Adatom controlled emergence of high hardness in biocompatible beta-Ti3Au intermetallic thin film surfaces

There is growing international interest in hard biocompatible thin film surface coatings to extend the lifetime of medical implants. Parameters of the physical vapour deposition technique can be utilized to fine tune the microstructure and resulting properties of the growing thin film surface by modifying the adatom mobility of the incoming species. This work investigates the evolution of high hardness and biocompatibility of sputter deposited beta-Ti3Au intermetallic thin film surfaces as a function of growth temperature and pressure. Titanium and gold are sputtered in an optimised 3:1 ratio over glass and Ti6Al4V substrates at varying pressures of 0.3 to 1.2 Pa and temperatures of 25 to 450°C. The microstructure and crystallinity of the deposited films improved with reduction in pressure from 1.2 to 0.3 Pa but development of the β-Ti3Au intermetallic compound occurred at temperatures above 350˚C. The density of the films also increased with reducing pressure, whereas improvement in their columnar structure was observed with increasing substrate temperature. These microstructural changes caused by adatom mobility variation, led to the emergence of superior mechanical surface hardness, reaching a peak value of 12.5 GPa for films grown at 0.3 Pa and 450°C. All thin film surfaces were highly biocompatible with ion leaching levels below 1 ppm, and films deposited at lower pressure exhibited much safer cytotoxic profiles against L929 mouse fibroblasts. This work demonstrates the emergence of high hardness and biocompatibility in Ti3Au thin film surfaces with potential as next generation medical implant coating materials

Antioxidant and Antiproliferative Properties of the Essential Oils of Satureja thymbra and Satureja parnassica and their Major Constituents

Aim: The biopotential of the essential oils of the Greek aromatic plants Satureja thymbra and Satureja parnassica were investigated, together with their major components carvacrol, thymol, γ-terpinene and p-cymene. Materials and Methods: Antioxidant and cancer cell cytotoxic properties were determined using 2,2-diphenyl-1-picrylhydrazyl and sulforhodamine B assays, respectively. The antiproliferative potential was studied against the MCF-7, A549, HepG2 and Hep3B cell lines. Results: S. thymbra oil possessed stronger antioxidant and antiproliferative capacity when tested on MCF-7 cells compared to S. parnassica oil. Thymol exhibited two-fold greater antioxidant potency than carvacrol, whereas γ-terpinene and p-cymene had no significant effect. Carvacrol was the most potent antiproliferative agent against A549 cells, while Hep3B cells were most sensitive to thymol. p-Cymene and γ-terpinene demonstrated negligible bioactivity. Conclusion: S. thymbra and S. parnassica essential oils exhibit significant but diverse antioxidant and antiproliferative activities, mainly attributed to their main components, carvacrol and thymol

An Evaluation of the Anti-Carcinogenic Response of Major Isothiocyanates in Non-Metastatic and Metastatic Melanoma Cells

Malignant melanoma is one of the most deadly types of solid cancers, a property mainly attributed to its highly aggressive metastatic form. On the other hand, different classes of isothiocy- anates, a class of phytochemicals, present in cruciferous vegetables have been characterized by considerable anti-cancer activity in both in vitro and in vivo experimental models. In the current study, we investigated the anti-cancer response of five isothiocyanates in an in vitro model of melanoma consisting of non-metastatic (A375, B16F-10) and metastatic (VMM1, Hs294T) malignant melanoma as well as non-melanoma epidermoid carcinoma (A431) and non-tumorigenic melanocyte-neighboring keratinocyte (HaCaT) cells. Our aim was to compare different endpoints of cytotoxicity (e.g., reactive oxygen species, intracellular glutathione content, cell cycle growth arrest, apoptosis and necrosis) descriptive of an anti-cancer response between non-metastatic and metastatic melanoma as well as non-melanoma epidermoid carcinoma and non-tumorigenic cells. Our results showed that exposure to isothiocyanates induced an increase in intracellular reactive oxygen species and glutathione contents between non-metastatic and metastatic melanoma cells. The distribution of cell cycle phases followed a similar pattern in a manner where non-metastatic and metastatic melanoma cells appeared to be growth arrested at the G2/M phase while elevated levels of metastatic melanoma cells were shown to be at sub G1 phase, an indicator of necrotic cell death. Finally, metastatic melanoma cells were more sensitive apoptosis and/or necrosis as higher levels were observed compared to non-melanoma epidermoid carcinoma and non-tumorigenic cells. In general, non-mela- noma epidermoid carcinoma and non-tumorigenic cells were more resistant under any experimental exposure condition. Overall, our study provides further evidence for the potential development of isothiocyanates as promising anti-cancer against non-metastic and metastatic melanoma cells, a property specific for these cells and not shared by non-melanoma epidermoid carcinoma or non-tumorigenic melanocyte cells

