Jong-Ki Kim

Enhanced proton treatment in mouse tumors through proton irradiated nanoradiator effects on metallic nanoparticles

The impact of protons on metallic nanoparticles (MNPs) produces the potent release of MNP-induced secondary electrons and characteristic x-rays. To determine the ability of secondary radiations to enhance proton treatment, the therapeutic irradiation of tumors was investigated in mice receiving 100-300 mg MNPs/kg intravenously prior to single dose, 10-41 Gy, proton irradiation. A proton beam was utilized to irradiate nanoparticles with a single Bragg peak set to occur inside a tumor volume (fully absorbed) or to occur after the beam had traversed the entire body. The dose-dependent increase in complete tumor regression (CTR) was 37-62% in the fully-absorbed irradiation group or 50-100% in the traversing irradiation group, respectively, compared with the proton-alone control mice (p < 0.01). One year survival was 58-100% versus 11-13% proton alone. The dose-dependent increase of intracellular reactive oxygen species level was 12-36% at 10 Gy compared with the proton-alone control cell. Therapeutic effective drug concentration that led to 100% CTR with a proton dose of 31 Gy was measured either 41 µg Au/g tissue or 59 µg Fe/g tissue. MNP-based proton treatment increased not only percent CTR and survival in vivo but also ROS generation in vitro, suggesting tumor dose enhancement from secondary radiation as one potent pathway of therapeutic enhancement.

Comment on ‘Therapeutic application of metallic nanoparticles combined with particle-induced x-ray emission effect’

Metallic nanoparticles (MNP) are able to release localized x-rays when activated with a high energy proton beam by the particle-induced x-ray emission (PIXE) effect. The exploitation of this phenomenon in the therapeutic irradiation of tumors has been investigated. PIXE-based x-ray emission directed at CT26 tumor cells in vitro, when administered with either gold (average diameter 2 and 13 nm) or iron (average diameter 14 nm) nanoparticles (GNP or SNP), increased with MNP solution concentration over the range of 0.1-2 mg ml(-1). With irradiation by a 45 MeV proton therapy (PT) beam, higher concentrations had a decreased cell survival fraction. An in vivo study in CT26 mouse tumor models with tumor regression assay demonstrated significant tumor dose enhancement, thought to be a result of the PIXE effect when compared to conventional PT without MNP (radiation-only group) using a 45 MeV proton beam (p < 0.02). Those receiving GNP or SNP injection doses of 300 mg kg(-1) body weight before proton beam therapy demonstrated 90% or 75% tumor volume reduction (TVR) in 20 days post-PT while the radiation-only group showed only 18% TVR and re-growth of tumor volume after 20 days. Higher complete tumor regression (CTR) was observed in 14-24 days after a single treatment of PT with an average rate of 33-65% for those receiving MNP compared with 25% for the radiation-only group. A lower bound of therapeutic effective MNP concentration range, in vivo, was estimated as 30-79 µg g(-1) tissue for both gold and iron nanoparticles. The tumor dose enhancement may compensate for an increase in entrance dose associated with conventional PT when treating large, solid tumors with a spread-out Bragg peak (SOBP) technique. The use of a combined high energy Bragg peak PT with PIXE generated by MNP, or PIXE alone, may result in new treatment options for infiltrative metastatic tumors and other diffuse inflammatory diseases.

Action of green tea catechin on bone metabolic disorder in chronic cadmium-poisoned rats.

The purpose of this study was to investigate the effects of green tea catechin on bone metabolic disorders and its mechanism in chronic cadmium-poisoned rats. Sprague-Dawley male rats weighing 100+/-10 g were randomly assigned to one control group and three cadmium-poisoned groups. The cadmium groups included a catechin free diet (Cd-0C) group, a 0.25% catechin diet (Cd-0.25C) group and a 0.5% catechin diet (Cd-0.5C) group according to their respective levels of catechin supplement. After 20 weeks, the deoxypyridinoline and crosslink values measured in urine were significantly increased in the Cd-0C group. Cadmium intoxication seemed to lead to an increase in bone resorption. In the catechin supplemented group (Cd-0.5C group), these urinary bone resorption marks, were decreased. The serum osteocalcin content in the cadmium-poisoned group was significantly increased as compared with the control group. In the catechin supplemented group serum osteocalcin content values were lower than the control group. The cadmium-intoxicated group (Cd-0C group), had lower bone mineral density than the control group (total body, vertebra, pelvis, tibia and femur). The catechin supplement increased bone mineral density to about the same as the control group. Bone mineral content showed a similar trend to total bone mineral density. Therefore, the bone mineral content of the Cd-0C group at the 20th week was significantly lower than the control group. The catechin supplemented group (Cd-0.5C group) was about the same as the control group. The cause of decreasing bone mineral density and bone mineral content by cadmium poisoning was due to the fast bone turnover rate, where bone resorption occurred at a higher rate than bone formation. The green tea catechin aided in normalizing bone metabolic disorders in bone mineral density, bone mineral content and bone calcium content caused by chronic cadmium intoxication.

