Towards Cancer Nanoradiopharmaceuticals-Radioisotope Nanocarrier System for Prostate Cancer Theranostics Based on Radiation-Synthesized Polymer Nanogels.

Despite the tremendous development of oncology, prostate cancer remains a debilitating malignancy. One of the most promising approaches to addressing this issue is to exploit the advancements of nanomedicine in combination with well-established nuclear medicine and radiotherapy. Following this idea, we have developed a radioisotope nanocarrier platform of electron-beam-synthesized nanogels based on poly(acrylic acid). We have developed a functionalization protocol, showing the very high (>97%) efficiency of the conjugation in targeting a ligand-bombesin derivative. This engineered peptide can bind gastrin-releasing peptide receptors overexpressed in prostate cancer cells; moreover, it bears a radioisotope-chelating moiety. Our nanoplatform exhibits very promising performance in vitro; the radiolabeled nanocarriers maintained high radiochemical purity of >90% in both the labeling buffer and human serum for up to 14 days. The application of the targeted nanocarrier allowed also effective and specific uptake in PC-3 prostate cancer cells, up to almost 30% after 4 h, which is a statistically significant improvement in comparison to carrier-free radiolabeled peptides. Although our system requires further studies for more promising results in vivo, our study represents a vital advancement in radionanomedicine-one of many steps that will lead to effective therapy for castration-resistant prostate cancer.

"To Be or Not to Be" of a Polymer Nanogel-Unravelling the Relationship of Product Properties vs. Synthesis Conditions Governing the Radiation Crosslinking of Poly(acrylic acid) Using GPC/SEC-MALLS.

In this paper, a state-of-the-art multi-detection gel permeation chromatography/size exclusion chromatography (GPC/SEC) system including multi-angle laser light scattering (MALLS) is applied to monitor radiation-induced synthesis of internally crosslinked nanostructures from poly(acrylic acid) (PAA). The aim is to demonstrate that this modern tool yields a more detailed picture of reaction mechanism and product structure than the techniques used to date. The prevailing intramolecular crosslinking narrows the molecular weight distribution from Mw/Mn = 3.0 to 1.6 for internally crosslinked structures. A clear trend from over 0.7 to 0.5 in the Mark-Houwink exponent and a decrease in Rg/Rh from 1.7 to 1.0 point to the formation of nanogels, more rigid and less permeable than the starting coils. Changes in the coil contraction factor (g' = [η]irradiated/[η]linear) as a function of the radical density revealed the existence of two modes in intramolecular crosslinking, the initial one (up to 0.075 radicals per monomer unit) where the compactness of products changes strongly with progressing crosslinking and a second one where further compacting is suppressed by the lower flexibility of the partially crosslinked chain segments. This indicates a transition from soft, still internally crosslinkable nanogels to more rigid structures, less prone to further intramolecular loop formation. Our findings provide means for the tailored design of new PAA nanomaterials.

Dextran Methacrylate Reactions with Hydroxyl Radicals and Hydrated Electrons in Water: A Kinetic Study Using Pulse Radiolysis.

Dextran methacrylate (Dex-MA) is a biodegradable polysaccharide derivative that can be cross-linked by ionizing radiation. It is therefore considered a potential replacement for synthetic hydrophilic polymers in current radiation technologies used for synthesizing hydrophilic cross-linked polymer structures such as hydrogels, mainly for medical applications. This work is focused on the initial steps of radiation-induced cross-linking polymerization of Dex-MA in water. Rate constants of two major transient water radiolysis products-hydroxyl radicals (•OH) and hydrated electrons (eaq-)-with various samples of Dex-MA (based on 6-500 kDa dextrans of molar degree of substitution or DS with methacrylate groups up to 0.66) as well as non-substituted dextran were determined by pulse radiolysis with spectrophotometric detection. It has been demonstrated that these rate constants depend on both the molecular weight and DS; reasons for these effects are discussed and reaction mechanisms are proposed. Selected spectral data of the transient species formed by •OH- and eaq--induced reactions are used to support the discussion. The kinetic data obtained in this work and their interpretation are expected to be useful for controlled synthesis of polysaccharide-based hydrogels and nanogels of predefined structure and properties.

