Biological Risk Assessment of Three Dental Composite Materials following Gas Plasma Exposure

Gas plasma is an approved technology that generates a plethora of reactive oxygen species, which are actively applied for chronic wound healing. Its particular antimicrobial action has spurred interest in other medical fields, such as periodontitis in dentistry. Recent work has indicated the possibility of performing gas plasma-mediated biofilm removal on teeth. Teeth frequently contain restoration materials for filling cavities, e.g., resin-based composites. However, it is unknown if such materials are altered upon gas plasma exposure. To this end, we generated a new in-house workflow for three commonly used resin-based composites following gas plasma treatment and incubated the material with human HaCaT keratinocytes in vitro. Cytotoxicity was investigated by metabolic activity analysis, flow cytometry, and quantitative high-content fluorescence imaging. The inflammatory consequences were assessed using quantitative analysis of 13 different chemokines and cytokines in the culture supernatants. Hydrogen peroxide served as the control condition. A modest but significant cytotoxic effect was observed in the metabolic activity and viability after plasma treatment for all three composites. This was only partially treatment time-dependent and the composites alone affected the cells to some extent, as evident by differential secretion profiles of VEGF, for example. Gas plasma composite modification markedly elevated the secretion of IL6, IL8, IL18, and CCL2, with the latter showing the highest correlation with treatment time (Pearson’s r > 0.95). Cell culture media incubated with gas plasma-treated composite chips and added to cells thereafter could not replicate the effects, pointing to the potential that surface modifications elicited the findings. In conclusion, our data suggest that gas plasma treatment modifies composite material surfaces to a certain extent, leading to measurable but overall modest biological effects.     

Singlet-Oxygen-Induced Phospholipase A2 Inhibition: A Major Role for Interfacial Tryptophan Dioxidation

Several studies have revealed that various diseases such as cancer have been associated with elevated phospholipase A₂ (PLA₂) activity. Therefore, the regulation of PLA₂ catalytic activity is undoubtedly vital. In this study, effective inactivation of PLA₂ due to reactive species produced from cold physical plasma as a source to model oxidative stress is reported. We found singlet oxygen to be the most relevant active agent in PLA₂ inhibition. A more detailed analysis of the plasma-treated PLA₂ identified tryptophan 128 as a hot spot, rich in double oxidation. The significant dioxidation of this interfacial tryptophan resulted in an N-formylkynurenine product via the oxidative opening of the tryptophan indole ring. Molecular dynamics simulation indicated that the efficient interactions between the tryptophan residue and phospholipids are eliminated following tryptophan dioxidation. As interfacial tryptophan residues are predominantly involved in the attaching of membrane enzymes to the bilayers, tryptophan dioxidation and indole ring opening leads to the loss of essential interactions for enzyme binding and, consequently, enzyme inactivation.     

Electroactivity of Superoxide Anion in Aqueous Phosphate Buffers Analyzed with Platinized Microelectrodes

The reactivity of platinized ultramicroelectrodes (Pt-black UMEs) towards superoxide anion O₂^.−, an unstable Reactive Oxygen Species (ROS), and its relatives, H₂O₂ and O₂, was studied. Voltammetric studies in PBS demonstrate that Pt-black UMEs provide: i) a well-resolved reversible redox signature for O₂^.− detected in both alkaline and physiological buffers (pH 12 and 7.4); ii) irreversible oxidation and reduction waves for H₂O₂ at pH 7.4. The oxygen reduction reaction (ORR) at Pt-black surfaces solely yields H₂O₂ (2 electrons/2 H⁺) at physiological pH. Consequently, Pt-black UMEs allow to sense different ROS including superoxide anion for future biomedical or physico-chemical investigations.     

微流控芯片毛细管电泳法检测细胞内活性氧与细胞凋亡关系

建立了微流控芯片毛细管电泳激光诱导荧光同时测定细胞内活性氧和凋亡信号的方法。先用AlexaFluor488 annexin V细胞凋亡试剂盒标记细胞凋亡的外翻磷脂酰丝氨酸,再用双氢罗丹明123(DHR123)标记细胞内活性氧,用PBS将细胞调整为终密度1.2×106cells/mL的细胞悬液。细胞群经反复冻融法破碎后,以20 mmol/L硼砂(pH 9.2)作电泳缓冲溶液,分离电压1.2 kV,进样时间60 s,1 min内可完成活性氧和细胞凋亡信号的同时测定。方法简单、快速,细胞内活性氧和DHR123的反应产物(Rh123)在0.5～3μmol/L浓度范围内线性关系良好,相关系数(r)为0.998,检出限(S/N=3)为0.058μmol/L,可用于细胞内活性氧的定量分析。测得HepG2肝癌细胞活性氧含量为0.16μmol/L,被阿霉素诱导凋亡后,细胞内活性氧含量升高至1.77μmol/L。     

