Comparative analysis of reciprocating and flat-side heat-treated rotary single-file systems: Design, metallurgical characteristics, and mechanical performance.

AIM: To compare two flat-side single-file rotary instruments with three single-file reciprocating systems through a multimethod assessment. METHODOLOGY: A total of 290 new NiTi single-file rotary (AF F One Blue 25/0.06 and Platinum V.EU 25/0.06) and reciprocating (One Files Blue R25, Reciproc Blue R25, Reciproc R25) instruments were selected, carefully examined for any major deformations, and evaluated regarding their macroscopic and microscopic design, nickel and titanium elements ratio, phase transformation temperatures, and mechanical performance (time/rotation to fracture, maximum torque, angle of rotation, microhardness, maximum bending, and buckling strengths). One-way anova post hoc Tukey, T-test, and nonparametric Mood's median tests were used for statistical comparisons (α = 5%). RESULTS: Tested instruments had identical blade counts and near-identical helical angles of approximately 24° (rotary instruments) and 151° (reciprocating instruments). The flat-side analysis revealed a few inconsistencies, such as discontinuity segments, different orientations, and gaps in the homogeneity of the bluish colour. Microscopically, flat-side instruments exhibited blade discontinuity and an incomplete S-shaped cross-section. The surface finish was smoother for One Files Blue and more irregular for both rotary instruments. There were distinct phase transformation temperatures amongst all instruments. All heat-treated instruments were in R-phase arrangement, and Reciproc was in R-phase plus austenite at test temperature (20°C). Compared with the reciprocating instruments, both flat-side instruments exhibited lower results in the cyclic fatigue tests using two different clockwise kinematics, maximum torque, angle of rotation, and maximum buckling strength (p < .05). The rotary systems also exhibited low flexibility (p < .05). AF F One Blue had the lowest microhardness, whilst Reciproc had the highest value. CONCLUSION: This multimethod investigation revealed that the flat-side rotary instruments underperformed the reciprocating instruments regarding cyclic fatigue (with two different clockwise kinematics), maximum torque, angle of rotation, maximum buckling strength, and flexibility. Manufacturing inconsistencies were also observed in some of the flat-side instruments, including discontinuity segments, different orientations, and in the homogeneity of their bluish colour given by the heat treatment.

Colonization versus encapsulation in cell-laden materials design: porosity and process biocompatibility determine cellularization pathways.

Seeding materials with living cells has been-and still is-one of the most promising approaches to reproduce the complexity and the functionality of living matter. The strategies to associate living cells with materials are limited to cell encapsulation and colonization, however, the requirements for these two approaches have been seldom discussed systematically. Here we propose a simple two-dimensional map based on materials' pore size and the cytocompatibility of their fabrication process to draw, for the first time, a guide to building cellularized materials. We believe this approach may serve as a straightforward guideline to design new, more relevant materials, able to seize the complexity and the function of biological materials. This article is part of the theme issue 'Bio-derived and bioinspired sustainable advanced materials for emerging technologies (part 1)'.

Self-Assembled Collagen Microparticles by Aerosol as a Versatile Platform for Injectable Anisotropic Materials.

Extracellular matrices (ECM) rich in type I collagen exhibit characteristic anisotropic ultrastructures. Nevertheless, working in vitro with this biomacromolecule remains challenging. When processed, denaturation of the collagen molecule is easily induced in vitro avoiding proper fibril self-assembly and further hierarchical order. Here, an innovative approach enables the production of highly concentrated injectable collagen microparticles, based on collagen molecules self-assembly, thanks to the use of spray-drying process. The versatility of the process is shown by performing encapsulation of secretion products of gingival mesenchymal stem cells (gMSCs), which are chosen as a bioactive therapeutic product for their potential efficiency in stimulating the regeneration of a damaged ECM. The injection of collagen microparticles in a cell culture medium results in a locally organized fibrillar matrix. The efficiency of this approach for making easily handleable collagen microparticles for encapsulation and injection opens perspectives in active tissue regeneration and 3D bioprinted scaffolds.

Water/ice phase transition: The role of zirconium acetate, a compound with ice-shaping properties.

