Multiple-field interstitial photodynamic therapy of subcutaneously transplanted cholangiocellular carcinoma RS-1 in rats
The aim of present study was to investigate an antitumor efficacy of multiple-field interstitial photodynamic therapy (iPDT) in vivo. Materials and Methods: The study was performed on 15 white random-bred rats with subcutaneously transplanted cholangiocellular carcinoma RS-1. Chlorine-based photosen...
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| Опубліковано в: : | Experimental Oncology |
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| Дата: | 2017 |
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Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України
2017
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| Цитувати: | Multiple-field interstitial photodynamic therapy of subcutaneously transplanted cholangiocellular carcinoma RS-1 in rats / D.A.Tzerkovsky // Experimental Oncology. — 2017 — Т. 39, № 2. — С. 117-120. — Бібліогр.: 17 назв. — англ. |
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Digital Library of Periodicals of National Academy of Sciences of Ukraine| _version_ | 1859800615144652800 |
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| author | Tzerkovsky, D.A. |
| author_facet | Tzerkovsky, D.A. |
| citation_txt | Multiple-field interstitial photodynamic therapy of subcutaneously transplanted cholangiocellular carcinoma RS-1 in rats / D.A.Tzerkovsky // Experimental Oncology. — 2017 — Т. 39, № 2. — С. 117-120. — Бібліогр.: 17 назв. — англ. |
| collection | DSpace DC |
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| description | The aim of present study was to investigate an antitumor efficacy of multiple-field interstitial photodynamic therapy (iPDT) in vivo. Materials and Methods: The study was performed on 15 white random-bred rats with subcutaneously transplanted cholangiocellular carcinoma RS-1. Chlorine-based photosensitizer (PS) Ce6CPPPS was administered via single injection at a dose of 2.5 mg/kg into the animal’s caudal vein. Photoirradiation (PI) of tumors was carried out 3 h after PS administration using 7 optical fibers SMA-905 with diode laser with 660 ± 5 nm wavelength at exposure doses of 150 and 300 J/cm²with 0.21 W/cm²fluency rate. The total power density was 360 mW and treatment time was 12 and 24 min. Antitumor efficacy of iPDT was assessed by evaluation of necrosis areas and depth of necrosis in experimental tumors. Results: The results have shown that interstitial PI with multi-field low power density enhanced the antitumor effect of PDT in the RS-1 model. Necrosis areas in tumor tissues after PI with exposure doses 150 and 300 J/cm²24 h and 96 h after treatment were 83.78 ± 4.25 and 100% (p = 0.00074); 56.79 ± 3.24 and 95.46 ± 1.64% (p < 0.00001), respectively. Conclusion: An analysis of the literature data and the results obtained in this study evidence on high effectiveness of the method of multiple-field.
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| first_indexed | 2025-12-07T15:12:52Z |
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Experimental Oncology 39, 117–120, 2017 (June) 117
MULTIPLE-FIELD INTERSTITIAL PHOTODYNAMIC THERAPY
OF SUBCUTANEOUSLY TRANSPLANTED CHOLANGIOCELLULAR
CARCINOMA RS-1 IN RATS
D.A. Tzerkovsky*
N.N. Alexandrov National Cancer Center of Belarus, Lesnoy 223040, Republic of Belarus
The aim of present study was to investigate an antitumor efficacy of multiple-field interstitial photodynamic therapy (iPDT) in vivo.
Materials and Methods: The study was performed on 15 white random-bred rats with subcutaneously transplanted cholangiocellular
carcinoma RS-1. Chlorine-based photosensitizer (PS) Ce6CPPPS was administered via single injection at a dose of 2.5 mg/kg into the
animal’s caudal vein. Photoirradiation (PI) of tumors was carried out 3 h after PS administration using 7 optical fibers SMA-905 with
diode laser with 660 ± 5 nm wavelength at exposure doses of 150 and 300 J/cm² with 0.21 W/cm² fluency rate. The total power density
was 360 mW and treatment time was 12 and 24 min. Antitumor efficacy of iPDT was assessed by evaluation of necrosis areas and depth
of necrosis in experimental tumors. Results: The results have shown that interstitial PI with multi-field low power density enhanced the
antitumor effect of PDT in the RS-1 model. Necrosis areas in tumor tissues after PI with exposure doses 150 and 300 J/cm2 24 h and
96 h after treatment were 83.78 ± 4.25 and 100% (p = 0.00074); 56.79 ± 3.24 and 95.46 ± 1.64% (p < 0.00001), respectively. Conclu-
sion: An analysis of the literature data and the results obtained in this study evidence on high effectiveness of the method of multiple-field.