Nanostrategies for Therapeutic and Diagnostic Targeting of Gastrin-Releasing Peptide Receptor.

Advances in nanomedicine bring the attention of researchers to the molecular targets that can play a major role in the development of novel therapeutic and diagnostic modalities for cancer management. The choice of a proper molecular target can decide the efficacy of the treatment and endorse the personalized medicine approach. Gastrin-releasing peptide receptor (GRPR) is a G-protein-coupled membrane receptor, well known to be overexpressed in numerous malignancies including pancreatic, prostate, breast, lung, colon, cervical, and gastrointestinal cancers. Therefore, many research groups express a deep interest in targeting GRPR with their nanoformulations. A broad spectrum of the GRPR ligands has been described in the literature, which allows tuning of the properties of the final formulation, particularly in the field of the ligand affinity to the receptor and internalization possibilities. Hereby, the recent advances in the field of applications of various nanoplatforms that are able to reach the GRPR-expressing cells are reviewed.

IAEA Contribution to Nanosized Targeted Radiopharmaceuticals for Drug Delivery.

The rapidly growing interest in the application of nanoscience in the future design of radiopharmaceuticals and the development of nanosized radiopharmaceuticals in the late 2000's, resulted in the creation of a Coordinated Research Project (CRP) by the International Atomic Energy Agency (IAEA) in 2014. This CRP entitled 'Nanosized delivery systems for radiopharmaceuticals' involved a team of expert scientist from various member states. This team of scientists worked on a number of cutting-edge areas of nanoscience with a focus on developing well-defined, highly effective and site-specific delivery systems of radiopharmaceuticals. Specifically, focus areas of various teams of scientists comprised of the development of nanoparticles (NPs) based on metals, polymers, and gels, and their conjugation/encapsulation or decoration with various tumor avid ligands such as peptides, folates, and small molecule phytochemicals. The research and development efforts also comprised of developing optimum radiolabeling methods of various nano vectors using diagnostic and therapeutic radionuclides including Tc-99m, Ga-68, Lu-177 and Au-198. Concerted efforts of teams of scientists within this CRP has resulted in the development of various protocols and guidelines on delivery systems of nanoradiopharmaceuticals, training of numerous graduate students/post-doctoral fellows and publications in peer reviewed journals while establishing numerous productive scientific networks in various participating member states. Some of the innovative nanoconstructs were chosen for further preclinical applications-all aimed at ultimate clinical translation for treating human cancer patients. This review article summarizes outcomes of this major international scientific endeavor.

On the Mechanism and Kinetics of Synthesizing Polymer Nanogels by Ionizing Radiation-Induced Intramolecular Crosslinking of Macromolecules.

Nanogels-internally crosslinked macromolecules-have a growing palette of potential applications, including as drug, gene or radioisotope nanocarriers and as in vivo signaling molecules in modern diagnostics and therapy. This has triggered considerable interest in developing new methods for their synthesis. The procedure based on intramolecular crosslinking of polymer radicals generated by pulses of ionizing radiation has many advantages. The substrates needed are usually simple biocompatible polymers and water. This eliminates the use of monomers, chemical crosslinking agents, initiators, surfactants, etc., thus limiting potential problems with the biocompatibility of products. This review summarizes the basics of this method, providing background information on relevant aspects of polymer solution thermodynamics, radiolysis of aqueous solutions, generation and reactions of polymer radicals, and the non-trivial kinetics and mechanism of crosslinking, focusing on the main factors influencing the outcomes of the radiation synthesis of nanogels: molecular weight of the starting polymer, its concentration, irradiation mode, absorbed dose of ionizing radiation and temperature. The most important techniques used to perform the synthesis, to study the kinetics and mechanism of the involved reactions, and to assess the physicochemical properties of the formed nanogels are presented. Two select important cases, the synthesis of nanogels based on polyvinylpyrrolidone (PVP) and/or poly(acrylic acid) (PAA), are discussed in more detail. Examples of recent application studies on radiation-synthesized PVP and PAA nanogels in transporting drugs across the blood-brain barrier and as targeted radioisotope carriers in nanoradiotherapy are briefly described.

