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.

3D-Printed Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate)-Cellulose-Based Scaffolds for Biomedical Applications.

While biomaterials have become indispensable for a wide range of tissue repair strategies, second removal procedures oftentimes needed in the case of non-bio-based and non-bioresorbable scaffolds are associated with significant drawbacks not only for the patient, including the risk of infection, impaired healing, or tissue damage, but also for the healthcare system in terms of cost and resources. New biopolymers are increasingly being investigated in the field of tissue regeneration, but their widespread use is still hampered by limitations regarding mechanical, biological, and functional performance when compared to traditional materials. Therefore, a common strategy to tune and broaden the final properties of biopolymers is through the effect of different reinforcing agents. This research work focused on the fabrication and characterization of a bio-based and bioresorbable composite material obtained by compounding a poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBH) matrix with acetylated cellulose nanocrystals (CNCs). The developed biocomposite was further processed to obtain three-dimensional scaffolds by additive manufacturing (AM). The 3D printability of the PHBH-CNC biocomposites was demonstrated by realizing different scaffold geometries, and the results of in vitro cell viability studies provided a clear indication of the cytocompatibility of the biocomposites. Moreover, the CNC content proved to be an important parameter in tuning the different functional properties of the scaffolds. It was demonstrated that the water affinity, surface roughness, and in vitro degradability rate of biocomposites increase with increasing CNC content. Therefore, this tailoring effect of CNC can expand the potential field of use of the PHBH biopolymer, making it an attractive candidate for a variety of tissue engineering applications.

An Overview of the Latest Progress in Internal Surface Finishing of the Additively Manufactured Metallic Components.

Fast progress in near-net-shape production of parts has attracted vast interest in internal surface finishing. Interest in designing a modern finishing machine to cover the different shapes of workpieces with different materials has risen recently, and the current state of technology cannot satisfy the high requirements for finishing internal channels in metal-additive-manufactured parts. Therefore, in this work, an effort has been made to close the current gaps. This literature review aims to trace the development of different non-traditional internal surface finishing methods. For this reason, attention is focused on the working principles, capabilities, and limitations of the most applicable processes, such as internal magnetic abrasive finishing, abrasive flow machining, fluidized bed machining, cavitation abrasive finishing, and electrochemical machining. Thereafter, a comparison is presented based on which models were surveyed in detail, with particular attention to their specifications and methods. The assessment is measured by seven key features, with two selected methods deciding their value for a proper hybrid machine.

Enamel Analysis by 3D Scanning after Three Orthodontic Clean-Up Procedures: An In-Vitro Test of a New Piezoelectric Tool.

(1) Background: To assess the clinical safety and efficacy of a new piezoelectric instrument for orthodontic clean-up; (2) Methods: An in-vitro comparative study on 75 teeth extracted for orthodontic reasons compared the tested method (Treatment 1) with two other procedures: One step finisher and polisher (Inverted cone One gloss Shofu Dental, Kyoto, Japan) (Treatment 2) and twelve-fluted tungsten carbide bur (123-603-00, Dentaurum, Pforzheim, Germany) and Sof-Lex discs Pop-On XT Kit (3M ESPE) (Treatment 3), with n:25 samples in each group. Clinical safety (enamel volume loss) and effectiveness (residual adhesive volume) were assessed using the structured light 3D scanner Atos Compact Scan (GOM GmbH) together with the support of Atos Professional software. The surfaces were scanned three times to assess: (i) the volume of the residual adhesive (RAV) after bracket removal; (ii) the volume of the relative residual adhesive (dAV) after the clean-up procedure; (iii) volume of the enamel loss (EVL); (3) Results: The mean RAV (mm3) was 0.239 ± 0.337; 0.069 ± 0.124, 0.120 ± 0.193 and the mean EVL (mm3) was 0.1870 ± 0.177, 0.187 ± 0.299 and 0.290 ± 0.205, for treatment 1, 2 and 3, respectively. The distribution was asymmetrical between groups in both cases; (4) Conclusions: The tested instrument proved to be effective and safe for post-orthodontic clean-up. With the increasing use of invisible aligners, the possibility of using an ergonomic and fast instrument is of benefit to both patient and practitioner.