Key Words: multiple-field interstitial photodynamic therapy, photosensitizer, cholangiocellular carcinoma.
Photodynamic therapy (PDT) is a modern and highly
effective option in the treatment of malignant tumors of the
skin, lung, bladder, brain, pancreas and other localiza-
tions [1–3]. PDT is a treatment method based on the
significant increase of the cytotoxicity of drugs with photo-
irradiation (PI) of the tumor tissue. According to numerous
studies, photochemical reactions include a direct interac-
tion of excited molecules with the help of PI, photosensi-
tizer (PS) on the substrate and forming transient radicals
that react with oxygen. Interaction initiates a complex cas-
cade of free radicals, such as singlet oxygen (1O2), hydroxyl
radical (OH), hydrogen peroxide (H2O2) and superoxide
anion radical (O2
–), causing the development of oxidative
stress syndrome. As a result, PDT effectively induces
tumor-cell apoptosis and necrosis (Necr). The two pos-
sible mechanisms might be: a) promoting mitochondria
to release cytochrome C and activate caspase-3, then
to initiate apoptosis; b) destroying microvessels, inhibition
of angiogenesis and the induction of ische mia and anoxia
of tumor cells, resulting in ischemic Necr [2, 4, 5].
PDT has rarely been used clinically in the treatment
of liver tumors, mainly because of effective accumula-
tion of PS in liver tissue [6, 7]. Distribution studies show
that PS are accumulated in high amounts in reticulo-
endothelial tissue, such as liver, spleen, and kidney, and
in particular, liver tissue contains high PS levels after
its administration [8, 9]. Therefore, superficial tumor
PI causes substantial liver Necr. The limited light penetra-
tion during superficial illumination makes it impossible
to treat deep-seated or larger solid tumors. In order
to prevent possible complications, research teams
recommended the use of an interstitial type of PI of liver
tumors.
The ability of interstitial photodynamic therapy (iPDT)
to cause a destruction of liver tumors has been demon-
strated in in vitro [10–12] and in vivo [13–15] studies, using
hematoporphyrin, photofrin, meta-tetra(hydroxyphenyl)
chlorine (mTHPC) and other photosensitizing agents.
In the present study, we investigated the effects
of Ce6CPPPS-mediated PDT using multi-field intersti-
tial PI on subcutaneously transplanted cholangiocel-
lular carcinoma RS-1 in rats.
MATERIALS AND METHODS
Experimental animals. The study was con-
ducted on 15 white randomly bred rats of both sexes
(220 ± 40 g) obtained from the vivarium of N.N. Ale-
xandrov National Cancer Center of Belarus. The animals
received a standard diet and had permanent access
to water. All manipulations were carried out according
to Methodic instructions for carrying out preclinical
investigations of the pharmacokinetics of pharmaco-
logic substances and drugs presented in the “Good
Laboratory Practice TKP 125-2008” (Health Ministry
of Republic Belarus, Minsk). Animal experiments were
performed according to the Rules of Ethic Commit-
tee. Before treatment, animals were anesthetized
by intramuscular introduction of a solution of droperidol
(5.0 mg/kg) and fentanil (0.05 mg/kg).
Experimental tumor model. As experimental
tumor model, we used rat cholangiocellular carcinoma
RS-1. RS-1 induced in rats with acetylaminefluorene
in 1956 was identified as hepatocholangioma and was
maintained by subcutaneous transplantation [16]. Be-
fore the treatment, the animals were anesthetized and
immobilized. For experiments, tumor homogenate was
implanted subcutaneously into the left inguinal area
by the injection of 0.5 ml of 20% tumor cells suspen-
sion in 0.6% Hank’s solution. The experiments were
performed 39–40 days after tumor implantation with
tumor diameter reaching 40–50 mm.
Submitted: March 03, 2017.
