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Gold nanorods (GNRs) produce heat upon irradiation with near-infrared light, enabling a tumor-targeted photothermal therapy. In this study, we prepared GNRs coated with sulfated hyaluronic acid (sHA) with a binding affinity for CD44 via electrostatic interactions to deliver GNRs to tumors efficiently and stably, and evaluated their usefulness for photothermal therapy. Cationic GNRs modified with trimethylammonium groups electrostatically interacted with native HA or sHA with varying degrees of sulfation to form complexes. While GNR/HA was unstable in saline, GNR/sHA maintained the absorbance peak in the near-infrared region, particularly for GNR/sHA with higher degrees of sulfation. GNR/sHA exhibited an intense photothermal effect upon irradiation with near-infrared light. Furthermore, in vitro and in vivo studies revealed that GNR coated with sHA containing approximately 1.2 sulfated groups per HA unit could accumulate in CD44-positive tumors via an HA-specific pathway. These findings indicate the effectiveness of GNR/sHA as a tumor-targeted photothermal therapeutic agent.
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Macrophage infiltration is a characteristic feature of atherosclerotic plaque progression. Since liposomes containing 1,2-distearoyl-sn-glycero-3-phosphoglycerol (DSPG) are efficiently phagocytosed by macrophages, we deduced that radiolabeled liposomes containing DSPG could potentially be used for nuclear imaging of vulnerable atherosclerotic plaques. Indium-111 (111In)-labeled liposomes containing different ratios of DSPG were developed with a high labeling efficiency. 111In-labeled liposomes with higher DSPG content showed higher uptake by macrophage-like RAW264 cells. A biodistribution study demonstrated rapid blood clearance and selective accumulation in the liver and spleen, especially in normal mice injected with 111In-labeled liposomes with higher DSPG content. Accumulation in atherosclerotic plaques was evaluated using 111In-labeled DSPG liposomes, which had the highest DSPG content among the studied liposomes. 111In-labeled DSPG liposomes accumulated in the plaques and the radioactive regions were mostly consistent with the distribution of macrophages. The target-to-non-target ratio of 111In-labeled DSPG liposomes was higher than that of 111In-labeled control liposomes without DSPG. These results suggest that 111In-labeled liposomes containing DSPG are useful for nuclear medical diagnosis of atherosclerotic plaques.
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Placa Aterosclerótica , Animais , Camundongos , Placa Aterosclerótica/diagnóstico por imagem , Lipossomos , Fosfatidilgliceróis , Distribuição Tecidual , MacrófagosRESUMO
INTRODUCTION: Systemic amyloidosis is a rare disease caused by the deposition of amyloid fibrils in various organs. Amyloid-targeted radiopharmaceuticals have been developed and applied to diagnose systemic amyloidosis peripherally; however, high-contrast imaging has not been achieved because of the high background signals in normal organs. To overcome this problem, we designed an amyloid-targeted radioiodinated probe 1 with a metabolizable linkage (ester bond) to release of radiolabeled metabolites (m-iodohippuric acid) in normal organs that could be rapidly excreted in the urine. METHODS: Compound 1 was synthesized by conjugating 2-(4-(methylamino)phenyl)benzo[d]thiazol-6-ol, an amyloid-targeting compound, with m-iodohippuric acid. [125I]1 was synthesized via iododestannylation using a tributyltin precursor. Mouse models of amyloid A (AA) amyloidosis, a type of systemic amyloidosis, were prepared by administering amyloid-enhancing factor to mice and used for in vitro autoradiography using organ sections and in vivo evaluation. RESULTS: [125I]1 was obtained with a radiochemical yield of 59% and radiochemical purity of over 95%. An in vitro autoradiographic study demonstrated that [125I]1 specifically binds to amyloid in the splenic tissue. Upon administration to normal mice, [125I]1 was distributed to organs throughout the body, followed by the rapid excretion of radioactivity in the urine as m-[125I]iodohippuric acid. Furthermore, ex vivo autoradiography showed that [125I]1 bound to the amyloid formed around the follicles in the spleens of AA amyloidosis model mice. CONCLUSION: These results suggest that the interposition of a metabolizable linkage between an amyloid-targeting moiety and a radiolabeled hippuric acid would be useful in the design of radiopharmaceuticals for high-contrast imaging of systemic amyloidosis.
