福本学（京大・原子炉）鈴木実、小野公二

共同通常

CO7-3 Study on Advanced Neutron Measurements Using a Small Size Neutron Scintillator

INTRODUCTION:
The Boron Neutron Capture
Therapy (BNCT) has been developed as one of thepromising radiotherapies. The neutron dose evaluation
for the BNCT is quite important. Optical fiber type detectors as one of the on-line and small neutron flux monitors have been developed. The conventional opticalfiber neutron detectors, however, show a continuous distribution without a characteristic shape, such as the fullenergy peak corresponding to the neutron induced reaction, in a pulse height spectrum due to large fluctuation ofcollected scintillation photons based on their poor lightcollection efficiency[1-3]. The sensitivity of these detectors depends on the detector signal gain. We, therefore, develop the advanced optical fiber type neutron detector using a small piece of Eu doped LiCaAlF6 scintillator. This detector can show an obvious neutron absorption peak and suppress the gamma-ray sensitivity.
In this report, we characterize the developed neutron detector at the Heavy Water Thermal Neutron Irradiation
Facility (HWTNIF) of Kyoto University Research Reactor (KUR).

counting rate [cps]

DEVELOPED DETECTOR: We fabricated the optical fiber type neutron detector using a small Eu:LiCaAlF 6
scintillator. The fabricated detector consists of a small
piece of Eu doped LiCaAlF6 scintillator, a plastic opticalfiber, a photomultiplier tube (PMT) and signal processing
circuits. A bulk Eu:LiCaAlF6 scintillator suffers frominfluence of gamma rays because of its relatively low /bratio. Figure 1 shows the pulse height spectra obtained
from a bulk Eu:LiCaAlF6 scintillator. Gamma-ray signals are confirmed to interfere the neutron absorption
peak.

100

252
Cf
60

Co
BG

10-1

100

200

300

252
Cf-252

Cf線源

400

500

pulse height [channel]
Fig. 1. Pulse height spectra obtained from a bulk
Eu:LiCaAlF6 scintillator.

Co-60
60
Co線源
BG

10-2

BG

10-3
0

50

100

150

波高値
Pulse [channel]
height (ch)Fig. 2. Pulse height spectra obtained from an optical fiber
detector with a small Eu:LiCaAlF6 scintillator.
EXPERIMENTS AT HWTNIF OF KUR: We characterize the fabricated detector at the HWTNIF of KUR.
The detector head was placed at various distances fromthe bismuth filter surface. An example of the pulse
height spectrum obtained from the fabricated detector inexperiments at the HWTMIF of KUR is shown in Fig. 3,
where the distance from the bismuth filter surface was
120 cm. A clear neutron absorption peak was obviously
observed.

100

50

0
0

10-2

0

10-1

[cps]
計数率
(cps)
rate
Count

Graduate School of Engineering, Nagoya University
1
Research Reactor Institute, Kyoto University

Figure 2 shows the pulse height spectra obtained froman optical fiber detector with a small LiCaAlF6 scintillator. The pulse height of signals induced only by gamma
rays can be suppressed compared with neutron induced
signals. This is because of differences in ranges ofcharged particles produced by neutrons and gamma rays.

Count rate (cps)

K. Watanabe, Y. Kawabata, M. Ushida, A. Uritani,
A. Yamazaki, Y. Sakurai1 and H. Tanaka1

100

200
300
400
Pulse height (ch)500

Fig. 3. Example of the pulse height spectrum obtained
from the fabricated detector in experiments at the HWTNIF of KUR.
REFERENCES:[1] M. Ishikawa et al., Appl. Rad. Isotopes, 61 (2004)
775-779.
[2] M. Ishikawa et al., Nucl. Instr. Meth. A, 551 (2005)
448-457.
[3] Y. Ito et al., Radiat. Prot. Dosim., 110 (2004)
619-622.

採択課題番号 24014
極少検出器を用いた中性子計測手法の高度化に関する研究（名大・工）瓜谷 章、渡辺賢一、山﨑 淳、川端勇矢、牛田雅人（京大・原子炉）櫻井良憲、田中浩基

共同通常

CO7-4 High Boron Content Liposomes and Their Promising Antitumor Effect for BNCT
H. Koganei, S. Tachikawa, M. Suzuki1, S. Masunaga1,
K. Ono1 and H. Nakamura
Faculty of Science, Gakushuin University
1
Research Reactor Institute, Kyoto University
INTRODUCTION: Boron neutron capture therapy(BNCT) functions as a double targeting therapy for cancer. Its therapeutic effect is realized by neutron beamirradiation and a boron delivery system (BDS). BNCT
uses the nuclear reaction of two species, boron-10 (10B)
and thermal neutrons. Although the low-energy thermal
neutrons (0.025 eV) are employed, the resulting α-particle and Li nuclei are high linear energy transfer(LET) particles that travel a short distance (approximately 5−9 μm) to destroy cells containing 10B. If 10B atoms
were selectively delivered to intracellular regions of tumor tissue, it would be possible to kill tumor cells selectively without seriously damaging adjacent healthy tissues.
In this study, we focused on lipophilic boron compounds embedded in a liposome bilayer. This strategy is
an attractive means to increase the overall incorporation
efficiency of boron containing species, as well as to raise
the gross boron content of liposomes [1]. We developed
high boron content liposomes by incorporating boron into
both the interior aqueous core and the membrane of liposomes. Indeed, this strategy yielded significant antitumor
effect on tumor-bearing mice after neutron irradiation, as
well as a reduction of the total liposome dose, revealingthat the current boronated liposome is one of the mostpromising candidates for practical use in BDSs for
BNCT.
EXPERIMENTS: DSPC (MC-8080) and DSPE-PEG(Sunbright DSPE-020CN) were purchased from Nippon
Oil and Fats (Tokyo, Japan). Cholesterol (Chol) was
purchased from Kanto Chemical (Tokyo, Japan).
10
B-enriched BSH and S–cyanoethyl protected 10B-enriched BSH were purchased from Stella Pharma Co.
(Osaka, Japan). Boron lipid (DSBL) was synthesizedaccording to the previously described procedures withmodification [2].
BSH-encapsulated DSBL-10% liposomes, which wereprepared from 10B-enriched DSBL, DSPC, Chol, and
DSPE-PEG (0.1:0.9:1:0.11, molar ratio) and 125 mM
BSH aqueous solution according to the REV method previously described [3], were injected into colon 26 tumor
bearing mice (female, 6−7 weeks old, 16−20 g, 5 mice ineach group) via the tail vein at doses of 15 and 30 mg 10B/kg (1500 and 3000 ppm of 10B concentration; 200 μL ofboronated liposome solution). The mice were placed inan acrylic mouse holder 36 h after i.v. injection. The mice
were irradiated in KUR. The antitumor effects of BNCT
were evaluated on the basis of the changes in tumor
volume of the mice. Mortality was monitored daily andtumor volume was measured at intervals of a few days.
RESULTS: The tumor growth curves are shown in
Figure 5. At doses of 15, 30, and 50 mg 10B/kg, tumor
growth in mice treated with BSH-encapsulating 10%
DSBL liposomes was significantly inhibited after thermal
neutron irradiation, and the tumor disappeared within
three weeks even when the dose of 15 mg 10B/kg was
injected. However, the inhibition was not observed inmice injected with BSH-encapsulating 10% DSBL liposomes (30 mg 10B/kg) without thermal neutron irradiation,or in hot and cold control mice [4].

Fig. 1. Tumor volumes in mice (Balb/c, female, 6 weeks
old, 14−20 g) bearing colon 26 solid tumor after i.v. injection of BSH-encapsulating 10% DSBL liposomes (15(-Δ-), 30 (-□-), 50 (-○-) mg 10B/kg) and thermal neutron
irradiation for 50 min (1 MW) at a rate of (1.5−1.8) x
1012 neutrons/cm2 36 h after injection. Tumor volumes ofmice without injection of liposomes with thermal neutron
irradiation (-●-), without thermal neutron irradiation(-▲-), or with injection the liposomes (30 mg 10B/kg)
without thermal neutron irradiation (-■-)”.
REFERENCES:[1] M. Ueno, et al., Bioorg. Med.Chem. 18 (2010) 3059.
[2] J.-D. Lee et al., Org. Lett. 9 (2007) 323.
[3] H. Nakamura et al., Org. Biomol. Chem. 10 (2012) 1374.
[4] H. Koganei et al., Bioconjugate Chem. 24 (2013) 124.