Few compounds feature ice-shaping properties. Zirconium acetate is one of the very few inorganic compounds reported so far to have ice-shaping properties similar to that of ice-shaping proteins, encountered in many organisms living at low temperature. When a zirconium acetate solution is frozen, oriented and perfectly hexagonal ice crystals can be formed and their growth follows the temperature gradient. To shed light on the water/ice phase transition while freezing zirconium acetate solution, we carried out differential scanning calorimetry measurements. From our results, we estimate how many water molecules do not freeze because of their interaction with Zr cations. We estimate the colligative properties of the Zr acetate on the apparent critical temperature. We further show that the phase transition is unaffected by the nature of the base which is used to adjust the pH. Our results provide thus new hints on the ice-shaping mechanism of zirconium acetate.

Water-mediated structuring of bone apatite.

It is well known that organic molecules from the vertebrate extracellular matrix of calcifying tissues are essential in structuring the apatite mineral. Here, we show that water also plays a structuring role. By using solid-state nuclear magnetic resonance, wide-angle X-ray scattering and cryogenic transmission electron microscopy to characterize the structure and organization of crystalline and biomimetic apatite nanoparticles as well as intact bone samples, we demonstrate that water orients apatite crystals through an amorphous calcium phosphate-like layer that coats the crystalline core of bone apatite. This disordered layer is reminiscent of those found around the crystalline core of calcified biominerals in various natural composite materials in vivo. This work provides an extended local model of bone biomineralization.

Comprehensive Assessment of Cyclic Fatigue Strength in Five Multiple-File Nickel-Titanium Endodontic Systems.

The resistance of nickel-titanium endodontic instruments against cyclic fatigue failure remains a significant concern in clinical settings. This study aimed to assess the cyclic fatigue strength of five nickel-titanium rotary systems, while correlating the results with the instruments' geometric and metallurgical characteristics. A total of 250 new instruments (sizes S1/A1, S2/A2, F1/B1, F2/B2, F3/B3) from ProTaper Gold, ProTaper Universal, Premium Taper Gold, Go-Taper Flex, and U-Files systems underwent mechanical testing. Prior to experimental procedures, all instruments were meticulously inspected to identify irregularities that could affect the investigation. Using a stereomicroscope, design characteristics such as the number of spirals, length, spirals per millimeter, and average helical angle of the active blade were determined. The surface finishing characteristics of the instruments were examined using a scanning electron microscope. Differential scanning calorimetry was employed to establish the instruments' phase transformation temperatures, while energy-dispersive X-ray spectroscopy was utilized to analyze the elemental composition of the alloy. The instruments were subjected to cyclic fatigue testing within a stainless steel non-tapered artificial canal featuring a 6 mm radius and 86 degrees of curvature. Appropriate statistical tests were applied to compare groups, considering a significance level of 0.05. The assessed design characteristics varied depending on the instrument type. The least irregular surface finishing was observed in U-Files and Premium Taper Gold files, while the most irregular surface was noted in Go-Taper Flex. All instruments exhibited near-equiatomic proportions of nickel and titanium elements, whereas ProTaper Universal and U-Files instruments demonstrated lower phase transformation temperatures compared to their counterparts. Larger-sized instruments, as well as ProTaper Universal and U-Files, tended to display lower cyclic fatigue strength results. Overall, the design, metallurgical, and cyclic fatigue outcomes varied among instruments and systems. Understanding these outcomes may assist clinicians in making more informed decisions regarding instrument selection.

Tunable biomimetic materials elaborated by ice templating and self-assembly of collagen for tubular tissue engineering.

Synthetic tubular grafts currently used in clinical context fail frequently, and the expectations that biomimetic materials could tackle these limitations are high. However, developing tubular materials presenting structural, compositional and functional properties close to those of native tissues remains an unmet challenge. Here we describe a combination of ice templating and topotactic fibrillogenesis of type I collagen, the main component of tissues' extracellular matrix, yielding highly concentrated yet porous tubular collagen materials with controlled hierarchical architecture at multiple length scales, the hallmark of native tissues' organization. By modulating the thermal conductivity of the cylindrical molds, we tune the macroscopic porosity defined by ice. Coupling the aforementioned porosity patterns with two different fibrillogenesis routes results in a new family of tubular materials whose textural features and the supramolecular arrangement of type I collagen are achieved. The resulting materials present hierarchical elastic properties and are successfully colonized by human endothelial cells and alveolar epithelial cells on the luminal side, and by human mesenchymal stem cells on the external side. The proposed straightforward protocol is likely to be adapted for larger graft sizes that address ever-growing clinical needs, such as peripheral arterial disease or tracheal and bronchial reconstructions.

