Microspheres with high specific activities of radionuclides are very interesting for internal radiotherapy treatments. This work focuses on the formulation and characterization of inorganic microspheres with a high content of holmium and therefore a high specific radioactivity of holmium-166. Two novel formulations of inorganic microspheres were obtained by dispersing solid holmium acetylacetonate microspheres (Ho₂(AcAc)₃-ms) in NaH₂PO₄ or NaOH solutions followed by 2 h incubation at room temperature. By exchange of acetylacetonate with phosphate or hydroxyl ions, holmium phosphate microspheres (HoPO₄-ms) and holmium hydroxide microspheres (Ho(OH)₃-ms) were formed respectively. The inorganic microspheres had a significantly smaller diameter (28.5 ± 4.4 μm (HoPO₄-ms) and 25.1 ± 3.5 μm (Ho(OH)₃-ms)) than those of Ho₂(AcAc)₃-ms (32.6 ± 5.2 μm). The weight percentage of holmium-165 in the microspheres increased significantly from 47% (Ho₂(AcAc)₃-ms) to 55% (HoPO₄-ms) and 73% (Ho(OH)₃-ms). After preparation of both HoPO₄-ms and Ho(OH)₃-ms, the stable holmium-165 isotope was partly converted by neutron activation into radioactive holmium-166 to yield radioactive microspheres. High specific activities were achieved ranging from 21.7 to 59.9 MBq/mg (¹⁶⁶HoPO₄-ms) and from 28.8 to 79.9 MBq/mg (¹⁶⁶Ho(OH)₃-ms) depending on the neutron activation time. The structure of both microspheres was preserved up to neutron activations of 6 h in a thermal neutron flux of 4.72 × 10¹⁶ n m⁻² s⁻¹. After activation, both microspheres revealed excellent stability in administration fluids (saline and phosphate buffer) having less than 0.05% of holmium released after 72 h incubation. Finally, the hemocompatibility of these inorganic microspheres was evaluated and it was shown that the microspheres did cause neither hemolysis nor depletion or inhibition of the coagulation factors of the intrinsic blood coagulation pathway meaning that the microspheres have a good hemocompatibility. Overall, this work shows that radioactive inorganic microspheres with high specific activities of holmium-166 can be prepared which potentially can be used for internal radionuclide therapy.

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We report a flexible single-cell isolation method by imaging-assisted hydrogel formation. Our approach consists of imaging-aided selective capture of cells of interest by encasing them into a polymeric hydrogel, followed by removal of unwanted cells and subsequent release of isolated cells by enzymatic hydrogel degradation, thus offering an opportunity for further analysis or cultivation of selected cells. We achieved high sorting efficiency and observed excellent viability rates (>98%) for NIH/3T3 fibroblasts and A549 carcinoma cells isolated using this procedure. The method presented here offers a mask-free, cost-efficient and easy-to-use alternative to many currently existing surface-based cell-sorting techniques, and has the potential to impact the field of cell culturing and isolation, e.g. single cell genomics and proteomics, investigation of cellular heterogeneity and isolation of best performing mutants for developing new cell lines.

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The aim of this study was the development of radioactive holmium phosphate microspheres (HoPO₄-MS) with a high holmium content and that are stable in human serum for selective internal radiation therapy (SIRT) of liver cancer. To this end, holmium acetylacetonate microspheres (HoAcAc-MS) were prepared (34.2 ± 1.0 µm in diameter, holmium content of 46.2 ± 0.8 and density of 1.7 g/cm³) via an emulsification and solvent evaporation method. The concentration of HoAcAc in the organic solvent, the temperature of emulsification and the stirring speed were varied for the preparation of the HoAcAc-MS to obtain microspheres with different diameters ranging from 11 to 35 µm. Subsequently, the AcAc ligands of the HoAcAc-MS were replaced by phosphate ions by simply incubating neutron irradiated HoAcAc-MS in a phosphate buffer solution (0.116 M, pH 4.2) to yield radioactive HoPO₄-MS. The obtained microspheres were analyzed using different techniques such as SEM-EDS, ICP-OES and HPLC. The prepared HoPO₄-MS (29.5 ± 1.2 µm in diameter and a density of 3.1 g/cm³) present an even higher holmium content (52 wt%) than the HoAcAc-MS precursor (46 wt%). Finally, the stability of the HoPO₄-MS was tested by incubation in human serum at 37 °C which showed no visible changes of the microspheres morphology and only 0.1% of holmium release was observed during the 2 weeks period of incubation. In conclusion, this study shows that stable radioactive HoPO₄-MS can be prepared with suitable properties to be used for cancer therapy.

