In vivo examination of enamel microcracks after orthodontic debonding: Is there a need for detailed analysis?

INTRODUCTION: Our aim was to assess changes in the number of enamel microcracks (EMCs) after removing metal brackets in teeth with and without visible EMCs before the bonding procedure. METHODS: Before bonding, 13 patients having teeth with visible EMCs and 13 subjects whose teeth were free of EMCs were included in the study. All patients were asked to complete a questionnaire with a detailed medical history at the beginning of treatment and after removing metal brackets. The number of teeth with visible EMCs and the number of premolars without EMCs were recorded for each subject twice, that is, before bonding and after debonding, together with the tooth sensitivity assessments elicited by compressed air and cold testing. RESULTS: The number of visible EMCs in premolars increased after removing metal brackets. EMCs were recorded in at least 25.0% of all evaluated teeth for the patients having teeth with and without visible EMCs at the beginning of treatment. However, the changes in the number of visible EMCs were not significantly different (P = 0.619) between the groups. For the subjects with visible EMCs, tooth sensitivity caused by cold was registered nearly 3 times more often after removing brackets compared with the patients without EMCs prior bonding. CONCLUSIONS: Formation of EMCs was noticed after debonding. Changes in the number appeared to be similar for the subjects with and without visible EMCs before bonding. Higher incidence of EMCs was associated with more frequent tooth sensitivity perceptions after removing brackets.

Tilted black-Si: ∼0.45 form-birefringence from sub-wavelength needles.

The self-organised conical needles produced by plasma etching of silicon (Si), known as black silicon (b-Si), create a form-birefringent surface texture when etching of Si orientated at angles of θi < 50 - 70° (angle between the Si surface and vertical plasma E-field). The height of the needles in the form-birefringent region following 15 min etching was d ∼ 200 nm and had a 100 µm width of the optical retardance/birefringence, characterised using polariscopy. The height of the b-Si needles corresponds closely to the skin-depth of Si ∼λ/4 for the visible spectral range. Reflection-type polariscope with a voltage-controlled liquid-crystal retarder is proposed to directly measure the retardance Δn × d/λ ≈ 0.15 of the region with tilted b-Si needles. The quantified form birefringence of Δn = -0.45 over λ = 400 - 700 nm spectral window was obtained. Such high values of Δn at visible wavelengths can only be observed in the most birefringence calcite or barium borate as well as in liquid crystals. The replication of b-Si into Ni-shim with high fidelity was also demonstrated and can be used for imprinting of the b-Si nanopattern into other materials.

Optical damage thresholds of microstructures made by laser three-dimensional nanolithography: publisher's note.

This publisher's note contains corrections to Opt. Lett.45, 13 (2020).OPLEDP0146-959210.1364/OL.45.000013.

Mesoscale laser 3D printing.

3D meso scale structures that can reach up to centimeters in overall size but retain micro- or nano-features, proved to be promising in various science fields ranging from micro-mechanical metamaterials to photonics and bio-medical scaffolds. In this work, we present synchronization of the linear and galvanometric scanners for efficient femtosecond 3D optical printing of objects at the meso-scale (from sub-µm to sub-cm spanning five orders of magnitude). In such configuration, the linear stages provide stitch-free structuring at nearly limitless (up to tens-of-cm) working area, while galvo-scanners allow to achieve translation velocities in the range of mm/s-cm/s without sacrificing nano-scale positioning accuracy and preserving the undistorted shape of the final print. The principle behind this approach is demonstrated, proving its inherent advantages in comparison to separate use of only linear stages or scanners. The printing rate is calculated in terms of voxels/s, showcasing the capability to maintain an optimal feature size while increasing throughput. Full capabilities of this approach are demonstrated by fabricating structures that reach millimeters in size but still retain sub-µm features: scaffolds for cell growth, microlenses, and photonic crystals. All this is combined into a benchmark structure: a meso-butterfly. Provided results show that synchronization of two scan modes is crucial for the end goal of industrial-scale implementation of this technology and makes the laser printing well aligned with similar approaches in nanofabrication by electron and ion beams.

Enamel microcracks in the form of tooth damage during orthodontic debonding: a systematic review and meta-analysis of in vitro studies.