Mechanical performance of biocompatible Ti-Au thin films grown on glass and Ti6Al4V substrates

Ti-Au intermetallic based material systems are being extensively studied to develop hard and wear resistant biocompatible thin film coatings over implant devices to extend their lifetime 1, 2. However, the measurement of these mechanical characteristics depends upon factors such as surface properties of the substrates and their temperature during thin film deposition. In this work, Ti-Au thin films were deposited by magnetron sputtering on both glass and Ti6Al4V substrates at two different temperatures. These films were studied for their mechanical properties by the nanoindentation technique in both load control and displacement control modes using a Berkovich tip. XRD patterns and cross section SEM images detail the microstructure while AFM images present the surface morphology of these Ti-Au thin films. Biocompatibility of the films is verified by cytotoxicity tests on L929 mouse fibroblast cells using Alamar blue reagent and the ions leaching in the film extracts is measured using the ICPOEMS technique. Standard deviation for hardness of films on glass substrates is ~4 times lower than that on Ti6Al4V substrates and is corelated to a corresponding increase in surface roughness from 2nm for glass to 40nm for Ti6Al4V substrates 3. Increasing substrate temperature leads to an increase in film hardness from 5.1 to 8.9GPa and is related to the development of a super hard β phase of the Ti3Au intermetallic. The standard deviation of this peak mechanical hardness value of 8.9GPa is reduced by 3 times when measured in displacement control mode compared to the value measured in load control mode due of the effect of nanoindentation tip penetration depth. All the Ti-Au thin films exhibit excellent cytotoxicity values above 95% and ion leaching below 100ppb. This work presents a comparative study to optimize hardness measurement of Ti-Au thin films, critical for a better understanding of these super hard biocompatible coatings

Evaluation of Bioactive Properties of Lipophilic Fractions of Edible and Non-Edible Parts of \u3ci\u3eNasturtium officinale\u3c/i\u3e (Watercress) in a Model of Human Malignant Melanoma Cells

Watercress is an enriched source of phenethyl isothiocyanate (PEITC), among other phytochemicals, with an antioxidant capacity. The aim of this study was to (i) chemically characterize and (ii) biologically evaluate the profile of the main health-promoting compounds contained in edible (i.e., mixture of leaves and lateral buds) and non-edible (i.e., stems) parts of watercress in an in vitro model of malignant melanoma consisting of human malignant melanoma (A375), non-melanoma (A431) and keratinocyte (HaCaT) cells. The extraction of the main constituents of watercress was performed by subjecting the freeze-dried edible and non-edible samples through different extraction protocols, whereas their concentration was obtained utilizing analytical methodologies. In addition, cell viability was evaluated by the Alamar Blue assay, whereas levels of oxidative stress and apoptosis were determined by commercially available kits. The edible watercress sample contained a higher amount of various nutrients and phytochemicals in the hexane fraction compared to the non-edible one, as evidenced by the presence of PEITC, phenolics, flavonoids, pigments, ascorbic acid, etc. The cytotoxicity potential of the edible watercress sample in the hexane fraction was considerably higher than the non-edible one in A375 cells, whereas A431 and HaCaT cells appeared to be either more resistant or minimally affected, respectively. Finally, levels of oxidative stress and apoptotic induction were increased in both watercress samples, but the magnitude of the induction was much higher in the edible than the non-edible watercress samples. Herein, we provide further evidence documenting the potential development of watercress extracts (including watercress waste by-products) as promising anti-cancer agent(s) against malignant melanoma cells

