Electron Beam Powder Bed Fusion of Ti-48Al-2Cr-2Nb Open Porous Scaffold for Biomedical Applications: Process Parameters, Adhesion, and Proliferation of NIH-3T3 Cells.

Titanium aluminide (TiAl)-based intermetallics, especially Ti-48Al-2Cr-2Nb, are a well-established class of materials for producing bulky components using the electron beam powder bed fusion (EB-PBF) process. The biological properties of Ti-48Al-2Cr-2Nb alloy have been rarely investigated, specifically using complex cellular structures. This work investigates the viability and proliferation of NIH-3T3 fibroblasts on Ti-48Al-2Cr-2Nb dodecahedral open scaffolds manufactured by the EB-PBF process. A process parameter optimization is carried out to produce a fully dense part. Then scaffolds are produced and characterized using different techniques, including scanning electron microscopy and X-ray tomography. In vitro viability tests are performed with NIH-3T3 cells after incubation for 1, 4, and 7 days. The results show that Ti-48Al-2Cr-2Nb represents a promising new entry in the biomaterial field.

Features of Vat-Photopolymerized Masters for Microfluidic Device Manufacturing.

The growing interest in advancing microfluidic devices for manipulating fluids within micrometer-scale channels has prompted a shift in manufacturing practices, moving from single-component production to medium-size batches. This transition arises due to the impracticality of lab-scale manufacturing methods in accommodating the increased demand. This experimental study focuses on the design of master benchmarks 1-5, taking into consideration critical parameters such as rib width, height, and the relative width-to-height ratio. Notably, benchmarks 4 and 5 featured ribs that were strategically connected to the inlet, outlet, and reaction chamber of the master, enhancing their utility for subsequent replica production. Vat photopolymerization was employed for the fabrication of benchmarks 1-5, while replicas of benchmarks 4 and 5 were generated through polydimethylsiloxane casting. Dimensional investigations of the ribs and channels in both the master benchmarks and replicas were conducted using an optical technique validated through readability analysis based on the Michelson global contrast index. The primary goal was to evaluate the potential applicability of vat photopolymerization technology for efficiently producing microfluidic devices through a streamlined production process. Results indicate that the combination of vat photopolymerization followed by replication is well suited for achieving a minimum rib size of 25 µm in width and an aspect ratio of 1:12 for the master benchmark.

On the Biomechanical Performances of Duplex Stainless Steel Graded Scaffolds Produced by Laser Powder Bed Fusion for Tissue Engineering Applications.

This experimental study aims to extend the know-how on biomechanical performances of duplex stainless steel (DSS) for tissue engineering applications to a graded lattice geometry scaffold based on the F53 DSS (UNS S32750 according to ASTM A182) produced by laser powder bed fusion (LPBF). The same dense-out graded geometry based on rhombic dodecahedral elementary unit cells investigated in previous work on 316L stainless steel (SS) was adopted here for the manufacturing of the F53 DSS scaffold (SF53). Microstructural characterization and mechanical and biological tests were carried out on the SF53 scaffold, using the in vitro behavior of the 316L stainless steel scaffold (S316L) as a control. Results show that microstructure developed as a consequence of different volume energy density (VED) values is mainly responsible for the different mechanical behaviors of SF53 and S316L, both fabricated using the same LPBF manufacturing system. Specifically, the ultimate compressive strength (σUC) and elastic moduli (E) of SF53 are three times and seven times higher than S316L, respectively. Moreover, preliminary biological tests evidenced better cell viability in SF53 than in S316L already after seven days of culture, suggesting SF53 with dense-out graded geometry as a viable alternative to 316L SS for bone tissue engineering applications.

Combined Effects of HA Concentration and Unit Cell Geometry on the Biomechanical Behavior of PCL/HA Scaffold for Tissue Engineering Applications Produced by LPBF.

