Deciphering the Broad Antimicrobial Activity of Melaleuca alternifolia Tea Tree Oil by Combining Experimental and Computational Investigations.

Tea Tree Oil (TTO) is an essential oil obtained from the distillation of Melaleuca alternifolia leaves and branches. Due to its beneficial properties, TTO is widely used as an active ingredient in antimicrobial preparations for topical use or in cosmetic products and contains about 100 different compounds, with terpinen-4-ol, γ-terpinene and 1,8-cineole (or eucalyptol) being the molecules most responsible for its biological activities. In this work, the antimicrobial activity of whole TTO and these three major components was evaluated in vitro against fungi, bacteria and viruses. Molecular dynamics simulations were carried out on a bacterial membrane model and a Coxsackievirus B4 viral capsid, to propose an atomistic explanation of their mechanism of action. The obtained results indicate that the strong antimicrobial activity of TTO is attributable to the induction of an altered membrane functionality, mediated by the incorporation of its components within the lipid bilayer, and to a possible ability of the compounds to bind and alter the structural properties of the viral capsid.

Oxidative Properties of Polystyrene Nanoparticles with Different Diameters in Human Peripheral Blood Mononuclear Cells (In Vitro Study).

With the ongoing commercialization, human exposure to plastic nanoparticles will dramatically increase, and evaluation of their potential toxicity is essential. There is an ongoing discussion on the human health effects induced by plastic particles. For this reason, in our work, we assessed the effect of polystyrene nanoparticles (PS-NPs) of various diameters (29, 44 and 72 nm) on selected parameters of oxidative stress and the viability of human peripheral blood mononuclear cells (PBMCs) in the in vitro system. Cells were incubated with PS-NPs for 24 h in the concentration range of 0.001 to 100 µg/mL and then labeled: formation of reactive oxygen species (ROS) (including hydroxyl radical), protein and lipid oxidation and cell viability. We showed that PS-NPs disturbed the redox balance in PBMCs. They increased ROS levels and induced lipid and protein oxidation, and, finally, the tested nanoparticles induced a decrease in PBMCs viability. The earliest changes in the PBMCs were observed in cells incubated with the smallest PS-NPs, at a concentration of 0.01 µg/mL. A comparison of the action of the studied nanoparticles showed that PS-NPs (29 nm) exhibited a stronger oxidative potential in PBMCs. We concluded that the toxicity and oxidative properties of the PS-NPs examined depended to significant degree on their diameter.

Lanthanum-containing bioparticles are associated with the influence of lanthanum on high phosphate mediated bone marrow stromal cells viability.

Lanthanum (La)-based binder appears effective in reducing serum inorganic phosphate (Pi) levels among chronic dialysis patients, yet concern remains about La accumulation in bone during long-term oral administration. In this study, the effect of lanthanum chloride (LaCl3) on bone marrow stromal cells (BMSCs) viability was investigated under high Pi situation. We found low concentration (10-9 M) of LaCl3 increased BMSCs viability, while high concentration (10-5 M) of LaCl3 not only inhibited BMSCs viability but also stimulated high Pi induced cell apoptosis. In addition, La-containing calcium phosphate (CaP) particles can be detected on cell surfaces and inside cells. We also found that inhibition of CaP formation by adding sodium pyrophosphate, a recognized inhibitor of hydroxyapatite formation, abrogated LaCl3 induced the BMSCs viability. For isolated La-containing CaP particles, the particle size increased and crystal phase switched with elevated concentration of LaCl3. These results demonstrated that La-containing CaP particles were associated with the process of LaCl3 mediated BMSCs viability and the physicochemical properties of these particles played an important role in modulating BMSCs function.

Evaluation of biological activity of quantum dots in a microsystem.