Synthesis and Properties of Targeted Radioisotope Carriers Based on Poly(Acrylic Acid) Nanogels.

Radiation crosslinking was employed to obtain nanocarriers based on poly(acrylic acid)-PAA-for targeted delivery of radioactive isotopes. These nanocarriers are internally crosslinked hydrophilic macromolecules-nanogels-bearing carboxylic groups to facilitate functionalization. PAA nanogels were conjugated with an engineered bombesin-derivative-oligopeptide combined with 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate chelating moiety, aimed to provide selective radioligand transport. 4-(4,6-Dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium (DMTMM) toluene-4-sulfonate was used as the coupling agent. After tests on a model amine-p-toluidine-both commercial and home-synthesized DOTA-bombesin were successfully coupled to the nanogels and the obtained products were characterized. The radiolabeling efficiency of nanocarriers with 177Lu, was chromatographically tested. The results provide a proof of concept for the synthesis of radiation-synthesized nanogel-based radioisotope nanocarriers for theranostic applications.

Thermally Conductive Shape Memory Polymer Composites Filled with Boron Nitride for Heat Management in Electrical Insulation.

The evaluation of a possible application of functional shrinkable materials in thermally conductive electrical insulation elements was investigated. The effectiveness of an electron beam and gamma radiation on the crosslinking of a selected high density polyethylene grade was analyzed, both qualitatively and quantitatively. The crosslinked polymer composites filled with ceramic particles were successfully fabricated and tested. On the basis of the performed investigation, it was concluded that the selected filler, namely a boron nitride powder, is suitable for the preparation of the crosslinked polymer composites with enhanced thermal conductivity. The shape memory effect was fully observed in the crosslinked samples with a recovery factor reaching nearly 99%. There was no significant influence of the crosslinking, stretching, and recovery of the polymer composite during shape memory phenomenon on the value of thermal conductivity. The proposed boron nitride filled polyethylene composite subjected to crosslinking is a promising candidate for fabrication of thermally shrinkable material with enhanced heat dissipation functionality for application as electrically insulating components.

Polymerization Reactions and Modifications of Polymers by Ionizing Radiation.

