Nanodiamond sensors for microbiology and cell biology

Nanodiamonds are nanoparticles (10 000 to 100 000 times smaller than a millimeter) milled from diamond and have various potential applications in microbiology and cell biology. Especially nanodiamonds that contain a specific fluorescent defect called 'NV center' are of great interest. They can be used to make very small magnetic signals visible with an optical microscope. This property can be used to detect magnetic molecules in cells, which are hard to detect otherwise and play an important role in processes like ageing, stress and signaling between cells. In my Phd research, I investigated the performance of these nanodiamond sensors in relation to their shape and size and how they interact with living bacteria

Effect of medium and aggregation on antibacterial activity of nanodiamonds

Fluorescent nanodiamonds are widely used as abrasives, optical or magnetic labels, in drug delivery or nanoscale sensing. They are considered very biocompatible in mammalian cells. However, in bacteria the situation looks different and results are highly controversial. This article presents a short review of the published literature and a systematic experimental study of different strains, nanoparticle sizes and surface chemistries. Most notably, particle aggregation behaviour and bacterial clumping are taken into consideration to explain reduced colony counts, which can be wrongly interpreted as a bactericidal effect. The experiments show no mechanism can be linked to a specific material property, but prove that aggregation and bacteriostatic effect of nanodiamond attachment play a significant role in the reported results

Nanosensors for diagnosis with optical, electric and mechanical transducers

Nanosensors with high sensitivity utilize electrical, optical, and acoustic properties to improve the detection limits of analytes. The unique and exceptional properties of nanomaterials (large surface area to volume ratio, composition, charge, reactive sites, physical structure and potential) are exploited for sensing purposes. High-sensitivity in analyte recognition is achieved by preprocessing of samples, signal amplification and by applying different transduction approaches. In this review, types of signals produced and amplified by nanosensors (based on transducers) are presented, to sense exceptionally small concentrations of analytes present in a sample. The use of such nanosensors, sensitivity and selectivity can offer different advantages in biomedical applications like earlier detection of disease, toxins or biological threats and create significant improvements in clinical as well as environmental and industrial outcomes. The emerging discipline of nanotechnology at the boundary of life sciences and chemistry offers a wide range of prospects within a number of fields like fabrication and characterization of nanomaterials, supramolecular chemistry, targeted drug supply and early detection of disease related biomarkers

Influence of the ceramic translucency on the relative degree of conversion of a direct composite and dual-curing resin cement through lithium disilicate onlays and endocrowns

INTRODUCTION: The goal of this study was to investigate the influence of the ceramic translucency, restoration type and polymerization time on the relative degree of conversion of a dual-curing resin cement and a conventional microhybrid resin composite using a high-power light-curing device. METHODS AND MATERIALS: Two 4.0 mm thick onlay (O) and two 7.5 mm thick endocrown (E) lithium disilicate restorations in high and low translucency (HT/LT) were fabricated on a decapitated molar. The pulp chamber was prepared to accommodate a 2 mm layer of a microhybrid resin composite (MHC) or dual-curing resin cement (DCC). Composite specimens were light-cured (n = 15; 1200 mW/cm2) without or through an onlay or endocrown restoration. Fourier-transform infrared spectroscopy (FTIR) absorbance curves were collected for the same composite specimen after 3 × 20, 3 × 40, 3 × 60 and 3 × 90 s of light-curing. The relative degree of conversion (DC%) was calculated and results analyzed using Kruskal-Wallis test and Friedman's ANOVA. Alpha was set at 0.05. RESULTS: After 3 × 60 s, the DC of MHC was significantly lower (p = 0.03; r = 0.61) under LT/EC restorations (Mdn: 77.8%) than HT/EC restorations (Mdn: 95.2%). DC of the DCC was not significantly affected by the ceramic translucency or restoration type. MHC had a significant higher DC than DCC under the HT/O, LT/O and HT/E restorations. There were no significant differences between MHC and DCC cured through LT/E restorations. CONCLUSION: DC for DCC was not significantly affected by the ceramic translucency or restoration type. DC for MHC was significantly lower for LT/EC than HT/EC restorations after 3 × 60s polymerization, but not different for the high translucent restorations and low translucent onlays. CLINICAL RELEVANCE: the use of light-curing microhybrid composite for bonding high translucent onlays and endocrowns and low translucent onlays seems feasible

Relaxometry for detecting free radical generation during Bacteria's response to antibiotics

Free radical generation plays a key role in killing bacteria by antibiotics. However, radicals are short-lived and reactive, and thus difficult to detect for the state of the art. Here we use a technique which allows optical nanoscale magnetic resonance imaging (MRI) to detect radical generation on the scale of single bacteria. We demonstrate that the radical generation in Staphylococcus aureus increases in the presence of UV irradiation as well as vancomycin and is dependent on the antibiotic's dose. With a method based on ensembles of nitrogen vacancy (NV) centers in diamond, we were able to follow the radical formation near individual bacteria over the whole duration of the experiment to reveal the dynamics of radical generation. Using this new approach, we observed free radical concentrations within nanoscale voxels around the diamond particles and determined its exact timing depending on the antibiotic dose. Since changes in the response to antibiotics emerge in only a few bacteria of the entire population, such a single-cell approach can prove highly valuable for research into drug resistance