The presented work aimed at systematic investigation of biological activity of CdSex S1- x /ZnS and CdSe/ZnS quantum dots (QDs), whose surface was modified with different ligands. For these studies, we used a microfluidic system combined with fluorescence microscopy techniques, which enabled analysis of cells' morphology, viability, and QDs uptake. PDMS and glass-based microfluidic system enabled the precise control of the cell environment, allowed to examine five replications of each tested QDs concentrations (statistically significant number), monitor multiple cellular events, and avoid manual preparation of QDs dilutions. We investigated the influence of the core composition and the type of surface modifiers on QDs toxicity. We also determined whether the examined nanoparticles penetrate into the cells. For all tested nanoparticles, the decrease of cells' viability was observed when increasing nanoparticles concentration. The decrease of live cells' number in microchambers and the accumulation of the nanoparticles around cultured cells were observed. The effect of hydrocarbon chain length of surface modifiers and QDs core composition on the cell viability was confirmed in our tests.

Cytotoxicity and Antibiofilm Activity of Silver-Polypropylene Nanocomposites.

The development of biocompatible nanomaterials that interface with human skin and tissue is critical for advancing prosthetics and other therapeutic medical needs. In this perspective, the development of nanoparticles with cytotoxicity and antibiofilm properties and biocompatibility characteristics are important. Metallic silver (Ag) exhibits good biocompatibility, but it is often challenging to integrate it into a nanocomposite without compromising its antibiofilm properties for optimal applications. In this study, new polymer nanocomposites (PNCs) with ultra-low filling content (0.0023-0.046 wt%) of Ag nanoplates were manufactured and tested. The cytotoxicity and antibiofilm activity of different composites with polypropylene (PP) matrix were examined. At first, PNCs surface were analyzed by means of AFM (atomic force microscopy) with phase contrast evaluation and FTIR (Fourier-transform infrared spectroscopy) to study the Ag nanoplates distribution. Subsequently, the cytotoxicity and growth properties of biofilms were assessed by MTT assay protocol and detection of nitric oxide radicals. Antibacterial and antibiofilm activities were measured against Gram-positive bacteria (Staphylococcus aureus) and Gram-negative bacteria (K. pneumoniae). The PNCs with silver exhibited antibiofilm activity although they did not inhibit regular planktonic bacterial growth. Moreover, the PNCs were not cytotoxic to mammalian cells and did not induce significant immune response. These features reveal the potential of the PNCs developed in this study for usage in fabrication of prosthetics and other smart structures for biomedical applications.

Assessment of TiO2 Nanoparticle Impact on Surface Morphology of Chinese Hamster Ovary Cells.

The process of nanoparticles entering the cells of living organisms is an important step in understanding the influence of nanoparticles on biological processes. The interaction of nanoparticles with the cell membrane is the first step in the penetration of nanoparticles into cells; however, the penetration mechanism is not yet fully understood. This work reported the study of the interaction between TiO2 nanoparticles (TiO2-NPs) and Chinese hamster ovary (CHO) cells using an in vitro model. The characterization of crystalline phases of TiO2 NPs was evaluated by transmission electron microscopy (TEM), X-ray diffraction (XRD) spectrum, and atomic force microscopy (AFM). Interaction of these TiO2 nanoparticles (TiO2- NPs) with the CHO cell membrane was investigated using atomic force microscopy (AFM) and Raman spectroscopy. The XRD analysis result showed that the structure of the TiO2 particles was in the rutile phase with a crystallite size of 60 nm, while the AFM result showed that the particle size distribution had two peaks with 12.1 nm and 60.5 nm. The TEM analysis confirmed the rutile phase of TiO2 powder. Our study showed that exposure of CHO cells to TiO2-NPs caused morphological changes in the cell membranes and influenced the viability of cells. The TiO2-NPs impacted the cell membrane surface; images obtained by AFM revealed an 'ultra structure' with increased roughness and pits on the surface of the membrane. The depth of the pits varied in the range of 40-80 nm. The maximal depth of the pits after the treatment with TiO2-NPs was 100% higher than the control values. It is assumed that these pits were caveolae participating in the endocytosis of TiO2-NPs. The research results suggest that the higher maximal depth of the pits after the exposure of TiO2-NPs was determined by the interaction of these TiO2-NPs with the cell's plasma membrane. Moreover, some of pits may have been due to plasma membrane damage (hole) caused by the interaction of TiO2-NPs with membrane constituents. The analysis of AFM images demonstrated that the membrane roughness was increased with exposure time of the cells to TiO2-NPs dose. The average roughness after the treatment for 60 min with TiO2-NPs increased from 40 nm to 78 nm. The investigation of the membrane by Raman spectroscopy enabled us to conclude that TiO2-NPs interacted with cell proteins, modified their conformation, and potentially influenced the structural damage of the plasma membrane.