採択課題番号 24023
中性子捕捉治療のための新規ホウ素薬剤の開発（学習院大・理）中村 浩之、立川 将士、小金井 逸人（京大・原子炉）鈴木 実、増永 慎一郎、小野 公二

共同通常

CO7-5Long-Term Result of BNCT for Different Types of Human Clear Cell Sarcoma
in Mouse Model

T. Fujimoto, T. Andoh1, Y. Sakurai2, M. Suzuki2,
H. Ichikawa1 and K. Ono2
Hyogo Cancer Center,
Department of Orthopaedic Surgery
1
Faculty of Pharmaceutical Sciences and Cooperative
Research Center of Life Sciences,
Kobe Gakuin University
2
Research Reactor Institute, Kyoto University
INTRODUCTION: Clear cell sarcoma (CCS) of tendons and aponeuroses is a rare neoplasm with no effective treatment other than surgery. Furthermore, this malignant tumor has a predilection for young adults and its
prognosis is poor [1]. Clearly, therefore, new therapeuticstrategies are required. In a previous study, we haveshown that the high accumulation of 10B both in cultured
human CCS cell lines and in CCS-bearing animal models,is potentially propitious in boron neutron capture therapy(BNCT) with the use of p-boronophenylalanine (BPA)[2,3]. We have recently also demonstrated theeffectiveness of short-term BNCT in the human
CCS-bearing nude mouse model [4]. Thus BNCT could
become a potential, new therapeutic option for thetreatment of human CCS. Consequently, in this study, weevaluated the long-term efficacy of BNCT for CCS, withthe use of nude mice intramuscularly transplanted withhuman CCS cell lines.
EXPERIMENTS:(1) Tumor cell line: Human CCS
cell lines, MP-CCS-SY [5], SU-CCS-1 [6] and HS-MM[7] were grown in RPMI 1640 and DMEM (for HS-MM)
with fetal bovine serum in a 5% CO2 humidified
incubator at 37˚C.
(2) BNCT for CCS-bearing animal: All animal experiments were carried out according to the regulations of theAnimal Care and Use Committee. Cells of CCS cell lines
were transplanted into the left femoral region of BALB/cnude mice. When CCS tumors grew to about 10-20mm indiameter, the animals were divided into 3 BNCT groups
and 3 control groups for each cell line. The animals, under anesthesia and through the femoral vein, were intravenously administered BPA-Fr (24mg 10B/kg; the BNCT
groups) or saline (the control groups), and then immediately placed in a chamber for thermal neutron irradiation
experiments. Thermal neutrons (1MW) were deliveredfrom the dorsum of the mouse, in the heavy water facility
at KURRI. LiF tiles were used to shield parts of the bodyother than the left leg. After the irradiation, the size of thetumor was measured as the maximum elliptical area ofthe tumor mass as follows: Maximum tumor area=(minoraxis)×(major axis), and the “Area ratio” was defined as
the area of each divided by the area of day 0. Tissue samples of the tumor mass resected under anesthesia from thecontrol groups on day 45, and from the BNCT groups onday 90 were fixed in 10% buffered-formalin solution andembedded in paraffin according to standard protocols.
Sections were then stained with hematoxylin-eosin (HE)for histological examination.
RESULTS: The doses absorbed (Gy) by the mice intramuscularly transplanted with the CCS cells were 7.2(HS-MM), 6.8 (MP-CCS-SY) and 7.1 (SU-CCS-1). After
irradiation, the size of the decreased time-dependently in
the BNCT groups until around day 20, and then regrew
time-dependently. In the control groups, the growth was
not suppressed by thermal neutron irradiation; the tumor
mass simply increased with time [Fig.1]. Histologicalexamination of the BNCT groups on day 90 revealed
regrowth of the CCS tumor of each cell line, with nodamage to normal surrounding tissue.

Fig.1. After the neutron irradiation, the size of the tumor
was measured as the maximum elliptical area of the tumor mass. BNCT groups (ΔMP BNCT; MP-CCS-SY,□HS BNCT; HS-MM, ○SU BNCT; SU-CCS-1) and control groups (▲MP Hot control; MP-CCS-SY, ■HS Hot
control; HS-MM, ●SU Hot control; SU-CCS-1).
REFERENCES:[1] F.M. Enzinger, Cancer, 18 (1965) 1163-1174.
[2] T. Fujimoto et al., Appl. Radiat. Isot., 69 (2011)
1713-1716.
[3] T. Andoh et al., Appl. Radiat. Isot., 69 (2011)1721-1724.
[4] T. Fujimoto et al., Appl. Radiat. Isot., 73 (2013)
96-100.
[5] H. Moritake et al., Cancer Genet. Cytogenet.,
135 (2002) 48-56.
[6] H. Sonobe et al., J. Pathology, 169 (1993) 317-322.
[7] A.L. Epstein et al., Cancer Res., 44 (1984) 1265-1274.

採択課題番号 24029
明細胞肉腫に対するホウ素中性子捕捉療法の検討
共同通常（京大・原子炉）鈴木実、櫻井良憲、小野公二（神戸学院大・薬）安藤 徹、市川秀喜（兵庫県立がんセンター・整形外科）藤本卓也

CO7-6

Application of B4C Nanoparticles for Boron Neutron Capture Therapy

N. Koshizaki, Y. Ishikawa1 and M. Suzuki2
Nanosystem Research Institute (NRI), National Institute of Advanced Industrial Science and Technology (AIST)
1
Kagawa University, Faculty of Engineering
2

Research Reactor Institute, Kyoto University

INTRODUCTION:
In carcinoma in situ (CIS) in the bladder, the cancer
cells are still only in the mucosa of the bladder, but arein flat sheets that look a bit like moss. CIS bladdercancer is called a high risk, early bladder cancer because if it is not treated it is very likely to spread into
the deeper layers of the bladder.
Nonradioactive isotope 10B atoms that absorb
low-energy (<0.5 eV) neutrons (thermal neutrons)
disintegrate into an alpha (4He) particle and a recoiled
lithium nucleus (7Li). These particles deposit high energy along their very short path (<10 µm). Thus, only
malignant cells with 10B are destroyed following thermal neutron irradiation. Theoretically, any normal cells
abutting the cancer cells are spared from high linear
energy transfer irradiation by 4He and 7Li particles.
We have been working on a nanoparticle fabrication
technique by irradiating laser light onto a plate placedor powder dispersed in liquid media. This technique
has several advantages over conventional nanoparticle
preparation in liquid phase, highly pure nanoparticles
with less use of surfactant molecules mostly toxic in
biological systems, and crystallized nanoparticle formation due to the transient high temperature process
induced by pulsed laser irradiation. Thus, these features are expected to be suitable for B4C nanoparticle
preparation by pulsed laser irradiation of B4C particles
dispersed in ethyl acetate under atmospheric pressure
and room temperature [1].
In the treatment of BNCT for CIS bladder cancer,
intravesical administration of B4C nanoparticles may
be appropriate drug delivery system as Bacille deCalmette et Guérin (BCG) vaccine treatment since carcinoma cells in CIS bladder cancer is exposed directly
by B4C nanoparticles applied to bladder.
We carried out a preliminary experiment investigating availability of B4C nanoparticles in the treatmentof BNCT for CIS bladder cancer.
EXPERIMENTS:
HeLa cells were used in this study. The cells weredispersed in the 96-well microplates at the concentrationof 10,000 cells per well the day before neutron irradiation.
In this experiment, B-10 rich nanoparticles (10B4C nanoparticles) were used in the BNCT treatment groups. The
treatment conditions were sorted into 5 groups as follows.
1. No treatment

採択課題番号 24030（産総研）越崎直人

2. 10B4C exposure without neutron irradiation
3. Neutron irradiation without 10B4C exposure.
4. BNCT with 10B4C nanoparticles (no rinse).
5. BNCT following 1 h-exposure with 10B4C and rinse
the cells with PBS.
In the treatment groups 4 and 5, 10B4C nanoparticles
were exposed for 1hour at the concentration of 10
g/well (0.31 cm2). Thermal neutron beam (Mixed mode)
irradiated the cells for 20 minutes. Cell viability was assayed by a colorimetric assay (WST-1assay).
RESULTS:
Thermal neutrons delivered to the cells were estimated as
1.0E+12 n/cm2. Results on viability in 5 treatmentgroups were shown in Fig. 1.

Fig. 1 In two control groups (groups 2 and 3, 10B4C
and IR), cell proliferation was not inhibited comparedwith no treatment group (PBS). In two BNCT groups(groups 4 and 5, 10B4C+IR and 10B4C (wash)+IR), decrease in cell viability was equally observed.
In the group 5, following rinse of the cells with PBS,
B4C nanoparticles attached to the cells was observed.
The observed nanoparticles were much lesser comparedwith those in the group 4, which suggested that BNCT
with much less 10B4C nanoparticles may be enough forsufficient cell toxicity.
DISCUSSION:BNCT with intravesical administration of 10B4C nanoparticles for CIS bladder cancer has three advantages
from a clinical viewpoint. One is that occurrence of systemic toxicity derived from the newly-developed boron
compound is not necessary to be considered. Second isthat the irradiated doses to rectum or small intestine
around the bladder is very low since no boron compound
distributed in these normal organs. Third is that 10B4C
nanoparticles are more economically prepared comparedwith other boron compounds, boronophenylalanine(BPA) and borocaptate sodium. We have planned to carryout in vivo BNCT experiments with 10B4C nanoparticles
using orthotropic bladder cancer model mice.
REFERENCES:[1] Y. Ishikawa et al., Appl. Phys. Lett., 91 (2007)
161110-161113.
10炭化ホウ素粒子を利用した硼素中性子捕捉療法の可能性（香川大・工）石川善恵（京大原子炉実験所）鈴木 実

共同通常

CO7-7Development of Novel Boron Compounds for BNCTS. Tanimori, M. Kirihata1, Y. Hattori1, M. Mukumoto1, N.
Kadono, M. Suzuki2, S. Masunaga2 and K. Ono2

H2N

10

HO

Graduate School of Life and Environmental Sciences,
Osaka Prefecture University
1
Research Organization for 21st Century, Osaka Prefecture University
2
Research Reactor Institute, Kyoto University

2-

CO2H

10

+

B

2Na

BPA

BSH

NH2

210

B

S

2CO2H

10

B

NH2

S

CO2H

n
2Na+

1a - c (1a: n = 2, 1b: n = 3, 1c: n = 6)

MATERIAL and METHOD: Cultures were inoculatedwith 1.0 x 106 cells/dish, and cells were grown for 24 h in
DMEM. The medium was replaced with the each medium containing each boron amino acid (final concentration was 1.0 mM in each case). The cells were cultured
for 24 h, and the medium was removed by aspiration. The
cells were washed with PBS, harvested by trypsinization, and then cell numbers were counted. After centrifugation, the tripsin was removed by aspiration, and to
the residual cells was added DMEM. The suspension of
the cell in DMEM (5.0 x 103 cells/mL, 1mL) was irradiated with thermal neutron for 0 - 90 min in column-shape
tube. The thermal neutron fluence was determined by

SH

OH

2Na+

Fig. 1.