Effects of green tea catechin on enzyme activities and gene expression of antioxidative system in rat liver exposed to microwaves

Abstract The purpose of this study was to investigate the effect of green tea catechin on enzyme activities and gene expression of antioxidative system in rat liver exposed to microwaves. Sprague-Dawley male rats 100±10 g body weight were randomly divided into control group and microwave exposed group: Microwave exposed group was further divided into three groups: catechin free diet (MW) group, 0.25% catechin (MW-0.25C) group and 0.5% catechin (MW-0.5C) group. The rats were irradiated with microwave at frequency of 2.45 GHz for 15 min and then the changes in the pattern of antioxidative defense system and gene expression were investigated for 16 days (the 2nd, 4th, 6th, 8th and 16th days), and compared with the control group. The activity of superoxide dismutase (SOD) in MW group was lower on the 4th day after irradiation and increased in the catechin supplementation group were on the 8th day, compared with control group. The activity of glutathone peroxidase (GSHpx) in MW group was lower than that in the control group on the 8th day after irradiation, but increased to the level of the control group on the 16th day and those of MW-0.25C and MW-0.5C groups showed the same level as the control group but that was higher than the control group from 6 days after irradiation. The content of thiobarbituric acid reactive substances (TBARS) in liver of MW group was increased to 1.3, 1.5, and 1.7 fold of the control group at 2, 4, and 6 days after irradiation respectively but that in MW-0.25C and MW-0.5C groups was increased to 1.1, 1.3 and 1,3 fold of the control group at 2, 4, and 6 days but recovered to the level of the control group at 16 days after irradiation. The level of SOD gene expression in MW group was lower than that in the control group but that of MW-0.25C and MW-0.5C group were higher than the MW group. The GSHpx gene expression in MW group was expressed lower than in the control group, but expressed at a higher level in the MW-0.25C and MW-0.5C groups. It is suggested that the damage of liver tissues was alleviated and function rapidly recovered to the normal level due to probable to the correction of imbalances in the antioxidative system with the administration of green tea catechin

Sonodynamically Induced Antitumor Effects of 5-Aminolevulinic Acid and Fractionated Ultrasound Irradiation in an Orthotopic Rat Glioma Model

The sonodynamically induced selective antitumor effects of 5-aminolevulinic acid (5-ALA) on a C6 glioma that was implanted in a rat brain were evaluated. One week after the inoculation of the brains with the C6 rat glioma cells, glioma development was monitored using a 1.5 T MRI. Brains both with and without intravenous administration of 5-ALA (60 mg/kg body weight) or Radachlorin (40 mg/kg body weight) were insonated by a 1 MHz ultrasound at a dose of 2.65 W/cm(2). Irradiation was performed in a fractionated manner to avoid any thermal effects in the tissue due to the focused ultrasound; 16 min of irradiation were followed by a 3 min recess, then 4 min of resumed irradiation. Mean tumor sizes, measured after the rats were sacrificed 2 weeks post treatment, were 122.48 ± 39.64 mm(3) in sham-operated rats, 87.42 ± 21.40 mm(3) in rats receiving ultrasound without 5-ALA, 10.50 ± 8.20 mm(3) in rats receiving ultrasound with 5-ALA, and 56.42 ± 12.48 mm(3) in rats receiving ultrasound with Radachlorin. The tumor size was significantly smaller in the therapy group receiving sonodynamic 5-ALA than in any of the other groups (p < 0.05). This experimental rat model showed that sonodynamic therapy can be useful in the treatment of deep-seated malignant gliomas.

