Scientific publications

Polymerization of carbazole dicarboxaldehydes and lysine derivatives by imine and acylhydrazone formation afforded peptide-derived molecular biodynamers. Characterization of their physicochemical properties revealed an interesting morphology change upon polymerization from monomers forming submicrometer spherical micelles to nanometer-sized rigid-rod-shaped polymeric particles. A combination of light-scattering methods, small-angle neutron scattering, and transmission electron microscopy enabled a detailed investigation of this morphological change. Moreover, we investigated by dynamic and static light scattering how the pH affects the fluorescence and size of the biodynamers. These morphological and pH-dependent changes are expected to open the door to a myriad of applications of molecular biodynamers.

Gold nanoparticles covered with a mixture of ligands of which one type contains solubilizing triethylene glycol residues and the other peripheral zinc(II)–dipicolylamine (DPA) complexes allowed the optical detection of hydrogenphosphate, diphosphate, and triphosphate anions in water/methanol 1:2 (v/v). These anions caused the bright red solutions of the nanoparticles to change their color because of nanoparticle aggregation followed by precipitation, whereas halides or oxoanions such as sulfate, nitrate, or carbonate produced no effect. The sensitivity of phosphate sensing depended on the nature of the anion, with diphosphate and triphosphate inducing visual changes at significantly lower concentrations than hydrogenphosphate. In addition, the sensing sensitivity was also affected by the ratio of the ligands on the nanoparticle surface, decreasing as the number of immobilized zinc(II)–dipicolylamine groups increased. A nanoparticle containing a 9:1 ratio of the solubilizing and the anion-binding ligand showed a color change at diphosphate and triphosphate concentrations as low as 10 μmol/L, for example, and precipitated at slightly higher concentrations. Hydrogenphosphate induced a nanoparticle precipitation only at a concentration of ca. 400 μmol/L, at which the precipitates formed in the presence of diphosphates and triphosphates redissolved. A nanoparticle containing fewer binding sites was more sensitive, while increasing the relative number of zinc(II)–dipicolylamine complexes beyond 25% had a negative impact on the limit of detection and the optical response. Transmission electron microscopy provided evidence that the changes of the nanoparticle properties observed in the presence of the phosphates were due to a nanoparticle crosslinking, consistent with the preferred binding mode of zinc(II)–dipicolylamine complexes with phosphate anions which involves binding of the anion between two metal centers. This work thus provided information on how the behavior of mixed monolayer-protected gold nanoparticles is affected by multivalent interactions, at the same time introducing a method to assess whether certain biologically relevant anions are present in an aqueous solution within a specific concentration range.

In recent years, extracellular vesicles (EVs) and outer membrane vesicles (OMVs) have become an extensive and diverse field of research. They hold potential as diagnostic markers, therapeutics and for fundamental biological understanding. Despite ongoing studies, numerous information regarding function, content and stability of EVs remains unclear. If EVs and OMVs ought to be used as therapeutics and in clinical environments, their stability is one of the most important factors to be considered. Especially for formulation development, EVs and OMVs need to be stable at higher temperatures. To the best of our knowledge, very little work has been published regarding heat stability of neither EVs nor OMVs. In the present study, we investigated B lymphoblastoid cell-derived EVs and OMVs derived from myxobacterial species Sorangiineae as model vesicles. We exposed the vesicles to 37 °C, 50 °C, 70 °C and 100 °C for 1 h, 6 h and 24 h, and also autoclaved them. Interestingly, physico-chemical analyses such as size, particle concentration and protein concentration showed minor alterations, particularly at 37 °C. Flow cytometry analysis emphasised these results suggesting that after heat impact, EVs and OMVs were still able to be taken up by macrophage-like dTHP-1 cells. These data indicate that both mammalian and bacterial vesicles show intrinsic stability at physiological temperature. Our findings are important to consider for vesicle formulation and for advanced bioengineering approaches.

A novel approach for electrochemical tuning of alcohol oxidase (AOx) and alcohol dehydrogenase (ADH) biocatalysis towards butanol-1 oxidation by incorporating enzymes in various designs of amperometric biosensors is presented. The biosensors were developed by using commercial graphene oxide-based screen-printed electrodes and varying enzyme producing strains, encapsulation approaches (layer-by-layer (LbL) or one-step electrodeposition (EcD)), layers composition and structure, operating conditions (applied potential values) and introducing mediators (Meldola Blue and Prussian Blue) or Pd-nanoparticles (Pd-NPs). Simultaneous analysis/screening of multiple crucial system parameters during the enzyme engineering process allowed to identify within a period of one month that four out of twelve proposed designs demonstrated a good signal reproducibility and linear response (up to 14.6 mM of butanol) under very low applied potentials (from −0.02 to −0.32 V). Their mechanical stability was thoroughly investigated by multi-analytical techniques prior to butanol determination in cell-free samples from an anaerobic butanol fermentation. The EcD-based biosensor that incorporates ADH, NAD+, Pd-NPs and Nafion showed no loss of enzyme activity after preparation and demonstrated capabilities towards low potential (−0.12 V) detection of butanol-1 in fermentation medium (4 mM) containing multiple electroactive species with almost 15 times enhanced sensitivity (0.2282 μA/mM ± 0.05) when compared to the LbL design. Furthermore, the ADH-Nafion bonding for the S. cerevisiae strain was confirmed to be 3 times higher than for E. coli.