Antibacterial and anticancer activity of ZnO with different morphologies: a comparative study.

ZnO nanoparticles (NPS) with different morphologies were synthesized, and the antibacterial and anticancer activity was studied, herein. The physicochemical characterization was carried out by X-ray diffraction (XRD), transmission electron microscopy (TEM), Fourier-transform infrared spectroscopy (FTIR) and UV-visible. To study the antibacterial and anticancer capability of ZnO NPS, Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) bacteria and HeLa cancer cells were exposed at different doses of ZnO NPS (7-250 µg/mL). TEM analysis confirmed the obtention of spherical, hexagonal and rod ZnO NPS with an average diameter of 20 ± 4 nm, 1.17 ± 0.3 µm and 1.11 ± 1.2 µm, respectively. XRD diffractograms showed the characteristic pattern of crystalline ZnO in wurtzite phase. FTIR and UV-vis spectra showed slight differences of the main absorption peaks, revealing that different ZnO NPS morphologies may cause shifts in spectra. Biological essays showed that the number of E. coli and S. aureus bacteria as well as HeLa cells decreases linearly by increasing the nanoparticle concentration. However, the best anticancer and antibacterial activity was shown by spherical ZnO NPS at 100 µg/mL. The better capability of spherical ZnO NPS than hexagonal and rod ZnO NPS is related with its small particle size. The present results suggest that the spherical ZnO NPS has a great potential as an antibacterial and anticancer agent.

Thymoquinone, an Active Compound of Nigella sativa: Role in Prevention and Treatment of Cancer.

BACKGROUND: Cancer is the leading cause of death worldwide and the current mode of cancer treatment causes side effects on normal cells and are still the key challenges in its' treatment. However, natural products or active compounds of medicinal plants have shown to be safe, affordable, and effective in diseases cure. METHODS: In this context, scientific studies evidence the health-promoting effects of natural products, which work through its anti-oxidant, anti-inflammatory, and anti-cancer activity. Thymoquinone (TM), a predominant active compound of Nigella sativa, has confirmed anti-neoplastic activity through its ability to regulate various genetic pathways. In addition, thymoquinone has established anti-cancerous effects through killing of various cancerous cells,and inhibiting the initiation, migration, invasion, and progression of the cancer. The anti-cancer effects of TM are chiefly mediated via regulating various cell signaling pathways such as VEGF, bcl2/bax ratio, p53, NF-kB, and oncogenes. RESULTS: The anti-cancer drugs have limitations in efficacy and also causes adverse side effects on normal cells. The combination of anti-cancer drugs and thymoquinone improves the efficacy of drugs which is evident by decrease resistance to drugs and regulation of various cell signaling pathways. Moreover, combination of anti-cancer drugs as well as thymoquinone shows synergistic effect on killing of cancer cells and cells viability. Thus, TM, in combination with anti-cancer drugs, can be a good strategy in the management of various types of cancer. CONCLUSION: In this review article, we deliver an outline of thymoquinone role in cancer inhibition and prevention of cancer-based on in vivo and in vitro studies. Further studies on thymoquinone based on clinical trials are highly required to explore the benefits of thymoquinone in cancer management.

The study of the interaction mechanism between bovine serum albumin and single-walled carbon nanotubes depending on their diameter and concentration in solid nanocomposites by vibrational spectroscopy.