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The last decade has witnessed great progress in understanding and manipulating self-assembly of block copolymers in solution. A wide variety of micellar structures can be created and many promising applications in bioscience have been reported. In particular, nano-fibrous micelles provide a great platform to mimic the filamentous structure of native extracellular matrix (ECM). However, the evaluation of this kind of filomicellar system with potential use in tissue engineering is virtually unexplored. The question behind it, such as if the block copolymer nano-fibrous micelles can regulate cellular response, has lingered for many years because of the difficulties in preparation and 3D manipulation of these tiny objects. Here, by using a combination approach of self-assembly of block copolymers and soft lithography, we establish a novel and unique nano-fibrous 2D platform of organized micelles and demonstrate that patterned micelles enable control over the cellular alignment behavior. The area density and orientation of fibrous micelles determine the alignment degree and directionality of cells, respectively. Furthermore, when cells were cultured on multi-directionally aligned micelles, a competitive response was observed. Due to the virtually infinite possibilities of functionalization of the micelle corona, our work opens a new route to further mimic the native fibrous networks with artificial micelles containing various functionalities.

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Pluronics P94 are block-copolymer showing prolonged circulation time and tumor-cell internalization in vitro, suggesting a potential for tumor accumulation and as a drug carrier. Here we report the results of the radiolabeled-P94 unimers (P94-111In-DTPA) on tumor uptake/retention and biodistribution after intravenous and intratumoral injection to tumor-bearing mice. Intravenous administration results in a high radioactive signal in the liver; while in tumor and other healthy tissues only low levels of radioactivity could be measured. In contrast, the intratumoral injection of P94 resulted in elevated levels of radioactivity in the tumor and low levels in other organs, including the liver. Independently from the injection route, the tumor tissue presented long retention of radioactivity. The minimal involvement of off-target tissues of P94, together with the excellent tracer retention over-time in the tumor designates Pluronic P94 copolymer as a highly promising carrier for anti-tumor drugs.@en

Chemotherapeutic drugs have multiple drawbacks, including severe side effects and suboptimal therapeutic efficacy. Nanomedicines assist in improving the biodistribution and target accumulation of chemotherapeutic drugs, and are therefore able to enhance the balance between efficacy and toxicity. Multiple types of nanomedicines have been evaluated over the years, including liposomes, polymer-drug conjugates and polymeric micelles, which rely on strategies such as passive targeting, active targeting and triggered release for improved tumor-directed drug delivery. Based on the notion that tumors and metastases are highly heterogeneous, it is important to integrate imaging properties in nanomedicine formulations in order to enable non-invasive and quantitative assessment of targeting efficiency. By allowing for patient pre-selection, such next generation nanotheranostics are useful for facilitating clinical translation and personalizing nanomedicine treatments.

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Neutron activation is widely applied for thepreparation of radioactive isotopes to be used in imaging and/or therapy. The type of diagnostic/therapeutic agents varies fromsmall chelates coordinating radioactive metal ions to complexnanoparticulate systems. Design of these agents often relies onconjugation of certain organic functionalities that determine theirpharmacokinetics, biodistribution, targeting, and cell-penetratingabilities, or simply on tagging them with an optical label. Theconjugation chemistry at the surface of nanoparticles and theirfinal purification often require laborious procedures that becomeeven more troublesome when radioactive materials are involved.This study represents a thorough investigation on the effects ofneutron activation on the organic moieties of functionalizednanoparticles, with special focus on166Ho2O3particles conjugated with PEG-fluorescein and PEG-polyarginine motives.Spectroscopic and thermogravimetric analyses demonstrate only a limited degradation of PEG-fluorescein upon irradiation of theparticles up to 10 h using a thermal neutronflux of 5×1016m−2s−1. Cell experiments show that the polyarginine-basedmechanisms of membrane penetration remain unaltered after exposure of the functionalized particles to the mixedfield ofneutrons and gammas present during activation. This confirms that radiation damage on the PEG-polyarginines is minimal.Intrinsic radiations from166Ho do not seem to affect the integrity of conjugated organic material. Thesefindings open up a newperspective to simplify the procedures for the preparation of functionalized metal-based nanosystems that need to be activated byneutron irradiation in order to be applied for diagnostic and/or therapeutic purposes.@en