 : 10BH

2a: cis, 2b: transBoron compounds.

averaging two gold foils symmetrically attached to thesurface of the column-shape tube along the direction ofincidence of thermal neutrons. After thermal neutron
exposure, 600 cells were placed in three Corning 60 mm
tissue culture dishes containing 3 mL DMEM to examine
colony formation. Seven days later, the colonies were
fixed with ethanol and stained with 0.1% crystal violet
for quantitative visualization by the naked eye.
1Surviving Fraction

INTRODUCTION: Boron neutron capture therapy(BNCT) is an attractive technique for cancer treatment.
Although many kinds of boron compounds such as aminoacid, nucleic acid and peptides have been reported asboron carrier for BNCT [1], only two compounds,
p-borono-L-phenylalanine (BPA) and mercapto-closoundecahydrododecaborate (BSH) are clinically used incure of cancer with BNCT. As part of our developing
studies on new boron delivery agents for BNCT, we havedesignedandsynthesized
thiododecaborate([B12H11]2--S-) unit containing L-amino acids (1a–c),which constitute a new class of tumor-seeking and watersoluble amino acids (Fig. 1). In vitro evaluation studies
on the cytotoxicity, killing effects by neutron irradiation,and micro distribution analysis performed previously by
our group suggested that 1a–c might be potential delivery
agents for BNCT [2]. On the other hand, recently, radio-labeled with 18F ,-cycloalkyl amino acids such as
1-aminocyclobutane-1-carboxylic acid (ACBC) arehighly noted as useful PET (positron emission tomography) tracers for diagnosis of cancer, since unusual aminoacids having ,- alkyl ring are incorporated selectively
and temporarily retained by cancer cells.
To develop practical materials utilizing 10B carrier, wehave newly synthesized dodecdaboratethio-unit containing , -cycloalkyl amino acids such as cis/trans[1-amino-3-(thiododecaboranyl)methyl]cyclobutane-1carboxylic acid (ACBC-BSH, 2a and 2b) bearing noasymmetric carbon atom, by extension of our reported
method. Here, we report the tumor cell killing effects of
2 against cultivated cancer cells.

B

0.1

CONT.
L-BPA
1a (n=2)
2a (cis)2b (trans)

0.01

0.001
0

1

2

3

4

5

6

Thermal Neutron Fluence (x 107 cm-2)

Fig. 2. Tumor cell killing effects of boron amino acids
against C6 Cell.
RESULTS: To confirm the usefulness of ACBC-BSH
2a and 2b for BNCT, we examined the tumor cell killing
effects of L-BPA, 1a, 2a, and 2b against C6 (rat glioma),cell. As shown in Fig. 2, ACBC-BSH 2a and 2b
showed higher killing effects than that of BPA and 1a.
This result suggests that ACBC-BSH 2 is useful for 10B
carrier on BNCT for glioma.
REFERENCES:[1] Soloway, A. H.; Tjarks, W.; Barnum, B. A.; Rong, F.
G.; Barth, R. F.; Codogni, I. M.; Wilson, J. G. Chem. Rev.,
1998, 98, 1515–1562.
[2] Hattori, Y.; Kusaka, S.; Mukumoto, M.; Uehara, K.;
Asano, T.; Suzuki, M.; Masunaga, S.; Ono, K.; Tanimori,S.; Kirihata, M. J. Med. Chem., 2012, 55, 6980-6984.

採択課題番号 24036
ホウ素中性子捕捉療法の新規ホウ素薬剤の開発研究
共同通常（阪府大院・生命環境）谷森 紳治、門野 尚之 （京大・原子炉）小野 公二、増永 慎一郎、
鈴木 実 （阪府大・21 機構）切畑 光統、椋本 麻里、服部 能英

CO7-8

BNCT for Malignant Melanoma and Head and Neck Cancer

N. Kamitani, K. Konishi, E. Yoden, S. Sasaoka1,
M. Uno2, N. Morita2, T. Harada2, T. Aihara3, Y. Sakurai4,
A. Maruhashi4, K. Ono4 and J. Hiratsuka
Kawasaki Medical School
Department of Radiology
1
Department of Dermatology
2
Department of Otolaryngology
3
University of Tsukuba Hospital
4
Research Reactor Institute, Kyoto University

INTRODUCTION: Advanced head and neck carcinoma (AHNC) and recurrent head and neck cancer(RHNC) are often radio-/chemo-resistant and show extensive growth, requiring a wide resection including surrounding normal tissues. To avoid severe impairment ofhead and neck structures, it is necessary to explore newtreatment for AHNC. Mishima first proposed employing
BNCT for malignant melanomas utilizing the specificmelanin synthesis activity of melanoma cells [1]. Kato etal. [2] began BNCT using both BSH and BPA for recurrent parotid gland carcinoma for the first time and reported excellent preliminary results. On the basis of theencouraging results of their pioneering clinical trial, ourmany years’ experience with melanoma BNCT and thetrend toward emphasizing the quality of life after treatment, we also began treating our patients with BNCT
using BPA alone [3-4]. This report is a summary oftreatment by BNCT using BPA alone in 2012.
16 cases are consulted to Kawasaki Medical School
Hospital as referral visit for BNCT in 2012. 10 patientswere treated, consisting 4 men, 6 women; a median ageof 60 years old (range 37-86 years). The number of headand neck recurrent carcinomas was 7, and that of cutaneous malignant melanomas was 3. 6 in head and neck recurrent cancer were squamous-cell carcinoma (SCC) recurrence, and one was the neck lymph node metastasis of
the malignant melanoma. The local recurrence was five,and the number of neck lymph node metastasis was one
among SCC. All cutaneous malignant melanomas treated
BNCT as first choice. Patients of head and neck recurrent
carcinoma have undergone the operation in the past except the one of malignant melanoma. The local recurrence was five, and the number of neck lymph node metastasis was one among SCC. All cutaneous malignant
melanomas treated BNCT as first choice. Patients of headand neck recurrent carcinoma have undergone the operation in the past except the one of malignant melanoma.
RESULT: Although the malignant melanoma faded, it
still remains. Neither a recurrence nor progression is accepted. Any adverse event is not occurred. One of thehead and neck recurrent carcinomas which is neck lymph
node metastasis of the malignant melanoma died in amonth after BNCT cause of rapid progression of cancer.

In two patients of head and neck cancer (SCC), the local
recurrence occurred within two months after BNCT.
The shortage of doses was able to be considered as
acause. And two of four patients which carried out partial
control accepted the recurrence outside the exposure field.
Although the partial control by BNCT is good, it does
not contribute to overall survival.
Conclusion: We are considering combined use withsystemic therapy may also be useful to find the better
result.

Details:
Age (median)male
FemaleHead and neck carcinoma

60 y.o.
(37-86 y.o.)
4
6
7

SCC

6
nasopharynx
hypopharynx
buccal mucosa
maxillary sinus
external auditory canal

1
1
1
1
2

malignant melanoma

1local recurrence (SCC)

5cervical lymph node metastasis
external auditory canalmalignant melanoma

2cutaneous malignant melanomas
heelupper lip
Result:local recurrence
metastasis

1
1
3
2
1
2
2

REFERENCES:[1] Y. Mishima, Pigment Cell Res., 1 (1973) 215-221.
[2] I. Kato et.al., Appl. Radiat. Isot., 61 (2004)
1069-1073.
[3] T.Aihara et al., Head Neck., 28 (2006) 850-855.
[4] T.Aihara et al., New Challenges in Neutron capture
therapy, (2010) 25-29.

採択課題番号 24037
中性子補足療法適応癌腫のプロトコル確率
共同通常（川崎医大）平塚純一、原田保、粟飯原輝人、宇野雅子、森田倫正、牧野英一、笹岡俊輔、小西圭、
神谷伸彦（京大・原子炉）小野公二、丸橋晃Study on the Semiconductor Device Error Irradiated with Thermal Neutrons

H. Tanaka, Y. Sakurai, J. Hori, M. Suzuki, S. Masunaga,
N. Kondo, M. Narabayashi, Y. Kinashi, K. Ono and
A. Maruhashi
Research Reactor Institute, Kyoto University

INTRODUCTION: Recently, the irradiation experiments of fast neutrons for semiconductor devices havebeen performed using cyclotron facility of ResearchCenter for Nuclear Physics, Osaka University [1]. However, irradiation test using thermal neutron for semiconductor device has not been systematically performed. If
the nuclide with large capture cross section of thermal
neutrons is contained in semiconductor devices, charged
particles are produced in semiconductor devises. Charged
particles create electron-hole pair toward those tracks that
become the current of noise in semiconductor devises.
There is fear that the current in semiconductor devices
triggers incorrect operation. Furthermore, the miniaturization of semiconductor device is proceeding and thecurrent of signal is becoming small. Thus, the sensitivity
for electronic noise becomes large. The aim of this study
is the investigation of the influence for semiconductor
device of thermal neutron irradiation.
EXPERIMENTS: The sample of semiconductor device was irradiated at Heavy Water Neutron Irradiation
Facility (HWNIF) of KUR [2]. To investigate the influence of thermal neutrons, the difference of the number of
error (bit flip of memory) for epithermal mode
(CO-0000F) and mix mode (OO-0000F) was recorded.
Fig. 1 shows the neutron spectrum of two irradiation
mode. Epithermal mode can produce the epithermal andfast neutrons without thermal neutrons using a cadmium
filter.
A number of semiconductor devices were irradiatedsimultaneously. The irradiated thermal neutron distribution was not uniform because of its large irradiation area.
In order to measure thermal neutron flux, gold wire wasset at the surface of each semiconductor device. Goldwire was taken from the surface of a semiconductor device to determine thermal neutron flux. The number of
error of each semiconductor device was recorded during
the irradiation of thermal neutron. Next, the number of
error was also recorded for epithermal and fast neutron
irradiation.
RESULTS: Table 1 shows the number of error of eachsemiconductor for each irradiation mode and thermal
neutron flux. The deviation of thermal neutron fluxes foreach semiconductor device was shown in this table. The
importance of the measurement of thermal neutron fluxes
was revealed according to these results. Fig. 2 shows thecomparison of the number of error, that was removed theinfluence of epithermal and fast neutrons, per thermal
neutron fluence. As shown in Fig. 2, the number of error
for sample D, E, F was larger than other samples. We
established the thermal neutron irradiation field for theinvestigation of error of semiconductor devices.