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Amiloidose , Compostos Radiofarmacêuticos , Camundongos , Animais , Compostos Radiofarmacêuticos/química , Ácido Iodoipúrico , Amiloidose/diagnóstico por imagem , Amiloide/metabolismoRESUMO
Brain tumors are an important cause of suffering and death. Glioblastoma are the most frequent primary tumors of the central nervous system in adults. They are associated with a very poor prognosis, since only 10% of GBM patients survive 5 years after diagnosis. Medulloblastoma are the most frequent brain malignancies in childhood; they affect the cerebellum in children under 10 years of age in 75% of cases. The current multimodal treatment comes at the expense of serious and often long-lasting side effects. Herein, we propose the synthesis of a library of novel alkoxyamines as anticancer drug candidates. The most efficient molecule, ALK4, was selected based on its ability to inhibit both survival and migration of GBM and MB cells in 2D cultures and in 3D tumor spheroids. A fluorescent derivative was used to show the early cytosolic accumulation of ALK4 in tumor cells. Spontaneous homolysis of ALK4 led to the release of alkyl radicals, which triggered the generation of reactive oxygen species, fragmentation of the mitochondrial network and ultimately apoptosis. To control its homolytic process, the selected alkoxyamine was bioconjugated to a peptide selectively recognized by matrix metalloproteases. This bioconjugate, named ALK4-MMPp, successfully inhibited survival, proliferation, and invasion of GBM and MB tumor micromasses. We further developed innovative brain and cerebellum organotypic models to monitor treatment response over time. It confirmed that ALK4-MMPp significantly impaired tumor progression, while no significant damage was observed on normal brain tissue. Lastly, we showed that ALK4-MMPp was well-tolerated in vivo by zebrafish embryos. This study provides a new strategy to control the activation of alkoxyamines, and revealed the bioconjugate ALK4-MMPp bioconjugate as a good anticancer drug candidate.
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Free radicals, such as hydroxyl radical, superoxide, and lipid-derived radical, have unpaired electrons, making them a highly reactive chemical species. They play important physiological roles, for example, in the removal of xenobiotic substances, such as bacteria and viruses, and in the production of chemical mediators, like prostaglandins and leukotrienes. However, excessive production of free radicals can cause structural defects in biomolecules like DNA and proteins, resulting in a loss of their normal functions. Hence, free radicals have been implicated in the onset and progression of various diseases such as cancer, atherosclerosis, and neurodegenerative diseases. However, there is very little clarity on the substantial amount, type, and location of free radicals in vivo, under pathological conditions. An investigation on the actual state of free radicals in vivo could lead to the diagnosis of pathological conditions and the elucidation of the mechanisms of their onset and progression; therefore, the development of in vivo radical detection methods is being widely pursued. Toward this end, nuclear medical imaging methods have recently attracted attention. In this study, we discuss the development of a nuclear medical imaging probe for the specific targeting of lipid radicals.
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Infarto da Artéria Cerebral Média , Estresse Oxidativo , Humanos , Radicais Livres/metabolismo , Superóxidos/metabolismo , LipídeosRESUMO
2,2,6,6-Tetramethylpiperidin-N-oxyl (TEMPO)-type nitroxides are susceptible to bioreduction, leading to a loss of radical properties. Although it has been reported that the steric and electronic environments around the N-O moiety of nitroxides affect the reduction, how the relative configuration of nitroxide derivatives alters it is unclear. In this study, we investigated the effect of diastereomers on the radical properties of C2- and C4-disubstituted TEMPO-type nitroxides. We succeeded in isolating the diastereomers of the studied nitroxides for the first time. In addition, we compared the reactivities of nitroxide derivatives with different substituents at the C2 and C4 positions toward ascorbate reduction. We found that the bulky substituents at both C2 and C4 and the electronic effect of C4 affected the reduction of the isomers. C2- and C4-disubstituted nitroxides were administered to mice for electron spin resonance imaging to assess bioreduction in the brain. Similar to the reactivity to reduction in vitro, a difference in the bioreduction of diastereomers was observed in brain tissues. Our research strongly indicates that bioreduction can be controlled by changing the relative configuration, which can be used in the design of nitroxide derivatives for biological applications.
