Combination of therapies is a common strategy in cancer treatment. Such combined therapies only have merit provided that there is superior therapeutic outcome with fewer side effects, compared to single therapies. Here, this work explores the possibility to combine chemotherapy with radionuclide therapy using polymeric micelles as a delivery vehicle. For this purpose, this work prepares poly(ε-caprolactone-b-ethylene oxide) (PCL-PEO) micelles and load them simultaneously with paclitaxel (PTX) and ¹⁷⁷Lu(III). This work chooses a 3D tumor spheroid composed of glioblastoma cells (U87) to evaluate the combined treatment. The diffusion of the micelles in the spheroid is investigated by confocal laser scanning microscopy (CLSM) and light-sheet fluorescence microscopy (LSFM). The results show that the micelles are able to penetrate deep into the spheroid within 24 h of incubation and mainly accumulated around or in the lysosomes once in the cell. Subsequently, this work evaluates the cell killing efficiency of the single treatments (PTX or ¹⁷⁷Lu(III)) versus combined treatment (PTX + ¹⁷⁷Lu(III)) by measuring the growth of the spheroids as well as by performing a cell-viability assay. The results indicate that the combined therapy achieves a superior therapeutic outcome with better cell growth inhibition and cell killing efficiency compared to the single treatments.

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A variety of polymer micelles are designed for the delivery of chemotherapeutic drugs to tumors. Although the promise of these nanocarriers is very high, in the clinic the effectivity is rather limited. Determining the in vivo fate of the micelles can greatly help to improve this treatment. Here, a simple and fast chelator-free method for radiolabeling of polymer micelles composed of different block copolymers is presented, which can allow evaluating the behavior of the nanocarriers in vivo using noninvasive nuclear imaging techniques (e.g., single photon computed tomography, SPECT). The radiolabeling method consists of adding the radioisotope ions, i.e., ¹¹¹In(III), resulting in a high radiolabeling efficiencies up to 90%. The results suggest that the radiolabeling efficiency depends on two important factors: the properties of the hydrophobic block in the block copolymer composing the micelle core, and the speciation of the radiometal salts. The formation of metal hydroxides and their precipitation in the core of the micelles appears to be a key factor for high stability. Moreover, the method can be applied to radiolabel the micelles in the presence of chemotherapeutic drugs. Finally, a SPECT study shows that the radiolabeled samples are stable in vivo without any evident loss of ¹¹¹In(III).

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In certain tumor and diseased tissues, reactive oxygen species (ROS), such as H₂O₂, are produced in higher concentrations than in healthy cells. Drug delivery and release systems that respond selectively to the presence of ROS, while maintaining their stability in “healthy” biological conditions, have great potential as on-site therapeutics. This study presents polymer micelles with 4-(methylthio)phenyl ester functionalities as a ROS-responsive reactivity switch. Oxidation of the thioether moieties triggers ester hydrolysis, exposing a hydrophylic carboxylate and leading to micellar disassembly. At 37 °C, the micelles fall apart on a timescale of days in the presence of 2 mM H₂O₂ and within hours at higher concentrations of H₂O₂ (60-600 mM). In the same time frame, the nanocarriers show no hydrolysis in oxidant-free physiological or mildly acidic conditions. This logic gate cascade behavior represents a step forward to realize drug delivery materials capable of selective response to a biomarker input.

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Four different MOFs were exposed to γrays by a cobalt-60 source reaching a maximum dose of 5 MGy. The results showed that the MIL-100 (Cr) and MIL-100 (Fe) did not exhibit obvious structural damage, suggesting their excellent radiation stability. MIL-101 (Cr) showed good radiation stability up to 4 MGy, but its structure started degrading with increasing radiation dose. Furthermore, the results showed that the structure of AlFu MOFs started to decompose at a gamma dose of 1 MGy, exhibiting a much lower tolerance to γradiation. At this radiation energy, the dominant interaction of the gamma-ray with MOFs is the Compton effect and the radiation stability of MOFs can be improved by prolific aromatic linkers, high linker connectivity, and good crystallinity. The results of this study indicate that MIL-100 and MIL-101 MOFs have a good potential to be employed in nuclear applications, where relatively high radiation doses play a role, for example, nuclear waste treatment and radionuclides production.