Diamond Color Centers in Diamonds for Chemical and Biochemical Analysis and Visualization

Beyond the sparkle, other properties of diamond havegained increasing attention in the past few decades amongchemists and physicists. Color centers-impurities formed byone or a few foreign atoms or vacancies in the diamondlattice-are one reason for this. While pure diamond istransparent, the presence of color centers causes changes incoloration. Color centers introduce additional electronic statesin the wide band gap of diamond, giving rise to transitions thatabsorb and emit light in the visible spectrum

Optical Detection of Intracellular Quantities Using Nanoscale Technologies

Optical probes that can be used to measure certain quantities with subcellular resolution give us access to a new level of information at which physics, chemistry, life sciences, and medicine become strongly intertwined. The emergence of these new technologies is owed to great advances in the physical sciences. However, evaluating and improving these methods to new standards requires a joint effort with life sciences and clinical practice. In this Account, we give an overview of the probes that have been developed for measuring a few highly relevant parameters at the subcellular scale: temperature, pH, oxygen, free radicals, inorganic ions, genetic material, and biomarkers. Luminescent probes are available in many varieties, which can be used for measuring temperature, pH, and oxygen. Since they are influenced by virtually any metabolic process in the healthy or diseased cell, these quantities are extremely useful to understand intracellular processes. Probes for them can roughly be divided into molecular dyes with a parameter dependent fluorescence or phosphorescence and nanoparticle platforms. Nanoparticle probes can provide enhanced photostability, measurement quality, and potential for multiple functionalities. Embedding into coatings can improve biocompatibility or prevent nonspecific interactions between the probe and the cellular environment. These qualities need to be matched however with good uptake properties, colloidal properties and eventually intracellular targeting to optimize their practical applicability. Inorganic ions constitute a broad class of compounds or elements, some of which play specific roles in signaling, while others are toxic. Their detection is often difficult due to the cross-talk with similar ions, as well as other parameters. The detection of free radicals, DNA, and biomarkers at extremely low levels has significant potential for biomedical applications. Their presence is linked more directly to physiological and clinical manifestations. Since existing methods for free radical detection are generally poor in sensitivity and spatiotemporal resolution, new reliable methods that are generally applicable can contribute greatly to advancing this topic in biology. Optical methods that detect DNA or RNA and protein biomarkers exist for intracellular applications, but are mostly relevant for the development of rapid point-of-care sample testing. To elucidate the inner workings of cells, focused multidisciplinary research is required to define the validity and limitations of a nanoparticle probe, in both physical and biological terms. Multifunctional platforms and those that are easily made compatible with conventional research equipment have an edge over other techniques in growing the body of research evidencing their versatility

Relaxometry for detecting free radical generation during Bacteria's response to antibiotics

Polyetheretherketone is one of the polymers developed as a material to produce a removable partial denture clasp. Clasp characteristics that affect retention strength are the clasp’s size and shape. This study was conducted to investigate the effect of the dimensions polyetheretherketone rods on the force needed to produce 0.5 mm deflection. This is a laboratory study. A total of 32 specimens of polyetheretherketone rods were made in 2 lengths (15 mm and 9 mm), 2 thicknesses (2,5 mm and 1,5 mm), and 2 shapes (taper and rectanguler), and were divided into 8 groups. The deflection measurement with a radius of 0.5 mm was carried out using a hydraulic universal testing machine with a speed of 5 mm/min. The most significant force was found in polyetheretherketone rods which were 9 mm long, 2.5 mm thick, both taper and rectangular, and significantly different from other rod sizes. The rod with a 2.5 mm thickness showed more force than the 1.5 mm. Furthermore, the tapered shape did not show significantly different force than the rectangular shape. The polyetheretherketone rod's length and thickness affect the force's magnitude for a deflection of 0.5 mm. © 2022,Journal of International Dental and Medical Research. All Rights Reserved

Fast, Broad-Band Magnetic Resonance Spectroscopy with Diamond Widefield Relaxometry

We present an alternative to conventional Electron Paramagnetic Resonance (EPR) spectroscopy equipment. Avoiding the use of bulky magnets and magnetron equipment, we use the photoluminescence of an ensemble of Nitrogen-Vacancy centers at the surface of a diamond. Monitoring their relaxation time (or T1), we detected their cross-relaxation with a compound of interest. In addition, the EPR spectra are encoded through a localized magnetic field gradient. While recording previous data took 12 min per data point with individual NV centers, we were able to reconstruct a full spectrum at once in 3 s, over a range from 3 to 11 G. In terms of sensitivity, only 0.5 μL of a 1 μM hexaaquacopper(II) ion solution was necessary