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Ácido Ascórbico , Óxidos N-Cíclicos , Camundongos , Animais , Espectroscopia de Ressonância de Spin Eletrônica/métodos , Óxidos de Nitrogênio , Marcadores de Spin , OxirreduçãoRESUMO
Nitroxides are known to undergo oxidation, reduction, and radical scavenging reactions due to their stable radicals. Nitroxides have a wide range of applications due to their reactivities, including radical detecting probes and catalysts. Because nitroxides are easily reduced by ascorbate, a reducing agent, in biological applications, it is critical to control their reactivity to use them as a probe to trace the target reaction. On the other hand, the phenyl group, which is present in many functional organic molecules, is useful for controlling the electronic and steric effects. However, there has been few systematic studies on the substituent effects of TEMPO-type nitroxides with phenyl rings in the vicinity of a radical (α-position). In this study, we synthesized three nitroxides with a phenyl group at the α-position of a TEMPO-type nitroxide and tested their redox properties. The results showed that the reduction reactivity and redox potential differed depending on the position of the phenyl group, implying that the phenyl group one carbon away from the α-carbon of the N-O moiety increases the degree of steric hindrance. This finding is expected to be the basis for the development of functional nitroxides.
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Carbono , Substâncias Redutoras , Óxidos N-Cíclicos/farmacologia , Óxidos de Nitrogênio , Oxirredução , Espectroscopia de Ressonância de Spin EletrônicaRESUMO
PURPOSE: To assess the risk factors for intraocular pressure (IOP) elevation during the early period post cataract surgery. STUDY DESIGN: Retrospective study. METHODS: This study involved 1587 eyes that underwent cataract surgery at the Baptist Eye Institute, Kyoto, Japan between April 2020 and May 2021. In all subjects, risk factors for early postoperative IOP elevation (i.e., an increase of IOP of 10 mmHg or more at 1-day postoperative compared with that at baseline, or a postoperative IOP of 28 mmHg or more) were analyzed by multivariate logistic regression analysis. RESULTS: Of the 1587 treated eyes in this study, 100 (6.3%) experienced early-postoperative IOP elevation. Of those 100 eyes, 78.0% were men, 27.0% had an axial length (AL) of ≥ 26.5 mm, 23.0% had a history of glaucoma treatment, 11.0% had poor mydriasis and 10.0% had intraoperative floppy iris syndrome (IFIS). Multivariate analysis findings revealed that male [odds ratio (OR) 4.36; 95% confidence interval (CI) 2.63-7.23; P < 0.001], AL of ≥ 26.5 mm (3.11; 1.83-5.30; P < 0.001), a history of glaucoma treatment (2.83; 1.63-4.91; P < 0.001), poorly mydriasis (2.63; 1.16-6.01; P = 0.02), IFIS (4.37; 1.78-10.74; P = 0.001) and baseline high IOP (1.09; 1.01-1.18; P = 0.03) were associated with increased IOP during the early period post cataract surgery. CONCLUSIONS: The findings in this study reveal that male sex, high myopia, a history of glaucoma treatment, poor mydriasis, IFIS and baseline high IOP are risk factors for IOP elevation during the early period post cataract surgery.
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Catarata , Glaucoma , Midríase , Facoemulsificação , Catarata/complicações , Feminino , Glaucoma/cirurgia , Humanos , Pressão Intraocular , Masculino , Midríase/complicações , Midríase/cirurgia , Estudos Retrospectivos , Fatores de RiscoRESUMO
Liposomes are highly biocompatible drug carriers in drug delivery systems (DDSs). Preferential accumulation of liposomes and acceleration of drug release at target tumor sites are essential for effective cancer therapy using liposomal formulations; however, conventional liposomes are unsuitable for on-demand drug release. We have previously reported that drug release can be accelerated via a bio-orthogonal inverse electron demand Diels-Alder (IEDDA) reaction between amphiphilic tetrazine (Tz)-containing liposomes and norbornene (NB) derivatives in vitro. In this study, we prepared HSTz-liposomes composed of hydrogenated soybean phosphatidylcholine (HSPC) and Tz compound (2-hexadecyl-N-(6-(6-(pyridin-2-yl)-1,2,4,5-tetrazin-3-yl)pyridin-3-yl)octadecanamide) with particle sizes of 60-80 nm and ζ-potentials of -5 to 0 mV. Similar to our previous report, the addition of 5-norbornene-2-carboxylic acid (NBCOOH) to HSTz-liposomes accelerated drug release from the liposomes in vitro. In the biodistribution study using colon26 tumor-bearing mice, the radiolabeled HSTz-liposomes were accumulated and retained in the tumor at 6-48 h post-injection, whereas the radioactivity in the blood almost disappeared at 48 h. Therefore, the timing of the injection of NBCOOH was selected to be 48 h after the injection of the HSTz-liposome to avoid the IEDDA reaction in the bloodstream. We investigated the in vivo drug release by evaluating the intratumoral localization of doxorubicin (DOX) encapsulated in HSTz-liposomes labeled with fluorescent lipids. In the tumors treated with HSTz-liposomes and NBCOOH, DOX was more widely dispersed in the tumor compared with fluorescent lipid, suggesting that the release of encapsulated drugs (DOX) from HSTz-liposomes was enhanced in the tumor tissue via the bio-orthogonal IEDDA reaction. Furthermore, the combination of DOX-encapsulated HSTz-liposomes with NBCOOH significantly suppressed tumor growth compared to conventional DOX-encapsulated liposomes. In conclusion, the bio-orthogonal IEDDA reactions in the liposomal membrane enabled the acceleration of drug release from HSTz-liposomes in vivo, suggesting a promising strategy for effective cancer therapy.