The results of the study of composites based on bovine serum albumin (BSA) and single-walled carbon nanotubes (SWCNT) are presented. Nanocomposites were created by evaporation of the water-albumin dispersion with nanotubes using diode laser with temperature control. Two types of nanotubes were used. SWCNT I were synthesized using the electric arc method, SWCNT II were synthesized using the gas phase method. SWCNT I had a diameter and length less than SWCNT II. The mechanism of interaction between BSA and SWCNT in solid nanocomposites is considered. An experimental and theoretical studies of the interaction between aspartic (Asp) and glutamic (Glu) amino acids located on the outer surface of BSA and nanotubes using of vibrational spectroscopy (Fourier-transform infrared (FTIR) and Raman spectroscopy) was carried out. The possibility of nanotubes functionalization by oxygen atoms of negative amino acid residues Asp and Glu, which are on the outer surface of BSA, is shown by molecular modeling. The formation of covalent bonds between BSA and SWCNT in nanocomposites with different concentrations of nanotubes (0.01, 0.1 and 1 g/l) was confirmed by vibrational spectra. The covalent interaction between BSA with SWCNT under the laser irradiation leads to the conformational changes in the secondary and tertiary structures of albumin. This is confirmed by a significant decrease in the intensity of the absorption bands in the high-frequency region. The calculation of the vibrational spectra of the three Glycine:Glycine, Glutamic acid:Threonine and Aspartic acid:Lysine complexes, which take into account hydrogen, ion-dipole and ion-ion bonds, showed that a disturbance in the intermolecular interaction between amino acid residues led to significant decrease in the intensity of absorption bands in the region of stretching vibrations bonds OH and NH. From the Raman spectra, it was found that a significant number of defects in SWCNT is caused by the covalent attachment of oxygen atoms to the graphene surface of nanotubes. An increase in the diameter of nanotubes (4 nm) has practically no effect on the absorption spectrum of nanocomposite, while measuring the concentration of SWCNT affects the FTIR spectra. This confirmed the hydrophobic interaction between BSA and SWCNT. Thus, it was shown that BSA solid nanocomposites with CNTs can interact either with the help of hydrophobic forces or with the formation of covalent bonds, which depends on the diameter of the used nanotubes. The viability of connective fibroblast tissue cells on nanocomposites with both types of SWCNT was demonstrated. It was found that nanocomposites based on SWCNT I provide slightly better compatibility of their structure with fibroblasts. It allows to achieve better cell adhesion to the nanocomposite surface. These criteria make extensive use of scaffold nanocomposites in biomedicine, depending on the requirements for their quality and application.

Gelatin-Coated Microfluidic Channels for 3D Microtissue Formation: On-Chip Production and Characterization.

Traditional two-dimensional (2D) cell culture models are limited in their ability to reproduce human structures and functions. On the contrary, three-dimensional (3D) microtissues have the potential to permit the development of new cell-based assays as advanced in vitro models to test new drugs. Here, we report the use of a dehydrated gelatin film to promote tumor cells aggregation and 3D microtissue formation. The simple and stable gelatin coating represents an alternative to conventional and expensive materials like type I collagen, hyaluronic acid, or matrigel. The gelatin coating is biocompatible with several culture formats including microfluidic chips, as well as standard micro-well plates. It also enables long-term 3D cell culture and in situ monitoring of live/dead assays.

ZnO Nanostructures and Electrospun ZnO-Polymeric Hybrid Nanomaterials in Biomedical, Health, and Sustainability Applications.

ZnO-based nanomaterials are a subject of increasing interest within current research, because of their multifunctional properties, such as piezoelectricity, semi-conductivity, ultraviolet absorption, optical transparency, and photoluminescence, as well as their low toxicity, biodegradability, low cost, and versatility in achieving diverse shapes. Among the numerous fields of application, the use of nanostructured ZnO is increasingly widespread also in the biomedical and healthcare sectors, thanks to its antiseptic and antibacterial properties, role as a promoter in tissue regeneration, selectivity for specific cell lines, and drug delivery function, as well as its electrochemical and optical properties, which make it a good candidate for biomedical applications. Because of its growing use, understanding the toxicity of ZnO nanomaterials and their interaction with biological systems is crucial for manufacturing relevant engineering materials. In the last few years, ZnO nanostructures were also used to functionalize polymer matrices to produce hybrid composite materials with new properties. Among the numerous manufacturing methods, electrospinning is becoming a mainstream technique for the production of scaffolds and mats made of polymeric and metal-oxide nanofibers. In this review, we focus on toxicological aspects and recent developments in the use of ZnO-based nanomaterials for biomedical, healthcare, and sustainability applications, either alone or loaded inside polymeric matrices to make electrospun composite nanomaterials. Bibliographic data were compared and analyzed with the aim of giving homogeneity to the results and highlighting reference trends useful for obtaining a fresh perspective about the toxicity of ZnO nanostructures and their underlying mechanisms for the materials and engineering community.