Objectives: To evaluate and compare the enamel microcracks (EMCs) characteristics (qualitative and quantitative) in the form of tooth damage before and after debonding from human teeth of in vitro studies. Eligibility criteria: Laboratorial studies evaluating EMCs characteristics before and after debonding metal and ceramic brackets from human teeth with intact buccal enamel. Information sources: An electronic search of four databases (all databases of the Cochrane Library, CA Web of Science, MEDLINE via PubMed, and Google Scholar) and additional manual searches were carried out, without language restrictions. Studies published between 2000 and 2017 years were selected. Reference lists of the included articles were screened, and authors were contacted when necessary. Risk of bias: The following six parameters were analyzed: blinding of examiner and outcome assessment, incomplete outcome data before bonding and after debonding, selective outcome reporting, and incomplete reporting of EMCs assessment. Included studies: Out of 430 potentially eligible studies, 259 were screened by title and abstract, 180 were selected for full-text analysis, 14 were included in the systematic review. Seven studies were selected for the meta-analysis. Synthesis of results: The results for EMCs characteristics were expressed as mean differences (MDs) with their 95 per cent confidence intervals (CIs), and calculated from random-effects meta-analyses. Debonding was associated with the increase in number (three studies, MD = 3.50, 95% CI, 2.13 to 4.87, P < 0.00001), length (seven studies, MD = 3.09 mm, 95% CI, 0.75-5.43, P < 0.00001), and width (three studies, MD = 0.39 µm, 95% CI, -0.01 to 0.79, P = 0.06) of EMCs. Considerable statistical heterogeneity was found for two forest plots evaluating the changes of number and length characteristics during debonding. Conclusions: There is weak evidence indicating length and width of EMCs increase following bracket removal and the scientific evidence concerning quantitative evaluation of the number parameter before and after debonding is insufficient. However, there is a strong evidence that after debonding the number of EMCs is likely to increase. Registration: No registration was performed.

Combination of additive and subtractive laser 3D microprocessing in hybrid glass/polymer microsystems for chemical sensing applications.

We present a novel hybrid glass-polymer micromechanical sensor by combining two femtosecond laser direct writing processes: laser illumination followed by chemical etching of glass and two-photon polymerization. This incorporation of techniques demonstrates the capability of combining mechanical deformable devices made of silica with an integrated polymer structure for passive chemical sensing application. We demonstrate that such a sensor could be utilized for investigating the elastic properties of polymeric microstructures fabricated via the two-photon polymerization technique. Moreover, we show that polymeric microstructure stiffness increases when immersed in organic liquids.

Does orthodontic debonding lead to tooth sensitivity? Comparison of teeth with and without visible enamel microcracks.

INTRODUCTION: Our aim was to assess the possible changes in sensitivity of teeth with and without visible enamel microcracks (EMCs) up to 1 week after the removal of metal brackets. METHODS: After debonding, 15 patients possessing teeth with visible EMCs and 15 subjects whose teeth were free of EMCs were enrolled in the study. For each experimental group, a control group was formed. The assessments of tooth sensitivity elicited by compressed air and cold testing were performed 5 times: just before debonding, immediately after debonding, and at 1, 3, and 7 days after debonding. Tooth sensitivity was recorded on a 100-mm visual analog scale. RESULTS: For the patients without visible EMCs, discomfort peaked immediately after debonding and started to decrease on day 1; at 1 week after debonding, the visual analog scale scores were lower than just before debonding and immediately after debonding. For the subjects possessing teeth with visible EMCs, the pattern of sensitivity dynamic was inherently the same. However, the patients with visible EMCs showed higher visual analog scale values at each time interval. CONCLUSIONS: Debonding leads to a short-term increase in tooth sensitivity. EMCs, a form of enamel damage, do not predispose to greater sensitivity perception in relation to bracket removal.

Wrinkled axicons: shaping light from cusps.

We propose a novel class of refractive optical elements by wrinkling the conical surface of a usual (conical) axicon, which leads to geometrical singularities (cusps). Such wrinkled axicons have been fabricated at the micron scale by using three-dimensional femtosecond-laser photopolymerization technique and we report on their experimental and numerical characterization. The beam shaping capabilities of these structures are discussed for both intensity and phase, which includes topological beam shaping that results from azimuthally modulated optical spin-orbit interaction.

Characterization of tissue engineered cartilage products: Recent developments in advanced therapy.

