In Vitro/In Vivo Toxicity Evaluation and Quantification of Iron Oxide Nanoparticles

Increasing biomedical applications of iron oxide nanoparticles (IONPs) in academic and commercial settings have alarmed the scientific community about the safety and assessment of toxicity profiles of IONPs. The great amount of diversity found in the cytotoxic measurements of IONPs points toward the necessity of careful characterization and quantification of IONPs. The present document discusses the major developments related to in vitro and in vivo toxicity assessment of IONPs and its relationship with the physicochemical parameters of IONPs. Major discussion is included on the current spectrophotometric and imaging based techniques used for quantifying, and studying the clearance and biodistribution of IONPs. Several invasive and non-invasive quantification techniques along with the pitfalls are discussed in detail. Finally, critical guidelines are provided to optimize the design of IONPs to minimize the toxicity.     

Multifunctional Properties of Multistage Spark Plasma Sintered HA–BaTiO3‐Based Piezobiocomposites for Bone Replacement Applications

This work reports the processing–microstructure–property correlation of novel HA–BaTiO₃‐based piezobiocomposites, which demonstrated the bone‐mimicking functional properties. A series of composites of hydroxyapatite (HA) with varying amounts of piezoelectric BaTiO₃ (BT) were optimally processed using uniquely designed multistage spark plasma sintering (SPS) route. Transmission electron microscopy imaging during in situ heating provides complementary information on the real‐time observation of sintering behavior. Ultrafine grains (≤0.50 μm) of HA and BT phases were predominantly retained in the SPSed samples. The experimental results revealed that dielectric constant, AC conductivity, piezoelectric strain coefficient, compressive strength, and modulus values of HA‐40 wt% BT closely resembles with that of the natural bone. The addition of 40 wt% BT enhances the long‐crack fracture toughness, compressive strength, and modulus by 132%, 200%, and 165%, respectively, with respect to HA. The above‐mentioned exceptional combination of functional properties potentially establishes HA‐40 wt% BT piezocomposite as a new‐generation composite for orthopedic implant applications.     

Sintering behavior and characteristics study of BaTiO3 with 50 wt% of B2O3-Bi2O3-SiO2-ZnO glass

The thermal analysis of B₂O₃-Bi₂O₃-SiO₂-ZnO (BBSZ) glass with different particle sizes and LiF addition was researched to study its temperature behavior. Next the composites with 50 wt% BaTiO₃–50 wt% BBSZ glass were prepared for shrinkage, microstructures and dielectric properties investigations. The differently treated BBSZ glass showed that the smaller glass particles clearly decreased its softening and crystallization temperatures. LiF addition had the same but much weaker effect.The composites showed two-stage shrinkage related to the softening of the glass and new phase generation of Bi₂₄Si₂O₄₀ at 385–450 °C, and Bi₄BaTi₄O₁₅ over 680 °C. The microstructures of the composites sintered at 720 °C showed Bi₄BaTi₄O₁₅, BaTiO₃ and Bi₂₄Si₂O₄₀ with residual ZnO phase. LiF addition increased the amount of Bi₄BaTi₄O₁₅, thus increasing the loss value. However the particle size of the glass did not effect to the dielectric properties of the composites showing permittivity of 248–256 and loss of 0.013 at 100 kHz.     

Modulatory Effects of Biosynthesized Gold Nanoparticles Conjugated with Curcumin and Paclitaxel on Tumorigenesis and Metastatic Pathways—In Vitro and In Vivo Studies

Background: Breast cancer is the most common cancer in women globally, and diagnosing it early and finding potential drug candidates against multi-drug resistant metastatic breast cancers provide the possibilities of better treatment and extending life. Methods: The current study aimed to evaluate the synergistic anti-metastatic activity of Curcumin (Cur) and Paclitaxel (Pacli) individually, the combination of Curcumin–Paclitaxel (CP), and also in conjugation with gold nanoparticles (AuNP–Curcumin (Au-C), AuNP–Paclitaxel (Au-P), and AuNP–Curcumin–Paclitaxel (Au-CP)) in various in vitro and in vivo models. Results: The results from combination treatments of CP and Au-CP demonstrated excellent synergistic cytotoxic effects in triple-negative breast cancer cell lines (MDA MB 231 and 4T1) in in vitro and in vivo mouse models. Detailed mechanistic studies were performed that reveal that the anti-cancer effects were associated with the downregulation of the expression of VEGF, CYCLIN-D1, and STAT-3 genes and upregulation of the apoptotic Caspase-9 gene. The group of mice that received CP combination therapy (with and without gold nanoparticles) showed a significant reduction in the size of tumor when compared to the Pacli alone treatment and control groups. Conclusions: Together, the results suggest that the delivery of gold conjugated Au-CP formulations may help in modulating the outcomes of chemotherapy. The present study is well supported with observations from cell-based assays, molecular and histopathological analyses.     