Proteotoxic stress-induced apoptosis in cancer cells: understanding the susceptibility and enhancing the potency.

Leiomyosarcoma (LMS) is aggressive cancer with few therapeutic options. LMS cells are more sensitive to proteotoxic stress compared to normal smooth muscle cells. We used small compound 2c to induce proteotoxic stress and compare the transcriptomic adaptations of immortalized human uterine smooth muscle cells (HUtSMC) and LMS cells SK-UT-1. We found that the expression of the heat shock proteins (HSPs) gene family is upregulated with higher efficiency in normal cells. In contrast, the upregulation of BH3-only proteins is higher in LMS cells. HSF1, the master regulator of HSP transcription, is sequestered into transcriptionally incompetent nuclear foci only in LMS cells, which explains the lower HSP upregulation. We also found that several compounds can enhance the cell death response to proteotoxic stress. Specifically, when low doses were used, an inhibitor of salt-inducible kinases (SIKs) and the inhibitor of IRE1α, a key element of the unfolded protein response (UPR), support proteotoxic-induced cell death with strength in LMS cells and without effects on the survival of normal cells. Overall, our data provide an explanation for the higher susceptibility of LMS cells to proteotoxic stress and suggest a potential option for co-treatment strategies.

Production of Dense Cu-10Sn Part by Laser Powder Bed Fusion with Low Surface Roughness and High Dimensional Accuracy.

Tin-bronze alloys with a tin content of at least 10 wt% have excellent mechanical properties, wear resistance, and corrosion resistance. Among these alloys, Cu-10Sn was investigated in this study for production with the laser powder bed fusion process with a 500W Yb:YAG laser. In particular, a design of experiment (DoE) was developed in order to identify the optimal process parameters to obtain full density, low surface roughness, and high dimensional accuracy. Samples were characterized with Archimedes' method and optical microscopy to determine their final density. It was shown that the first method is fast but not as reliable as the second one. A first mechanical characterization was performed through microhardness tests. Finally, a set of process parameters was identified to produce fully dense samples with low surface roughness and high accuracy. The results showed that the volumetric energy density could represent an approach that is too simplified, therefore limiting the direct correlation with the physical aspects of the process.

Enhancing Proteotoxic Stress in Leiomyosarcoma Cells Triggers Mitochondrial Dysfunctions, Cell Death, and Antitumor Activity in vivo.

Leiomyosarcomas are rare and aggressive tumors characterized by a complex karyotype. Surgical resection with or without radiotherapy and chemotherapy is the standard curative treatment. Unfortunately, a high percentage of leiomyosarcomas recurs and metastasizes. In these cases, doxorubicin and ifosfamide represent the standard treatment but with low response rates. Here, we evaluated the induction of proteotoxic stress as a possible strategy to kill leiomyosarcoma cells in a therapeutic perspective. We show that aggressive leiomyosarcomas coexist with high levels of proteotoxic stress. As a consequence, we hypothesized that leiomyosarcoma cells are vulnerable to further increases of proteotoxic stress. The small compound 2c is a strong inducer of proteotoxic stress. In leiomyosarcoma cells, it triggers cell death coupled to a profound reorganization of the mitochondrial network. By using stimulated emission depletion microscopy, we have unveiled the existence of DIABLO/SMAC clusters that are modulated by 2c. Finally, we have engineered a new version of 2c linked to polyethylene glycol though a short peptide, named 2cPP. This new prodrug is specifically activated by proteases present in the tumor microenvironment. 2cPP shows a strong antitumor activity in vivo against leiomyosarcomas and no toxicity against normal cells.

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.