Legislative requirements for the quality of pharmacological agents underwent certain evolution when new type of therapies emerged. This relates to cell based medicines, such as tissue engineered cartilage products (TECP) which are increasingly developed as new modalities for widely prevalent orthopaedic disorders. Although quality measures for TECP are subject to the same general regulatory quality requirements, combination of cellular and scaffold substances requires definition of specific characteristics in vitro that are highly relevant to potency and efficacy of the newly designed medicinal product. One of the specific issues in designing cell based medicines is the fact that the biological activity of active substance, or cells, usually is altered after seeding them on a three-dimensional scaffold. Newly acquired features of the TECP are influenced by chemical, physical and mechanical characteristics of the scaffolds. A vast array of analytical methods has been employed to measure efficacy and potency of TECP in cartilage regeneration studies in vitro. Designing specific physical characteristics of scaffolds may become essential part influencing pharmacological activity of cell based medicinal products, and discern TECP from typical pharmacological products. As an example, increasingly growing popularity of three-dimensional printing that utilizes direct laser writing technique provides an opportunity to improve efficacy of the final TECP. This review is intended to provide brief summary of current approaches used to characterize cells and scaffolds in vitro before and after combination into TECP. Validating TECP as pharmacological agents with unique biological and physical characteristics may broaden their clinical application.

Plasmon assisted 3D microstructuring of gold nanoparticle-doped polymers.

3D laser lithography of a negative photopolymer (zirconium/silicon hybrid solgel SZ2080) doped with gold nanoparticles (Au NPs) is performed with a 515 nm and 300 fs laser system and the effect of doping is explored. By varying the laser-generated Au NP doping concentration from 4.8 · 10(-6) wt% to 9.8 · 10(-3) wt% we find that the fabricated line widths are enlarged by up to 14.8% compared to structures achieved in pure SZ2080. While implicating a positive effect on the photosensitivity, the doping has no adverse impact on the mechanical quality of intricate 3D microstructures produced from the doped nanocompound. Additionally, we found that SZ2080 increases the long term (∼months) colloidal stability of Au NPs in isopropanol. By discussing the nanoparticle-light interaction in the 3D polymer structures we provide implications that our findings might have on other fields, such as biomedicine and photonics.

Assessment of human gingival fibroblast interaction with dental implant abutment materials.

The biocompatibility of dental implant abutment materials depends on numerous factors including the nature of the material, its chemical composition, roughness, texture, hydrophilicity and surface charge. The aim of the present study was to compare the viability and adhesion strength of human gingival fibroblasts (HGFs) grown on several dental materials used in implant prosthodontics. Surfaces of the tested materials were assessed using an optical imaging profiler. For material toxicity and cellular adhesion evaluation, primary human gingival fibroblast cells were used. To evaluate the strength of cellular adhesion, gingival fibroblasts were cultured on the tested materials and subjected to lateral shear forces by applying 300 and 500 rpm shaking intensities. Focal adhesion kinase (FAK) expression and phosphorylation in cells grown on the specimens were registered by cell-based ELISA. There was a tendency of fibroblast adhesion strength to decrease in the following order: sandblasted titanium, polished titanium, sandblasted zirconium oxide, polished zirconium oxide, gold-alloy, chrome-cobalt alloy. Higher levels of total as well as phospho-FAK protein were registered in HGFs grown on roughened titanium. Material type and surface processing technique have an impact on gingival fibroblast interaction with dental implant abutment materials.

Molecular changes in endometrium origin stromal cells during initiation of cardiomyogenic differentiation induced with Decitabine, Angiotensin II and TGF- ß1.

Stem cells' differentiation toward cardiac lineage is a complex process dependent on various alterations in molecular basis and regulation pathways. The aim of the study is to show that endometrium-derived stromal cells - menstrual, endometrial and endometriotic, could be an attractive source for examination of the mechanisms underlying cardiomyogenesis. After treatment with Decitabine, Angiotensin II and TGF-ß1, cells demonstrated morphological dedifferentiation into early cardiomyocyte-like cells and expressed CD36, CD106, CD172a typically used to sort for human pluripotent stem cell-derived cardiomyocytes. RT-qPCR revealed changed cells' genetic profiles, as majority of cardiac lineage differentiation related genes and cardiac ion channels (calcium, sodium, potassium) coding genes were upregulated after 6 and 13 days of exposure. Additionally, analysis of expression of various signaling proteins (FOXO1, PDGFB, TGFBR1, mTOR, VEGFA, WNT4, Notch1) coding genes showed differences between cell cultures as they seem to employ distinct signaling pathways through differentiation initiation. Early stages of differentiation had biggest impact on cardiomyogenesis related proteins (Nkx-2.5, EZH2, FOXO3a, H3K9Ac) levels, as we noticed after conducting Western blot and as expected, early cardiac transcription factor Nkx-2.5 was highly expressed and localized in nucleus of differentiating cells. These findings led us to assess endometrium origin stromal cells' potential to differentiate towards cardiomyogenic lineage and better understand the regulation of complex differentiation processes in ex vivo model systems.