Photon activated therapy (PAT) using monochromatic Synchrotron x-rays and iron oxide nanoparticles in a mouse tumor model: feasibility study of PAT for the treatment of superficial malignancy

BackgroundX-rays are known to interact with metallic nanoparticles, producing photoelectric species as radiosensitizing effects, and have been exploited in vivo mainly with gold nanoparticles. The purpose of this study was to investigate the potential of sensitizing effect of iron oxide nanoparticles for photon activated therapy.MethodsX-rays photon activated therapy (PAT) was studied by treating CT26 tumor cells and CT26 tumor-bearing mice loaded with 13-nm diameter FeO NP, and irradiating them at 7.1 keV near the Fe K-edge using synchrotron x-rays radiation. Survival of cells was determined by MTT assay, and tumor regression assay was performed for in vivo model experiment. The results of PAT treated groups were compared with x-rays alone control groups.ResultsA more significant reduction in viability and damage was observed in the FeO NP-treated irradiated cells, compared to the radiation alone group (p < 0.04). Injection of FeO NP (100 mg/kg) 30 min prior to irradiation elevated the tumor concentration of magnetite to 40 μg of Fe/g tissue, with a tumor-to-muscle ratio of 17.4. The group receiving FeO NP and radiation of 10 Gy showed 80% complete tumor regression (CTR) after 15–35 days and relapse-free survival for up to 6 months, compared to the control group, which showed growth retardation, resulting in 80% fatality. The group receiving radiation of 40 Gy showed 100% CTR in all cases irrespective of the presence of FeO NP, but CTR was achieved earlier in the PAT-treated group compared with the radiation alone group.ConclusionsAn iron oxide nanoparticle enhanced therapeutic effect with relatively low tissue concentration of iron and 10 Gy of monochromatic X-rays. Since 7.1 keV X-rays is attenuated very sharply in the tissue, FeO NP-PAT may have promise as a potent treatment option for superficial malignancies in the skin, like chest wall recurrence of breast cancer.

An Antithrombotic Fucoidan, Unlike Heparin, Does Not Prolong Bleeding Time in a Murine Arterial Thrombosis Model: A Comparative Study of Undaria pinnatifida sporophylls and Fucus vesiculosus

The antithrombotic activities and bleeding effects of selected fucoidans (source from either Undaria pinnatifida sporophylls or from Fucus vesiculosus) have been compared with heparin in the ferric chloride‐induced arterial thrombus mouse model. Thrombosis was induced by applying 5% ferric chloride for 3u2009min on the carotid artery region of Balb/c mouse. Five minutes prior to thrombus induction, mice were infused through the tail vein with either saline (control) or polysaccharides. Either fucoidan or heparin was dosed at 0.1, 1.25, 2.5, 5.0, 10, 25, or 50u2009mg/kg intravenously (i.v.) The carotid blood flow was monitored until more than 60u2009min post‐thrombus induction. Mouse tail transection bleeding time was measured up to 60u2009min after making a cut in the mouse tail. Both antithrombotic and bleeding effects were observed in a dose‐dependent manner for both fucoidans and heparin. Thrombus formation was totally (reflected by Doppler flow meter) inhibited at either 5 or 50u2009mg/kg of unfractionated Undaria fucoidan or a low‐molecular‐weight Undaria fucoidan fraction, respectively, without prolonging the time‐to‐stop bleeding compared with the control (pu2009<u20090.01). The total inhibition of thrombus formation was observed for unfractionated Fucus fucoidan at 25u2009mg/kg where the time‐to‐stop bleeding was still significantly prolonged, by as much as 8u2009±u20091.7u2009min (pu2009<u20090.02). In contrast the heparin‐treated group showed total inhibition of thrombus formation even at a small dose of 0.8u2009mg/kg (400u2009IU) at which bleeding continued until 60u2009min. In conclusion algal fucoidans are highly antithrombotic without potential haemorrhagic effects compared with heparin in the arterial thrombus model, but this property differs from algal species to species, and from the molecular structure of fucoidans. Copyright

Inactivation of multidrug resistant (MDR)- and extensively drug resistant (XDR)-Mycobacterium tuberculosis by photodynamic therapy