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Lipossomos , Neoplasias , Animais , Antibióticos Antineoplásicos , Linhagem Celular Tumoral , Doxorrubicina/análogos & derivados , Doxorrubicina/química , Camundongos , Neoplasias/tratamento farmacológico , Norbornanos , Polietilenoglicóis , Distribuição TecidualRESUMO
INTRODUCTION: Systemic amyloidosis is a group of diseases characterized by the deposition of amyloid protein in multiple organs throughout the body and causing their dysfunction. As amyloid deposition is observed at an early phase and is highly specific to systemic amyloidosis, noninvasive detection of amyloid is considered useful for the early diagnosis of systemic amyloidosis. In this study, we designed and synthesized a novel radiolabeled amyloid imaging probe, sodium (E)-4-amino-3-((4-(6-iodobenzothiazol-2-yl)phenyl)diazenyl)naphthalene-1-sulfonate (1), which combines two amyloid-binding compounds, thioflavin-T and Congo-red, and evaluated its effectiveness in diagnosing amyloidosis. METHODS: A tributyltin precursor was synthesized through a 5-step reaction from 2-amino-6-bromobenzothiazole, and [125I]1 was synthesized by an iododestannylation reaction with a tributyltin precursor. Mouse models of amyloid A (AA) amyloidosis, a type of systemic amyloidosis, were prepared by intraperitoneal injection of amyloid-enhancing factor into mice. An in vitro autoradiographic study was performed using spleen sections from normal mice and AA amyloidosis mice. Furthermore, [125I]1 was intravenously injected into mice, and its distribution was evaluated. Finally, an ex vivo autoradiographic study was performed using AA amyloidosis mice. RESULTS: [125I]1 was obtained with a radiochemical yield of 66% and a radiochemical purity of over 95%. In vitro autoradiography revealed specific binding of [125I]1 to thioflavin-S-stained regions in the spleen. Normal mice showed relatively rapid clearance of [125I]1 from the organs, whereas radioactivity was retained in the spleen, where amyloid deposition was observed in model mice. Furthermore, ex vivo autoradiography showed a heterogeneous distribution of [125I]1, which was co-localized with thioflavin-S-stained regions in the spleen of model mice. CONCLUSION: These results indicate the potential of radioiodinated 1 as a nuclear imaging probe for diagnosing AA amyloidosis.