Three-dimensional laser micro-sculpturing of silicone: towards bio-compatible scaffolds.

Possibility to form three-dimensional (3D) micro-structures in silicone elastomer (polydimethylsiloxane; PDMS) doped with different photo-initiators was systematically investigated using direct laser writing with femtosecond laser pulses at different exposure conditions. Accuracy of the 3D structuring with resolution of ~5 µm and a fabrication throughput of ~720 µm(3)/s, which is exceeding the previously reported values by ~ 300(×), was achieved. Practical recording velocities of ~ 1 mm/s were used in PDMS with isopropyl-9H-thioxanthen-9-one (ISO) and thioxanthen-9-one (THIO) photo-initiators which both have absorption at around 360 nm wavelength. The 3D laser fabrication in PDMS without any photo-initiator resulting in a fully bio-compatible material has been achieved for the first time. Rates of multi-photon absorption and avalanche for the structuring of silicone are revealed: the two-photon absorption is seeding the avalanche of a radical generation for subsequent cross-linking. Direct writing enables a maskless manufacturing of molds for soft-lithography and 3D components for microfluidics as well as scaffolds for grafts in biomedical applications.

Black silicon: substrate for laser 3D micro/nano-polymerization.

We demonstrate that black silicon (b-Si) made by dry plasma etching is a promising substrate for laser three-dimensional (3D) micro/nano-polymerization. High aspect ratio Si-needles, working as sacrificial support structures, have flexibility required to relax interface stresses between substrate and the polymerized micro-/nano- objects. Surface of b-Si can be made electrically conductive by metal deposition and, at the same time, can preserve low optical reflectivity beneficial for polymerization by direct laser writing. 3D laser polymerization usually performed at the irradiation conditions close to the dielectric breakdown is possible on non-reflective and not metallic surfaces. Here we show that low reflectivity and high metallic conductivity are not counter- exclusive properties for laser polymerization. Electrical conductivity of substrate and its permeability in liquids are promising for bio- and electroplating applications.

Evaluation of enamel micro-cracks characteristics after removal of metal brackets in adult patients.

The purpose of this study was to evaluate and compare enamel micro-crack characteristics of adult patients before and after removal of metal brackets. After the examination with scanning electron microscopy (SEM), 45 extracted human teeth were divided into three groups of equal size: group 1, the teeth having enamel micro-cracks, group 2, the teeth without initial enamel micro-cracks, and group 3, control group to study the effect of dehydration on existing micro-cracks or formation of new ones. For all the teeth in groups 1 and 2, the same bonding and debonding procedures of metal brackets were conducted. The length and width of the longest enamel micro-crack were measured for all the teeth before and after removal of metal brackets. The changes in the location of the micro-cracks were also evaluated. In group 3, teeth were subjected to the same analysis but not bonded. The mean overall width of micro-cracks after removal of metal brackets was 3.82 µm greater than before bonding procedure (P < 0.05). Also, a significant difference was noticed between the width of micro-cracks in first zone (cervical third) and third zone (occlusal third) after debonding procedure (P < 0.05). New enamel micro-cracks were found in 6 of 15 (40 per cent) examined teeth. Greatest changes in the width of enamel micro-cracks after debonding procedure appear in the cervical third of the tooth. On the basis of this result, the dentist must pay extra care and attention to this specific area of enamel during removal of metal brackets in adult patients.

Anti-Reflective Coatings Produced via Atomic Layer Deposition for Hybrid Polymer 3D Micro-Optics.

The increasing demand for optics quality requires the lowest optical power loss, which can occur from unwanted reflections. Laser direct writing (LDW) allows for the fabrication of complex structures, which is particularly advantageous in micro-optic applications. This research demonstrates the possibility of forming an anti-reflective coating on hybrid polymer micro-lenses fabricated by employing LDW without changing their geometry. Such coating deposited via atomic layer deposition (ALD) decreased the reflection from 3.3% to 0.1% at a wavelength of 633 nm for one surface of hybrid organic-inorganic SZ2080™ material. This research validates the compatibility of ALD with LDW 3D multiphoton lithography synergistically, expanding its applications on optical grade sub-100 µm scale micro-optics.

Biocompatibility enhancement via post-processing of microporous scaffolds made by optical 3D printer.

