Influence of the Coarse Grain Structure of a Titanium Alloy Ti-4Al-3V Formed by Wire-Feed Electron Beam Additive Manufacturing on Strain Inhomogeneities and Fracture.

In this work, based on the multilevel approach, the features of the structure and properties of titanium alloy, formed during high-performance additive manufacturing by wire-feed electron beam technology, were studied. Methods of non-destructive X-ray control and tomography, along with optical and scanning electron microscopy, were used to study the structure at different scale levels of the sample material. The mechanical properties of the material under stress were revealed via the simultaneous observation of the peculiarities of deformation development, using a Vic 3D laser scanning unit. Using microstructural and macrostructural data, as well as fractography, the interrelations of structure and material properties caused by the technological features of the printing process and the composition of used welding wire were revealed.

Al-Al3Ni In Situ Composite Formation by Wire-Feed Electron-Beam Additive Manufacturing.

The regularities of microstructure formation in samples of multiphase composites obtained by additive electron beam manufacturing on the basis of aluminum alloy ER4043 and nickel superalloy Udimet-500 have been studied. The results of the structure study show that a multicomponent structure is formed in the samples with the presence of Cr23C6 carbides, solid solutions based on aluminum -Al or silicon -Si, eutectics along the boundaries of dendrites, intermetallic phases Al3Ni, AlNi3, Al75Co22Ni3, and Al5Co, as well as carbides of complex composition AlCCr, Al8SiC7, of a different morphology. The formation of a number of intermetallic phases present in local areas of the samples was also distinguished. A large amount of solid phases leads to the formation of a material with high hardness and low ductility. The fracture of composite specimens under tension and compression is brittle, without revealing the stage of plastic flow. Tensile strength values are significantly reduced from the initial 142-164 MPa to 55-123 MPa. In compression, the tensile strength values increase to 490-570 MPa and 905-1200 MPa with the introduction of 5% and 10% nickel superalloy, respectively. An increase in the hardness and compressive strength of the surface layers results in an increase in the wear resistance of the specimens and a decrease in the coefficient of friction.

Features of the Macro-, Micro-, and Fine Structure of the Nickel Superalloy Product Material Formed by the Method of Electron Beam Additive Manufacturing.

In the present work, the products in the form of vertical walls were made of heat-resistant nickel-based superalloy ZhS32 via the method of electron beam additive technology. Unidirectional printing strategy was applied. The effect of heat input and 3D printing strategy on the macrostructure, dimensions, and morphology of microstructure elements was established. It was shown that the additive product material has a directed macrostructure. The only exclusion was the final layer with a thickness of no more than 3.5 mm. The directed macrostructure consisted of dendrites oriented predominantly along the crystallographic direction {001} of the primary dendrite arms. The misorientation of the dendrite axes did not exceed 9 degrees. The angle between the predominant dendrite growth direction and the normal to the substrate was 23 degrees. The average primary dendrite arms' spacing increased monotonically from 16 µm at 5 mm from the substrate to 23 µm in the final layers of the product material (the overall height was 41 mm). It was found that the average size of γ' (Ni3Al)-phase precipitations in the form of nanoscale and submicrocrystalline cuboids varied in the range of 76 to 163 nm depending on the distance from the substrate. The size of γ'-phase precipitations reached a maximum at about 30 mm from the substrate, while in the final layers of the product material, the average cuboid size did not exceed 135 nm. Extreme dependence of the size of γ'-phase precipitations on the height of the product followed from a combination of a given monotonic decrease in heat input and heat accumulation in the product material as it formed, as did additional heat removal by means of radiation during formation of the final layer of the product without re-melting. Chemical elements of the austenitic steel substrate material were not detected in the product material more than 8 mm from the substrate. There were no macrodefects, such as voids, in the entire volume of the product material.

Aluminum Bronze/Udimet 500 Composites Prepared by Electron-Beam Additive Double-Wire-Feed Manufacturing.

Novel composite CuA19Mn2/Udimet-500 alloy walls with different content of the Udimet 500 were built using electron-beam double-wire-feed additive manufacturing. Intermixing both metals within the melted pool resulted in dissolving nickel and forcing out the aluminum from bronze. The resulting phases were NiAl particles and grains, M23C6/NiAl core/shell particles and Cu-Ni-Al solid solution. Precipitation of these phases resulted in the increased hardness and tensile strength as well as reduced ductility of the composite alloys. Such a hardening resulted in improving the wear resistance as compared to that of source aluminum bronze.

Antiviral photodynamic therapy: Inactivation and inhibition of SARS-CoV-2 in vitro using methylene blue and Radachlorin.