Fig.1. Neutron spectrum of CO-0000F and OO-0000F
mode at the surface of gamma ray shield of HWNIF.
Table 1. the number of error of each semiconductor
for each irradiation mode and thermal neutron fluxes
Thermal neutron
The number of error
Flux [n/cm/s] OO-0000F CO-0000F
Sample A
1.0E+853
21
Sample B
1.0E+839
17
Sample C
9.1E+711054
Smaple D
2.0E+87378
293
Sample E
2.1E+83959
110
Sample F
2.1E+83537
92
Sample G
1.2E+839
124000
Number of the error per thermal neutron
fluence(a.u)

CO7-9

3500

3000
2500
2000
1500
1000

500
0

A

B

C

D

E

F

G

Fig.2. The comparison of the number of error for eachsample.
REFERENCES:[1]Y. Iwamoto, M. Fukuda, Y. Sakamoto, A. Tamii, et al.,Nuclear Technology, 173(2), 210-217(2007)[2] Y. Sakurai, T. Kobayashi, Nucl. Instrum. Meth. A453,
569-596(2000)

採択課題番号 24043
中性子照射による半導体デバイスエラーに関する研究（京大・原子炉）田中浩基、櫻井良憲、堀順一

一般通常CO7-10

Tissue Changes by BNCT of the Oral Cancer Tissue at Having Used
Hyaluronan Conjugated PEG Liposome

Y. Ito, Y. Ariyoshi, Y. Kimura, Y. Nakajima, Y. Takei,
T. Ueno and M. Suzuki1

images of tumor tissue proliferation were observed.
In the future, methods to increase the effectiveness ofkilling tumor cells should be considered.

Department of Dentistry & Oral Surgery, Division of
Medicine for Function & Morphology of Sensory Organs,
Osaka Medical College
1
Research Reactor Institute, Kyoto University
INTRODUCTION: In Boron neutron capture therapy(BNCT), selective and highly concentrated boron accumulation in tumor cells is important. A study was carriedout in our division using various liposomes [1, 2].
Hyaluronan is a ligand of CD44 and RHAMM, which isexcessively expressed in tumor cells, and Hyaluronan-conjugated PEG Liposome (HA PEG Liposome) isan active targeting candidate for tumor cells.
In this study, the effect of neutron irradiation was investigated by histological observation, in order to consider BSH transport using HA PEG Liposomes in micewith oral squamous cancer cells.
EXPERIMENTS: Tumor bearing mice were preparedby injecting cultured SAS cells into the right thigh of
BALB/c mice. When tumor size reached approximately
10 mm, HA-PEG-Liposomal BSH was intravenously
administered from the tail vein. This injection was carriedout at the point in time when boron concentration in thetumor tissue could be maintained at a comparatively
higher condition than the surrounding tissue than that ofprevious experiments, in other words, two days before ofneutron irradiation. For reference, a control group whichused liposomes without boron was also set. After neutron
irradiation, the mice were allowed to grow for 8 weeks.
At the 8 weeks, all mice were euthanized, and tissue
samples were taken. Formalin fixation was carried out on
the samples, and the samples were set as a formalin fixedparaffin sections according to the standard method, HE
dyed, and observed by a microscope.
RESULTS: Regarding tumor size observation after
irradiation, among the group administered BSH, significant tumor shrinkage effect was observed in some specimens and no effect was observed in other specimens. No
tumor shrinkage effect was observed in the control group.
Variation in the results in the administered group is assumed to be due to technical error in intravenous injection, etc.
In specimens where tumor shrinkage effect was observed, the site of the tumor was macroscopically ulcer-like in appearance and no tumor was observed. Histologically, the ulcer sites were observed on the epithelial
surface, and tumor residue was observed in the connective tissue layer under the ulcer. In the control group,

Fig. 1. Macroscopic photo of a mouse before neutron
irradiation

Fig. 2. Macroscopic photo at the 8 weeks after neutron
irradiation, boron injected group

Fig. 3. Macroscopic photo at the 8 th week after neutron
irradiation, boron non-injected group

Fig. 4. HE tissue image, ulcer-bearing section in the epithelium, and tumor cells are observed in the connective
tissue layer under the ulcer. On the ulcer side of the tumor,
necrotic tissue and fibrin deposition was observed.
(bar=2mm)
REFERENCES:[1] T. Shimahara et al., Bulletin of the Osaka Medical
College. 55(2009): 21-29[2] Y. Kimura et al., Bulletin of the Osaka Medical
Col-lege. 56(2010): 65-7

採択番号 24046
口腔癌に対する硼素中性子補足療法の基礎的研究
共同通常（京大・原子炉）鈴木実 （大阪医科大学・口腔外科）伊藤雄一、有吉靖則、木村吉宏、中島世市郎、武井祐子、植野高章

CO7-11Development of Water-Soluble Gd Metallofullerenes/PEG-b-Polyamine
Complexed Nanoparticles for Neutron Capture Therapy

Y. Nagasaki1-3, T. Yaguchi1, Z. Gao1, S. Sumitani1, Y.
Horiguchi1, S. Noda,4 Y. Nakanishi,4 H. Shinohara,4 M.
Suzuki5and K. Ono5
1

Institute of Materials Science, Graduate School of Pureand Applied Sciences, University of Tsukuba,
2
Master’s School of Medical Sciences, Graduate Schoolof Comprehensive Human Sciences, University of Tsukuba
3
Satellite Laboratory, International Center for Materials
Nanoarchitectonics (WPI-MANA), National Institute ofMaterials Sci-ence (NIMS)
4
Graduate School of Science Division of Material Science, Nagoya University
5
Radiation Oncology Research Laboratory, Research
Reactor Institute, Kyoto University
INTRODUCTION: Gadolinium element is one ofpromising candidates for neutron capture therapy (NCT)
because of high neutron capture cross section. The essential toxicity of Gd ion is one of the problems to solve.
Gadolinium chelates with specific ligand such as diethylenetriaminepentaacetic
acid(DTPA),
and
1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid
(DOTA), have been proposed to reduce toxicity. However, these Gd chelates still show toxicity and are notenough for NCT, because high dose is required for NCT
treatment. Gadolinium endohedral metallofullerene(Gd@C82) are emerging compounds and promising as
new biomaterials to suppress the toxicity of Gd by fullerene cage. In order to use Gd@C82 compounds in biological fluid, delivatization of Gd@C82 has been done inmost cases, which causes unwanted bioactivity. Thus, wehave been used fullerenes without delivatization. To solubilize fullerenes in aqueous media, poly(ethylene glycol)-block-poly(2-(N,N-diethylamino)ethyl methacrylate(PEG–b–PAMA) were employed in our previous work[1].
The result of the NCT experiment in vitro showed that
Gd@82/PEG–b–PAMA complexed nanoparicle was effective NCT agent. To develop the more safety water-soluble Gd@C82 nanoparticles as NCT agent, radical
containing amphiphilic block copolymer (PEG–b–PMNT)
was used as dispersion agent [2]. PEG–b–PMNT is low
toxic polymer (LD50 > 600 mg/Kg), and it acts as reactive
oxygen species (ROS) scavenger to suppress theunwanted immune response induced by ROS after neutron
irradiation. In this report, we investigate the NCT by
Gd@C82/PEG–b–PMNT
complexed
nanoparti-cles(GdNPs). To confirm the tumor treatment, tumor

採択課題番号24047

sizes of the mice were measured after neutron irradiation.
EXPERIMENTS:
Preparation of Gd@C82 nanoparticles: 10 mg of
Gd@C82 was dissolved in 50 mL of DMF and sonicatedfor 1 h. 50 mg of PEG–b–PMNT was added to the mixture and the sonication continued for 3 h. The mixture
was transferred to a dialysis tube and dialyzed against 2 L
of water. The external water was changed 5 times at t = 3,
15, 39, 63 and 87 h. The solution was condensed by ultrafiltration and dispersed in PBS for in vivo experiment.
Preparation of tumor bearing mice (BLAB/c, male,
5weeks): Tumors were prepared in mice legs by hypodermic injection of colon-26 cells (1,000,000 cells per
mouse). This procedure was carried out a week before theneutron irradiation.
Neutron irradiation: The administration of the GdNPs
were carried out via tail vein (1 mg of Gd@C82 permouse). After 2 d, the mice were irradiated thermal neutrons for 120 min at a rate of 9.4 − 9.8 × 1012 neutrons/cm2). After the neutron irradiation, the tumor sizesof mice were measured for 26 d.
RESULTS:
Evaluation of obtained Gd@C82 nanoparticles: The sizeof the GdNP was evaluated by DLS. The result showedthat the average size of GdNP was ca. 40 nm and thedispersion property of the particles is quite stable enoughto keep the dispersion for over a year.
Neutron Irradiation: The tumor grew up to 6 – 7 cm3after 26 d without GdNPs and no thermal neutron irradiation (n = 4). On the other hand, the growth was effectively suppressed in the mice treated with GdNPs with
the irradiation of thermal neutron (average size was 2 cm3,
n = 4). The suppression of tumor growth was also observed in the mice with the irradiation of thermal neutron
without GdNPs (average size was approx. 4 cm3, n = 5),
the suppression efficacy of tumor growth of the micewith GdNPs with the irradiation was higher than that of
the mice without GdNPs with the irradiation (p-value =0.144). Our results demonstrate that the rational material
design of GdNPs holds promise for the future of GdNCT.
REFERENCES:[1] Y. Horiguchi, S. Kudo, Y. Nagasaki, Sci.
Technol. Adv. Mater., 12 (2011) 044607.
[2] T. Yoshitomi, D. Miyamoto, Y. Nagasaki:
Biom-acromolecules, 10 (2009) 596.