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Amiloidose/diagnóstico , Benzotiazóis/química , Vermelho Congo/química , Desenvolvimento de Medicamentos , Compostos Radiofarmacêuticos/química , Animais , Autorradiografia , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Feminino , Radioisótopos do Iodo , Camundongos , Camundongos Endogâmicos ICR , Estrutura Molecular , Compostos Radiofarmacêuticos/síntese química , Relação Estrutura-AtividadeRESUMO
Significance: The imbalance in redox homeostasis is known as oxidative stress, which is relevant to many diseases such as cancer, arteriosclerosis, and neurodegenerative disorders. Overproduction of reactive oxygen species (ROS) is one of the factors that trigger the redox state imbalance in vivo. The ROS have high reactivity and impair biomolecules, whereas antioxidants and antioxidant enzymes, such as ascorbate and glutathione, reduce the overproduction of ROS to rectify the redox imbalance. Owing to this, redox monitoring tools have been developed to understand the redox fluctuations in oxidative stress-related diseases. Recent Advances: In an attempt to monitor redox substances, including ROS and radical species, versatile modalities have been developed, such as electron spin resonance, chemiluminescence, and fluorescence. In particular, many fluorescent probes have been developed that are selective for ROS. This has significantly contributed to understanding the relevance of ROS in disease onset and progression. Critical Issues: To date, the dynamics of ROS and radical fluctuation in in vivo redox states remain unclear, and there are a few methods for the in vivo detection of redox fluctuations. Future Directions: In this review, we summarize the development of radiolabeled probes for monitoring redox-relevant species by nuclear medical imaging that is applicable in vivo. In the future, translational research is likely to be advanced through the development of highly sensitive and in vivo applicable detection methods, such as nuclear medical imaging, to clarify the underlying dynamics of ROS, radicals, and redox substances in many diseases. Antioxid. Redox Signal. 36, 797-810.
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Antioxidantes , Estresse Oxidativo , Antioxidantes/metabolismo , Diagnóstico por Imagem , Oxirredução , Espécies Reativas de OxigênioRESUMO
PURPOSE: Nasal screening is performed to avoid the complications of postoperative surgical site infections (SSI), especially those due to antimicrobial-resistant species such as methicillin-resistant Staphylococcus aureus (MRSA). This study examined the relationship between bacterial isolates from the conjunctiva and the nasal cavity. METHODS: All patients were diagnosed with ocular surface infections, and the organisms in the conjunctiva and the nasal cavity were isolated. We investigated the relationship of the following antimicrobial-resistant bacteria between the conjunctiva and the nose: MRSA, methicillin-resistant CNS (MRCNS), levofloxacin-resistant (LVFX-R) Corynebacterium spp. Data were analyzed using Fisher's exact test, and the odds ratio was examined. RESULTS: This study included 188 eyes of 188 subjects (87 males and 101 females; mean age 58.5 years, range 11-97 years). MRSA (4 eyes), MRCNS (29 eyes), and LVFX-R Corynebacterium spp. (41 eyes) were identified from the conjunctiva, and MRSA (6 eyes), MRCNS (38 eyes), and LVFX-R Corynebacterium spp. (41 eyes) were identified from the nasal cavity. There was a significant relationship detected between the conjunctiva and the nose for MRSA, MRCNS, and LVFX-R Corynebacterium spp. MRSA displayed high sensitivity (0.750, 95% confidence interval [CI]; 0.301 to 0.987) and specificity (0.984, 95% CI; 0.953 to 0.996) in nasal cavity cultures, and the odds ratio was 181.00 times (95% CI; 18.41 to 2320). CONCLUSION: This study showed a significant relationship between conjunctival and nasal cultures of MRSA, MRCNS, and LVFX-R Corynebacterium spp., suggesting that nasal cavity culture is a potentially useful screening method for detecting resistant bacteria, especially MRSA, in the conjunctiva.
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Anti-Infecciosos , Staphylococcus aureus Resistente à Meticilina , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Bactérias , Criança , Túnica Conjuntiva/microbiologia , Feminino , Humanos , Levofloxacino , Masculino , Resistência a Meticilina , Pessoa de Meia-Idade , Adulto JovemRESUMO
PURPOSE: To investigate the trend of seasonal variation of intraocular pressure (IOP) in patients with normal-tension glaucoma over a 20-year period by retrospectively analyzing the Kyoto Prefectural University of Medicine Glaucoma Registry database as real-world data. DESIGN: Retrospective cohort study. METHODS: Data points (n = 49,007) were extracted retrospectively from the medical records of 1774 patients with normal-tension glaucoma (665 male patients and 1109 female patients; mean ± SD age was 59.8 ± 14.4 years; and mean ± SD observation period was 5.6 ± 4.4 years) seen over the 20-year period. We first calculated the mean IOP from all available data of each month from January 1997 through December 2016. The data were then categorized into 5 groups of 4 consecutive years each (1997-2000, 2001-2004, 2005-2008, 2009-2012, and 2013-2016) and the mean IOP of each month within the group was calculated. Seasonal variations of IOP over the 20-year study period and in the 5 consecutive groups were then investigated via nonlinear multiple regression analysis. RESULTS: A continuous decrease of IOP was detected throughout the 20-year period (P < .001), with distinct seasonal variation. The annual mean ± SD IOP was highest (13.9 ± 2.7 mm Hg) in the oldest group (1997-2000), with a gradual decrease in each subsequent group, finally becoming lowest (12.3 ± 2.7 mm Hg) in the most recent group (2013-2016) (P < .001), and all of them were accompanied by distinct seasonal variation (P < .001). CONCLUSIONS: Based on the Kyoto Prefectural University of Medicine Glaucoma Registry real-world longitudinal data, our findings revealed a continuous decrease and distinct seasonal variation of IOP in patients with normal-tension glaucoma throughout the 20-year study period.