This experimental study aims at filling the gap in the literature concerning the combined effects of hydroxyapatite (HA) concentration and elementary unit cell geometry on the biomechanical performances of additively manufactured polycaprolactone/hydroxyapatite (PCL/HA) scaffolds for tissue engineering applications. Scaffolds produced by laser powder bed fusion (LPBF) with diamond (DO) and rhombic dodecahedron (RD) elementary unit cells and HA concentrations of 5, 30 and 50 wt.% were subjected to structural, mechanical and biological characterization to investigate the biomechanical and degradative behavior from the perspective of bone tissue regeneration. Haralick's features describing surface pattern, correlation between micro- and macro-structural properties and human mesenchymal stem cell (hMSC) viability and proliferation have been considered. Experimental results showed that HA has negative influence on scaffold compaction under compression, while on the contrary it has a positive effect on hMSC adhesion. The unit cell geometry influences the mechanical response in the plastic regime and also has an effect on the cell proliferation. Finally, both HA concentration and elementary unit cell geometry affect the scaffold elastic deformation behavior as well as the amount of micro-porosity which, in turn, influences the scaffold degradation rate.

Improved biomechanical behavior of 316L graded scaffolds for bone tissue regeneration produced by laser powder bed fusion.

Graded lattice scaffolds based on rhombic dodecahedral (RD) elementary unit cell geometry were manufactured in 316L stainless steel (SS) by laser powder bed fusion (LPBF). Two different strategies based on varying strut thickness layer-by-layer in the building direction were adopted to obtain the graded scaffolds: a) decreasing strut size from core to edge to produce the dense-in (DI) structure and b) increasing strut size in the same direction to produce the dense-out (DO) structure. Both graded structures (DI and DO) were constructed with specular symmetry with respect to the central horizontal axis. Structural, mechanical, and biological characterizations were carried out to evaluate feasibility of designing appropriate biomechanical performances of graded scaffolds in the perspective of bone tissue regeneration. Results showed that mechanical behavior is governed by graded geometry, while printing parameters influence structural properties of the material such as density, textures, and crystallographic phases. The predominant failure mechanism in graded structures initiates in correspondence of thinner struts, due to high stress concentrations on strut junctions. Biological tests evidenced better proliferation of cells in the DO graded scaffold, which in turn exhibits mechanical properties close to cortical bone. The combined control of grading strategy, printing parameters and elementary unit cell geometry can enable implementing scaffolds with improved biomechanical performances for bone tissue regeneration.

Assessment of the Remineralizing Potential of Biomimetic Materials on Early Artificial Caries Lesions after 28 Days: An In Vitro Study.

This study aimed to evaluate the loss of mineral content in the enamel surface in early artificial lesions and to assess the remineralizing potential of different agents by means of SEM coupled with energy-dispersive X-ray analysis (EDX). The analysis was performed on the enamel of 36 molars divided into six equal groups, in which the experimental ones (3-6) were treated using remineralizing agents for a 28-day pH cycling protocol as follows: Group 1, sound enamel; Group 2, artificially demineralized enamel; Group 3, CPP-ACP treatment; Group 4, Zn-hydroxyapatite treatment; Group 5, NaF 5% treatment; and Group 6, F-ACP treatment. Surface morphologies and alterations in Ca/P ratio were evaluated using SEM-EDX and data underwent statistical analysis (p < 0.05). Compared with the sound enamel of Group 1, the SEM images of Group 2 clearly showed loss of integrity, minerals, and interprismatic substances. Groups 3-6 showed a structural reorganization of enamel prisms, interestingly comprising almost the entire enamel surface. Group 2 revealed highly significant differences of Ca/P ratios compared with other groups, while Groups 3-6 showed no differences with Group 1. In conclusion, all tested materials demonstrated a biomimetic ability in remineralizing lesions after 28 days of treatment.

Influence of Trabecular Geometry on Scaffold Mechanical Behavior and MG-63 Cell Viability.

In a scaffold-based approach for bone tissue regeneration, the control over morphometry allows for balancing scaffold biomechanical performances. In this experimental work, trabecular geometry was obtained by a generative design process, and scaffolds were manufactured by vat photopolymerization with 60% (P60), 70% (P70) and 80% (P80) total porosity. The mechanical and biological performances of the produced scaffolds were investigated, and the results were correlated with morphometric parameters, aiming to investigate the influence of trabecular geometry on the elastic modulus, the ultimate compressive strength of scaffolds and MG-63 human osteosarcoma cell viability. The results showed that P60 trabecular geometry allows for matching the mechanical requirements of human mandibular trabecular bone. From the statistical analysis, a general trend can be inferred, suggesting strut thickness, the degree of anisotropy, connectivity density and specific surface as the main morphometric parameters influencing the biomechanical behavior of trabecular scaffolds, in the perspective of tissue engineering applications.

