Adsorption and Distribution of Triplex-hybridizing Bioconjugated Gold Nanoclusters inside Spermatozoa- A Study using Confocal Microscopy and Fluorescence Lifetime Imaging.

The study of the interactions between biofunctionalized gold nanoclusters (Au NCs) and spermatozoa is highly relevant to evaluate the potential of Au NCs as imaging probes and transfection agents in the reproductive biology. In this work, confocal laser scanning microscopy (CLSM) was used to investigate the distribution of Au NCs bioconjugated with peptide (nuclear localisation sequence, NLS) and oligonucleotide (locked nucleic acid, LNA) ligands in bovine spermatozoa. Fluorescence lifetime imaging (FLIM) was employed to detect changes in the NC´s chemical environment. We observed a pronounced regio-selective accumulation of the bioconjugates in spermatozoa with high concentration at the equatorial segment. Furthermore, 3D-CLSM showed successful non-endosomal cellular uptake of the conjugates by intact sperm cells and the distribution of the bioconjugates was found to be influenced by the ligand types. Interestingly, the FLIM data showed differences in lifetime depending on membrane integrity. Furthermore, ligand-dependent changes in lifetime between NC bioconjugates carrying peptide and oligonucleotide ligands were found, probably attributed to specific interactions with sperm cell compartments.

Platinum-Iridium Alloy Nanoparticle Coatings Produced by Electrophoretic Deposition Reduce Impedance in 3D Neural Electrodes.

Platinum-based neural electrodes, frequently alloyed with Ir or W, are routinely used to treat neurological disorders. However, their performance is impaired by an increase in impedance that compromises long-term implant functionality. Though there are multiple coating techniques available to address this issue, electrode, and base material often exhibit a compositional mismatch, which impairs mechanical stability and may lead to toxicological side effects. In this work, we coated Pt wire electrodes with ligand-free electrostatically stabilized colloidal Pt90Ir10, Pt90W10, and Pt50W50 alloy nanoparticles (NPs) matching electrode compositions using electrophoretic deposition (EPD) with direct-current (DC) and pulsed-DC fields in aqueous medium. The generated alloy NPs exhibit a solid solution structure as evidenced by HR-TEM-EDX and XRD, though additional WOx phases were identified in the Pt50W50 samples. Consequently, coating efficiency was also impaired in the presence of high W mass fractions in the alloy NPs. Characterization of the NP coatings by cyclic voltammetry and impedance spectroscopy yielded a significant reduction of the impedance in the Pt90Ir10 sample in comparison to the control coated with Pt NPs. The electrochemical surface area (ECSA) of the PtW alloy coatings, on the other hand, was significantly reduced.

Comparing Direct and Pulsed-Direct Current Electrophoretic Deposition on Neural Electrodes: Deposition Mechanism and Functional Influence.

Electrophoretic deposition (EPD) of platinum nanoparticles (PtNPs) on platinum-iridium (Pt-Ir) neural electrode surfaces is a promising strategy to tune the impedance of electrodes implanted for deep brain stimulation in various neurological disorders such as advanced Parkinson's disease and dystonia. However, previous results are contradicting as impedance reduction was observed on flat samples while in three-dimensional (3D) structures, an increase in impedance was observed. Hence, defined correlations between coating properties and impedance are to date not fully understood. In this work, the influence of direct current (DC) and pulsed-DC electric fields on NP deposition is systematically compared and clear correlations between surface coating homogeneity and in vitro impedance are established. The ligand-free NPs were synthesized via pulsed laser processing in liquid, yielding monomodal particle size distributions, verified by analytical disk centrifugation (ADC). Deposits formed were quantified by UV-vis supernatant analysis and further characterized by scanning electron microscopy (SEM) with semiautomated interparticle distance analyses. Our findings reveal that pulsed-DC electric fields yield more ordered surface coatings with a lower abundance of particle assemblates, while DC fields produce coatings with more pronounced aggregation. Impedance measurements further highlight that impedance of the corresponding electrodes is significantly reduced in the case of more ordered coatings realized by pulsed-DC depositions. We attribute this phenomenon to the higher active surface area of the adsorbed NPs in homogeneous coatings and the reduced particle-electrode electrical contact in NP assemblates. These results provide insight for the efficient EPD of bare metal NPs on micron-sized surfaces for biomedical applications in neuroscience and correlate coating homogeneity with in vitro functionality.

Co-doping of iron and copper ions in nanosized bioactive glass by reactive laser fragmentation in liquids.

Bioactive glass (BG) is a frequently used biomaterial applicable in bone tissue engineering and known to be particularly effective when applied in nanoscopic dimensions. In this work, we employed the scalable reactive laser fragmentation in liquids method to produce nanosized 45S5 BG in the presence of light-absorbing Fe and Cu ions. Here, the function of the ions was twofold: (i) increasing the light absorption and thus causing a significant increase in laser fragmentation efficiency by a factor of 100 and (ii) doping the BG with bioactive metal ions up to 4 wt%. Our findings reveal an effective downsizing of the BG from micrometer-sized educts into nanoparticles having average diameters of <50 nm. This goes along with successful element-specific incorporation of the metal ions into the BG, inducing co-doping of Fe and Cu ions as verified by energy-dispersive X-ray spectroscopy (EDX). In this context, the overall amorphous structure is retained, as evidenced by X-ray powder diffraction (XRD). We further demonstrate that the level of doping for both elements can be adjusted by changing the BG/ion concentration ratio during laser fragmentation. Consecutive ion release experiments using inductively-coupled plasma mass spectrometry (ICP-MS) were conducted to assess the potential bioactivity of the doped nanoscopic BG samples, and cell culture experiments using MG-63 osteoblast-like cells demonstrated their cytocompatibility. The elegant method of in situ co-doping of Fe and Cu ions during BG nanosizing may provide functionality-advanced biomaterials for future studies on angiogenesis or bone regeneration, particularly as the level of doping may be adjusted by ion concentrations and ion type in solution.

Mechanical Stability of Nano-Coatings on Clinically Applicable Electrodes, Generated by Electrophoretic Deposition.

The mechanical stability of implant coatings is crucial for medical approval and transfer to clinical applications. Here, electrophoretic deposition (EPD) is a versatile coating technique, previously shown to cause significant post-surgery impedance reduction of brain stimulation platinum electrodes. However, the mechanical stability of the resulting coating has been rarely systematically investigated. In this work, pulsed-DC EPD of laser-generated platinum nanoparticles (PtNPs) on Pt-based, 3D neural electrodes is performed and the in vitro mechanical stability is examined using agarose gel, adhesive tape, and ultrasonication-based stress tests. EPD-generated coatings are highly stable inside simulated brain environments represented by agarose gel tests as well as after in vivo stimulation experiments. Electrochemical stability of the NP-modified surfaces is tested via cyclic voltammetry and that multiple scans may improve coating stability could be verified, indicated by higher signal stability following highly invasive adhesive tape stress tests. The brain sections post neural stimulation in rats are analyzed via laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS). Measurements reveal higher levels of Pt near the region stimulated with coated electrodes, in comparison to uncoated controls. Even though local concentrations in the vicinity of the implanted electrode are elevated, the total Pt mass found is below systemic toxicologically relevant concentrations.