Long‐Term Stability,Regeneration and Recycling of Imidazolium‐based Ionic Liquids

Ionic liquids (ILs) are discussed in many current research papers extensively in terms of their potential use in the chemical industry, as process aids and novel materials. The long‐term stability of the IL is for industrial applications as important as to know which species arise during the degradation due to thermal, mechanical, chemical or electrochemical stress. The investigation of the long‐term stability of two selected ILs over several months under process‐like conditions is presented with a subsequent analysis by LC‐MS to identify the resulting decomposition products. Knowledge about the occurring species and their analytical quantification are basis for the selection of appropriate processes for the separation of the decomposition products and the development of recycling processes for ILs. Particularly melt crystallization processes are suitable for separating structurally similar decomposition products that typically occur in the IL degradation.     

Nanocomposites of Surface‐Modified BaTiO3 Nanoparticles Filled Ferroelectric Polymer with Enhanced Energy Density

We report nanocomposites of increased dielectric permittivity, enhanced electric breakdown strength and high‐energy density based on surface‐modified BaTiO₃ (BT) nanoparticles filled poly(vinylidene fluoride) polymer. Polyvinylprrolidone (PVP) is used as the surface modification agent and homogeneous nanocomposite films have been prepared by solution casting processing. The dielectric permittivity of the nanocomposite with treated BT is higher than those with untreated BT and reaches the maximum value of 77 (1 kHz) at BT concentration of 55 vol%. The electric breakdown strength of the nanocomposite is greatly enhanced to 336 MV/m at BT concentration of 10 vol% and the calculated energy density is 6.8 J/cm³. The results indicate that using PVP as surface modification agent can greatly enhance the dielectric permittivity and electric breakdown strength of the ceramic–polymer nanocomposite and achieve high‐energy density for energy storage and power capacitor applications.     

Densification and biocompatibility of sintering 3.0 mol% yttria-tetragonal ZrO2 polycrystal ceramics with x wt% Fe2O3 and 5.0 wt% mica powders additive

The densification and biocompatibility of sintered 3.0 mol% yttria-tetragonal zirconia polycrystal (3Y-TZP) ceramics, with X wt% Fe₂O₃ and 5.0 wt% mica powders (denoted by 3Y-TZP: X-5.0 wt% mica) have been studied. When the pellets of 3Y-TZP: X-5.0 wt% mica were sintered at 1300 °C for 1 h, the relative shrinkage increases from 19.20–19.43% with the X increased from 0.3 to 1.0. The relative shrinkage of pellets containing 1.0 wt% Fe₂O₃ (X=1.0) increased from 19.43–19.59% when sintering temperatures were raised from 1300 °C to 1450 °C. X-ray diffraction results show that the pellets of 3Y-TZP: X-5.0 wt% mica sintered at 1400 °C for 1 h only contained single phase of tetragonal ZrO₂ (t-ZrO₂). When the sintering temperature was higher than 1400 °C, the Vickers microhardness was greatest in the pellets with X=0.5. Within pellets with the same Fe₂O₃ content, the dominant wavelength (λ_d) was only slightly different for pellets sintered at 1300 °C and those sintered at 1450 °C. The results of the materials were evaluated in vitro cytotoxicity tests reveals that the powders and sintered pellets are safe materials. The oral mucosa irritation tests did not find erythema or histopathological change including normal epithelium, and was free from leucocyte infiltration, vascular congestion and oedema.     

Increased Expression of Galectin-3 in Skin Fibrosis: Evidence from In Vitro and In Vivo Studies

Skin fibrosis is a hallmark of a wide array of dermatological diseases which can greatly impact the patients’ quality of life. Galectin-3 (GAL-3) has emerged as a central regulator of tissue fibrosis, playing an important pro-fibrotic role in numerous organs. Various studies are highlighting its importance as a skin fibrotic diseases biomarker; however, there is a need for further studies that clarify its role. This paper aims to ascertain whether the expression of GAL-3 is increased in relevant in vitro and in vivo models of skin fibrosis. We studied the role of GAL-3 in vitro using normal human dermal fibroblasts (NHDF) and fibrocytes. In addition, we used a skin fibrosis murine model (BALB/c mice) and human biopsies of healthy or keloid tissue. GAL-3 expression was analyzed using real time PCR, Western blot and immunostaining techniques. We report a significantly increased expression of GAL-3 in NHDF and fibrocytes cell cultures following stimulation with transforming growth factor β1 (TGFβ1). In vivo, GAL-3 expression was increased in a murine model of systemic sclerosis and in human keloid biopsies. In sum, this study underlines the involvement of GAL-3 in skin fibrosis using several models of the disease and highlights its role as a relevant target.     