Multimethod analysis of a novel multi-coloured heat-treated NiTi rotary system: design, metallurgy, mechanical properties, and shaping ability.

INTRODUCTION: This study aimed to compare a new multi-colored rotary system with four heat-treated rotary instruments using the multimethod approach. METHODS: Three-hundred instruments of RCS Rainbow, Rotate, RaCe EVO, OneCurve, and ProTaper Ultimate systems were evaluated regarding their design (stereomicroscopy, scanning electron microscopy, 3D surface scanning), metallurgy, and mechanical performance (cyclic fatigue, torsional resistance, bending and buckling resistance, cutting ability). Unprepared surfaces after canal preparation of maxillary molars were evaluated using micro-CT. Kruskal-Wallis and one-way ANOVA post hoc Tukey tests were used for statistical comparisons (α=5%). RESULTS: Instruments exhibited variations in active blade length, number of spirals, and cross-sectional designs. RCS Rainbow showed specific phase transformation temperatures, highest bending (400.5gf) and buckling (286gf) resistance values, and lowest mean angle of rotation (529º) (P<.05). OneCurve exhibited superior cutting ability (8.4mm) and longer time to fracture (112s). RaCe EVO displayed the lowest time to fracture (51s), maximum torque (1.2N.cm), buckling (174gf), and bending resistance (261gf) values (P<.05). ProTaper Ultimate showed the highest torque (1.6N.cm) and angle of rotation (611º) (P<.05), while its bending load (262gf) was comparable to RaCe EVO (P>.05). Rotate instrument showed intermediate values in the mechanical tests. No difference was observed regarding the unprepared canal surfaces (p>0.05) CONCLUSIONS: RCS Rainbow demonstrates a trade-off between flexibility and other mechanical properties. Its dimensions exceeded those of other instruments, affording it higher torque resistance, yet concurrently reducing its flexibility, angle of rotation, and cutting ability. OneCurve stands out as a well-balanced choice by integrating geometric design and mechanical performance.

Silica-sepiolite nanoarchitectures.

Silica-sepiolite heterostructured materials have been prepared as novel nanoarchitectures by generation of SiO2 nanoparticles (NPs) on the surface of the sepiolite fibrous clay mineral. The synthetic approach implies the use of organo-sepiolites dispersed in isopropanol to which is incorporated a selected silicon alcoxysilane, such as tetramethoxysilane (TMOS), that then is slowly hydrolyzed to procure the formation of a viscous gel under ultrasound irradiation. Once the sol-gel reaction is achieved the intermediate silica-sepiolite organo-heterostructures can be submitted to a thermal treatment for the removal of the organic matter, which finally gives rise to the silica-sepiolite nanoarchitectures. Influence of different experimental variables, such as nature of both alkoxysilane precursor and organo-sepiolite as well as their relative ratio in the reaction media, in the characteristics of both intermediate silica-sepiolite organo-heterostructures and final nanoarchitectures has been explored. Both type of heterostructured materials have been characterized by means of diverse experimental techniques such as CHN chemical analysis, TG-DTA, XRD, FTIR, 29Si NMR, FE-SEM and TEM. Special attention has been devoted to the analysis of changes in the morphological and textural features of the SiO2-sepiolite samples before and after the thermal treatment carried out for removing the organic matter and consolidation of the silica network. This study describes the resulting nanoarchitectures as sepiolite microfibers covalently assembled to silica nanoparticles exhibiting specific surface areas ca. to 350 m2/g, practically without microporous contribution. Preliminary tests regarding the use of the SiO2-sepiolite nanoarchitectures as nanofillers in polymer nanocomposites have been also investigated in order to show one of their potential fields of application. Mechanical properties of epoxy resin nanocomposites have been determined and discussed considering the different nature of the external surface of the intermediate organo-heterostructures and the final inorganic nanoarchitectures.

Multifocus optical microscopy applied to the study of archaeological metals.