*Correspondence: E-mail: tzerkovsky@mail.ru
Abbreviations used: iPDT — interstitial photodynamic therapy; mTH-
PC — meta-tetra(hydroxyphenyl)chlorine; Necr — necrosis; PDT —
photodyna mic therapy; PI — photoirradiation; PS — photosensitizer.
Exp Oncol 2017
39, 2, 117–120
118 Experimental Oncology 39, 117–120, 2017 (June)
Photosensitizer. Chlorin e6 conjugated with poly-
vinyl pyrrolidone (Ce6CPPPS) produced by Scientific
Pharmaceutical Center of RUE “Belmedpreparaty”,
Minsk, Republic of Belarus) was injected into the tail
vein at a standard dose of 2.5 mg/kg.
Interstitial photodynamic therapy. PI of tumors was
carried out 3 h after PS administration using diode laser
with 660 ± 5 nm wavelength (“PDT Diode laser”, Minsk,
Republic Belarus) at exposure doses of 150 and 300 J/cm²
with 0.21 W/cm² fluency rate. The total power density was
360 mW and treatment time was 12 and 24 min. 7 optical
fibers SMA-905 (“Fotonika Plus”, Ukraine) for PI were
introduced into the tumor at a distance of 1 cm from each
other at a depth of 1–1.5 cm (Fig. 1).
Fig. 1. Multiple-field iPDT: a — photo-irradiation session; b — sub-
cutaneously transplanted tumor during the photo-irradiation session
The distribution of the total power density for each
fiber is shown in Table 1 and Fig. 2.
Table 1. Distribution of the total power density (P = 360 mW) to optical fibers
Optical fiber No. 1 2 3 4 5 6 7
Power density, mW 43.2 36.0 50.4 99.4 43.2 42.4 45.4
Power density, % 12.0 10.0 14.0 27.6 12.0 11.8 12.6
Antitumor efficacy was evaluated 24 and 96 h after
the treatment by quantification of RS-1 tumor necrosis
area (NecrA) by vital staining of tumor bearing animals
with 0.6% Evans blue solution. The animals were sac-
rificed by chloroform; the tumors were removed, fixed
in 10% formalin solution and frozen. Transverse tumor
sections 2–3 mm thick were made. NecrA due to di-
rect effect on tumor cells or structural and functional
disorders in microcirculation remained unstained. The
percentage of tumor necrotic unstained parts was
evaluated using “ImageJ” (NIH, Bethesda, USA).
Statistical processing of the results. The values
obtained were processed using standard statistical
methods of Origin Stat 7.0 software. Statistical signifi-
cance of differences was relevant at p < 0.05.
RESULTS
Early post-treatment changes. In the study, the
sensitivity of subcutaneously implanted RS-1 to iPDT
was investigated. According to our findings, all animals
subjected to iPDT at an exposure dose 300 J/cm2 pre-
sented the necrotic changes of maximal size (100%).
In the control group, this index showing the emergence
of spontaneous central Necr was insignificant (14.88 ±
4.33%). Post-treatment necrotic changes after iPDT were
assessed using the vital staining technique with Evans
blue. Fig. 3 shows the data on NecrA in histotopographic
sections of RS-1 in untreated control and 24 h after iPDT
at the exposure doses of 150 and 300 J/cm2.
Fig. 4 shows the data on NecrA in histotopographic
sections of RS-1 in untreated control and 96 h after iPDT
at the exposure doses of 150 and 300 J/cm2.
Percentage of Necr in groups of animals treated with
iPDT at the exposure doses of 150 and 300 J/cm2 24 and
96 h after treatment is shown in Table 2.
Depth of tumor Necr in groups of animals treated with
iPDT at exposure doses of 150 and 300 J/cm2 24 and
96 h after treatment is shown in Table 3.