Ionizing radiation has become the most effective way to modify natural and synthetic polymers through crosslinking, degradation, and graft polymerization. This review will include an in-depth analysis of radiation chemistry mechanisms and the kinetics of the radiation-induced C-centered free radical, anion, and cation polymerization, and grafting. It also presents sections on radiation modifications of synthetic and natural polymers. For decades, low linear energy transfer (LLET) ionizing radiation, such as gamma rays, X-rays, and up to 10 MeV electron beams, has been the primary tool to produce many products through polymerization reactions. Photons and electrons interaction with polymers display various mechanisms. While the interactions of gamma ray and X-ray photons are mainly through the photoelectric effect, Compton scattering, and pair-production, the interactions of the high-energy electrons take place through coulombic interactions. Despite the type of radiation used on materials, photons or high energy electrons, in both cases ions and electrons are produced. The interactions between electrons and monomers takes place within less than a nanosecond. Depending on the dose rate (dose is defined as the absorbed radiation energy per unit mass), the kinetic chain length of the propagation can be controlled, hence allowing for some control over the degree of polymerization. When polymers are submitted to high-energy radiation in the bulk, contrasting behaviors are observed with a dominant effect of cross-linking or chain scission, depending on the chemical nature and physical characteristics of the material. Polymers in solution are subject to indirect effects resulting from the radiolysis of the medium. Likewise, for radiation-induced polymerization, depending on the dose rate, the free radicals generated on polymer chains can undergo various reactions, such as inter/intramolecular combination or inter/intramolecular disproportionation, b-scission. These reactions lead to structural or functional polymer modifications. In the presence of oxygen, playing on irradiation dose-rates, one can favor crosslinking reactions or promotes degradations through oxidations. The competition between the crosslinking reactions of C-centered free radicals and their reactions with oxygen is described through fundamental mechanism formalisms. The fundamentals of polymerization reactions are herein presented to meet industrial needs for various polymer materials produced or degraded by irradiation. Notably, the medical and industrial applications of polymers are endless and thus it is vital to investigate the effects of sterilization dose and dose rate on various polymers and copolymers with different molecular structures and morphologies. The presence or absence of various functional groups, degree of crystallinity, irradiation temperature, etc. all greatly affect the radiation chemistry of the irradiated polymers. Over the past decade, grafting new chemical functionalities on solid polymers by radiation-induced polymerization (also called RIG for Radiation-Induced Grafting) has been widely exploited to develop innovative materials in coherence with actual societal expectations. These novel materials respond not only to health emergencies but also to carbon-free energy needs (e.g., hydrogen fuel cells, piezoelectricity, etc.) and environmental concerns with the development of numerous specific adsorbents of chemical hazards and pollutants. The modification of polymers through RIG is durable as it covalently bonds the functional monomers. As radiation penetration depths can be varied, this technique can be used to modify polymer surface or bulk. The many parameters influencing RIG that control the yield of the grafting process are discussed in this review. These include monomer reactivity, irradiation dose, solvent, presence of inhibitor of homopolymerization, grafting temperature, etc. Today, the general knowledge of RIG can be applied to any solid polymer and may predict, to some extent, the grafting location. A special focus is on how ionizing radiation sources (ion and electron beams, UVs) may be chosen or mixed to combine both solid polymer nanostructuration and RIG. LLET ionizing radiation has also been extensively used to synthesize hydrogel and nanogel for drug delivery systems and other advanced applications. In particular, nanogels can either be produced by radiation-induced polymerization and simultaneous crosslinking of hydrophilic monomers in "nanocompartments", i.e., within the aqueous phase of inverse micelles, or by intramolecular crosslinking of suitable water-soluble polymers. The radiolytically produced oxidizing species from water, •OH radicals, can easily abstract H-atoms from the backbone of the dissolved polymers (or can add to the unsaturated bonds) leading to the formation of C-centered radicals. These C-centered free radicals can undergo two main competitive reactions; intramolecular and intermolecular crosslinking. When produced by electron beam irradiation, higher temperatures, dose rates within the pulse, and pulse repetition rates favour intramolecular crosslinking over intermolecular crosslinking, thus enabling a better control of particle size and size distribution. For other water-soluble biopolymers such as polysaccharides, proteins, DNA and RNA, the abstraction of H atoms or the addition to the unsaturation by •OH can lead to the direct scission of the backbone, double, or single strand breaks of these polymers.

Amniotic Stem Cells Cultured on Thermoresponsive Polymers Allow Obtaining a Full Cell Sheet.

Amniotic stem cells promote adhesion and migration of epithelial cells. Obtaining a full sheet containing amniotic stem cells seems to be the best solution for the treatment of burn wounds. The main advantage of this method is obtaining a full sheet of cells by lowering the temperature below the transition temperature, which does not affect extracellular matrix. The purpose of this work was to produce a skin substitute-a full sheet consisting of amniotic mesenchymal stem cells-and compare with well-known fibroblast sheet. Amniotic membrane cells revealed better tendency to full sheet detachment than fibroblasts. Confluence after 24 hours was always higher on polymer-coated dishes than on normal polypropylene dishes. Also viability was higher than on the control culture dish, while the number of apoptotic cells was always highest on polypropylene (control). Ile-Lys-Val-ala-Val (IKVAV) 0.28 addition to poly (poly [ethylene glycol] ethyl methacrylate) (PTEGMA) caused best cell confluence and highest percentage of cells in mitosis phase of cell cycle, but also worst cell detachment was observed in both cell types on PTEGMA IKVAV 0.28. Viability of cells transferred in cell sheet form onto a new culture dish was higher than when detached as suspension enzymatically. Additionally, percentage of apoptotic cells transferred in cell sheet form onto a new culture dish was always lower than when detached as suspension enzymatically. Culturing of PTEGMA, PTEGMA IKVAV 0.28 and PTEGMA IKVAV 0.14 have a stimulating effect on number of cells in mitosis in amniotic cell culture even after cell sheet transfer onto a new dish, whereas such effect with fibroblast was not observed.