Biogenic Silver Nanoparticles as a Stress Alleviator in Plants: A Mechanistic Overview

Currently, the growth and yield of crops are restrained due to an increase in the occurrence of ecological stresses globally. Biogenic generation of nanomaterials is an important step in the development of environmentally friendly procedures in the nanotechnology field. Silver-based nanomaterials are significant because of their physical, chemical, and biological features along with their plentiful applications. In addition to useful microbes, the green synthesized Ag nanomaterials are considered to be an ecologically friendly and environmentally biocompatible method for the enhancement of crop yield by easing stresses. In the recent decade, due to regular droughts, infrequent precipitation, salinity, and increased temperature, the climate alternation has changed certain ecological systems. As a result of these environmental changes, crop yield has decreased worldwide. The role of biogenic Ag nanomaterials in enhancing methylglyoxal detoxification, antioxidant defense mechanisms, and generating tolerance to stresses-induced ROS injury has been methodically explained in plants over the past ten years. However, certain studies regarding stress tolerance and metal-based nanomaterials have been directed, but the particulars of silver nanomaterials arbitrated stresses tolerance have not been well-reviewed. Henceforth, there is a need to have a good understanding of plant responses during stressful conditions and to practice the combined literature to enhance tolerance for crops by utilization of Ag nanoparticles. This review article illustrates the mechanistic approach that biogenic Ag nanomaterials in plants adopt to alleviate stresses. Moreover, we have appraised the most significant activities by exogenous use of Ag nanomaterials for improving plant tolerance to salt, low and high temperature, and drought stresses.     

Redox Reactions of Biologically Active Molecules upon Cold Atmospheric Pressure Plasma Treatment of Aqueous Solutions

Cold atmospheric pressure plasma (CAPP) is widely used in medicine for the treatment of diseases and disinfection of bio-tissues due to its antibacterial, antiviral, and antifungal properties. In agriculture, CAPP accelerates the imbibition and germination of seeds and significantly increases plant productivity. Plasma is also used to fix molecular nitrogen. CAPP can produce reactive oxygen and nitrogen species (RONS). Plasma treatment of bio-tissue can lead to numerous side effects such as lipid peroxidation, genotoxic problems, and DNA damage. The mechanisms of occurring side effects when treating various organisms with cold plasma are unknown since RONS, UV-Vis light, and multicomponent biological tissues are simultaneously involved in a heterogeneous environment. Here, we found that CAPP can induce in vitro oxidation of the most common water-soluble redox compounds in living cells such as NADH, NADPH, and vitamin C at interfaces between air, CAPP, and water. CAPP is not capable of reducing NAD⁺ and 1,4-benzoquinone, despite the presence of free electrons in CAPP. Prolonged plasma treatment of aqueous solutions of vitamin C, 1,4-hydroquinone, and 1,4-benzoquinone respectively, leads to their decomposition. Studies of the mechanisms in plasma-induced processes can help to prevent side effects in medicine, agriculture, and food disinfection.     

Metabolite profiling and in-vitro colon cancer protective activity of Cycas revoluta cone extract

Abstract

The methanolic extract of Cycas revoluta cone (MECR) was analyzed by GC-MS and UHPLC for metabolite profiling and was evaluated for anti-colon cancer property by using in vitro assays like Cell Viability Assay, Colony Formation Assay, ROS Determination, Flowcytometry, DAPI staining assay, Tunel assay. GC-MS and HPLC analysis confirmed the presence of different phytochemicals in the extract of Cycas revoluta cone. In-vitro studies showed MECR extract showed significant anti-colon cancer activity by reducing proliferation and inducing apoptosis in colon cancer cell (HCT-8) line, but no such activity was seen in normal colon cell (CCD-18Co) line. The investigation confirms that MECR may be a promising candidate in colon cancer protection.     

The CO2 Reforming of Natural Gas in a Pulsed Corona Discharge Reactor

The conversion of CO ₂ and (CH ₄+CO ₂ ) mixtures to CO, at room temperature and atmospheric pressure conditions, in pulsed corona discharges, was investigated. Conversion of pure CO ₂ was 16.8% at 10 cm ³ -min ^–1 flow rate, which corresponds to 75 mol-min ^–1 rate of conversion. The CO ₂ conversion was improved to 38% (85 mol-min ^–1 by feeding the reactor with CH ₄+CO ₂ gas mixture (1:1 ratio), simultaneously with CH ₄ conversion of 46% (102.7 mol-min ^–1 ) at 10 cm ³ -min ^–1 flow rate of feed gases and 9 W power conditions. Rate of CO production is increased from 110 to 180 mol-min ^–1 with the variation of feed gas (CH ₄+CO ₂ mixture, 1:1 ratio) flow rate from 10 to 40 cm ³ -min ^–1 at 9W, which corresponds to energy efficiency of 2.5 to 4.1%. Highest energy yield of 25 g/kWh for CH ₄ conversion, 29 g/kWh for CO ₂ conversion, and 33 g/kWh for CO production were achieved.     