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The focus of this thesis is on the utilization of biodegradable drug nanocarriers combined with ionizing radiation in cancer treatment. Polymeric micelles based on PCL-PEO block copolymers were selected as the main platform for delivering various therapeutic substances due to their size, degradability and easy preparation. This thesis can roughly be divided into two main parts. In the first part we combined the external radiation beams with PCL-PEO micelles which are applied as a nanoplatform for chemotherapeutic drugs and photosensitizer and investigated the possibility that using the radiation power as a trigger for drug release from the micelles. In the second part, we focus more on the cooperation of radionuclides and PCL-PEO micelles. So far we developed a chelator-free method to radiolabel micelles for determining their in vivo behavior, as well as evaluated the possibility to combine chemotherapy with radionuclide therapy using the micelles as a nanoplatform. In both parts we attempted to unravel mechanisms behind the observed phenomena to be able to adjust the nano-carriers accordingly. @en

Polymeric micelles, due to their easy preparation and versatile properties, have been widely applied as one of the most popular carriers for chemotherapeutic agents. Such micelles primarily prevent the leakage of drugs during transportation and thus protect healthy tissue. Controlled drug release, which releases the drugs at the site of interest using internal or external stimuli as triggers, can further improve the safety of the drug delivery process. In this paper, we investigate whether ionizing radiation can be used to initiate release, focusing on using Cerenkov light as a possible trigger. For this purpose, micelles composed of the degradable polymer poly(ϵ-caprolactone-b-ethylene glycol) (PCL-PEO) were first loaded with the photosensitizer chlorin e6 (Ce6) and subsequently exposed to gamma or X-ray radiation of varying radiation doses. The results reveal that Ce6 was released from the micelles under radiation, regardless of the energy of incident photons, showing that Cerenkov light was not the driving force behind the observed release. SANS measurements showed that the volume fraction of the micelles containing Ce6 was reduced after exposure to radiation. This change in volume fraction suggests that the number of micelles was reduced, which was probably responsible for the release of Ce6. The exact mechanism, however, remains unclear. Subsequently, the PCL-PEO micelles were loaded with Ce6 and one of the following drugs: doxorubicin (Dox), docetaxel (DTX), and paclitaxel (PTX). Under radiation exposure, Dox, which is quite stable in single-loaded micelles, shows an enhanced release profile in the presence of Ce6, while DTX and PTX remained in the micelles, regardless of the presence of Ce6.

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Gamma radiation triggered secondary crosslinking of dextran hydrogels leads to macroscopic hydrogel contraction. The authors use stable polymer hydrogels, prepared through azide-alkyne crosslinking, containing surplus alkyne groups. γ-irradiation of these gels leads to more alkyne crosslinking, enabling controlled increase of crosslink density, which in turn leads to an increase of hydrogel stiffness and macroscopic hydrogel contraction. Gel contraction scales linearly with the applied radiation dose. The same mechanism is applied to achieve γ-radiation triggered release of the small molecule cargo, akin to wringing out a sponge. γ-irradiation of touching hydrogel objects leads to gel fusion and the formation of a self-supporting gel connection, demonstrating the reactivity of the excess alkyne groups. They envision applications in gel gluing and the construction of complex gel architectures, as well as in responsive materials for controlled release.

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We present a new light cleavable polymer containing o-nitrobenzene thioacetal groups in the main chain. By conjugation to a PEG block, we synthesized block copolymers capable of forming nanoparticles in aqueous solution. We studied drug encapsulation and release using the model drug Nile Red. Irradiation with UV-A light (365 nm) leads to efficient degradation of the polymers and associated burst release of the payload. Unlike other thioacetal and thioketal polymers, these polymers are stable to reactive oxygen species (ROS), preventing non-triggered release. Moreover, the nanocarriers showed low cytotoxicity in cell viability experiments.

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It is well-known that energy-rich radiation induces water splitting, eventually yielding hydrogen peroxide. Synthetic applications, however, are scarce and to the best of our knowledge, the combination of radioactivity with enzyme-catalysis has not been considered yet. Peroxygenases utilize H2O2 as an oxidant to promote highly selective oxyfunctionalization reactions but are also irreversibly inactivated in the presence of too high H2O2 concentrations. Therefore, there is a need for efficient in situ H2O2 generation methods. Here, we show that radiolytic water splitting can be used to promote specific biocatalytic oxyfunctionalization reactions. Parameters influencing the efficiency of the reaction and current limitations are shown. Particularly, oxidative inactivation of the biocatalyst by hydroxyl radicals influences the robustness of the overall reaction. Radical scavengers can alleviate this issue, but eventually, physical separation of the enzymes from the ionizing radiation will be necessary to achieve robust reaction schemes. We demonstrate that nuclear waste can also be used to drive selective, peroxygenase-catalyzed oxyfunctionalization reactions, challenging our view on nuclear waste in terms of sustainability.

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A great number of fluorescent probes have been developed for detecting singlet oxygen (¹O₂), which is considered to be one of the most effective reactive oxygen species (ROS), especially in clinical applications. The commercially available fluorescent probe Singlet Oxygen Sensor Green (SOSG) is widely used due to its reported high selectivity to ¹O₂. In this study, we carried out systemic experiments to determine the activation of SOSG in the presence of ionizing radiation. The results show that the SOSG probe exhibits a pronounced fluorescence increase as a function of radiation dose delivered by gamma-rays as well as X-rays, in conditions where the formation of singlet oxygen is not expected. Furthermore, scavenger tests indicate that hydroxyl radicals may be involved directly or indirectly in the activation process of SOSG although the exact mechanism remains unknown.

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