Fig. 2. Scheme of fibers location in the tumor and tumor cell
death mechanism
Experimental Oncology 39, 117–120, 2017 (June) 119
Table 3. Depth of Necr in histotopographic sections of subcutaneously
transplanted RS-1 tumor of rats after treatment
Groups Number
of sections, n
Depth of tumor
Necr# (L), сm Mean NecrA (Sн), %
Control 16 – 14.88 ± 4.33
[min = 4.43; maх = 20.25]
iPDT* 150 J/сm2
16 2.69 ± 0.16 83.78 ± 4.25
[min = 53.46; maх = 100]
iPDT* 300 J/сm2
14 2.39 ± 0.11 100
[min = 100; maх = 100]
iPDT** 150 J/сm2
15 2.15 ± 0.11 56.79 ± 3.24
[min = 36.00; maх = 83.63]
iPDT** 300 J/сm2
15 2.84 ± 0.15 95.46 ± 1.64
[min = 81.79; maх = 100]
Note: #L iPDT 150 J/сm2 24 h vs L iPDT 150 J/сm2 96 h, p = 0.009; L iPDT
300 J/сm2 24 h vs L iPDT 300 J/сm2 96 h, p = 0.022.
The results obtained suggest that iPDT with multi-field
low power density PI enhances the effect on the RS-1 rats
tumor model. The use of exposure dose of 300 J/cm2 sig-
nificantly increases NecrA in tumor tissues compared with
150 J/cm2 24 h (p = 0.00074) and 96 h (p < 0.00001).
DISCUSSION
Malignant primary and metastatic liver tumors are
a serious problem for modern oncology. The main
treatment for this type of tumors is surgical removal and
chemotherapy. In spite of obvious achievements of the
medical science of the last decades, the results of treat-
ment of patients with liver tumors remain disappointing.
With poor prognosis, development of new therapeutic
modalities is desirable, especially for patients with non-
resectable, non-transplantable, or recurrent liver tumors.
Local tumor ablation therapies such as radiofrequency
ablation, ethanol injection, cryotherapy, and PDT are
potentially useful palliative approaches. iPDT is an effec-
tive and promising method of treatment of liver tumors.
At the moment, we found only a few articles, confirm-
ing the effectiveness of iPDT in the treatment of primary
and metastatic liver tumors in laboratory animals [13, 14].
Experimental studies, using hematoporphyrin derivative
and photofrin, have shown iPDT to be capable of inducing
tumor destruction within the liver, despite limitations like
non-selective uptake and limited light penetration [13].
van Hillegersberg et al. [13] reported about high antitumor
efficacy of iPDT (λ = 625 nm; 200 mW/cm2; 100–1600 J)
with intravenously administered photofrin at a dose
of 5 mg/kg in the treatment of colon carcinoma CC531,
implanted in liver of Wag/Rij rats. The NecrA in the tumor
and surrounding normal liver tissues depended on the
absorbed dose of PI and increased with its increase
(p < 0.001). The authors noted that application of an ex-
posure dose of 800 J allows to achieve complete regres-
sion of tumors in 66.7% of animals. The authors have
proved the fact that the use of iPDT allows achieving
a good therapeutic results with a minimal risk of damage
to normal liver tissues. Rovers et al. [14] performed iPDT
(λ = 652 nm, 0.1 W, 15 J) with intravenously administered
of mTHPC at a dose 0.3 mg/kg of body weight on Wag/Rij
rats with metastatic liver orthotopic model CC531 colon
adenocarcinoma. Authors reported that iPDT resulted
in complete tumor remission in 87% of animals [14].