Assessment of cell viability in four novel endodontic sealers.

OBJECTIVE: The aim of this study was to evaluate the viability of human periodontal ligament (PDL) cells on MTA-Fillapex, GuttaFlow 2, TotalFill Sealer, and BioRoot™ RCS in comparison to conventional epoxy resin-based (AH Plus) and zinc oxide-eugenol-based (Roth's 801) sealers. MATERIALS AND METHODS: Sealers were divided into two groups, and five coverslips for each material per group were prepared. In the first group, PDLs were added immediately after the preparation of sealers (Fresh Group), and in the second, PDLs were added after 24 h. PDLs were cultured for 72 h and afterward, counted using standard hematocytometry. A Mann-Whitney U-test and Kruskal-Wallis test were used for the statistical analysis. The level of significance was set at 5%. Furthermore, cell morphology was assessed by confocal microscopy. RESULTS: The number of viable cells for the 24 h-set groups was higher than the freshly mixed in all sealers except Roth's 801. In both groups, GuttaFlow 2 presented the highest number of viable cells. In a descending order of cells' survival, TotalFill, BioRoot, and MTA-Fillapex are following and the conventional sealers, AH Plus and Roth's 801, seem not to exhibit the biological properties of the others. Cells grown on GuttaFlow 2, TotalFill, and BioRoot were observed to be well-formed. In contrast, MTA-Fillapex exhibited untypical morphology. No cells were detected on the surfaces of AH Plus, as well as Roth's 801. CONCLUSIONS: All novel sealers presented increased cell viability in comparison to conventional sealers. GuttaFlow 2 exhibited the highest cell viability.

Enhancing Saccharomyces cerevisiae reactive oxygen species and ethanol stress tolerance for high-level production of protopanoxadiol.

Protopanaxadiol (PPD) is an active compound in Panax ginseng. Recently, an optimized PPD synthesis pathway contained a ROS releasing step (a P450-type PPD synthase, PPDS) was introduced into Saccharomyces cerevisiae. Here reported a synergistic effect of PPDS-CPR (CPR, cytochrome P450 reductase) uncoupling and ethanol stress on ROS releasing, which reduced cells viability. To build a robust strain, a cell wall integrity associated gene SSD1 was high-expressed to improve ethanol tolerance, and ROS level decreased for 24.7%. Then, regulating the expression of an oxidative stress regulation gene YBP1 decreased 75.2% of ROS releasing, and improved cells viability from 71.3±1.3% to 88.3±1.4% at 84h. Increased cells viability enables yeast to produce more PPD through feeding additional ethanol. In 5L fermenter, PPD production of W3a-ssPy reached to 4.25±0.18g/L (19.48±0.28mg/L/OD600), which is the highest yield reported so far. This work makes the industrial production of PPD possible by microbial fermentation.

The effect of polymeric membrane surface on HaCaT cell properties.

The control of the surface properties is an important issue for applicability of polymer membranes interacting with cells. In this work, the influence of surface roughness and stiffness of two polymer membranes on viability and mechanical properties of keratinocytes was studied. Terpolimer polyglicolide, polycaprolactone and polylactide, (PGA-PCL-PLA) and copolymer polycaprolactone, polyglicolide (PGA-PCL) substrates were used for membranes fabrication. Surface modification - the hydrolysis of the obtained membranes was carried out. The analysis of membranes' surface properties revealed that RMS surface roughness and roughness factor of PGA-PCL-PLA membrane decreased after hydrolysis while its stiffness increased. In contrast, the PGA-PCL membrane stiffness was only slightly affected by NaOH treatment. Immortalized human keratinocytes (HaCaT) were grown under standard conditions on the surface of the studied membranes and characterized by means of atomic force microscopy and fluorescence microcopy. The results showed the substrate-dependent effect on cells' properties.

Microfluidic Synthesis and Biological Evaluation of Photothermal Biodegradable Copper Sulfide Nanoparticles.