Beads for Cell Immobilization: Comparison of Alternative Additive Manufacturing Techniques.

The attachment or entrapment of microbial cells and enzymes are promising solutions for various industrial applications. When the traps are beads, they are dispersed in a fluidized bed in a vessel where a pump guarantees fresh liquid inflow and waste outflow without washing out the cells. Scientific papers report numerous types of cell entrapment, but most of their applications remain at the laboratory level. In the present research, rigid polymer beads were manufactured by two different additive manufacturing (AM) techniques in order to verify the economy, reusability, and stability of the traps, with a view toward a straightforward industrial application. The proposed solutions allowed for overcoming some of the drawbacks of traditional manufacturing solutions, such as the limited mechanical stability of gel traps, and they guaranteed the possibility of producing parts of constant quality with purposely designed exchange surfaces, which are unfeasible when using conventional processes. AM proved to be a viable manufacturing solution for beads with complex shapes of two different size ranges. A deep insight into the production and characteristics of beads manufactured by AM is provided. The paper provides biotechnologists with a manufacturing perspective, and the results can be directly applied to transit from the laboratory to the industrial scale.

Remineralization Efficacy of Four Remineralizing Agents on Artificial Enamel Lesions: SEM-EDS Investigation.

Dental remineralization represents the process of depositing calcium and phosphate ions into crystal voids in demineralized enamel, producing net mineral gain and preventing early enamel lesions progression. The aim of the present study was to qualitatively and quantitatively compare the remineralizing effectiveness of four commercially available agents on enamel artificial lesions using Scanning Electron Microscopy (SEM) combined with Energy Dispersive Spectroscopy (EDS) techniques. Thirty-six extracted third molars were collected and randomly assigned to six groups (n = 6), five of which were suspended in demineralizing solution for 72 h to create enamel artificial lesions, and one serving as control: G1, treated with a mousse of casein phosphopeptide and amorphous calcium−phosphate (CPP-ACP); G2, treated with a gel containing nano-hydroxyapatite; G3, treated with a 5% SF varnish; G4, treated with a toothpaste containing ACP functionalized with fluoride and carbonate-coated with citrate; G5, not-treated artificial enamel lesions; G6, not demineralized and not treated sound enamel. G1−G4 were subjected to pH cycling over a period of seven days. Analyses of the specimens' enamel surfaces morphology were performed by SEM and EDS. Data were statistically analyzed for multiple group comparison by one-way ANOVA/Tukey's test (p < 0.05). The results show that the Ca/P ratio of the G5 (2.00 ± 0.07) was statistically different (p < 0.05) from G1 (1.73 ± 0.05), G2 (1.76 ± 0.01), G3 (1.88 ± 0.06) and G6 (1.74 ± 0.04), while there were no differences (p > 0.05) between G1, G2 and G6 and between G4 (2.01 ± 0.06) and G5. We concluded that G1 and G2 showed better surface remineralization than G3 and G4, after 7 days of treatment.

Improving the radiopacity of Fe-Mn biodegradable metals by magnetron-sputtered W-Fe-Mn-C coatings: Application for thinner stents.

In this exploratory work, micrometric radiopaque W-Fe-Mn-C coatings were produced by magnetron sputtering plasma deposition, for the first time, with the aim to make very thin Fe-Mn stents trackable by fluoroscopy. The power of Fe-13Mn-1.2C target was kept constant at 400 W while that of W target varied from 100 to 400 W producing three different coatings referred to as P100, P200, P400. The effect of the increased W power on coatings thickness, roughness, structure, corrosion behavior and radiopacity was investigated. The coatings showed a power-dependent thickness and W concentration, different roughness values while a similar and uniform columnar structure. An amorphous phase was detected for both P100 and P200 coatings while γ-Fe, bcc-W and W3C phases found for P400. Moreover, P200 and P400 showed a significantly higher corrosion rate (CR) compared to P100. The presence of W, W3C as well as the Fe amount variation determined two different micro-galvanic corrosion mechanisms significantly changing the CR of coatings, 0.26 ± 0.02, 59.68 ± 1.21 and 59.06 ± 1.16 µm/year for P100, P200 and P400, respectively. Sample P200 with its most uniform morphology, lowest roughness (RMS = 3.9 ± 0.4 nm) and good radiopacity (∼6%) appeared the most suitable radiopaque biodegradable coating investigated in this study.