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Glaucoma , Pressão Intraocular , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Estações do Ano , Tonometria OcularRESUMO
The aim of this study was to establish a drug delivery system (DDS) for marked therapy of tumors using a thermoresponsive polymer, polyoxazoline (POZ). The effectiveness of the following was investigated: (i) the delivery of gold nanorods (GNRs) to tumor tissues, (ii) heat production of GNR upon irradiation with near-infrared (NIR) light, and (iii) high accumulation of an intravenously injected radiolabeled POZ as a drug carrier in tumors by sensing heat produced by GNRs. When the GNR solution was irradiated with NIR light (808 nm), the solution temperature was increased both in a GNR-concentration-dependent manner and in a light-dose-dependent manner. POZ, with a lower critical solution temperature of 38 °C, was aggregated depending on the heat produced by the GNR irradiated by NIR light. When it was intratumorally pre-injected into colon26-tumor-bearing mice, followed by NIR light irradiation (GNR+/Light+ group), the tumor surface temperature increased to approximately 42 °C within 5 min. Fifteen minutes after irradiation with NIR light, indium-111 (111In)-labeled POZ was intravenously injected into tumor-bearing mice, and the radioactivity distribution was evaluated. The accumulation of POZ in the tumor was significantly (approximately 4-fold) higher than that in the control groups (GNR+/without NIR light irradiation (Light-), without injection of GNR (GNR-)/Light+, and GNR-/Light- groups). Furthermore, an in vivo confocal fluorescence microscopy study, using fluorescence-labeled POZ, revealed that uptake of POZ by the tumor could be attributed to the heat produced by GNR. In conclusion, we successfully established a novel DDS in which POZ could be efficiently delivered into tumors by using the heat produced by GNR irradiated with NIR light.
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BACKGROUND: The application of gold nanorods (GNRs) in photothermal therapy is a promising avenue for cancer treatment. The aim of this study was to develop a GNR-based targeted photothermal therapy for melanoma. METHODS: We utilized the electrostatic interaction between cationic GNRs and an anionic polymer chondroitin sulfate A (CSA), which has an affinity for binding to melanoma cells, to construct an anionic binary GNR-CSA complex (GNR-CS) at an optimal theoretical charge ratio of the trimethylammonium groups of GNR: carboxyl and sulfate groups of CSA = 1:2.5. The cytotoxicity to normal cells and erythrocyte agglutination activity of GNR-CS were evaluated. After the cellular uptake of GNR-CS by melanoma cells (B16-F10) was investigated, the photothermal performance of GNR-CS against B16-F10 cells was evaluated in vitro. RESULTS: The particle size and zeta potential of GNR-CS were approximately 35 nm and -20 mV, respectively. GNR-CS showed little cytotoxicity to normal cells and low erythrocyte agglutination activity, indicating good biocompatibility. Compared with negatively-charged GNR, GNR-CS was highly taken up by B16-F10 cells even if it was negatively charged. Cellular uptake was significantly suppressed upon treatment with excess CSA, suggesting the involvement of a CSA-specific uptake pathway. Furthermore, irradiation of the GNR-CS solution with near-infrared (NIR) light increased its temperature in light-intensity and GNR-concentration dependent manners. GNR-CS exhibited significant and GNR-dose dependent cytotoxicity in melanoma cells in combination with NIR light irradiation. CONCLUSION: GNRs coated with CSA have the potential as a medicine in targeted photothermal therapy for melanoma.