Preparation and characterisation of the magneto-electric xBiFeO3–(1 − x)BaTiO3 ceramics

The solid solutions of BiFeO₃–BaTiO₃ have been prepared via solid state with a view to obtaining magnetoelectric properties, i.e. ferroelectric and magnetic activity in the same range of temperatures. Optimum calcination and sintering strategy for obtaining pure perovskite phase, dense ceramics (>97% relative density) and homogeneous microstructures have been determined. The sample of composition 0.7BiFeO₃–0.3BaTiO₃ reported in the present work is pseudo-cubic at room temperature. The permittivity is ɛ_r ≈ 150 at the room temperature and shows a broad ferro-para phase transition at around 175 °C where ɛ_r ≈ 1600. This diffuse maximum of the permittivity, similar to that in relaxors, is due to the chemical inhomogeneity in both A and B sites of the perovskite unit cell ABO₃. Higher losses, tan δ > 1, appear above 200 °C and other different conduction mechanisms start to be active particularly at temperatures higher than 400 °C, when the ceramic becomes conductive. The magnetic properties show a succession of transitions from weak ferro/ferrimagnetism-to-antiferromagnetism and antiferromagnetism-to-paramagnetism at T_N1 ≈ 10 K and T_N2 ≈ 265 K. Below T_N2 the ceramic 0.7BiFeO₃–0.3BaTiO₃ can present magnetoelectric coupling, due to the fact that is simultaneously ferroelectric and antiferromagnetic.     

Microstructural,mechanical and marine water tribological properties of plasma-sprayed graphene nanoplatelets reinforced Al2O3- 40 wt% TiO2 coating

Graphene nanoplatelets (GNPs) as reinforcement in the ceramic matrix is rising continuously due to their outstanding mechanical and lubricative properties. Herein, different compositions of GNPs (0.5–2 wt%) reinforced alumina-titania coatings were prepared using atmospheric plasma spraying. The relative density of AT coating increased from 83% to 94% with just (1.5 wt%) addition of GNP. Consequently, mechanical properties i.e. hardness and elastic modulus were improved by ~77% and ~69% respectively. Fracture toughness also increased from 2.65 ± 0.95 MPa.m^1/2 to 5.85 ± 1.07 MPa.m^1/2. Furthermore, the seawater wear test, using a ball-on-disc tribometer revealed that the wear rate of AT coating decreased from ~11 × 10^?14 m³/Nm to ~4 × 10^?14 m³/Nm, whereas the coefficient of friction reduced from 0.33 ± 0.05–0.16 ± 0.03. The mechanisms involved to improve these properties, viz. GNP sandwiching, crack bridging, crack arrest, etc. GNP’s multi-layers facilitated long-term lubricity and enhanced the wear resistance properties of the coatings.     

Biochemical Analysis of Leukocytes after In Vitro and In Vivo Activation with Bacterial and Fungal Pathogens Using Raman Spectroscopy

Biochemical information from activated leukocytes provide valuable diagnostic information. In this study, Raman spectroscopy was applied as a label-free analytical technique to characterize the activation pattern of leukocyte subpopulations in an in vitro infection model. Neutrophils, monocytes, and lymphocytes were isolated from healthy volunteers and stimulated with heat-inactivated clinical isolates of Candida albicans, Staphylococcus aureus, and Klebsiella pneumoniae. Binary classification models could identify the presence of infection for monocytes and lymphocytes, classify the type of infection as bacterial or fungal for neutrophils, monocytes, and lymphocytes and distinguish the cause of infection as Gram-negative or Gram-positive bacteria in the monocyte subpopulation. Changes in single-cell Raman spectra, upon leukocyte stimulation, can be explained with biochemical changes due to the leukocyte’s specific reaction to each type of pathogen. Raman spectra of leukocytes from the in vitro infection model were compared with spectra from leukocytes of patients with infection (DRKS-ID: DRKS00006265) with the same pathogen groups, and a good agreement was revealed. Our study elucidates the potential of Raman spectroscopy-based single-cell analysis for the differentiation of circulating leukocyte subtypes and identification of the infection by probing the molecular phenotype of those cells.     