INTRODUCTION: Recently, the COVID-19 pandemic has spread globally, necessitating the development of new methods for its prevention and treatment. The purpose of this study was to evaluate the antiviral activity of photodynamic therapy (PDT) against SARS-CoV-2 in vitro. METHODS: Vero E6 cells and SARS-CoV-2 isolated in Russia were used for PDT with methylene blue (MB) and Radachlorin. A continuous laser with wavelength λ = 662 nm in doses of 16 J/cm2 and 40 J/cm2 laser irradiation was used for PDT of a viral suspension and SARS-CoV-2-infected cells. The direct cytopathogenic effect of SARS-CoV-2 was evaluated via light microscopy to calculate the TCID50 in the samples and perform statistical analysis. RESULTS: Viral suspensions of SARS-CoV-2 that had a TCID50 greater than 103 were inactivated by PDT in the presence of MB and Radachlorin. Vero E6 cells were protected from 104 TCID50 of SARS-CoV-2 by PDT post infection. The range of protective concentrations was 1.0-10.0 µg/ml and 0.5-5.0 µg/ml for MB and Radachlorin, respectively. Additionally, it was found that MB and Radachlorin also possess significant antiviral activity even without PDT. The 50 % inhibitory concentration (IC50) against 102 TCID50 of SARS-CoV-2 was found to be 0.22 and 0.33 µg/mL with the addition of MB and Radachlorin, respectively, to cells concomitantly with virus, whereas in the case of applying the photosensitizers at 3.5 h post infection, the IC50 was 0.6 and 2.0 µg/mL for MB and Radachlorin, respectively. CONCLUSION: PDT shows high antiviral activity against SARS-CoV-2 when combined with MB and Radachlorin in vitro.

Impairments of Antigen-Presenting Cells in Pulmonary Tuberculosis.

The phenotype and functional properties of antigen-presenting cells (APC), that is, circulating monocytes and generated in vitro macrophages and dendritic cells, were investigated in the patients with pulmonary tuberculosis (TB) differing in lymphocyte reactivity to M. tuberculosis antigens (PPD-reactive versus PPD-anergic patients). We revealed the distinct impairments in patient APC functions. For example, the monocyte dysfunctions were displayed by low CD86 and HLA-DR expression, 2-fold increase in CD14(+)CD16(+) expression, the high numbers of IL-10-producing cells, and enhanced IL-10 and IL-6 production upon LPS-stimulation. The macrophages which were in vitro generated from peripheral blood monocytes under GM-CSF were characterized by Th1/Th2-balance shifting (downproduction of IFN-γ coupled with upproduction of IL-10) and by reducing of allostimulatory activity in mixed lymphocyte culture. The dendritic cells (generated in vitro from peripheral blood monocytes upon GM-CSF + IFN-α) were characterized by impaired maturation/activation, a lower level of IFN-γ production in conjunction with an enhanced capacity to produce IL-10 and IL-6, and a profound reduction of allostimulatory activity. The APC dysfunctions were found to be most prominent in PPD-anergic patients. The possible role of APC impairments in reducing the antigen-specific T-cell response to M. tuberculosis was discussed.

In Vitro biocompatibility evaluation of a root canal filling material that expands on water sorption.

INTRODUCTION: CPoint is a polymeric endodontic point that takes advantage of water-induced, non-isotropic radial expansion to adapt to canal irregularities. This study evaluated the effects of CPoint on the viability and mineralization potential of odontoblast-like cells. METHODS: The biocompatibility of CPoint and commercially available gutta-percha points was evaluated by using a rat odontoblast-like cell line (MDPC-23). Cell viability was evaluated with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, flow cytometry, and confocal laser scanning microscopy. The mineralization potential of MDPC-23 cells, in the presence of the root-filling materials, was evaluated by examining the changes in osteogenic gene marker expression (quantitative real-time polymerase chain reaction), alkaline phosphatase activity, alizarin red S assay, and transmission electron microscopy. RESULTS: CPoint showed higher initial cytotoxicity compared with gutta-percha and Teflon (P < .05), which became nonsignificant after 4 immersion cycles. Significant differences were also found between eluents from CPoint and gutta-percha at 1:1 concentration (P < .05) but not at 1:10 or 1:100 concentration. Both materials induced minimal apoptosis-induced alteration in plasma membrane permeability, as evidenced by flow cytometry and confocal laser scanning microscopy. Compared with the Teflon negative control, CPoint and gutta-percha groups showed up-regulation of most osteogenic gene markers except for dentin sialophosphoprotein, which was down-regulated. Alkaline phosphatase activity and alizarin red assay for CPoint and gutta-percha were both significantly higher than for Teflon but not significantly different from each other (P > .05). Transmission electron microscopy showed discrete nodular electron-dense mineralization foci in all 3 groups. CONCLUSIONS: The in vitro biocompatibility of CPoint is comparable to gutta-percha with minimal adverse effects on osteogenesis after elution of potentially toxic components.