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Melanoma , Nanotubos , Fotoquimioterapia , Linhagem Celular Tumoral , Sulfatos de Condroitina , Ouro , Humanos , Melanoma/tratamento farmacológico , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/farmacologia , Fototerapia , Terapia FototérmicaRESUMO
Melanoma is a highly malignant skin cancer that frequently metastasizes to the lung, bone, and brain at an early phase. Therefore, noninvasive detection of metastasized melanoma could be beneficial to determine suitable therapeutic strategies. We previously reported a biocompatible ternary anionic complex composed of plasmid DNA (pDNA), polyethyleneimine (PEI), and γ-polyglutamic acid (γ-PGA) based on an electrostatic interaction, which was highly taken up by melanoma cells (B16-F10), even if it was negatively charged. Here, we developed a radiolabeled γ-PGA complex by using indium-111 (111In)-labeled polyamidoamine dendrimer (4th generation; G4) instead of pDNA and iodine-125 (125I)-labeled PEI instead of native PEI, and evaluated its effectiveness as a melanoma-targeted imaging probe. This ternary complex was synthesized at a theoretical charge ratio; carboxyl groups of 111In-diethylenetriaminepentaacetic acid (DTPA)-G4 : amino groups of 125I-PEI : carboxyl groups of γ-PGA was 1 : 8 : 16, and the size and zeta potential were approximately 29 nm and -33 mV, respectively. This complex was taken up by B16-F10 cells with time. Furthermore, a biodistribution study, using normal mice, demonstrated its accumulation in the liver, spleen, and lung, where macrophage cells are abundant. Almost the same level of radioactivity derived from both 111In and 125I was observed in these organs at an early phase after probe injection. Compared with the normal mice, significantly higher lung-to-blood ratios of radioactivity were observed in the B16-F10-lung metastatic cancer model. In conclusion, the radiolabeled γ-PGA complex would hold potentialities for nuclear medical imaging of lung metastatic melanoma.
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Dendrímeros/administração & dosagem , Neoplasias Pulmonares/diagnóstico , Nanopartículas/administração & dosagem , Ácido Pentético/administração & dosagem , Polietilenoimina/administração & dosagem , Ácido Poliglutâmico/análogos & derivados , Animais , Linhagem Celular Tumoral , Dendrímeros/farmacocinética , Radioisótopos de Índio , Radioisótopos do Iodo , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/secundário , Masculino , Melanoma/metabolismo , Melanoma/patologia , Camundongos Endogâmicos BALB C , Ácido Pentético/farmacocinética , Polietilenoimina/farmacocinética , Ácido Poliglutâmico/administração & dosagem , Ácido Poliglutâmico/farmacocinética , Neoplasias Cutâneas/metabolismo , Neoplasias Cutâneas/patologia , Distribuição TecidualRESUMO
It is well known that lipid carbon radicals (lipid radicals) are the origin of lipid peroxidation and are involved in various diseases such as cancer. Therefore, the in vivo detection of lipid radicals would be expected to lead to early diagnosis of these diseases. However, there are no methods for measuring lipid radicals in vivo. Nitroxides are known to be highly reactive with lipid radicals, but they tend to be reduced in vivo. Focusing on the excellent detection sensitivity of nuclear medical imaging, we have developed a radioiodinated nitroxide derivative with resistance to bioreduction for the in vivo detection of lipid radicals. The desired compound was obtained successfully and was highly stable against bioreduction while maintaining high reactivity toward lipid radicals. The I-125 labeling was efficacious with radiochemical yields of 84-87% and radiochemical purities of >99%. A cellular uptake assay showed that the radioiodinated compound was significantly taken up by cells under lipid radical-producing conditions compared to that in the absence of lipid radical production. A biodistribution study indicated that the radioiodinated compound accumulated more in organs where lipid peroxidation was promoted than the methoxyamine derivative, which lost reactivity to lipid radicals. These results indicated that the developed probe became trapped in cells or organs by reacting with lipid radicals. Thus, the radioiodinated nitroxide is a candidate probe for in vivo detection of lipid radicals.
