Slawomir Kadlubowski

Synthesis of poly(acrylic acid) nanogels by preparative pulse radiolysis

Abstract Nanogels are sub-micron size, water-swellable crosslinked structures of hydrophilic polymers. In this work a radiation-based synthesis method that has been previously tested for neutral polymers is applied for production of nanogels of a synthetic polyelectrolyte—poly(acrylic acid) (PAA). In this technique dilute, deoxygenated PAA solution (pH 2) circulating in a closed loop is subjected to pulse irradiation with fast electrons. In each pulse many tens of radicals are instantaneously formed on every macromolecule. One of the major reaction paths of these radicals is intramolecular recombination leading to the formation of nanogels. It is demonstrated that radiation-induced reactions in our system show a typical feature of intramolecular crosslinking, i.e . a strong decrease in dimensions of a polymer coil without an accompanying decrease in molecular weight. In accordance with expectations based on earlier observations on non-polar polymers, intramolecular recombination of PAA-derived radicals proceeds according to non-classical kinetics. A model of non-homogeneous kinetics with time-dependent rate constant has been applied to describe this behaviour and the relationship between kinetic parameters and initial average number of radicals per chain is briefly discussed. The weight-average molecular weight of the products is influenced by side reactions, mainly degradation (chain breakage) and intermolecular crosslinking.

Nano-, micro- and macroscopic hydrogels synthesized by radiation technique

Radiation techniques, due to the additive-free initiation and easy process control, are very suitable tools for synthesis of biomaterials, especially hydrogels. In our group, a number of techniques have been elaborated allowing for targeted synthesis of gels of various size ranges, from internally crosslinked individual macromolecules, via microgels to macroscopic hydrogels. An example of a mature technology of this kind are hydrogel wound dressings, being now produced on large scale in Poland and other countries. Current research projects include: hydrogel-based system for anticancer therapy due to local drug delivery, systems for encapsulation of living cells, new approach to the synthesis of polymeric material for intervertebral discs implant, temperature-sensitive membranes, hydrogel phantoms of 3D radiation dosimeter for radiotherapy, degradation-resistant nanogels and microgels for biomedical purposes (e.g. synovial fluid substitute), hydrogel-based dietary products and adjustment of the molecular weight of biopolymers.

Radiation-synthesized protein-based drug carriers: Size-controlled BSA nanoparticles

Nanotechnology has broadened the options for the delivery of agents of biotechnological and clinical relevance. Currently, attention has been driven towards the development of protein-based nanocarriers due to high the biocompatibility and site-specific delivery. In this work we report radiation-synthesized bovine serum albumin nanoparticles as an attempt to overcome limitations of available albumin particles, as a novel route for the development of crosslinked protein nanocarriers for the administration of chemotherapic agents or radiopharmaceuticals. Albumin containing phosphate buffer solutions were irradiated using γ-irradiation at distinct cosolvent concentrations-ethanol or methanol. Nanoparticle size was followed by DLS and bityrosine crosslinking formation using fluorescence measurements and SDS-PAGE. In addition, computational experiments were performed to elucidate the mechanism and pathways for the nanoparticle formation. The synthesis of BSA nanoparticles using γ-irradiation in the presence of a cosolvent allowed the formation of the nanoparticles from 7 to 70 nm without the use of any chemical crosslinker as confirmed by SDS-PAGE and DLS analysis. The combination of cosolvent and γ-irradiation allowed a fine tuning with regard to protein size.

Synthesis of papain nanoparticles by electron beam irradiation - A pathway for controlled enzyme crosslinking.

Crosslinked enzyme aggregates comprise more stable and highly concentrated enzymatic preparations of current biotechnological and biomedical relevance. This work reports the development of crosslinked nanosized papain aggregates using electron beam irradiation as an alternative route for controlled enzyme crosslinking. The nanoparticles were synthesized in phosphate buffer using various ethanol concentrations and electron beam irradiation doses. Particle size increase was monitored using dynamic light scattering. The crosslinking formation by means of bityrosine linkages were measured by fluorescence spectra and the enzymatic activity was monitored using Na-Benzoyl-dl-arginine p-nitroanilide hydrochloride as a substrate. The process led to crosslinked papain nanoparticles with controlled sizes ranging from 6 to 11nm depending upon the dose and ethanol concentration. The irradiation atmosphere played an important role in the final bioactivity of the nanoparticles, whereas argon and nitrous oxide saturated systems were more effective than at atmospheric conditions in terms of preserving papain enzymatic activity. Highlighted advantages of the technique include the lack of monomers and crosslinking agents, quick processing with reduced bioactivity changes, and the possibility to be performed inside the final package simultaneously with sterilization.

Tetrazolium salts-Pluronic F-127 gels for 3D radiotherapy dosimetry

This work is a follow-up study for a recently-proposed 3D radiochromic gel dosimeter that contains a tetrazolium salt and a physical gel matrix made of poly(ethylene oxide)-block-poly(propylene oxide)-block-poly(ethylene oxide) (Pluronic F-127). Several tetrazolium salts were examined in this work, including tetrazolium violet, blue tetrazolium chloride, nitro blue tetrazolium chloride (NBT), tetranitro blue tetrazolium chloride (tNBT) and thiazolyl blue tetrazolium bromide (TBTB). The salt-containing gel dosimeters were compared with the first Pluronic gel composition that contained 2,3,5-triphenyltetrazolium chloride (TTC) as the radiation-sensitive component (dose sensitivity of 0.0023 (Gy cm)-1). The Pluronic gels with NBT and tNBT outperformed the other gels, including the TTC-containing gel, with respect to their dose sensitivity and low dose-response. The NBT gels were found to have better stability over time than tNBT gels. Sensitization of the gels to ionizing radiation was examined by addition of tert-butyl alcohol and sodium formate. The best composition was 0.0818% NBT (1 mM), 25% Pluronic F-127 and 0.136  ×  10-2% sodium formate. This gel dosimeter was insensitive to changes in dose rate for photons of different energies. The mean dose sensitivity amounted to 0.0047  ±  0.1  ×  10-4 (Gy cm)-1. A diversion in the dose-response was observed for the gel irradiated with electrons. Additional characteristics of the NBT gel were a linear-dose range and a dynamic-dose range between  <1 and  ⩾150 Gy and a dose threshold of  <1 Gy. The dose distribution registered for the NBT-Pluronic gel was stable after irradiation for over 7 d with no visible diffusion of the irradiated part, which is analogous to the original TTC-Pluronic gel.