Cytotoxic activity of dendritic cells as a possible mechanism of negative regulation of T lymphocytes in pulmonary tuberculosis.

The PD-1/B7-H1-mediated induction of T cell apoptosis/anergy as a possible mechanism of immune response failure was studied in 76 patients with pulmonary tuberculosis (TB) with normal and low-proliferative response to antigens of M. tuberculosis (purified protein derivative (PPD)). It was revealed that dendritic cells (DCs), generated in vitro from patient blood monocytes with GM-CSF + IFN-α, were characterized by increased B7-H1 expression, upproduction of IL-10, and reducing of allostimulatory activity in mixed lymphocyte culture (MLC). Moreover, DCs of patients with TB were able to enhance T cell apoptosis and to block T-cell division in MLC. It was shown that neutralizing anti-PD1 antibodies significantly decreased the proapoptogenic/tolerogenic effect of DCs. Correlation analysis revealed a direct relationship between IL-10 production and level of B7-H1 expression in the general group of investigated patients. It was demonstrated that generation of healthy donor DCs in the presence of IL-10 led to an increase in the number of DCs-expressed B7-H1 molecule, DC proapoptogenic activity, and a decrease in their allostimulatory activity. Obviously, the revealed phenomenon of the PD-1/B7-H1-mediated pro-apoptogenic activity of DCs is clinically significant since the cytotoxic/tolerogenic potential of DCs is more pronounced in patients with PPD anergy.

Intrafibrillar silicification of collagen scaffolds for sustained release of stem cell homing chemokine in hard tissue regeneration.

Traditional bone regeneration strategies relied on supplementation of biomaterials constructs with stem or progenitor cells or growth factors. By contrast, cell homing strategies employ chemokines to mobilize stem or progenitor cells from host bone marrow and tissue niches to injured sites. Although silica-based biomaterials exhibit osteogenic and angiogenic potentials, they lack cell homing capability. Stromal cell-derived factor-1 (SDF-1) plays a pivotal role in mobilization and homing of stem cells to injured tissues. In this work, we demonstrated that 3-dimensional collagen scaffolds infiltrated with intrafibrillar silica are biodegradable and highly biocompatible. They exhibit improved compressive stress-strain responses and toughness over nonsilicified collagen scaffolds. They are osteoconductive and up-regulate expressions of osteogenesis- and angiogenesis-related genes more significantly than nonsilicified collagen scaffolds. In addition, these scaffolds reversibly bind SDF-1α for sustained release of this chemokine, which exhibits in vitro cell homing characteristics. When implanted subcutaneously in an in vivo mouse model, SDF-1α-loaded silicified collagen scaffolds stimulate the formation of ectopic bone and blood capillaries within the scaffold and abrogate the need for cell seeding or supplementation of osteogenic and angiogenic growth factors. Intrafibrillar-silicified collagen scaffolds with sustained SDF-1α release represent a less costly and complex alternative to contemporary cell seeding approaches and provide new therapeutic options for in situ hard tissue regeneration.

Quaternary ammonium silane-functionalized, methacrylate resin composition with antimicrobial activities and self-repair potential.

The design of antimicrobial polymers to address healthcare issues and minimize environmental problems is an important endeavor with both fundamental and practical implications. Quaternary ammonium silane-functionalized methacrylate (QAMS) represents an example of antimicrobial macromonomers synthesized by a sol-gel chemical route; these compounds possess flexible Si-O-Si bonds. In present work, a partially hydrolyzed QAMS co-polymerized with 2,2-[4(2-hydroxy 3-methacryloxypropoxy)-phenyl]propane is introduced. This methacrylate resin was shown to possess desirable mechanical properties with both a high degree of conversion and minimal polymerization shrinkage. The kill-on-contact microbiocidal activities of this resin were demonstrated using single-species biofilms of Streptococcus mutans (ATCC 36558), Actinomyces naeslundii (ATCC 12104) and Candida albicans (ATCC 90028). Improved mechanical properties after hydration provided the proof-of-concept that QAMS-incorporated resin exhibits self-repair potential via water-induced condensation of organic modified silicate (ormosil) phases within the polymerized resin matrix.

Effects of an experimental calcium aluminosilicate cement on the viability of murine odontoblast-like cells.