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Antioxidantes , Radioisótopos do Iodo , Óxidos N-Cíclicos , Espectroscopia de Ressonância de Spin Eletrônica , Radicais Livres , Lipídeos , Óxidos de Nitrogênio , Distribuição TecidualRESUMO
Bio-orthogonal inverse electron demand Diels-Alder (IEDDA) reactions between liposomes containing a tetrazine-based (Tz) compound and 2-norbornene (2-NB) could be a novel trigger for accelerating drug release from the liposomes via temporary membrane destabilization, as shown in our previous report. Herein, we evaluated the in vitro drug release using NB derivatives with carboxyl groups [5-norbornene-2-carboxylic acid (NBCOOH) and 5-norbornene-2,3-dicarboxylic acid (NB(COOH)2)] to investigate the effects of substituents at the NB backbone on the drug release rate. First, POTz-liposome composed of a Tz compound (2-hexadecyl-N-(6-(6-(pyridin-2-yl)-1,2,4,5-tetrazin-3-yl)pyridin-3-yl)octadecanamide) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine (POPC) were prepared. The mass spectrometry analysis revealed the binding of NB derivatives to the Tz compound via the IEDDA reaction after the POTz-liposome reacted with the NB derivatives. Indium-111-labeled diethylenetriaminepentaacetic acid (111In-DTPA) was encapsulated inside the liposomes, and the drug release rate was quantified by measuring radioactivity. At 24 h after incubation with 2-NB, NBCOOH, and NB(COOH)2, the release rates of 111In-DTPA from POTz-liposome were 21.0, 80.8, and 23.3%, respectively, which were significantly higher than those of POTz-liposome that was not treated with NB derivatives (4.2%), indicating the involvement of the IEDDA reaction for prompting drug release. Additionally, a thermodynamic evaluation using Langmuir monolayers was conducted to explore the mechanism of the accelerated drug release. An increase in membrane fluidity and a reduction in intermolecular repulsion between POPC and the Tz compound were observed after the reaction with NB derivatives, especially for NBCOOH. Thus, the IEDDA reaction in the liposomal membrane could be a potent trigger for accelerating the release of encapsulated drugs by regulating membrane fluidity and intermolecular repulsion in the liposomal membrane.
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Elétrons , Lipossomos , Reação de Cicloadição , Fluidez de Membrana , MembranasRESUMO
Thus far, no accurate measurement technology has been developed to detect lipid alkyl radicals (lipid radicals), which cause lipid peroxidation. Therefore, we aimed to develop a nuclear medical imaging probe that can be taken up in the lipophilic site in cells such as biological membranes, by reacting specifically with the lipid radicals generated there. We designed and synthesized 4-(4-[125I]iodobenzamido)-2,2,6,6-tetramethylpiperidine-1-oxyl, which shows high reactivity to lipid radicals with a high radiochemical yield and purity. Intracellular retention was found to increase significantly when lipid radicals were produced.
RESUMO
Time courses of the redox status in the brains of mice after X-ray or carbon-ion beam irradiation were observed by magnetic resonance redox imaging (MRRI). The relationship between radiation-induced oxidative stress on the cerebral nervous system and the redox status in the brain was discussed. The mice were irradiated by 8-Gy X-ray or carbon-ion beam (C-beam) on their head under anesthesia. C-beam irradiation was performed at HIMAC (Heavy-Ion Medical Accelerator in Chiba, NIRS/QST, Chiba, Japan). MRRI measurements using a blood-brain-barrier-permeable nitroxyl contrast agent, MCP or TEMPOL, were performed using 7-T scanner at several different times, i.e., 5-10â¯h, 1, 2, 4, and 8 day(s) after irradiation. Decay rates of the nitroxyl-enhanced T1-weighted MR signals in the brains were estimated from MRRI data sets, and variation in the decay rates after irradiation was assessed. The variation in decay rates of MCP and TEMPOL after X-ray or C-beam irradiation was similar, but different variation patterns were observed between X-ray and C-beam. The apparent decay rate of both MCP and TEMPOL decreased due to the temporal reduction of blood flow in the brain several hours after X-ray and/or C-beam irradiation. After decreasing, the apparent decay rates of nitroxyl radicals in the brain gradually increased during the following days after X-ray irradiation or rapidly increased 1 day after C-beam irradiation. The sequential increase in nitroxyl decay rates may have been due to the oxidative atmosphere in the tissue due to ROS generation. X-ray and C-beam irradiation resulted in different redox responses, which may have been due to time-varying oxidative stress/injury, in the mouse brain. The C-beam irradiation effects were more acute and larger than those of X-ray irradiation.