Providing a 3D environment that mimics the native extracellular matrix is becoming increasingly important for various applications such as cell function studies, regenerative medicine, and drug discovery. Among the most critical parameters to consider are the scaffold's complicated micro-scale geometry and material properties. Therefore, stereolithography based on photopolymerization is an emerging technique because of its ability to selectively form volumetric structures from liquid resin through localized polymerization reactions. However, one of the most important parameters of the scaffold is biocompatibility, which depends not only on the material but also on the exposure conditions and post-processing, which is currently underestimated. To investigate this systematically, microporous scaffolds with pore sizes of 0.05 mm3 corresponding to a porosity of 16,4% were fabricated using the stereolithography printer Asiga PICO2 39 UV from the widely used resins FormLabs Clear and Flexible. The use of various polymers is usually limited for cells because, after wet chemical development, the non-negligible amount of remaining monomers intertwined in the photopolymerized structures is significantly toxic to cells. Therefore, the aim of this research was to find the best method to remove monomers from the 3D scaffold by additional UV exposure. For this purpose, a Soxhlet extractor was used for the first time, and the monomers were immersed in different alcohols. A Raman microspectroscopy was also used to investigate whether different post-processing methods affect DC (cross-linking) to find out if this specifically affects the biocompatibility of the scaffolds. Finally, mesenchymal stem cells from rat dental pulp were examined to confirm the increased biocompatibility of the scaffolds and their ability to support cell differentiation into bone tissue cells.

Teeth Microcracks Research: Towards Multi-Modal Imaging.

This perspective is an overview of the recent advances in teeth microcrack (MC) research, where there is a clear tendency towards a shift from two-dimensional (2D) to three-dimensional (3D) examination techniques, enhanced with artificial intelligence models for data processing and image acquisition. X-ray micro-computed tomography combined with machine learning allows 3D characterization of all spatially resolved cracks, despite the locations within the tooth in which they begin and extend, and the arrangement of MCs and their structural properties. With photoluminescence and micro-/nano-Raman spectroscopy, optical properties and chemical and elemental composition of the material can be evaluated, thus helping to assess the structural integrity of the tooth at the MC site. Approaching tooth samples having cracks from different perspectives and using complementary laboratory techniques, there is a natural progression from 3D to multi-modal imaging, where the volumetric (passive: dimensions) information of the tooth sample can be supplemented by dynamic (active: composition, interaction) image data. Revelation of tooth cracks clearly shows the need to re-assess the role of these MCs and their effect on the structural integrity and longevity of the tooth. This provides insight into the nature of cracks in natural hard materials and contributes to a better understanding of how bio-inspired structures could be designed to foresee crack propagation in biosolids.

Polarisation Control in Arrays of Microlenses and Gratings: Performance in Visible-IR Spectral Ranges.

Microlens arrays (MLAs) which are increasingly popular micro-optical elements in compact integrated optical systems were fabricated using a femtosecond direct laser write (fs-DLW) technique in the low-shrinkage SZ2080TM photoresist. High-fidelity definition of 3D surfaces on IR transparent CaF2 substrates allowed to achieve ∼50% transmittance in the chemical fingerprinting spectral region 2-5 µm wavelengths since MLAs were only ∼10 µm high corresponding to the numerical aperture of 0.3 (the lens height is comparable with the IR wavelength). To combine diffractive and refractive capabilities in miniaturised optical setup, a graphene oxide (GO) grating acting as a linear polariser was also fabricated by fs-DLW by ablation of a 1 µm-thick GO thin film. Such an ultra-thin GO polariser can be integrated with the fabricated MLA to add dispersion control at the focal plane. Pairs of MLAs and GO polarisers were characterised throughout the visible-IR spectral window and numerical modelling was used to simulate their performance. A good match between the experimental results of MLA focusing and simulations was achieved.

Closely packed hexagonal conical microlens array fabricated by direct laser photopolymerization.

We apply femtosecond laser direct writing in photopolymers for manufacturing of conical microlenses and closely packed arrays thereof. We demonstrate the fabrication of high optical quality axicons of 15 µm in radius, having 150°, 160°, and 170° cone angles. Their optical properties and performance are modeled using the finite-difference time-domain method and compared with experimentally measured data. Additionally, optimization of the laser direct writing parameters regarding these types of micro-objects is presented. Possible applications of closely packed arrays of axicon microlenses are discussed, having potential attractivity in the fields of modern microscopy, light-based material processing, particle manipulation in microfluidic, and optofluidic applications.