Screening,Synthesis, and In Vitro Evaluation of Vinyl Sulfones as Inhibitors of Complement‐Dependent Cytotoxicity in Neuromyelitis Optica

Neuromyelitis optica (NMO) is a demyelinating autoimmune disease of the optic nerve and spinal cord triggered by binding of NMO‐specific immunoglobulin G (NMO‐IgG) auto‐antibodies to the water channel aquaporin‐4 (AQP4) in astrocytes. To find potential NMO therapeutics, a screening system was established and used to identify inhibitors of NMO‐IgG‐mediated complement‐dependent cytotoxicity (CDC). The screening of approximately 400 compounds yielded potent hit compounds with inhibitory effects against CDC in U87‐MG cells expressing human AQP4. Derivatives of the hit compounds were synthesized and evaluated for their inhibition of CDC. Of the small molecules synthesized, (E)‐1‐(2‐((4‐methoxyphenyl)sulfonyl)vinyl)‐[4‐[(3‐trifluoromethyl)phenyl] methoxy]benzene ( 5 c ) showed the most potent activity in both stably transfected U87‐MG cells and mice‐derived astrocytes. The results of this study suggest that 5 c , which targets NMO‐IgG‐specific CDC, may be useful as a research tool and a potential candidate for therapeutic development for the treatment of NMO.     

The In Vitro Cytotoxicity of Eremothecium oil and Its Components—Aromatic and Acyclic Monoterpene Alcohols

The microscopic fungi Eremothecium ashbyi and E. gossypii are known for their ability to synthetize essential oil, which has a composition similar to that of rose oil. The development of Eremothecium oil technology enables the production of rose-scented products, which are demanded by pharmaceutical, food, and perfumery industries. This study focuses on assessing the in vitro cytotoxicity of Eremothecium oil, in comparison with that of rose oil, using a combination of methods and two cell types (3T3 mouse fibroblast cell line and bone-marrow-derived mesenchymal stromal cells (BM-MSCs)). The Eremothecium oil samples possessed cytotoxic effects that varied among strains and batches. The revealed cytotoxicity level may be used to tailor the qualitative and quantitative composition of Eremothecium oil to achieve a particular quality in its end products. These results require further analysis using other cell types and assays based on measuring other cell functions.     

Phorbol 12-Myristate 13-Acetate Induced Toxicity Study and the Role of Tangeretin in Abrogating HIF-1α-NF-κB Crosstalk In Vitro and In Vivo

Phorbol 12-myristate 13-acetate (PMA) is a potent tumor promoter and highly inflammatory in nature. Here, we investigated the toxic effects of PMA on different model system. PMA (10 μg) caused chromosomal aberrations on the Allium cepa root tip and induced mitotic dysfunction. Similarly, PMA caused embryonic and larval deformities and a plummeted survivability rate on zebrafish embryo in a dose-dependent manner. Persistently, PMA treatment on immortalized human keratinocyte human keratinocyte (HaCaT) cells caused massive inflammatory rush at 4 h and a drop in cell survivability at 24 h. Concomitantly, we replicated a cutaneous inflammation similar to human psoriasis induced by PMA. Herein, we used tangeretin (TAN), as an antagonist to counteract the inflammatory response. Results from an in vivo experiment indicated that TAN (10 and 30 mg/kg) significantly inhibited PMA stimulated epidermal hyperplasia and intra-epidermal neutrophilic abscesses. In addition, its treatment effectively neutralized PMA induced elevated reactive oxygen species (ROS) generation on in vitro and in vivo systems, promoting antioxidant response. The association of hypoxia-inducible factor 1-alpha (HIF-1α)-nuclear factor kappa-light-chain-enhancer of activated b cells (NF-κB) crosstalk triggered by PMA enhanced PKCα-ERK1/2-NF-κB pathway; its activation was also significantly counteracted after TAN treatment. Conclusively, we demonstrated TAN inhibited the nuclear translocation of HIF-1α and NF-κB p65. Collectively, TAN treatment ameliorated PMA incited malignant inflammatory response by remodeling the cutaneous microenvironment.     

Long‐Term Properties of Injection‐Molded Micro‐Parts: Influence of Part Dimensions and Cooling Conditions on Aging Behavior

A variety of polymer parts used in microsystems technology is produced by injection molding. For dimensioning and design of these products, both the material properties and changes during the life cycle have to be taken into account. The aging behavior of parts with decreasing dimensions or of parts processed under different cooling conditions is discussed in terms of their morphological and mechanical long‐term properties. The results indicate that a decreasing part size leads to higher post‐crystallization and higher thermo‐oxidative degradation. This leads to size‐dependent changes in mechanical properties. Adjusted process conditions by application of thermal low‐conductive mold materials can favor the process‐related properties and thus the long term properties can be improved.