高い腫瘍選択性を有するボロン含有ナノ粒子の創製と共同通常
中性子捕捉療法への展開（筑波大・数理）長崎幸夫、矢口達也、角谷省吾、高振宇、堀口諭吉（名古屋大学）野田祥子、
中西勇介、篠原久典（京大・原子炉）鈴木実、小野公二

CO7-12

Pilot Biodistribution Study of Poly-ion Complex MediatedGadolinium Delivery System to Cancer Model In vivo

H. Yanagie1,2, N. Dewi1, Y. Sakurai2, M. Yanagawa3,
T. Iizuka3, T. Nagasaki4, M. Suzuki5, Y. Sakurai5,
S. Masunaga5, K. Ono5 and H. Takahashi1,21Department of Nuclear Engineering & Management,
The University of Tokyo
2
Cooperative Unit of Medicine & Engineering, The University of Tokyo Hospital3
Department of Veternary Surgery, The University of Tokyo
4
Department of Bioengineering, Osaka City University,
5
Research Reactor Institute, Kyoto University
INTRODUCTION:
Tumour cell destruction in gadolinium (Gd) neutron-capture therapy (GdNCT) is due to the nuclear reaction between Gd atoms and thermal neutrons. It is necessary for effective neutron capture therapy to accumulate Gd atoms in the tumour cells without affecting adjacent healthy cells.
We have developed novel efficient gene transfection
system, comprising the plasmid/polycation complex core
and the outer polyanion-coating. We used Polyethyleneimine (PEI) as polycationic polymer, and Polyethylene
glycol derivatives having carboxylic acid [1,2]. We had
also applied this poly-ion complex as heavy ion delivery
systems. In this study, we prepare Gadoteridol entrapped poly-ion complex for selective intravenous or
intratumoural injection for mouse colon cancer model
applicating to GdNCT, and evaluate the poly-ion complex as selective Gd delivery carrier to cancer tissues.
EXPERIMENTS:(1) Cell: Mouse colon cancer cell line: Colon 26(2) Polyethyleneimine: ExGen 500 (Fermentas Ltd.) wasused. ExGen 500 is linear shaped 22 kDa PEI. The efficient gene transfer activity of ExGen 500 is related to its
capacity for condensing DNA, interacting with anionicproteoglycans of the cell membrane, protecting DNA andinducing endosomal swelling and rupture before DNA
degradation.
(3) PEG-C: Polyethylene glycol derivatives having carboxylic acid was synthesized (MW of PEG was 8940,
17.7 carboxylic acid groups were binding per PEG molecule).
(4) JTS-1: pH dependent fusogenic peptide was kindly
gifted by Professor Leaf Huang, Department of Pharmacy,
University of Pittsburgh, PA, USA.
(5) Gadoteridol: (±) -10 - (2-hydroxypropyl) -1,4,7,
10- tetraazacyclo-dodecane -1,4,7- triacetatogadolinium［III］( C17H29GdN4O7 ) (MW: 558.69, 1396.5 mg/5ml) .
(6) Gadolinium Delivery in the Colon 26 tumours①Colon 26 cells (2 x 106) were injected subcutaneously
into the back of female BALB/c mice. At 10–14 days
after injection, when the estimated tumour weight
reached about 500 mg, the recipient animals were injected intravenously or intratumorally with either Gd/PEI,
Gd/PEI/PEG-C/JTS-1, or Gd solution.
②One, and three days after injections, the Gd concentrations of the tumor nodules, blood, and normal organs
were measured. The Gd concentrations of tissues were
determinated by ICP- Mass Spectroscopy of Juntendo
University.
RESULTS:The Gd concentration in tumour by intratumoural (IT)delivery with Gd /P EI was 53.7 ppm, 22.5 ppm, after 2hours and 12 hours, respectively. The Gd concentration intumour by IT delivery with Gd /P EI/P E G - C/J T S -1
was 21.9 ppm, 12.3 ppm, after 2 hours and 12 hours, respectively. The Gd concentration in tumour by IT delivery with Gd so l u tio n was 17.4 ppm, 6.8 ppm, after 2hours, 12 hours, respectively.
The Gd concentrations in Colon 26 tumour on delivery
with poly-ion complex (Gd /P EI ) was 3 times superior tosimple Gd so l u tio n after 2 hours IT injection.
The Gd concentration in tumour by intravenous (IV)delivery with Gd /P EI was 4.0 ppm, 1.2 ppm, after 2hours, 12 hours, respectively. The Gd concentration intumour by IV delivery with Gd /P EI /P EG - C /J T S-1
was 9.8 ppm, 2.7 ppm, after 2 hours, 12 hours, respectively. The Gd concentration in tumour by IV delivery
with Gd so l ut io n was 1.1 ppm, 0.0 ppm, after 2 hours,
12 hours, respectively.
The Gd concentrations in Colon 26 tumour on delivery
with poly-ion complex (Gd /P EI ) was 4 times superior tosimple Gd so l u tio n after 2 hours IV injection, and also
9 times superior to control groups in the usage ofpoly-ion complex (Gd /P EI / P EG -C /J T S -1 ) .
CONCLUSION:
Poly-ion complex (Gd /P EI /P EG - C/J T S -1 ) can beapplied to the Gd delivery systems with the retention activity and dispersion activity. We are ongoing to evaluate
the suppressive activity with these complexes by thermal
neutron irradiation.
REFERENCES:[1] M. Sakae et al., Biomed Pharmacother., 62 (2008)
448-453.
[2] H. Yanagie et al., IGCC 2013 Abstract., (2010) P4-7

採択課題番号 24050 中性子捕捉療法の一般外科領域癌への展開に向けた基礎的研究 共同通常
（東大・原子力国際専攻）高橋浩之、柳衛宏宣、Novriana Dewi（東大・獣医）柳川将志、飯塚智也（東大・心臓外科）櫻井由里子、毛利きくえ（大阪市大・バイオ）長崎 健、李 家暐、朱 宇
翔、河崎 陸（京大・原子炉）小野公二、増永慎一郎、鈴木 実、櫻井良憲

CO7-13

Pilot Clinical Study of Boron Neutron Capture Therapy
for Recurreced Hepatic Cancer

H. Yanagie1,2, S. Higashi3, K. Seguchi3, I. Ikushima4, K.
Oyama5, Y. Nonaka6, S. Maruyama5, R. Hatae5, M. Suzuki7, S. Masunaga7, T. Kinashi7, Y. Sakurai7, H. Tanaka7,
A. Maruhashi7, K. Ono7, J. Nakajima2,8, M. Ono2,8, M.
Eriguchi5 and H. Takahashi1,2
1

The University of Tokyo, Department of Nuclear Engineering & Management
2
Cooperative Unit of Medicine & Engineering, The University of Tokyo Hospital, Tokyo
3
Kojinkai Medical City East Hospital, Department ofSurgery, Miyazaki
4
Miyakonojyo Metropolitan Hospital, Department of Radiology, Miyazaki
5Japan Anti-Tuberculosis Association, Shin-Yamate Hospital, Department of Surgery, Tokyo
6
Keiaikai Hoyo Hospital, Department of Surgery, Iwate
7
Kyoto University, Research Reactor Institute, Osaka
8
The University of Tokyo Hospital, Department of Cardiothoracic Surgery, Tokyo, JAPANINTRODUCTION:
Applications of boron neutron-capture therapy (BNCT)
has been increased clinically in patients with a lot ofcancers in hole body. The main two 10Boron compounds(sodium mercaptoundecahydro dodeca- borate : 10BSH, 10B-p borono- phenylalanine (10BPA) and
its fructose complex ) are used to clinical trials. Tumour
cell destruction in BNCT is due to the nuclear reaction
between 10Boron and thermal neutrons. For effective
BNCT therapy, it is necessary to accumulate 10B atoms in
the tumour cells without affecting adjacent healthy cells.
Most of hepatocellular carcinomas (HCC) are thought
to be incurable, and limited surgical operation, chemotherapy, or radiation therapies are available for a prolonged survival. Suzuki et al. had reported that the intra-arterial administration of a boron compound withIPSO is technically an application of chemoembolization,
which has been widely used for the treatment of liver
tumours. They also reported the clinical results of thefirst patient with multiple hepatocellular carcinomas(HCCs) treated with BNCT. Higashi et al. prepared along term inseparable, water-in-oil-water emulsion(WOW) containing 8-60 mg of epirubicin for use in arterial injection therapy to treat patients with HCC. The
WOW was prepared by membrane emulsification technique using a controlled pore glass.
We started the pilot clinical studies of BNCT to recurrence breast cancer, hepatic cancer, and gastrointestinal
cancers. In this paper, we present pilot clinical study inpatients of hepatic cancer.