van Duijnhoven et al. [17] investigated effects of iPDT
(λ = 652 nm, 200 mW, 16 J/сm2) with mTHPC (0.3 mg/kg)
on liver metastases of orthotopic model of CC531 colon
adenocarcinoma. Authors reported that iPDT was effec-
tive in causing photodynamically-induced photochemical
Necr of tumors, but it did not affect the growth rate of no-
nilluminated tumors in the liver. Immunological staining
Fig. 3. Histotopographic sections of subcutaneously transplanted rats cholangiocellular carcinoma RS-1 in control group (a);
24 h after iPDT at the exposure doses of 150 J/cm2 (b) and 300 J/cm2 (c) (blue — viable tumor tissue, red — photochemical Necr)
Table 2. NecrA in histotopographic sections of subcutaneously transplanted RS-1 tumor of rats after treatment
Groups Number of sections, n Mean tumors area# (Sc), сm2 p Mean NecrA& (Sн), % p
Control 16 11.78 ± 1.12
[min = 6.33; maх = 14.32] – 14.88 ± 4.33
[min = 4,43; maх = 20,25] –
iPDT* 150 J/сm2
16 13.57 ± 0.94
[min = 9.33; maх = 20.81] 0.23 83.78 ± 4.25
[min = 53.46; maх = 100] < 0.00001
iPDT* 300 J/сm2
14 9,74 ± 0,53
[min = 5.39; maх = 11.33] 0.11 100
[min = 100; maх = 100] < 0.00001
iPDT** 150 J/сm2
15 9.53 ± 0.71
[min = 5.81; maх = 13.61] 0.10 56.79 ± 3.24
[min = 36.00; maх = 83.63] < 0.00001
iPDT** 300 J/сm2
15 13.43 ± 1.32
[min = 6.98; maх = 22.39] 0.35 95.46 ± 1.64
[min = 81.79; maх = 100] < 0.00001
Note: Tables 2 and 3: *NecrA in the tumor was evaluated 24 h after iPDT. **NecrA in the tumor was evaluated 96 h after iPDT. #Sc iPDT 150 J/сm2 24 h vs Sc iPDT
150 J/сm2 96 h, p =0.0018; Sc iPDT 300 J/сm2 24 h vs Sc iPDT 300 J/сm2 96 h, p = 0.0015; Sc iPDT 150 J/сm2 24 h vs Sc iPDT 300 J/сm2 24 h, p =
0.0013; Sc iPDT 150 J/сm2 96 h vs Sc iPDT 300 J/сm2 96 h, p = 0.015. &Sн iPDT 150 J/сm2 24 h vs Sн iPDT 150 J/сm2 96 h, p = 0.00002; Sн iPDT
300 J/сm2 24 h vs Sн iPDT 300 J/сm2 96 h, p = 0.01; Sн iPDT 150 J/сm2 24 h vs Sн iPDT 300 J/сm2 24 h, p = 0.00074; Sн iPDT 150 J/сm2 96 h vs Sн iPDT
300 J/сm2 96 h, p < 0.00001.
120 Experimental Oncology 39, 117–120, 2017 (June)
of tumors showed natural killer cells to be significantly
lower in tumors treated with interstitial PI than in control
tumors (p < 0.05). Otake et al. [15] published the results
of their study on the efficacy of iPDT (λ = 630 nm, 160 mW,
47–90 J/cm2) with 5-ALA at a dose of 500 mg/kg ad-
ministered intravenously 3 h before PI in the treatment
of chemically induced hepatocellular carcinoma in rats.
The authors concluded that in all treated tumors, Necr
was evident at 24 h after 5ALA-iPDT [15]. Several teams
of scientists made successful attempts for the clinical
testing of iPDT for patients with unresectable primary and
metastatic malignant liver tumors [6, 7]. Vogl et al. [7]
presented the results of several phase I clinical trials based
on the use of iPDT in the treatment of the liver metastases.
In the first study authors reported about 5 patients with
liver metastasis of colorectal cancer treated with PS SQN
400 (mTHPBC) at a dose of 6 mg/kg and PI (λ = 740 nm;
60 J/cm2) 120 h after SQN 400 admi nistration. Depending
on the location and size of lesions, the authors have used
from 4 to 7 special fibers installed in the tumor site under
the center CT control. After 3 months in all cases CT study
recorded photodynamically-induced Necr with no evi-
dence of damage to the normal liver parenchyma, and
after 6 months in 50% of the metastases continued growth
was not revealed [7]. In the second study, the authors
have included 4 patients with liver metastasis of colorectal
cancer (n = 3) and melanoma (n = 1). PS LS 11 (Talaporfin
sodium) was injected intravenously at a standard dose
of 40.0 mg/m2. At 4 h post-injection, a tumor was pho-
toirradiated with a “PDT laser” with 25 mm optical fibers
(λ = 630 nm; 400 mW/cm2; 100 J/cm2). The average size
of Necr, confirmed the results of CT studies, was 14 mm.
At the control observation after 6 weeks in 2 cases com-
plete regression of tumor was recorded [7].
However, a moderate number of studies in this field
points on the need for further research to determine
antitumor efficacy of iPDT in the treatment of this pathol-
ogy. In further experimental studies, we plan to study the
antitumor efficacy of iPDT in the treatment of other tumor
strains. If positive results will be obtained, we will be able
to recommend the developed method for approbation
in clinical oncology.