Surface processing for iron-based degradable alloys: A preliminary study on the importance of acid pickling.

The formation of a heterogeneous oxidized layer, also called scale, on metallic surfaces is widely recognized as a rapid manufacturing event for metals and their alloys. Partial or total removal of the scale represents a mandatory integrated step for the industrial fabrication processes of medical devices. For biodegradable metals, acid pickling has already been reported as a preliminary surface preparation given further processes, such as electropolishing. Unfortunately, biodegradable medical prototypes presented discrepancies concerning acid pickling studies based on samples with less complex geometry (e.g., non-uniform scale removal and rougher surface). Indeed, this translational knowledge lacks a detailed investigation on this process, deep characterization of treated surfaces properties, as well as a comprehensive discussion of the involved mechanisms. In this study, the effects of different acidic media (HCl, HNO3, H3PO4, CH3COOH, H2SO4 and HF), maintained at different temperatures (21 and 60 °C) for various exposition time (15-240 s), on the chemical composition and surface properties of a Fe-13Mn-1.2C biodegradable alloy were investigated. Changes in mass loss, morphology and wettability evidenced the combined effect of temperature and time for all conditions. Pickling in HCl and HF solutions favor mass loss (0.03-0.1 g/cm2) and effectively remove the initial scale.

Biomechanical performances of PCL/HA micro- and macro-porous lattice scaffolds fabricated via laser powder bed fusion for bone tissue engineering.

The present experimental study aims to extend know-how on resorbable polycaprolactone/hydroxyapatite (PCL/HA, 70/30 wt%) scaffolds, produced by Laser Powder Bed Fusion (LPBF) technology, to geometrically complex lattice structures and micro porous struts. Using optimized LPBF printing parameters, micro- and macro-porous scaffolds for bone tissue regeneration were produced by regularly repeating in space Diamond (DO) and Rhombic Dodecahedron (RD) elementary unit cells. After production, scaffolds were submitted to structural, mechanical, and biological characterization. The interaction of scaffolds with human Mesenchymal Stem Cells (hMSCs) allowed studying the degradative processes of the PCL matrix. Biomechanical performances and biodegradation of scaffolds were compared to literature results and bone tissue data. Mechanical compression test, biological viability up to 4 days of incubation and degradation rate evidenced strong dependence of scaffold behavior on unit cell geometry as well as on global geometrical features.

Topological, Mechanical and Biological Properties of Ti6Al4V Scaffolds for Bone Tissue Regeneration Fabricated with Reused Powders via Electron Beam Melting.

Cellularized scaffold is emerging as the preferred solution for tissue regeneration and restoration of damaged functionalities. However, the high cost of preclinical studies creates a gap between investigation and the device market for the biomedical industry. In this work, bone-tailored scaffolds based on the Ti6Al4V alloy manufactured by electron beam melting (EBM) technology with reused powder were investigated, aiming to overcome issues connected to the high cost of preclinical studies. Two different elementary unit cell scaffold geometries, namely diamond (DO) and rhombic dodecahedron (RD), were adopted, while surface functionalization was performed by coating scaffolds with single layers of polycaprolactone (PCL) or with mixture of polycaprolactone and 20 wt.% hydroxyapatite (PCL/HA). The mechanical and biological performances of the produced scaffolds were investigated, and the results were compared to software simulation and experimental evidence available in literature. Good mechanical properties and a favorable environment for cell growth were obtained for all combinations of scaffold geometry and surface functionalization. In conclusion, powder recycling provides a viable practice for the biomedical industry to strongly reduce preclinical costs without altering biomechanical performance.

Solid-State Phase Transformations in Thermally Treated Ti-6Al-4V Alloy Fabricated via Laser Powder Bed Fusion.

Laser Powder Bed Fusion (LPBF) technology was used to produce samples based on the Ti-6Al-4V alloy for biomedical applications. Solid-state phase transformations induced by thermal treatments were studied by neutron diffraction (ND), X-ray diffraction (XRD), scanning transmission electron microscopy (STEM) and energy-dispersive spectroscopy (EDS). Although, ND analysis is rather uncommon in such studies, this technique allowed evidencing the presence of retained ß in α' martensite of the as-produced (#AP) sample. The retained ß was not detectable by XRD analysis, nor by STEM observations. Martensite contains a high number of defects, mainly dislocations, that anneal during the thermal treatment. Element diffusion and partitioning are the main mechanisms in the α ↔ ß transformation that causes lattice expansion during heating and determines the final shape and size of phases. The retained ß phase plays a key role in the α' → ß transformation kinetics.