The continuous synthesis of biodegradable photothermal copper sulfide nanoparticles has been carried out with the aid of a microfluidic platform. A comparative physicochemical characterization of the resulting products from the microreactor and from a conventional batch reactor has been performed. The microreactor is able to operate in a continuous manner and with a 4-fold reduction in the synthesis times compared to that of the conventional batch reactor producing nanoparticles with the same physicochemical requirements. Biodegradation subproducts obtained under simulated physiological conditions have been identified, and a complete cytotoxicological analysis on different cell lines was performed. The photothermal effect of those nanomaterials has been demonstrated in vitro as well as their ability to generate reactive oxygen species.

Luffa echinata Roxb. Induced Apoptosis in Human Colon Cancer Cell (SW-480) in the Caspase-dependent Manner and Through a Mitochondrial Apoptosis Pathway.

BACKGROUND: Luffa echinata Roxb. (LER) (Cucurbitaceae) showed tremendous medicinal importance and are being used for the treatment of different ailments. OBJECTIVE: In this study, the antiproliferative properties and cell death mechanism induced by the extract of the fruits of LER were investigated. MATERIALS AND METHODS: MTT and LDH assay were used to test the antiproliferative and cytotoxicity of LER extract, respectively. The intracellular ROS were measured by a fluorometric assay. The expression of several apoptotic-related proteins in SW-480 cells treated by LER was evaluated by Western blot analysis. RESULTS: The methanolic extract of LER fruits inhibited the proliferation of human colon cancer cells (SW-480) in both dose- and time-dependent manners. The LER-treated cells showed obvious characteristics of cell apoptosis, including cell shrinkage, destruction of the monolayer, and condensed chromatin. In addition, treatments of various concentrations of LER extracts caused the release of lactate dehydrogenase as a dose-dependent manner via stimulation of the intracellular metabolic system. LER induced apoptosis, DNA fragmentation, and cellular ROS accumulation in SW-480 cells. Treatment of LER on SW-480 cells promoted the expression of caspases, Bax, Bad, and p53 proteins and decreased the levels of Bcl-2 and Bcl-XL. CONCLUSIONS: These results indicated that treatment with LER-induced cell death in mitochondrial apoptosis pathway by regulating pro-apoptotic proteins via the up regulation of the p53 protein. These findings highlight the potentials of LER in the treatment of human colon cancer. SUMMARY: LER induced apoptosis, DNA fragmentation, and cellular ROS accumulation in SW-480 cells. Treatment of LER on SW-480 cells promoted the expression of caspases, Bax, Bad, and p53 proteins and decreased the levels of Bcl-2 and Bcl-XL.

Cytotoxicity of TiO2 nanoparticles to mussel hemocytes and gill cells in vitro: Influence of synthesis method, crystalline structure, size and additive.

Increasing the production and applications of TiO2 nanoparticles (NPs) has led to grow concerns about the consequences for the environment. In this study, we investigated the effects of a set of TiO2 NPs on the viability of mussel hemocytes and gill cells using neutral red and thiazolyl tetrazolium bromide assays. For this, we compared the cytotoxicity of TiO2 NPs (0.1-100 mg Ti/L) produced by different techniques: rutile NPs (60 nm) produced by milling and containing disodium laureth sulfosuccinate (DSLS), rutile NPs (10, 40 and 60 nm) produced by wet chemistry and anatase/rutile NPs (∼100 nm) produced by plasma synthesis. The commercially available P25 anatase/rutile NPs (10-20 nm) were also tested. Exposures were performed in parallel with their respective bulk forms and the cytotoxicity of the additive DSLS was also tested. Z potential values in distilled water indicated different stabilities depending on the NP type and all NPs tested formed agglomerates/aggregates in cell culture media. In general, TiO2 NPs showed a relatively low and dose-dependent toxicity for both cell models with the two assays tested. NPs produced by milling showed the highest effects, probably due to the toxicity of DSLS. Size-dependent toxicity was found for NPs produced by wet chemistry (10 nm > 40 nm and 60 nm). All TiO2 NPs tested were more toxic than bulk forms excepting for plasma produced ones, which were the least toxic TiO2 tested. The mixture bulk anatase/rutile TiO2 was more toxic than bulk rutile TiO2. In conclusion, the toxicity of TiO2 NPs varied with the mode of synthesis, crystalline structure and size of NPs and can also be influenced by the presence of additives in the suspensions.