Pore collapse upon drying is a well-known phenomenon in ultrafiltration PES membranes. Here we demonstrate how alteration of membrane surface chemistry can be used to control the extent of pore collapse and ultimately to tailor membrane properties. Commercial hollow-fiber PES membranes were modified via surface-initiated ATRP to obtain different polymer-grafted membranes and were subsequently dried to facilitate pore collapse. The different polymer grafts could be used for controlling the water flux and solute rejection characteristics of the membranes. Controlled membrane pore collapse could be exploited to obtain higher rejection of sodium chloride, magnesium sulfate and calcein. Calcein as the largest solute showed almost full rejection (98.9 ± 0.3%) on the membrane. The chemical nature of the grafted polymer was directly reflected in the water flux-to-rejection ratio and the extent of pore collapse.

Abstract Engineered nano–bio cellular interfaces driven by vertical nanostructured materials are set to spur transformative progress in modulating cellular processes and interrogations. In particular, the intracellular delivery—a core concept in fundamental and translational biomedical research—holds great promise for developing novel cell therapies based on gene modification. This study demonstrates the development of a mechanotransfection platform comprising vertically aligned silicon nanotube (VA-SiNT) arrays for ex vivo gene editing. The internal hollow structure of SiNTs allows effective loading of various biomolecule cargoes; and SiNTs mediate delivery of those cargoes into GPE86 mouse embryonic fibroblasts without compromising their viability. Focused ion beam scanning electron microscopy (FIB-SEM) and confocal microscopy results demonstrate localized membrane invaginations and accumulation of caveolin-1 at the cell–NT interface, suggesting the presence of endocytic pits. Small-molecule inhibition of endocytosis suggests that active endocytic process plays a role in the intracellular delivery of cargo from SiNTs. SiNT-mediated siRNA intracellular delivery shows the capacity to reduce expression levels of F-actin binding protein (Triobp) and alter the cellular morphology of GPE86. Finally, the successful delivery of Cas9 ribonucleoprotein (RNP) to specifically target mouse Hprt gene is achieved. This NT-enhanced molecular delivery platform has strong potential to support gene editing technologies.

Abstract Engineered nano–bio cellular interfaces driven by 1D vertical nanostructures (1D-VNS) are set to prompt radical progress in modulating cellular processes at the nanoscale. Here, tuneable cell–VNS interfacial interactions are probed and assessed, highlighting the use of 1D-VNS in immunomodulation, and intracellular delivery into immune cells—both crucial in fundamental and translational biomedical research. With programmable topography and adaptable surface functionalization, 1D-VNS provide unique biophysical and biochemical cues to orchestrate innate and adaptive immunity, both ex vivo and in vivo. The intimate nanoscale cell–VNS interface leads to membrane penetration and cellular deformation, facilitating efficient intracellular delivery of diverse bioactive cargoes into hard-to-transfect immune cells. The unsettled interfacial mechanisms reported to be involved in VNS-mediated intracellular delivery are discussed. By identifying up-to-date progress and fundamental challenges of current 1D-VNS technology in immune-cell manipulation, it is hoped that this report gives timely insights for further advances in developing 1D-VNS as a safe, universal, and highly scalable platform for cell engineering and enrichment in advanced cancer immunotherapy such as chimeric antigen receptor-T therapy.

Multimaterial additive manufacturing is an enabling tool for exploring difficult to access structure–property relationships. In this work, a recently developed multimaterial printing approach, solution mask liquid lithography, is used to produce porous polymer–polymer composites inspired by tough, hierarchical structures found in nature. The results demonstrate that varying the size and packing of pores in the core structure leads to significant enhancement in crack deflection. Finite element analysis reveals that this enhancement is linked to geometry-dependent stress distribution.

Aqueous rechargeable batteries are sustainable energy storage devices with the potential to replace the current state-of-the-art organic phase secondary batteries. Electrode materials for secondary batteries are often based on composite structures, which combine an electronically conducting scaffold with an ionic conductor, whose properties define battery capacity. Optimal integration of these components can be challenging: here we describe a novel approach to prepare electrode materials based on growth at the liquid-liquid interface. This is illustrated with the synthesis of a carbon nanotube/Prussian blue nanocomposite as free-standing transparent thin films, which are applied as cathodes for aqueous rechargeable potassium batteries. Prussian blue is synthesized through an acid-induced decomposition of ferricyanide, promoted by an interfacial electron transfer from an organic phase donor (1,1′-dimethylferrocene) under ambient conditions. The interfacial synthesis yields selective growth of cubic Prussian blue crystals on the carbon nanotube walls, enhancing interaction between the ionic and electronically conducting components, and resulting in a self-assembled film at the liquid/liquid interface. The films are readily transferred to flexible membranes and applied as cathodes in an aqueous rechargeable K+ battery. Coin-cell devices with activated carbon anodes gave a capacity of 47.6 mAh g−1 at 0.25 A g−1 with an energy density of 33.75 Wh kg−1

We discuss the complex interplay between host and guest dynamics for a polymer in soft confinement by a droplet-phase microemulsion. Intermediate scattering functions obtained by neutron spin echo spectroscopy are first analysed by means of an effective diffusion coefficient. From its dependence on the absolute of the scattering vector q we concluded a sophisticated model for the systems dynamics taking both polymer and microemulsion contributions into account. Global fitting of this model to the intermediate scattering functions at all measured q-values and all investigated confinement sizes eventually allows for a precise disentangling of the pure polymer dynamics in confinement from the overlaying microemulsion dynamics. Validity of our approach is further supported by numerical random walk calculations.