Studies on cultural metal artifacts can benefit greatly from microscopy techniques. The examination of microstructural features can provide relevant information about ancient manufacturing techniques, as well as about corrosion/degradation processes. In the present work, advantages of the use of multifocus imaging techniques in optical microscopy for the study of archaeological metals are presented. An archaeometallurgical study of a large collection of bronzes demonstrates the possibility of a microstructural study with no need for sample removal, which is a great advantage in the study of cultural objects. In addition, the study of mounted samples illustrates the advantages of the multifocus technique in the examination of particular corrosion features, with the possibility of three-dimensional reconstructions.

Circulating Tumor Cells in Cancer Diagnostics and Prognostics by Single-Molecule and Single-Cell Characterization.

Circulating tumor cells (CTCs) represent an interesting source of biomarkers for diagnosis, prognosis, and the prediction of cancer recurrence, yet while they are extensively studied in oncobiology research, their diagnostic utility has not yet been demonstrated and validated. Their scarcity in human biological fluids impedes the identification of dangerous CTC subpopulations that may promote metastatic dissemination. In this Perspective, we discuss promising techniques that could be used for the identification of these metastatic cells. We first describe methods for isolating patient-derived CTCs and then the use of 3D biomimetic matrixes in their amplification and analysis, followed by methods for further CTC analyses at the single-cell and single-molecule levels. Finally, we discuss how the elucidation of mechanical and morphological properties using techniques such as atomic force microscopy and molecular biomarker identification using nanopore-based detection could be combined in the future to provide patients and their healthcare providers with a more accurate diagnosis.

Porous yet dense matrices: using ice to shape collagen 3D cell culture systems with increased physiological relevance.

Standard in vitro cell cultures are one of the pillars of biomedical sciences. However, there is increasing evidence that 2D systems provide biological responses that are often in disagreement with in vivo observations, partially due to limitations in reproducing the native cellular microenvironment. 3D materials that are able to mimic the native cellular microenvironment to a greater extent tackle these limitations. Here, we report Porous yet Dense (PyD) type I collagen materials obtained by ice-templating followed by topotactic fibrillogenesis. These materials combine extensive macroporosity, favouring the cell migration and nutrient exchange, as well as dense collagen walls, which mimic locally the extracellular matrix. When seeded with Normal Human Dermal Fibroblasts (NHDFs), PyD matrices allow for faster and more extensive colonisation when compared with equivalent non-porous matrices. The textural properties of the PyD materials also impact cytoskeletal and nuclear 3D morphometric parameters. Due to the effectiveness in creating a biomimetic 3D environment for NHDFs and the ability to promote cell culture for more than 28 days without subculture, we anticipate that PyD materials could configure an important step towards in vitro systems applicable to other cell types and with higher physiological relevance.

Biomimetic Silk Macroporous Materials for Drug Delivery Obtained via Ice-Templating.

Silk from Bombyx mori is one of the most exciting materials in nature. The apparently simple arrangement of its two major components─two parallel filaments of silk fibroin (SF) coated by a common sericin (SS) sheath─provides a combination of mechanical and surface properties that can protect the moth during its most vulnerable phase, the pupal stage. Here, we recapitulate the topology of native silk fibers but shape them into three-dimensional porous constructs using an unprecedented design strategy. We demonstrate, for the first time, the potential of these macroporous silk foams as dermal patches for wound protection and for the controlled delivery of Rifamycin (Rif), a model antibiotic. The method implies (i) removing SS from silk fibers; (ii) shaping SF solutions into macroporous foams via ice-templating; (iii) stabilizing the SF macroporous foam in a methanolic solution of Rif; and (iv) coating Rif-loaded SF foams with a SS sheath. The resulting SS@SF foams exhibit water wicking capacity and accommodate up to ∼20% deformation without detaching from a skin model. The antibacterial behavior of Rif-loaded SS@SF foams against Staphylococcus aureus on agar plates outperforms that of SF foams (>1 week and 4 days, respectively). The reassembly of natural materials as macroporous foams─illustrated here for the reconstruction of silk-based materials─can be extended to other multicomponent natural materials and may play an important role in applications where controlled release of molecules and fluid transport are pivotal.

A Multimethod Assessment of a New Customized Heat-Treated Nickel-Titanium Rotary File System.