INTRODUCTION: Quick-setting calcium aluminosilicate cement with improved washout resistance is a potential substitute for calcium silicate cements in endodontics. This study examined the effect of an experimental calcium aluminosilicate cement (Quick-Set; Primus Consulting, Bradenton, FL) on the viability of odontoblast-like cells. METHODS: The biocompatibility of Quick-Set and white ProRoot MTA (WMTA; Dentsply Tulsa Dental Specialties, Tulsa, OK) cements and their eluents was evaluated using a murine dental papilla-derived odontoblast-like cell line (MDPC-23); 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was used to examine the effects of the 2 hydraulic cements on mitochondrial metabolic activity. Flow cytometry and confocal laser scanning microscopy were used to identify the effects of the 2 cements on cell death-induced plasma membrane permeability to fluorescent dyes and DNA stains. RESULTS: After the first week of immersion in culture medium, Quick-Set and WMTA were more cytotoxic than the Teflon-negative control (P < .05), and the cells exhibited more apoptosis/necrosis than Teflon (P < .05). After the second week of immersion, the 2 cements were as biocompatible as Teflon (P > .05), with cells exhibiting minimal apoptosis/necrosis. Eluents from the set cements at 1:1 dilution were significantly more cytotoxic that eluents at 1:10 or 1:100 dilution (P < .05). CONCLUSIONS: Quick-Set and WMTA exhibited similar cytotoxicity profiles. They possess negligible in vitro toxicologic risks after time-dependent elution of toxic components.

Effects of calcium silicate-based materials on the flexural properties of dentin.

INTRODUCTION: Prolonged exposure of root dentin to calcium hydroxide alters the fracture resistance of dentin. Calcium silicate-based materials (CSMs) used in endodontics release calcium hydroxide on setting. This study examined whether prolonged contact of dentin with CSMs adversely affects its mechanical properties. METHODS: Dentin beams prepared from extracted human molars (7 × 3 × 0.3 mm) were divided into 3 groups on the basis of the material to which dentin was exposed (Biodentine, MTA Plus, and untreated control beams). Three-point flexure to failure was performed for each beam at designated exposure times (24 hours, 1, 2, and 3 months; n = 10). Data were analyzed with 2-factor repeated-measures analyses of variance to determine the effects of material and aging time on flexural modulus, flexural strength, and modulus of toughness (α = 0.05). RESULTS: For flexural modulus, there was no significant difference for material (P = .947) or aging time (P = .064) when compared with baseline control. For flexural strength, significant differences were associated with aging time (P < .001) but not with material (P = .349). Flexural strength of dentin exposed to Biodentine decreased significantly after 2 and 3 months, whereas that exposed to MTA Plus decreased significantly after 3 months of aging (P < .05). For modulus of toughness, significant declines were observed for both material (P < .004) and aging time (P < .001). CONCLUSIONS: Both CSMs alter material toughness more than the strength and stiffness of dentin after aging in 100% relative humidity. Because dentin toughness is attributed to its collagen matrix, the amount of collagen extracted from mineralized dentin and changes in collagen ultrastructure should be further examined after exposure of dentin to CSMs.

T Cell Functional Disturbances in Patients with Pulmonary Tuberculosis.

The investigations of 38 patients with pulmonary tuberculosis (PT) revealed combined T cell and monocyte functional disturbances. Indeed, the percentages of CD4(+) and CD8(+) T lymphocytes, proliferative response and IL-2 production, as well as the percentages of HLA DR(+) monocytes and IL-1beta production were significantly decreased in PT patients as compared with normal individuals. Herewith the absolute T lymphocyte number did not undergo the pronounced changes. The decrease of T cell proliferative response was not mediated through immunosuppressive action of monocytes or T lymphocytes since removing of "adherent" cells from patient's peripheral blood mononuclear cells (PBMC) or pretreatment of PBMC with indomethacin and cyclophosphan failed to recover mitogenic reactivity in vitro. The patient's sera also did not significantly influence on PBMC proliferation. The decrease of IL-2 production and the stimulation of T cell proliferative response via TcR-CD3 complex, i.e. through the classic pathway of activation, indicated the anergy of T lymphocyte in tuberculosis patients. Furthermore, T lymphocytes were characterized by enhanced apoptosis. It should be noted, that patient's sera (especially in the patients with an initially high apoptosis) promoted significant anti-apoptotic activity. It is likely that this mechanism may be an explanation, why absolute T lymphopenia is absent during tuberculosis infection. Our findings suggest, that T lymphocyte dysfunctions in patients with PT are caused by impairments of T cell activation process, which lead to predominance of "negative" response (induction anergy, apoptosis) and to a lesser degree connected with direct suppressive mechanisms mediated by monocytes, T lymphocytes or serum factors.