On the Mechanisms of the Effects of Ionizing Radiation on Diblock and Random Copolymers of Poly(Lactic Acid) and Poly(Trimethylene Carbonate).

This article demonstrates that ionizing radiation induces simultaneous crosslinking and scission in poly(trimethylene carbonate-co-d-lactide) diblock and random copolymers. Copolymer films were electron-beam (EB) irradiated up to 300 kGy under anaerobic conditions and subsequently examined by evaluation of their structure (FT-IR, NMR), molecular weight, intrinsic viscosities, and thermal properties. Radiation chemistry of the copolymers is strongly influenced by the content of ester linkages of the lactide component. At low lactide content, crosslinking reaction is the dominant one; however, as the lactide ratio increases, the ester linkages scission becomes more competent and exceeds the crosslinking. Electron paramagnetic resonance (EPR) measurements indicate that higher content of amorphous carbonate units in copolymers leads to a reduction in free radical yield and faster radical decay as compared to lactide-rich compositions. The domination of scission of ester bonds was confirmed by identifying the radiolytically produced alkoxyl and acetyl radicals, the latter being more stable due to its conjugated structure.

Hydroxyl radical-induced crosslinking and radiation-initiated hydrogel formation in dilute aqueous solutions of carboxymethylcellulose.

Ionizing radiation causes chain scission of polysaccharides in the absence of crosslinking agents. It has been demonstrated before that degradation of carboxyalkylated polysaccharides may be prevented, despite presence of strong electrostatic repulsing forces between chains, at very high polymer concentration in water (paste-like state) when physical proximity promotes recombination of radiation-generated polymer radicals. In such conditions, crosslinking dominates over chain scission and covalent, macroscopic gels can be formed. In an approach proposed in this work, neutralizing the charges on carboxymethylcellulose (CMC) by lowering the pH results in retracting the electrostatic repulsion between chain segments and thus allows for substantial reduction of polymer concentration required to achieve gelation due to domination of crosslinking reactions. Electron-beam irradiation of aqueous solutions of low pH containing 0.5-2% CMC results in hydrogel formation with 70% yield, while both concentration and dose determine their swelling properties. Time-resolved studies by laser flash photolysis clearly indicate strong pH influence on decay kinetics of CMC radicals.

Determination of Propagation Rate Coefficient for the Polymerization of N-Vinylpyrrolidone in Aqueous Solution by Pulsed Electron Polymerization and Size Exclusion Chromatography.

A new method for determination of a propagation rate coefficient in radical polymerization, pulsed electron polymerization-size exclusion chromatography (PEP-SEC), has been tested on N-vinylpyrrolidone in water and shown to yield results very similar to those obtained by the well-established pulsed laser polymerization-size exclusion chromatography (PLP-SEC). A potential advantage of PEP-SEC is its applicability to studying polymerizations in nontransparent systems and lack of any additives. Series of nanosecond pulses of high-energy electrons from an accelerator generate radicals which initiate polymerization. Further analysis of the samples and data processing are the same as in PLP-SEC. The described technique can be potentially developed into a method complementary and/or comparative to PLP-SEC.

Poly(epsilon-caprolactone) biomaterial sterilized by E-beam irradiation.