Chemical and Biological Characterization of the Anticancer Potency of \u3ci\u3eSalvia fruticosa\u3c/i\u3e in a Model of Human Malignant Melanoma

Malignant melanoma is one of the most aggressive types of skin cancer with an increasing incidence worldwide. Thus, the development of innovative therapeutic approaches is of great importance. Salvia fruticosa (SF) is known for its anticancer properties and in this context, we aimed to investigate its potential anti-melanoma activity in an in vitro model of human malignant melanoma. Cytotoxicity was assessed through a colorimetric-based sulforhodamine-B (SRB) assay in primary malignant melanoma (A375), non-malignant melanoma epidermoid carcinoma (A431) and non-tumorigenic melanocyte neighbouring keratinocyte (HaCaT) cells. Among eight (8) different fractions of S. fruticosa extracts (SF1-SF8) tested, SF3 was found to possess significant cytotoxic activity against A375 cells, while A431 and HaCaT cells remained relatively resistant or exerted no cytotoxicity, respectively. In addition, the total phenolic (Folin–Ciocalteu assay) and total flavonoid content of SF extracts was estimated, whereas the antioxidant capacity was measured via the inhibition of tert-butyl hydroperoxide-induced lipid peroxidation and protein oxidation levels. Finally, apoptotic cell death was assessed by utilizing a commercially available kit for the activation of caspases - 3, - 8 and - 9. In conclusion, the anti-melanoma properties of SF3 involve the induction of both extrinsic and intrinsic apoptotic pathway(s), as evidenced by the increased activity levels of caspases - 8, and - 9, respectively

Marine-Derived Surface Active Agents: Health-Promoting Properties and Blue Biotechnology-Based Applications

Surface active agents are characterized for their capacity to adsorb to fluid and solid-water interfaces. They can be classified as surfactants and emulsifiers based on their molecular weight (MW) and properties. Over the years, the chemical surfactant industry has been rapidly increasing to meet consumer demands. Consequently, such a boost has led to the search for more sustainable and biodegradable alternatives, as chemical surfactants are non-biodegradable, thus causing an adverse effect on the environment. To these ends, many microbial and/or marine-derived molecules have been shown to possess various biological properties that could allow manufacturers to make additional health-promoting claims for their products. Our aim, in this review article, is to provide up to date information of critical health-promoting properties of these molecules and their use in blue-based biotechnology (i.e., biotechnology using aquatic organisms) with a focus on food, cosmetic and pharmaceutical/biomedical applications

Toxicity Profiling of Biosurfactants Produced by Novel Marine Bacterial Strains

Surface active agents (SAAs), currently used in modern industry, are synthetic chemicals produced from non-renewable sources, with potential toxic impacts on humans and the environment. Thus, there is an increased interest for the identification and utilization of natural derived SAAs. As such, the marine environment is considered a promising source of biosurfactants with low toxicity, environmental compatibility, and biodegradation compared to their synthetic counterparts. MARISURF is a Horizon 2020 EU-funded project aiming to identify and functionally characterize SAAs, derived from a unique marine bacterial collection, towards commercial exploitation. Specifically, rhamnolipids produced by Marinobacter MCTG107b and Pseudomonas MCTG214(3b1) strains were previously identified and characterized while currently their toxicity profile was assessed by utilizing well-established methodologies. Our results showed a lack of cytotoxicity in in vitro models of human skin and liver as indicated by alamar blue and propidium iodide assays. Additionally, the use of the single gel electrophoresis assay, under oxidative stress conditions, revealed absence of any significant mutagenic/anti-mutagenic potential. Finally, both 2,2’-azino-bis (3-ethylbenzothiazoline-6-sulphonicacid) (ABTS) and 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) cell-free assays, revealed no significant anti-oxidant capacity for neither of the tested compounds. Consequently, the absence of significant cytotoxicity and/or mutagenicity justifies their commercial exploitation and potential development into industrial end-user applications as natural and environmentally friendly biosurfactants