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Copyright © Experimental Oncology, 2017
Fig. 4. Histotopographic sections of subcutaneously transplanted rats cholangiocellular carcinoma RS-1 in control group (a);
96 h after iPDT at the exposure doses of 150 J/cm2 (b) and 300 J/cm2 (c) (blue — viable tumor tissue, red — photochemical Necr)
|
| id | nasplib_isofts_kiev_ua-123456789-137629 |
| institution | Digital Library of Periodicals of National Academy of Sciences of Ukraine |
| issn | 1812-9269 |
| language | English |
| last_indexed | 2025-12-07T15:12:52Z |
| publishDate | 2017 |
| publisher | Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України |
| record_format | dspace |
| spelling | Tzerkovsky, D.A. 2018-06-17T14:19:39Z 2018-06-17T14:19:39Z 2017 Multiple-field interstitial photodynamic therapy of subcutaneously transplanted cholangiocellular carcinoma RS-1 in rats / D.A.Tzerkovsky // Experimental Oncology. — 2017 — Т. 39, № 2. — С. 117-120. — Бібліогр.: 17 назв. — англ. 1812-9269 https://nasplib.isofts.kiev.ua/handle/123456789/137629 The aim of present study was to investigate an antitumor efficacy of multiple-field interstitial photodynamic therapy (iPDT) in vivo. Materials and Methods: The study was performed on 15 white random-bred rats with subcutaneously transplanted cholangiocellular carcinoma RS-1. Chlorine-based photosensitizer (PS) Ce6CPPPS was administered via single injection at a dose of 2.5 mg/kg into the animal’s caudal vein. Photoirradiation (PI) of tumors was carried out 3 h after PS administration using 7 optical fibers SMA-905 with diode laser with 660 ± 5 nm wavelength at exposure doses of 150 and 300 J/cm²with 0.21 W/cm²fluency rate. The total power density was 360 mW and treatment time was 12 and 24 min. Antitumor efficacy of iPDT was assessed by evaluation of necrosis areas and depth of necrosis in experimental tumors. Results: The results have shown that interstitial PI with multi-field low power density enhanced the antitumor effect of PDT in the RS-1 model. Necrosis areas in tumor tissues after PI with exposure doses 150 and 300 J/cm²24 h and 96 h after treatment were 83.78 ± 4.25 and 100% (p = 0.00074); 56.79 ± 3.24 and 95.46 ± 1.64% (p < 0.00001), respectively. Conclusion: An analysis of the literature data and the results obtained in this study evidence on high effectiveness of the method of multiple-field. en Інститут експериментальної патології, онкології і радіобіології ім. Р.Є. Кавецького НАН України Experimental Oncology Original contributions Multiple-field interstitial photodynamic therapy of subcutaneously transplanted cholangiocellular carcinoma RS-1 in rats Article published earlier |
| spellingShingle | Multiple-field interstitial photodynamic therapy of subcutaneously transplanted cholangiocellular carcinoma RS-1 in rats Tzerkovsky, D.A. Original contributions |
| title | Multiple-field interstitial photodynamic therapy of subcutaneously transplanted cholangiocellular carcinoma RS-1 in rats |
| title_full | Multiple-field interstitial photodynamic therapy of subcutaneously transplanted cholangiocellular carcinoma RS-1 in rats |
| title_fullStr | Multiple-field interstitial photodynamic therapy of subcutaneously transplanted cholangiocellular carcinoma RS-1 in rats |
| title_full_unstemmed | Multiple-field interstitial photodynamic therapy of subcutaneously transplanted cholangiocellular carcinoma RS-1 in rats |
| title_short | Multiple-field interstitial photodynamic therapy of subcutaneously transplanted cholangiocellular carcinoma RS-1 in rats |
| title_sort | multiple-field interstitial photodynamic therapy of subcutaneously transplanted cholangiocellular carcinoma rs-1 in rats |
| topic | Original contributions |
| topic_facet | Original contributions |
| url | https://nasplib.isofts.kiev.ua/handle/123456789/137629 |
| work_keys_str_mv | AT tzerkovskyda multiplefieldinterstitialphotodynamictherapyofsubcutaneouslytransplantedcholangiocellularcarcinomars1inrats |