We investigated the effects of photodynamic therapy (PDT) on anti-tuberculosis (TB) activity by measuring inactivation rates, expressed as D-value, of MDR- and XDR-Mycobacterium tuberculosis (M. tb) clinical strains in vitro. Approximately 10(6) colony forming unit per milliliter (CFU/ml) of the bacilli were irradiated with various doses of laser light after exposure to photosensitizers. Survival of M. tb was measured by enumerating CFU in 7H10 medium to measure D-values. No inactivation of M. tb was observed when exposed to photosensitizers (radachlorin or DH-I-180-3) only or laser light only (P>0.1). Treatment with a combination of photosentizer and laser inactivated M. tb although there was a significant difference between the types of photosensitizers applied (P<0.05). Linear inactivation curves for the clinical M. tb strains were obtained up to laser doses of 30 J/cm(2) but prolonged irradiation did not linearly inactivate M. tb, yielding sigmoid PDT inactivation curves. D-values of M. tb determined from the slope of linear regression lines in PDT were not significantly different and ranged from 10.50 to 12.13 J/cm(2) with 670 nm laser irradiation at 100 mW/cm(2) of the fluency rate, except for a drug-susceptible strain among the clinical strains tested. This suggests that PDT inactivated M. tb clinical strains regardless of drug resistance levels of the bacilli. Intermittent and repeated PDT allowed acceleration of the inactivation of the bacilli as a way to avoid the sigmoid inactivation curves. In conclusion, PDT could be alternative as a new option for treatment for MDR- and XDR-tuberculosis.

Molecular modeling study of the norfloxacin-DNA complex.

Abstract Molecular modeling and molecular dynamics were performed to investigate the interaction of norfloxacin with the DNA oligonucleotide 5′-d(ATACGTAT)2. Eight quinolone-DNA binding structures were built by molecular modeling on the basis of experimental results. A 100ps molecular dynamics calculation was carried out on two groove binding models and six partially intercalating models. The resulting average structures were compared with each other and to free DNA structure as a reference. The favorable binding mode of norfloxacin to a DNA substrate was pursued by structural assess including steric hindrance, presence of hydrogen-bonding, non-bonding energies of the complex and presence of abnormal structural distortion. Although two of the intercalative models showed the highest binding energy and the lowest non-bonding interaction energy, they presented structural features which contrast with experimental results. On the other hand, one groove binding model demonstrated the most acceptable structure when the experimental observation was accounted. In this model, hydrogen bonding of the carbonyl and carboxyl group of the norfloxacin rings with the DNA bases was present, and norfloxacin binds to the amine group of the guanine base which protrudes toward the minor groove of B-DNA.

Enhanced production of reactive oxygen species by gadolinium oxide nanoparticles under core-inner-shell excitation by proton or monochromatic X-ray irradiation: implication of the contribution from the interatomic de-excitation-mediated nanoradiator effect to dose enhancement.

Abstract Core–inner-valence ionization of high-Z nanoparticle atomic clusters can de-excite electrons through various interatomic de-excitation processes, thereby leading to the ionization of both directly exposed atoms and adjacent neutral atoms within the nanoparticles, and to an enhancement in photon–electron emission, which is termed the nanoradiator effect. To investigate the nanoradiator-mediated dose enhancement in the radio-sensitizing of high-Z nanoparticles, the production of reactive oxygen species (ROS) was measured in a gadolinium oxide nanoparticle (Gd-oxide NP) solution under core–inner-valence excitation of Gd with either 50xa0keV monochromatic synchrotron X-rays or 45xa0MeV protons. This measurement was compared with either a radiation-only control or a gadolinium-chelate magnetic resonance imaging contrast agent solution containing equal amounts of gadolinium as the separate atomic species in which Gd–Gd interatomic de-excitations are absent. Ionization excitations followed by ROS measurements were performed on nanoparticle-loaded cells or aqueous solutions. Both photoexcitation and proton impact produced a dose-dependent enhancement in the production of ROS by a range of factors from 1.6 to 1.94 compared with the radiation-only control. Enhanced production of ROS, by a factor of 1.83, was observed from Gd-oxide NP atomic clusters compared with the Gd-chelate molecule, with a Gd concentration of 48xa0μg/mL in the core-level photon excitation, or by a factor of 1.82 under a Gd concentration of 12xa0μg/mL for the proton impact at 10xa0Gy (pxa0<xa00.02). The enhanced production of ROS in the irradiated nanoparticles suggests the potential for additional therapeutic dose enhancements in radiation treatment via the potent Gd–Gd interatomic de-excitation-driven nanoradiator effect.