RESULT [Case 1]
In accordance with the clinical results of Higashi andcolleagues, water-in- oil-in-water (WOW) emulsion has
been used as the carrier of anti-cancer agents on intra-arterial injections in clinical trials. We would like toapply BNCT for the treatment of HCC in order to increase the selection of therapies available for HCC patients. We developed a 10BSH containing WOW emulsion
using a double emulsification technique.
A 63-year-old man with multiple HCCs was enrolled
as the first patient in a pilot study for treating BNCT with
10
BSH containing WOW emulsion. The patient had been
performed right hepatectomy in 6 years ago. Hepatic
arterial chemotherapies with epirubicin containing WOW
emulsion were performed in the recurrence stages. The
multiple tumours in the left liver lobe were treated with
BNCT by selective intra-arterial infusion of 10BSH containing WOW emulsion. The pre-BNCT dosimetry was
performed using SERA(mean tumour fluence is 12Gy-Eq
on 56 minutes BNCT (Maximum 19Gy-Eq on tumour),and maximum fluence of normal mucosa is 5.0 Gy-Eq).
The tumour size was remained stable during 3 months
after BNCT. No adverse effect as a result of BNCT was
observed during the treatment and follow-up period. The
BNCT-treated tumours showed regrowth 3 months after
BNCT, so the patient has continued the repeated hepatic
arterial chemotherapy of epirubicin containing WOW
emulsion.
The present results showed that 10B-containing WOW
emulsion can be applied as a novel intra-arterial boron
carrier for BNCT for HCC.
A

C

B

Figure1. A : Mcro-photo.
10
BSH-containing WOW
Emulsion, B : The shemaof the positioning of patient, C : Two dimensionaldose distributions(Gy-Eq)
for tumour in the left lobe
resulting from irradiation
with frontal epithermal
neutron beams (surface is
30Gy-Eq)

採択課題番号 24051 中性子捕捉療法の一般外科領域癌への展開に向けた臨床的研究 共同通常
（東大・原子力国際専攻）高橋浩之、柳衛宏宣（宏仁会メディカルシティ東部病院）東 秀史、
瀬口浩司（結核予防会 新山手病院）江里口正純、丸山正二、小山和行（啓愛会 宝陽病院）野中泰政（京大・原子炉）小野公二、増永慎一郎、鈴木 実、櫻井良憲

CO7-14Development of Boronated Liposome for Boron Neutron Capture Therapy

M. Shirakawa1, K. Nakai1,2, Y. Yamamoto1, F. Yoshida1,2,
T. Yamamoto1,2, A. Matsumura1,2, H. Tanaka3, M. Suzuki3and S. Masunaga3
1

Department of Graduate School of Comprehensive Human Sciences, University of Tsukuba
2
Department of Neurosurgery, Faculty of Medicine, University of Tsukuba
3Particle Radiation Oncology Research Center, Research
Reactor Institute, Kyoto Universityii)5%PBL lipsome group revealed tumor growth inhibition. There are significant difference after 10 days fromthe irradiation compared with other groups. (student T
test P<0.05 ) (Fig.2) In 5% PBL liposome group, there
was one complete remission.

INTRODUCTION: Boron Neutron Capture Therapy(BNCT) has cell selective radiation therapy theoretically. Therapeutic effect of the boron compound is basedon alpha particles produced by the 10B (n, α) 7Li reaction
and tumor selectivity [1]. We developed novel boronatedliposome for BNCT, analyzed the neutron capture effectfor the cancer cell line and tumor bearing mouse model[2].
EXPERIMENTS: Synthesized the new boronated lipid
compound (PBL). Boronated lipid and other lipids(DSPC: cholesterol: PBL = 1:1:0.12) were dissolved inorganic solvent and assembled as a liposome of 100 μm
diameter using lipid film methods and extruder.
i) In Vitro Colony forming assay using V79 and 379A
was performed with three groups of samples; (1) Boronated liposome without medium change, (2) Boronatedliposome with medium change, (3) no boron argentsgroups. Each cell suspesions were irradiated at KUR.
Irradiation time of each groups were 15min, 30min, and45min. After 1week incubation, the colonies were counted.
ii) In Vivo tumor growth inhibitory test with animal experimental tumor bearing model. 107 of CT26 colon cancer cells were injected subcutaneously to right thigh of
BALB/c mice. 2week after the injection, 5% PBL liposome was administrated via tail vein. The Boron dose
was 10mg/kg, volume of 100µl. Concentration of theboron was 2000ppm.
Mice were devided four groups; (1) 5% PBL lipsomegroup, (2) BSH water solutions, (3) Neutron only and (4)no treatment groups.
RESULTS:i) Boronated liposome and neutron capture effect In Vitro.
(2) Medium wash group and (3) no boron agent (neutrononly) groups had same dose response. It means that the
5% PBL liposome was not accumulating into cells fromthe surrounding medium. (1) Without wash group hadcell killing effect compared with (2) and (3). It meanssurrounding medium containing 5% PBL liposome could
effect to cell. (Fig.1)

採択課題番号 24052

Fig. 1. Cytotoxicity reaction by thermal neutron irradiation with PBL modified liposome. The V79 379A cells
were irradiated with 5.6Gy for 15 minute, 12.1Gy for
30minute and 18.4Gy for 45minute. Data are represented
as ratio of control. (mean± S.D.).

Fig. 2. Tumor growth inhibitory effect by thermal neutron irradiation with PBL modified liposome. The tumor-bearing mice were irradiated with 14.2 Gy for 2
hours. (mean± S.D.)

REFERENCES:[1] K. Maruyama, et al., J. Controlled. Release. 98(2004) 195-207.
[2] M. Shirakawa et al., App. Rad. Isot.,67 (2009)
S88-90.

新規化合物および細胞内濃度変調による中性子捕捉反応による 共同通常
細胞生存試験および腫瘍増殖抑制効果（筑波大学・医学医療系）中井 啓、山本哲哉、吉田文代、白川 真、山本陽平、上前洋二、佐藤英介（京大・原子炉）増永慎一郎、櫻井良憲、田中浩基

CO7-15The Benefit of High LET-Radiation to Glioma Stem-Like Cells

Y. Hirota, S. Masunaga1, N. Kondo1, T. Fujishiro2,
S. Kawabata2, A. Fujimori3, H. Hirakawa3, H Yajima3,
K. Ono1, T. Kuroiwa2 and S. Miyatake2
Graduate School of Neurosurgery, Osaka Medical College
1
Research Reactor Institute, Kyoto University
2
Department of Neurosurgery, Osaka Medical College
3
National Institute of Radiological science, Heavy-ion
Radiobiology Research group, Research Center for
Charged Particle Therapy

PURPOSE:
Radiation therapy with surgery and chemotherapy is thestandard treatment for glioblastoma multiforme (GBM)[1]. However, almost half of GBM patients cannot survive one year after diagnosis, and the prognosis of patients with GBM has not been improved over the past
decades. Recently, there have been some reports showing
the presence of glioma stem cells (GSCs) in malignant
gliomas which are regarded as highly radio-resistant tolow linear energy transfer (LET) photons[1, 2]. On theother hand, we have applied boron neutron capture therapy (BNCT) for GBM. This is a unique tumor-selective
particle radiotherapy using neutron irradiation, especiallythermal neutron irradiation. Boron-10 (10B) releases alpha(4He) and 7Li particles by 10B(n,α)7Li reaction. The key
players of anti-tumor effects in BNCT are these high linear energy transfer (LET) particles. With BNCT, goodresults have been achieved for patients with GBM andrecurrent malignant glioma[3, 4]. Here we analyzed thebenefit of high LET particles to GSCs.

MATERIALS AND METHODS:
Glioma stem-like cells (GSLCs) were induced from
GBM cell line A172 in stem cell-culture medium [5]. The
phenotype of these GSLCs and wild type cell lines wereconfirmed by western blot analysis and fluorescence-activated cell sorting (FACS) using stem cellmarkers. These cells were irradiated with 60Co gamma
rays or neutron beams. Radio-sensitivity was assessed bya colony-forming assay[6] and the number of DNA double strand breaks (DSBs) using histone gamma-H2AX
foci detection assay[7, 8].

RESULTS:
In stem cell-cultured medium, GSLCs could form
neurosphere-like spheroid cells. And GSLCs expressedneural stem cell markers more frequently in western blot
analysis and the ratio of CD133 positive cells increased
day by day. GSLCs were radio-resistant to gamma-rays incomparison with parental cultured cell lines, but neutron
beams could overcome the resistance. Twenty-four hours
after irradiation with gamma-rays, the number of gamma-H2AX foci in GSLCs was significantly less than thatof parental cells, while there was no apparent differencein the number of these foci between GSLCs and parental
cultured cell lines following neutron beam irradiation. In
addition, neutron beam irradiation induced larger gamma-H2AX foci than those observed after gamma-ray irradiation in both types of A172 cells.

CONCLUSION:
Neutron beams can induce elastic scattering and nitrogenneutron capture reaction, and produce proton particle(H+). This particle is high LET radiation and it could
overcome radioresistance of GSLCs with unrepairable
DSBs. So we could demonstrate that high LET radiation
may be able to overcome GSCs that are resistant to low
LET radiation. It is necessary to further investigate theusefulness of high LET radiation to control GSCs, andhigh LET radiation therapy such as BNCT has a very
important role in further treatment for therapy-resistant
GBM.

REFERENCES:[1]S. Bao et al, Nature 444 (2006) 756-760.
[2]C. Charn-jung et al, Biochemical and Biophysical Research Communications, 380 (2009) 236-242.
[3]S. Miyatake et al, Journal of Neuro-oncology, 91(2009) 199-206.
[4]S. Kawabata et al, Journal of Radiation Research, 50(2009) 51-60.
[5]Q. Lei et al, Cancer Letters 279 (2009) 13-21.
[6]A. Maria et al, International Journal of Radiation Oncology Biology Physics, 79 (2011) 262-268.
[7]A. Osma et al, International Journal of Radiation Oncology Biology Physics, 75 (2009) 1216-1225.
[8]Y. Kinashi et al, Radiation Oncology, 6 (2011) 106.