The effects of ionizing radiation (electron beam) on poly(epsilon-caprolactone) (PCL) were studied by analyzing changes in viscosity-average and weight-average molecular weight and radius of gyration, and by performing sol-gel analysis and swelling tests. Samples were irradiated under various conditions: solid and molten PCL in the presence or absence of air. The overall efficiency of crosslinking is higher for samples irradiated in the molten state than in the solid state, and is reduced in the presence of oxygen. Based on three kinds of experiments (molecular weight dependence on the dose in the pre-gelation region, sol-gel analysis, and swelling study), radiation-chemical yields of intermolecular crosslinking and scission were determined and are discussed in terms of the mechanism of radiation-induced reactions in PCL. Properties of the gels formed by high-dose irradiation and mechanical properties of irradiated PCL were analyzed. Irradiation causes an increase in the compression modulus of PCL. This process occurs at the pre-gelation stage and continues in the gel-containing system. We have demonstrated, for the first time, that irradiation of solid PCL is accompanied by a pronounced post-effect, which manifests itself by changes in the average molecular weight. EPR data indicate that this effect, at least in part, is caused by the presence of long-lived radicals trapped in the crystalline regions. Irradiation with the sterilizing dose does not cause a statistically significant change in the biocompatibility of PCL after subsequent storage for 79 d, as determined by preliminary osteoblast vitality tests.

[Investigation of local reaction of muscular tissue after injection of polyvinylpyrrolidone preparation]. / Badania miejscowej reakcji tkanki miesniowej po iniekcji preparatu poliwinylopirolidonu.

Natural preparations for replenishing of hyaluronic acid of zoogenous origin used till now, are characterized with quit low biocompatibility and also too short effect of their action. Recently worked out synthetic polyvinylpyrrolidone preparation PVP, contains modification constituting internally netting of microgels to improve polymer bioresistance. The introduce modification can, however influence PVP biocompatibility after deposition into tissues of the living organism. The aim of research was evaluation of the local reaction of muscular tissue after PVP infection. The results of research concerned a control group where normal saline was used for infections. The research was carried out on 18 Wistar rats and included macroscopic and histologic observations made in the 3rd, 5th, 7th, 14th and 30th day after PVP injection into thigh muscle. The local reaction of muscular tissue was macroscopically characterized with inflammatory reaction till the 7th day after PVP injection, in later terms the observed changes disappeared. Microscopic research showed that PVP till the 7th day after PVP injection caused rather strong diffuse non-specific inflammatory process, yet without essential participation of neutrophils leading to producing of loose intra-muscular. Connective tissue in a later term. The carried out tests showed presence of PVP in muscular tissue till the 30th day after injection.

[Tissue reaction after injection of polyvinylpyrrolidone (PVP) preparation into knee joint. Experiment]. / Odczyn tkankowy po iniekcji preparatu poliwinylopirolidonu (PVP) do stawu kolanowego. Badania doswiadczalne.

Internally netted miscogelatinated preparation on the basis of polyvinylpyrrolidone PVP desioned for viscosuplementation of joint fluid was worked out. Netted structure of microgel grains presents larger resistance to the degrading action of free radicals than analogical linear polymer chains. Application of zoogeous preparations of hyaluronic acid results in short-term effects of their usage in evoking reaction foreign protein. Replenishment of joint fluid with preparation with higher biostability from biocompatible synthetic polymer-polyvinylpyrrolidone could improve the function of synovial through restoration of its proper viscosity and protection of the joint for a longer period of time. The aim of the experiment was determination of bioresistance and reaction of microgel PVP on the tissues of synovial joint. The tests were carried out on 10 white New Zealand rabbits after injection PVP into the knee joint for 3, 7, 14 and 30 days and submitted to macroscopic and histological evaluation. The results of tests were compared with the data obtained after injection of normal saline. Macroscopically, there were no changes in the limits of articular capsule and cartilage; there was only slight and enlargement of synovial membrane in the first 7 days after PVP injection. In histological tests it was observed that reaction in the knee joint after PVP injection was characterised it single inflammatory chains without essential participation of neutrophils observed only in synovial membrane and limited to places were tested preparation was seen. Microgel PVP was present in diverticula of synovial membrane to 30th day after injection.