This study aimed to compare three endodontic rotary systems. The new Genius Proflex (25/0.04), Vortex Blue (25/0.04), and TruNatomy (26/0.04v) instruments (n = 41 per group) were analyzed regarding design, metallurgy, and mechanical performance, while shaping ability (untouched canal walls, volume of removed dentin and hard tissue debris) was tested in 36 anatomically matched root canals of mandibular molars. The results were compared using one-way ANOVA, post hoc Tukey, and Kruskal−Wallis tests, with a significance level set at 5%. All instruments showed symmetrical cross-sections, with asymmetrical blades, no radial lands, no major defects, and almost equiatomic nickel−titanium ratios. Differences were noted in the number of blades, helical angles, cross-sectional design, and tip geometry. The Genius Proflex and the TruNatomy instruments had the highest and lowest R-phase start and finish temperatures, as well as the highest and lowest time and cycles to fracture (p < 0.05), respectively. The TruNatomy had the highest flexibility (p < 0.05), while no differences were observed between the Genius Proflex and the Vortex Blue (p > 0.05). No differences among tested systems were observed regarding the maximum torque, angle of rotation prior to fracture, and shaping ability (p > 0.05). The instruments showed similarities and differences in their design, metallurgy, and mechanical properties. However, their shaping ability was similar, without any clinically significant errors. Understanding these characteristics may help clinicians to make decisions regarding which instrument to choose for a particular clinical situation.

Multimethod Assessment of Design, Metallurgical, and Mechanical Characteristics of Original and Counterfeit ProGlider Instruments.

A multimethod study was conducted to assess the differences between original (PG-OR) and counterfeit (PG-CF) ProGlider instruments regarding design, metallurgical features, and mechanical performance. Seventy PG-OR and PG-CF instruments (n = 35 per group) were evaluated regarding the number of spirals, helical angles, and measuring line position by stereomicroscopy, while blade symmetry, cross-section geometry, tip design, and surface were assessed by scanning electron microscopy. Energy-dispersive X-ray spectroscopy and differential scanning calorimetry were used to identify element ratio and phase transformation temperatures, while cyclic fatigue, torsional, and bending testing were employed to assess their mechanical performance. An unpaired t-test and nonparametric Mann−Whitney U test were used to compare instruments at a significance level of 5%. Similarities were observed in the number of spirals, helical angles, blade symmetry, cross-sectional geometries, and nickel−titanium ratios. Measuring lines were more reliable in the original instrument, while differences were noted in the geometry of the tips (sharper tip for the original and rounded for the counterfeit) and surface finishing with PG-CF presenting more surface irregularities. PG-OR showed significantly more time to fracture (118 s), a higher angle of rotation (440°), and a lower maximum bending load (146.3 gf) (p < 0.05) than PG-CF (p < 0.05); however, maximum torque was similar for both instruments (0.4 N.cm) (p > 0.05). Although the tested instruments had a similar design, the original ProGlider showed superior mechanical behavior. The results of counterfeit ProGlider instruments were unreliable and can be considered unsafe for glide path procedures.

Gelatin renaturation and the interfacial role of fillers in bionanocomposites.

This work describes a systematic study of gelatin-sepiolite structural bionanocomposites to show how the renaturation level of the biopolymer is highly dependent on the type of mineral particle used. The aim of the work is to prove that chemical interactions between both components (hydrogen and covalent bonding) determine the organization level of the biopolymer which in turn results in drastic differences in the elastic properties of the prepared bionanocomposites. To assess this, several systematic modifications were introduced into the silicate structure and surface, generating four derivatives. Two derivatives prepared by thermal treatments, monohydrated sepiolite and protoenstatite, and two chemically modified sepiolites, amino and epoxy terminated, were prepared and used as the inorganic (or hybrid) phase in the bionanocomposites. The thermal and chemical modifications performed on the sepiolite surface induced a dramatic decrease in the renaturation level as determined by DSC and FTIR techniques. On the other hand, untreated sepiolite induced a higher renaturation level in the polypeptide, probably due to the alignment of the collagen-like triple helix along sepiolite external surface channels. The measured mechanical properties of the studied compositions confirm that the renaturation level of gelatin is a key factor in understanding the elastic properties of bionanocomposites. These results suggest that mineral particles introduced in the polypeptide matrix provide an effective control over the matrix crystallinity giving rise to tunable mechanical properties of the final bionanocomposite.