新規硼素化合物の有用性と腫瘍幹細胞の克服
採択課題番号 24054（大阪医大）宮武伸一、弘田祐己、川端信司、平松亮、頼経英倫那（岡山大）道上宏之、森亜希子、王飛霏（熊本大）富澤一仁（京大原子炉）小野公二

共同通常

CO7-16

A Biodistribution Study of Carboran Sugar

M. Takagaki, I. Snajdr1, R. Satapathy1, NS. Hosmane1
and S. Masunaga2
College of Nursing, Aino Univ
1
Dept Chem & Biochem, Northern Illinois Univ
2
Research Reactor Institute, Kyoto University

Carboran sugar (carboranyl-thio-d-glucose: TDG)
has been chemically modified via novel approachand reduction of its IC50 has been achieved to bevery low 5.3x10-2 M that is almost half of that of
BSH (2.75x10-2 M) [1].
In this study, biodistribution of TDG derivative has
been preliminary investigated via -autoradiography. C6 tumor cells were implanted via
stereotactic maneuvers into Wister’s rat brain. 2
weeks after the implantation, 100mg/kg bodyweight of the compound was injected into
peritoneal cavity. 3hs after injection, whole brain
was removed and rapidly frozen in liquid nitrogen.
Frozen sections were mounted onto the solid-state
track detectors: Kodak LR 115 and were exposedby thermal neutrons. The detector were then etchedin 10% NaOH solution at 60℃ to emerge - and/or
recoil 7Li particles tracks of 10B(n,)7Li that could
be numerically evaluated via an ordinary lightmicroscope.
Figure 1 showed the -track autoradiography of thecompound in a rat C6 brain tumor.

The biodistribution of the compound was very
similar with that of BSH. Perhaps it distributes inthe brain via non-permeable fashion of blood brain
barrier, but blood tumor barrier.
Although our preliminary in-vitro-BNCT studies of
the compound showed that the surviving fraction
was smaller than that of BPA, its tumor cell killing
effect is still low for clinical purpose as BPA.
Further modification of its ligand has beenaddressed in this study. Perhaps the compound
combines the best of BSH and BPA.
REFERENCE[1] I. Snajdr, et al. 2013 European J. Chemistry (ibid)

Fig. 1.-track autoradiography of boron biodistribution
in C6 rat brain tumor.

採択課題番号 24057
悪性脳腫瘍のための熱外中性子捕捉療法の基礎的研究（藍野大）高垣政雄（東邦大・医）東丸貴信（京大・原子炉）増永慎一郎

共同通常CO7-17

Clinical Studies on BNCT for 5 Cases of Head and Neck Cancer

I. Kato, N. Yamamoto1, Y. Fujita2, M. Ohmae3, M. Suzuki7, S. Masunaga7, M. Nakazawa, T. Sumi, A. Maruhashi7,
Y. Imahori5, M. Kirihata6 and K. Ono7
Dept. of Oral &Max.fac. Surg. II, Grad. Sch. of Dent.,
Osaka Univ
1
Dept. of Oral &Max.fac. Surg., Saiseikai-Senri Hpsp
2
Dept. of Oral &Max.fac.Surg., Higashiosaka General
Hosp
3
Oral &Max. fac. Surg., Rinku,General Med. Center
4
Dept. of Oral &Max.fac. Surg., Rinku General Medical
Center5
CICS Inc
6
Graduate School of Agriculture and Life Science, Osaka
Prefecture University
7
Research Reactor Institute, Kyoto University
INTRODUCTION: We had first reported that six patients with head and neck cancer (HNC) had been treatedwith BNCT [1]. We also report long term (more than
5-year) clinical outcomes of our 26 patients with recurrent HNC treated with BNCT [2]. We summarized 5 patients with HNM who had treated with BNCT at KUR inlast year in Table 1.
PURPOSES: The purpose of this study was to estimate
safety and effectiveness of BNCT for patients with advanced/ recurrent HNC for which there were no othertreatment options.
RESULTS: We also report here latest clinical outcomesof 35 patients with recurrent HNC
All cases are advanced such as 17 (53%) out of 35 patients had developed regional lymph node metastases.
Distant metastases were developed in 10 cases (29%)
during treatment. (1)10B concentration of tumor/normal
tissue ratios (T/N ratio) of FBPA-PET studies were SCC:
1.8-6.0, sarcoma: 2.5-4.0, parotid tumor: 2.5-3.7. (1) Regression rates were CR: 13cases (51%), PR: 13cases(37%), PD: 3cases (9%), NE (not evaluated):1case. Response rate was 88%. (2) Mean Survival time was
24.2months. 2-year overall survival rate (OS) and 6-year

OS were 42% and 36%, respectively. (3)BNCT improved
QOL, PS and survival periods. (4)Survival periods after
BNCT were 1-84 months. (5) Adverse events were brain
necrosis, osteomyelitis and transient mucositis and alopecia and so on.
Case 1: A 40 year-old female with recurrent adenoid
cystic carcinoma of nasal cancer (rT0N3M0, ACC), who
had got a skull-base operation and irradiated RT 60Gy
post operatively in 2004. About 5-year later, she had got
another surgical operation because of intra-orbital recurrence. In 2011 she had developed another recurrence justafter she had got endoscopic surgery for a solitary lungmetastasis. FBPA-PET study revealed 10B concentrationof T/B ratio was 2.0. Then she has been treated with BPA(500mg/kg) mediated BNCT at KUR in June 2012 andshe has been disease free survival for so far 11-month.
Case 5: A 65-year old woman with SCC at Lt-WK(T4N0M0) had got surgery with microscopic forearmreconstruction in March 2011 and she had got surgery of
Rt-RND and postoperative radiotherapy (54Gy) in June,
Lt-RND in December. She had got bilateral selective
intra-artery chemotherapy (CDDP+TXT) after having
developed recurrence in February 2012. Again she had
developed cervical LN metastases (L-Level II:4cm,
R-Level V:1.5cm) so she had weekly treated with [Cetuximub (400mg/m2,250mg/m2)+Paclitaxel:60mg/m2]x7.
FBPA-PET study resulted that T/B ratio=4.0. Just before
BNCT, the left of level II LN had grown to 5cm withundefined margin which was infiltrated into para-pharyngeal area. The LN was necrotic and skin was
ruptured and discharged cancer milk. She treated BNCT
in February, 2013. After BNCT the LN had completely
disappeared and had covered with normal skin. Then she
has been disease free survival for so far 5-month.
REFERENCES:[1] I. Kato, et al., Appl. Radiat. Isot., 61 (2004) 1069-1073.
[2] R. Barth, et al., Radiat. Oncol., in press.

Table 1. Treatment Summary of 5 Cases
Case
No.

Pt‘s Initial
(Age)

1

Y・M (40)

Clinical Diag.
(Histopathol. Diag.)
Rt-op.OKK, op. lt-Lung meta
(ACC)

2

A・H（51）

Rt-Op. ZK, Rt-LN meta.(SCC)

3

K・M (71)

Rt-Op.OKK (SCC)

4

K・Y(83)

Rt-0p.OGK (SCC)

5

A・K(60)

Lt-Op-WK、Lt-LN meta

10B-conc.
Blood(ppm)

T-max of thermal neutron (D)
T/B
ratio

Fluence(E+11n/cm-2)

History of
RT: (Gy)

T-Peak Gy-Eq

2.0

18.0

50

26（1.6 ㎝）

25.7
28.0
34.0
28.0
23.0

Irradiationtime(min.)(May , 2013)% Reduction (Period)
Prognosis (Survival)

6.9/18.0

55

CR(11M)・Alive(11M)

6.4/10.0

90

CR(10M)・Alive(10M)

6.8/15.0

56

CR（4M）Alive(（4M）

Total-RBE-Dose Eq (Gy-Eq)
T-deepest
Gy-E
24(4cm)Skin/Mu
cosa

2.2

23.0

63

23（4.0 ㎝）

22(4cm)

6.0

13.0

50

44（1.5 ㎝）

28(5cm)4060(2.5cm)

44（5 ㎝）

2.8/1133

CR（3M）Alive(（3M）

45

44（1.5 ㎝）

28（5 ㎝）

6.8/1256

CR（3M）Alive(（3M）

7.0
4.0

15.0

採択課題番号 24088
頭頸部悪性腫瘍におけるホウ素中性子捕捉療法の臨床的研究
共同通常（阪大・2 口外）加藤逸郎、岩井聡一、墨 哲郎、中澤光博、由良義明（阪大・工）村田 勲（慶大）岡本正人（長崎大）梅田博昭、柳本惣市（りんくう医療セ）大前政利（東大阪総合）千足浩久（市立池田）大西徹郎（田中クリニック）田中 善（京大・原子炉）田中浩基、鈴木 実、櫻井良
憲、増永慎一郎、丸橋晃、小野公二

CO7-18

B4C Particle as a Boron Compound for BNCT

T. Iwagami, I. Kato, N. Yamamoto1, Y. Ishikawa2,
N. Koshizaki3, H. Tanaka4, Y. Sakurai4, S. Masunaga4,
M. Suzuki4, A. Maruhashi4, K. Ono4 and Y. Yura
Graduate School of Denistry, Osaka University
1
Department of Oral Surgery, Saiseikai Senri Hospital2
National Institute of Advanced Industrial Science andTechnology
3
Graduate School of Engineering, Hokkaido University
4
Research Reactor Institute, Kyoto University

INTRODUCTION: B4C nano particles are producedby liquid phase laser irradiation method [1]. The diameterof the particle is 200 nm. Sonoporation is a low ultrasound which makes small transient holes in the cell
membrane and introduces external materials such as drug
and gene into the cell [2]. In this study, we investigatedwhether sonoporation could be used to introduce B4C
particles into the oral squamous cell carcinoma (SCC)
cells.

