A new gradient copolymer has been synthesized by the living cationic ring-opening polymerization of hydrophilic 2-ethyl-2-oxazoline with lipophilic 2-(4-dodecyloxyphenyl)-2-oxazoline (EtOx-grad-DPOx). The prepared copolymer is capable of assembling in water to yield polymeric nanoparticles that are successfully loaded with an anticancer agent, curcumin. Self-assembly of the copolymer was found to be tuned by the polarity as well as the hydrogen bonding ability of solvents. Solvent took distinctive role in the preparation of unloaded and curcumin-loaded nanoparticles. The stability of the nanoparticles was increased by curcumin loading promoted by curcumin-polymer interactions. Further, the chemical stability of curcumin in water is largely enhanced inside the polymeric nanoparticles. Curcumin-loaded (EtOx-grad-DPOx) copolymer nanoparticles showed excellent stability in the biological medium, low cytotoxicity, and concentration dependent uptake by U87 MG and HeLa cells, which indicate the possibility of their efficient application in drug delivery.

Oxidative phosphorylation and glycolysis are important features, by which cells could bypass oxidative stress. The level of oxidative stress, and the ability of cells to promote oxidative phosphorylation or glycolysis, significantly determined proliferation or cell demise. In the present work, we have employed selective mitochondrial probe MitoTracker™ Orange CMTM/Ros (MTO) to estimate the level of oxidative stress in cancer cells at different stressed conditions. MTO is partially sensitive to decrease of mitochondrial membrane potential and to reactive oxygen species (ROS) generated in mitochondria. We have demonstrated, that fluorescence lifetime of MTO is much more sensitive to oxidative stress than intensity-based approaches. This method was validated in different cancer cell lines. Our approach revealed, at relatively low ROS levels, that Gö 6976, a protein kinase C (PKC) α inhibitor, and rottlerin, an indirect PKCδ inhibitor, increased mitochondrial ROS level in glioma cell. Their involvement in oxidative phosphorylation and apoptosis was investigated with oxygen consumption rate estimation, western blot and flow-cytometric analysis. Our study brings new insight to identify feeble differences in ROS production in living cells.

There seems to be general agreement that oxidative stress is involved in many pathological conditions including Parkinson’s, Alzheimer’s, and other neurodegenerative diseases, and overall aging. Cerium oxide nanoparticles, also known as nanoceria (CeO2–NPs), have shown promise as catalytic antioxidants, based on their ability to switch between Ce3+ and Ce4+ valence states. In the present work we have synthesized and characterized CeO2–NPs, examined the effect of CeO2–NPs on amyloidogenesis of insulin, and analyzed the impact of CeO2–NPs on oxidative stress and biocompatibility in vitro in three types of invasive cancer cells, and in vivo in the preclinical model of the chorioallantoic membrane (CAM) of quail embryos. The different experimental techniques revealed a high stability and homogeneity of the “naked” CeO2–NPs synthesized by precipitation from a reversal microemulsion. The CeO2–NPs were 5–6 nm in diameter (TEM) and monodispersed and have a ζ +46.9 mV ζ potential in Milli-Q water. We demonstrated for the first time that CeO2–NPs affect insulin fibrillation in a dose-dependent manner. The inhibiting, IC50, and disassembling, DC50, concentrations were calculated to be ∼100 ± 3.5 and ∼200 ± 5.5 μg/mL, respectively. Furthermore, CeO2–NPs demonstrated reliable biocompatibility and sufficient uptake by glioma and breast cancer cells. The presence of a high concentration of CeO2–NPs within the cells resulted only in local changes in metabolic activity and generation of oxidative stress at a low level. Moreover, high biocompatibility with CeO2–NPs was shown in vivo in the CAM.

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Self-assembled nanostructures of amphiphilic gradient copoly(2-oxazoline)s have recently attracted attention as promising delivery systems for the effective delivery of hydrophobic anticancer drugs. In this study, we have investigated the effects of increasing hydrophobic side chain length on the selfassembly of gradient copolymers composed of 2-ethyl-2-oxazoline as the hydrophilic comonomer and various 2-(4-alkyloxyphenyl)-2oxazolines as hydrophobic comonomers. We show that the size of the formed polymeric nanoparticles depends on the structure of the copolymers. Moreover, the stability and properties of the polymeric assembly can be affected by the loading of hypericin, a promising compound for photodiagnostics and photodynamic therapy (PDT). We have found the limitation that allows rapid or late release of hypericin from polymeric nanoparticles. The nanoparticles entering the cells by endocytosis decreased the hypericininduced PDT, and the contribution of the passive process (diffusion) increased the probability of a stronger photoeffect. A study of fluorescence pharmacokinetics and biodistribution revealed differences in the release of hypericin from nanoparticles toward the quail chorioallantoic membrane, a preclinical model for in vivo studies, depending on the composition of polymeric nanoparticles. Photodamage induced by PDT in vivo well correlated with the in vitro results. All formulations studied succeeded in targeting hypericin at cancer cells. In conclusion, we demonstrated the promising potential of poly(2-oxazoline)-based gradient copolymers for effective drug delivery and sequential drug release needed for successful photodiagnostics and PDT in cancer therapy.

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polymers confer distinct mechanical and biological properties to single bacteria and to mature biofilms. When individual bacteria attach to a surface, intracellular levels of cyclic-di-GMP increase. Cyclic-di-GMP is required to change gene expression to initiate the transition to the biofilm state. What specific cues control cyclic-di-GMP production were previously unknown -we show that this is controlled by mechanical shear stress, which is primarily impacted by bacterial motility and the EPS coating on bacteria. This opens up the possibility of making a new class of anti-biofilm surface, by using a 2D fluid that cannot sustain a lateral shear stress and thereby preventing activation of the cyclic-di-GMP signal. P. aeruginosa biofilm infections in the cystic fibrosis (CF) lung often last for decades, ample time for the infecting strain(s) to evolve. Production of alginate is well-known to tend to increase during CF infections. More recently, it is becoming recognized that CF infections also evolve to increase PSL production. Alginate chemically protects biofilms, but also makes them softer and weaker. Here, we show that PSL stiffens and strengthens biofilms, and that increased PSL production in biofilms grown from CF clinical isolates completely rescues the mechanical weakening caused by alginate.

Due to the simple one-step preparation method and a promising application in biomedical research, amphiphilic gradient copoly(2-oxazoline)s are gaining more and more interest compared to their analogous block copolymers. In this work, the curcumin solubilization ability was tested for a series of amphiphilic gradient copoly(2-oxazoline)s with different lengths of hydrophobic side-chains, consisting of 2-ethyl-2-oxazoline as a hydrophilic monomer and 2-(4-alkyloxyphenyl)-2-oxazoline as a hydrophobic monomer. It is shown that the length of the hydrophobic side-chain in the copolymers plays a crucial role in the loading of curcumin onto the self-assembled nanoparticles. The kinetic stability of self-assembled nanoparticles studied using FRET shows a link between their integrity and cellular uptake in human glioblastoma cells. The present study demonstrates how minor changes in the molecular structure of gradient copoly(2-oxazoline)s can lead to significant differences in the loading, stability, cytotoxicity, cellular uptake, and pharmacokinetics of nano-formulations containing curcumin. The obtained results on the behavior of the complex of gradient copoly(2-oxazoline)s and curcumin may contribute to the development of effective next-generation polymeric nanostructures for biomedical applications.