Proteotoxic Stress and Cell Death in Cancer Cells.

To maintain proteostasis, cells must integrate information and activities that supervise protein synthesis, protein folding, conformational stability, and also protein degradation. Extrinsic and intrinsic conditions can both impact normal proteostasis, causing the appearance of proteotoxic stress. Initially, proteotoxic stress elicits adaptive responses aimed at restoring proteostasis, allowing cells to survive the stress condition. However, if the proteostasis restoration fails, a permanent and sustained proteotoxic stress can be deleterious, and cell death ensues. Many cancer cells convive with high levels of proteotoxic stress, and this condition could be exploited from a therapeutic perspective. Understanding the cell death pathways engaged by proteotoxic stress is instrumental to better hijack the proliferative fate of cancer cells.

GSK3ß is a key regulator of the ROS-dependent necrotic death induced by the quinone DMNQ.

Signaling pathways controlling necrosis are still mysterious and debated. We applied a shRNA-based viability screen to identify critical elements of the necrotic response. We took advantage from a small molecule (G5) that makes covalent adducts with free thiols by Michael addition and elicits multiple stresses. In cells resistant to apoptosis, G5 triggers necrosis through the induction of protein unfolding, glutathione depletion, ER stress, proteasomal impairments, and cytoskeletal stress. The kinase GSK3ß was isolated among the top hits of the screening. Using the quinone DMNQ, a ROS generator, we demonstrate that GSK3ß is involved in the regulation of ROS-dependent necrosis. Our results have been validated using siRNA and by knocking-out GSK3ß with the CRISPR/Cas9 technology. In response to DMNQ GSK3ß is activated by serine 9 dephosphorylation, concomitantly to Akt inactivation. During the quinone-induced pro-necrotic stress, GSK3ß gradually accumulates into the nucleus, before the collapse of the mitochondrial membrane potential. Accumulation of ROS in response to DMNQ is impaired by the absence of GSK3ß. We provide evidence that the activities of the obligatory two-electrons reducing flavoenzymes, NQO1 (NAD(P)H quinone dehydrogenase 1) and NQO2 are required to suppress DMNQ-induced necrosis. In the absence of GSK3ß the expression of NQO1 and NQO2 is dramatically increased, possibly because of an increased transcriptional activity of NRF2. In summary, GSK3ß by blunting the anti-oxidant response and particularly NQO1 and NQO2 expression, favors the appearance of necrosis in response to ROS, as generated by the quinone DMNQ.

Design of Additively Manufactured Structures for Biomedical Applications: A Review of the Additive Manufacturing Processes Applied to the Biomedical Sector.

Additive manufacturing (AM) is a disruptive technology as it pushes the frontier of manufacturing towards a new design perspective, such as the ability to shape geometries that cannot be formed with any other traditional technique. AM has today shown successful applications in several fields such as the biomedical sector in which it provides a relatively fast and effective way to solve even complex medical cases. From this point of view, the purpose of this paper is to illustrate AM technologies currently used in the medical field and their benefits along with contemporary. The review highlights differences in processes, materials, and design of additive manufacturing techniques used in biomedical applications. Successful case studies are presented to emphasise the potentiality of AM processes. The presented review supports improvements in materials and design for future researches in biomedical surgeries using instruments and implants made by AM.

Proposal of an innovative benchmark for accuracy evaluation of dental crown manufacturing.

An innovative benchmark representing a dental arch with classic features corresponding to different kinds of prepared teeth is proposed. Dental anatomy and general rules for tooth preparation are taken into account. This benchmark includes tooth orientation and provides oblique surfaces similar to those of real prepared teeth. The benchmark is produced by additive manufacturing (AM) and subjected to digitization by a dental three-dimensional scanner. The evaluation procedure proves that the scan data can be used as reference model for crown restorations design. Therefore this benchmark is at the basis for comparative studies about different CAD/CAM and AM techniques for dental crowns.