Fig. 1. Cell viability after neutron
irradiation with B4C in the medium

EXPERIMENTS: SAS cells derived from oral SCC
were used. Cells were exposed to thermal neutron at
Kyoto University Reactor (KUR) [3]. An ultrasound machine, Sonitron 2000V, and a microbubble, SV-25, wereused [4]. The cell viability was examined by MTT assay.
The cell surface was observed using a scanning electron
microscope.
RESULTS: After sonoporation in the presence of microbubbles, small holes (1-2µm) were observed on thecell surface by a scanning electron microscope. The cell
viability in the groups treated with B4C-mediated BNCT
was lower than that in the group treated with neutron
alone (Fig.1). B4C-mediated BNCT in combination withsonoporation (ultrasound in the presence of microbubble)
was most effective. When cells were mixed with B4C,
washed in PBS and exposed to thermal neutron, cell killing effect of B4C was lost, but it remained in combination with sonoporation (Fig.2).
CONCLUSION: B4C particle can be used as a boron
compound for BNCT. Sonoporation may introduce B4C
particle into oral SCC cells.

Fig. 2. Cell viability after neutron
irradiation without B4C in themediumREFERENCES:[1] Y. Ishikawa et al., Journal of Nanoscience Nanotechnology. 8 (2010) 5467-5470.
[2] J M. Escoffre et al., Current Gene Therapy. 13 (2013)
2-14.
[3] Y. Fujita et al., Radiation Oncology. 4 (2009) 63.
[4] M. Shintani et al., Virology Journal. 8 (2011) 446.

採択課題番号 24089 口腔悪性腫瘍におけるホウ素中性子捕捉療法に関する基礎的研究 共同通常
（阪大・歯）岩上隆紀、加藤逸郎、由良義明(済生会千里病院)山本直典（京大・原子炉）田中浩基、
櫻井良憲、増永慎一郎、鈴木実、丸橋晃、小野公二

CO7-19

Microdosimetry of Neutron Field with Low-Enriched Uranium
at Kyoto University Reactor

S. Endo,Y. Onizuka1, T. Kajimoto, M. Takada2,
H. Tanaka3 and Y. Sakurai3
Graduate School of Engineering, Hiroshima University
1
Junshin-Gakuen College
2
National Institute of Radiological Science3
Research Reactor Institute, Kyoto University

1CO0001
OO0001

0.1

RESULTS:
The microdosimetric single event spectrum of yf(y) and yd(y) for epithermal neutron mode:
CO0000 and mixed neutron mode: OO0000 are shown in
Fig. 1 (a) and (b), respectively. The broad structure
below 20 keV/m of OO0000 mode is due to the (n,)reaction by thermal neutrons. Similarly, peak structurearound 200 keV/m of mixed mode is due to the17O(n,) reaction by thermal neutrons. These structure
relatively small contribution for epithermal neutron mode.
These results are consistent with each other.
REFERENCES:[1] Y. Onizuka, S. Endo, M. Ishikawa, M. Hoshi, M.
Takada, T. Kobayashi, Y. Sakurai, H. Utsumi, S. Uehara,
N. Hayabuchi, N. Maeda, T. Takatsuji, and K. Fujikawa,“Microdosimetry of epithermal neutron field attheKyoto University Reactor,” Radiat. Prot. Dosim. 99,
383–385 , 2002.

0.01

0.001

CO0000(a)
0.0001
1

10

100

1000

y (keV/m)

0.7
CO0001(b) OO0001

0.6

OO00000.5

0.4

yf (y)

EXPERIMENTS: Microdosimetric single event spectrum for the BNCT clinical irradiation filed (epithermal
neutron mode: CO0000 and mixed neutron mode:
OO0000) [3] has been measured with tissue equivalent
proportional counter (TEPC) with a condition of a 1 msite size. In order to take wide lineal energy range, signals from TEPC were divided into two types of amplifier-gain (high-gain and low-gain). Pulse heights were analyzed by two USB-MCAs (Kromek Co LTD, Kspec).
Pulse height distribution of two types of gain have beenconnected and microdosimetric single event spectrum
have been obtained.

yf (y)OO0000INTRODUCTION: Kyoto University Reactor was shut
down in 2006. After that, the fuel has been changed fromhigh-enriched uranium of 93% to low-enriched uraniumof 20%. The reactor has been restarted from 2010. The
neutron energy spectrum has not hopefully unchanged bythis change. However, there are lacking of measurements. Our group had accumulated data of microdosimetric studies at BNCT beam therapeutic beam before
the changes of the fuel enrichment [1,2]. In this study,
microdosimetric measurements are carried out to estimate
BNCT neutron beam for the low-enriched fuel operationand compared with previous spectra.

0.3CO0000

0.2

0.1

1

10

100

1000

y (keV/m)

Fig. 1(a) yf(y) and (b) yd(y) single event spectrum[2] S. Endo, Y. Onizuka, M. Ishikawa, M. Takada, Y. Sakurai, T. Kobayashi, K. Tanaka, M. Hoshi, K. Shizuma,
Microdosimetry of neutron field for boron neutron
capture therapy at Kyoto University reactor, Radiat.
Prot. Dos. 110, 641-644, 2004.
[3] Sakurai, Y., Kobayashi, T., Ono, K. and Kanda, K.
The Irradiation Characteristics of the Upgraded Heavy
Water Facility of the Kyoto University Reactor. In: Advances in Neutron Capture Therapy, Vol. I, Medical
Physics, Eds B. Larsson, J. Crawford, and R. Weinreich,(Amsterdam: Elsevier) pp. 316–320 (1997).

採択課題番号 24098 低濃縮燃料移行後の BNCT 医療照射場のマイクロドシメトリ（純真学園大）鬼塚昌彦、（広島大・工）遠藤 暁、梶本 剛、（放医研）高田正志（京大・原子炉）田中浩基、櫻井良憲

共同通常

CO7-20

Combination therapy of C6 Glial Tumor Cell by BNCT and PDT Sensitizedby a Boron Lich Porphyrin Derivative

N. Miyoshi, H. Tanaka1, M. Sakurai1, K. Ono1, A. A.
Shtil2, V. Ol'shevskaya3, V. N. Kalini3 and A. V. Zaitsev 3
Department of Tumor Pathology, Faculty of Medicine,
University of Fukui
1
Research Reactor Institute, Kyoto University
2
Russian Blokhim Cancer Center, Russian Federation
3
Nesmeyanov Institute of Organoelement Compounds,
Russian Federation

Fig.1. Chemical structure and the absorption spectrum of
the capture molecule.
Molecular weight =1564.63,C48H60B44F16N4Na4={5,10,15,20-terakis[4-(1-caba-closo-dodecabrane-1-yl)tet
rafluorophenyl]17,18-dihydrophyrin}} tetrasodium.
Two observations of the cell damages, 100% - survival
fraction = damaged fraction (%) and the necrotic fraction
were obtained from the flow cytometory analysis asshown in Figure 2.

INTRODUCTION: The distribution of BNCT as oneof clinical cancer treatments will be spread by the progress of the sensitizers and the compact instruments infuture. We had developed a sensitizer with lich of F(16)and B(44) elements in one molecules by collaboration
research of Russian groups [1]. The molecule has both
effects of photosensitizing and the other capture of neutron, which is easy water-soluble (>10mM) and the photosensitizing effect is very high even at ppm order of theconcentration. We had been studied photodynamic therapy (PDT) of cancer for 30 years using many porphyrinderivatives. There had be not seen the reports about the
combination studies yet using one sensitizer.
EXPERIMENTS: (1) Uptake Conditions: 2x106 C6 rat
glial tumor cells/ ml were incubated in the 10M Compound-B (Figure-1: MW=1564.63) of the medium solution for 14.5hr at CO2 incubator. The Boron-10 incorporated in the cells was 3.15 ppm. (2) Irradiation Dose andthe Sequence: The neutron beam dose was 0.575Gy for
10 min. The laser dose at 665 nm was 115.2 J/cm2 for 10
min. The photodynamic irradiation was done within 2 hr
after the BNCT treatment. (3) Damaged Cell Analysis:
The cell number at the 36 hr incubation after the combination treatments were measured by the flow cytometory
analysis (FACS Canto-II type, BD Ltd. Co.) using double
fluorescent staining (Apoptosis Kit: annexine-V/FITC
and PI, MBL Co.) 15 min before the analysis.
RESULTS: The Compound-B in Fig. 1 was used for the
combination therapy of BNCT and PDT at low concentration of 3.15 ppm.

採択課題番号 24103

Fig.2. Cell damaged and Necrotic Cell Fractions (%) in
the Different Treated Groups.
The cell damaged fraction was correlated with the necrotic fraction, especially BNCT and the combination
with PDT. It was considered that both treatments of
BNCT and combination was more effective treatment ofcancer cell comparing PDT because the necrotic fractionof combination of 51% was the highest value comparing
with the total (41%) value with PDT (21%) and BNCT(27%), respectively. It is expected that there will be presented a synergic effect between BNCT and PDT treatments.
REFERENCES:[1] D. Kaluzhny et al., PLoS One, 6 (11) (2011) e27151.

BNCT と光線力学治療の併用療法に両用の増感剤を使用した
共同通常
癌培養細胞共同実験（福井大・医）三好憲雄（京大・原子炉）田中浩基、櫻井正憲、小野公二（ロシア国立がんセンター）
Alexander A. Shtil（ロシア国立有機材料物質研究所）Valentina A. Ol'shevskaya