Recent advances in ice templating: from biomimetic composites to cell culture scaffolds and tissue engineering.

Ice templating - or freeze casting - has flourished in multiple domains as a straightforward process to shape solutions and particle suspensions into macroporous materials. Longtime used as a process to shape colloidal suspensions into lightweight ceramics, the use of ice templating has evolved to fabricate materials that mimic the architecture of biological tissues such as nacre and bone. Recently, the technique has been used to shape biopolymers for cell culture systems and tissue engineering applications and eventually to allow the fabrication of biomaterials containing living cells. Here we review how ice templating has progressed to cope with intrinsically labile biological matter and how these advances may shape the future 3D cell culture, tissue engineering and ultimately, cryobiology.

Biomimetic Tough Gels with Weak Bonds Unravel the Role of Collagen from Fibril to Suprafibrillar Self-Assembly.

Biological tissues rich in type I collagen exhibit specific hierarchical fibrillar structures together with remarkable mechanical toughness. However, the role of collagen alone in their mechanical response at different structural levels is not fully understood. Here, it is proposed to rationalize such challenging interplay from a materials science perspective through the subtle control of this protein self-assembly in vitro. It is relied on a spray-processing approach to readily use the collagen phase diagram and set a palette of biomimetic self-assembled collagen gels in terms of suprafibrillar organization. Their mechanical responses unveil the involvement of mechanisms occurring either at fibrillar or suprafibrillar scales. Noticeably, both modulus at early stage of deformations and tensile toughness probe the suprafibrillar organization, while durability under cyclic loading and stress relaxation reflect mechanisms at the fibril level. By changing the physicochemical environment, the interfibrillar interactions are modified toward more biomimetic mechanical responses. The possibility of making tissue-like materials with versatile compositions and toughness opens perspectives in tissue engineering.

Preservation of biomaterials and cells by freeze-drying: Change of paradigm.

Freeze-drying is the most widespread method to preserve protein drugs and vaccines in a dry form facilitating their storage and transportation without the laborious and expensive cold chain. Extending this method for the preservation of natural biomaterials and cells in a dry form would provide similar benefits, but most results in the domain are still below expectations. In this review, rather than consider freeze-drying as a traditional black box we "break it" through a detailed process thinking approach. We discuss freeze-drying from process thinking aspects, introduce the chemical, physical, and mechanical environments important in this process, and present advanced biophotonic process analytical technology. In the end, we review the state of the art in the freeze-drying of the biomaterials, extracellular vesicles, and cells. We suggest that the rational design of the experiment and implementation of advanced biophotonic tools are required to successfully preserve the natural biomaterials and cells by freeze-drying. We discuss this change of paradigm with existing literature and elaborate on our perspective based on our new unpublished results.

Sepiolite-Hydrogels: Synthesis by Ultrasound Irradiation and Their Use for the Preparation of Functional Clay-Based Nanoarchitectured Materials.

Sepiolite and palygorskite fibrous clay minerals are 1D silicates featuring unique textural and structural characteristics useful in diverse applications, and in particular as rheological additives. Here we report on the ability of grinded sepiolite to generate highly viscous and stable hydrogels by sonomechanical irradiation (ultrasounds). Adequate drying of such hydrogels leads to low-density xerogels that show extensive fiber disaggregation compared to the starting sepiolite-whose fibers are agglomerated as bundles. Upon re-dispersion in water under high-speed shear, these xerogels show comparable rheological properties to commercially available defibrillated sepiolite products, resulting in high viscosity hydrogels that minimize syneresis. These colloidal systems are thus very interesting as they can be used to stabilize many diverse compounds as well as nano-/micro-particles, leading to the production of a large variety of composites and nano/micro-architectured solids. In this context, we report here various examples showing how colloidal routes based on sepiolite hydrogels can be used to obtain new heterostructured functional materials, based on their assembly to solids of diverse topology and composition such as 2D and 1D kaolinite and halloysite aluminosilicates, as well as to the 2D synthetic Mg,Al-layered double hydroxides (LDH).