Cross-Contamination Quantification in Powders for Additive Manufacturing: A Study on Ti-6Al-4V and Maraging Steel.

Metal additive manufacturing is now taking the lead over traditional manufacturing techniques in applications such as aerospace and biomedicine, which are characterized by low production volumes and high levels of customization. While fulfilling these requirements is the strength of metal additive manufacturing, respecting the tight tolerances typical of the mentioned applications is a harder task to accomplish. Powder bed fusion (PBF) is a class of additive manufacturing in which layers of metal powder are fused on top of each other by a high-energy beam (laser or electron beam) according to a computer-aided design (CAD) model. The quality of raw powders for PBF affects the mechanical properties of additively manufactured parts strongly, and therefore it is crucial to avoid the presence of any source of contamination, particularly cross-contamination. In this study, the identification and quantification of cross-contamination in powders of Ti-6Al-4V and maraging steel was performed using scanning electron microscopy (SEM) and energy-dispersive spectroscopy (EDS) techniques. Experimental results showed an overall good reliability of the developed method, opening the way for applications in machine learning environments.

Theoretical and experimental evaluation of piezo-optic parameters and photoelastic constant in tetragonal PWO.

The tetragonal PbWO4 (PWO) is one of the most important scintillating crystals, being used both in the Compact Muon Solenoid (CMS) experiment at the European Organization for Nuclear Research (CERN) and in the PANDA project at the Facility for Antiproton and Ion Research (FAIR). Light yield and other relevant scintillation properties depend, among many factors, also on the crystal mechanical quality. Accordingly, a detailed knowledge of crystal piezo-optic properties is a mandatory step toward understanding elasto-optic behavior and performing crystal quality control. In this paper, we evaluate for the first time, to the best of our knowledge, by means of both photoelastic and x-ray measurements, some components of the piezo-optic tensor; moreover, when the crystal is acted upon by a uniaxial stress, we obtain an evaluation for the rotation angle of the optic plane under stress as well as the photoelastic constant. These parameters are necessary to detect the residual stresses within the crystal, if any, and to give an overall quality measure. Such a methodology is in general suitable for any tetragonal crystals.

Highly Textured FeCo Thin Films Deposited by Low Temperature Pulsed Laser Deposition.

The effect of the deposition temperature (Tdep) on the crystallographic orientation of pulsed laser-deposited FeCo/MgO(100) thin film was determined by means of X-ray reflectivity and high resolution trasmission electron microscopy analysis and was correlated with the magnetic anisotropy properties measured by angle dependent hysteresis loops. Highly textured films with a bcc structure and very smooth surface were obtained even at room temperature, the film being [100] and [110] oriented, at Tdep=25 °C and 150 °C, respectively. The cubic symmetry is reflected in the angular dependence of remanent magnetization, showing a 4-fold character, whose in-plane distribution is consistent with the different crystallographic orientations of the films. The high structural quality, even at room temperature, is reflected in a high value of the saturation magnetization and low coercivity, matching the requirements for technological applications.

Evaluation of an automated system for root canal irrigation: a scanning electron microscopy study.

This study evaluated a new automated system using alternative irrigants for root canal cleaning treatments. This method relies on a system inserting an enzymatic solution based on Trypsin flowing inside the pulp chambers and root canals, completely avoiding traditional endodontic instrumentation. Sixty freshly extracted human molar teeth were randomly divided into 4 groups to assess 3 regimens (R1-3) differing in 0.25% Trypsin/EDTA and 5% Sodium Hypochlorite (NaOCl) solutions administration. Scanning electron microscopy observations and scores taking into account changes in dentin tubules were used to assess treatment effects in pulp chambers and roots. Significant changes in root cleaning ability relative to administration timing were observed, with the best results found in R3, with scheduled alternated cycles of Trypsin/EDTA and NaOCl inside the tooth. The non-invasive root canal method demonstrates good teeth cleaning ability independent of root morphology. This equipment may provide lower discomfort levels for patients undergoing endodontic treatment.