Photoinduced Dynamics and Toxicity of a Cancer Drug in Proximity of Inorganic Nanoparticles under Visible Light

Drug sensitization with various inorganic nanoparticles (NPs) has proved to be a promising and an emergent concept in the field of nanomedicine. Rose bengal (RB), a notable photosensitizer, triggers the formation of reactive oxygen species under green‐light irradiation, and consequently, it induces cytotoxicity and cell death. In the present study, the effect of photoinduced dynamics of RB upon complexation with semiconductor zinc oxide NPs is explored. To accomplish this, we successfully synthesized nanohybrids of RB with ZnO NPs with a particle size of 24 nm and optically characterized them. The uniform size and integrity of the particles were confirmed by high‐resolution transmission electron microscopy. UV/Vis absorption and steady‐state fluorescence studies reveal the formation of the nanohybrids. ultrafast picosecond‐resolved fluorescence studies of RB–ZnO nanohybrids demonstrate an efficient electron transfer from the photoexcited drug to the semiconductor NPs. Picosecond‐resolved Förster resonance energy transfer from ZnO NPs to RB unravel the proximity of the drug to the semiconductor at the molecular level. The photoinduced ROS formation was monitored using a dichlorofluorescin oxidation assay, which is a conventional oxidative stress indicator. It is observed that the ROS generation under green light illumination is greater at low concentrations of RB–ZnO nanohybrids compared with free RB. Substantial photodynamic activity of the nanohybrids in bacterial and fungal cell lines validated the in vitro toxicity results. Furthermore, the cytotoxic effect of the nanohybrids in HeLa cells, which was monitored by MTT assay, is also noteworthy.     

Removal of lipopolysaccharide and reactive oxygen species using sialic acid immobilized polysulfone dialyzer

Sialic acid (N‐acetylneuraminic acid, NANA) was covalently immobilized onto the surface of a polysulfone (PSF) hollow fiber membrane. Prior to the immobilization, the surface of PSF was treated with ozone, followed by grafting with acrylic acid, and then the esterification of NANA. The surface concentration of NANA was determined by 2‐thiobarbituric acid (TBA) test. Hemocompatibility, the capability of suppressing oxidative stress, and clearance of lipopolysaccharide (LPS) from the resulting hollow fiber membrane were evaluated. The results show that by immobilizing NANA onto PSF hollow fiber, the adhesion of platelet was reduced, while both APTT and PT were little affected. Furthermore, oxidative stress was suppressed by NANA‐immobilized PSF hollow fibers. The level of LPS was also greatly reduced. Copyright © 2009 John Wiley & Sons, Ltd.     

The effect of covalent immobilization of sialic acid on the removal of lipopolysaccharide and reactive oxygen species for polyethylene terephthalate

Polyethylene terephthalate (PET) was aminolyzed with 1,6‐diaminohexane (DAH) and then sialic acid (NANA) was immobilized via amidation onto the surface. The surface concentration of NANA was determined by 2‐thiobarbituric acid (TBA) test. The hemocompatibility of the resulting PET fabrics was evaluated based on complete blood count (CBC), coagulating times, and protein adsorption. The ability to remove lipopolysaccharide (LPS) was also determined. In addition, the effect of contacting NANA‐immobilizing PET on the suppression of reactive oxygen species (ROS) production was measured by the chemiluminescence (CL) method. The results show that by immobilizing NANA onto PET, the adhesion of platelet (PLt) was reduced, and oxidative stress was suppressed. The level of LPS was also greatly reduced. Copyright © 2010 John Wiley & Sons, Ltd.     

Non‐Thermal Plasma in Contact with Water: The Origin of Species

Non‐thermal atmospheric pressure plasma has attracted considerable attention in recent years due to its potential for biomedical applications. Determining the mechanism of the formation of reactive species in liquid treated with plasma is thus of paramount importance for both fundamental and applied research. In this work, the origin of reactive species in plasma‐treated aqueous solutions was investigated by using spin‐trapping, hydrogen and oxygen isotopic labelling and electron paramagnetic resonance (EPR) spectroscopy. The species originating from molecules in the liquid phase and those introduced with the feed gas were differentiated by EPR and ¹H NMR analysis of liquid samples. The effects of water vapour and oxygen admixtures in the feed gas were investigated. All the reactive species detected in the liquid samples were shown to be formed largely in the plasma gas phase. It is suggested that hydrogen peroxide (determined by UV/Vis analysis) is formed primarily in the plasma tube, whereas the radical species ?OOH, ?OH and ?H are proposed to originate from the region between the plasma nozzle and the liquid sample.     

Determination of Cobalt(II)-Hydrogen Peroxide-Induced DNA Oxidative Damage and Preventive Antioxidant Activity by CUPRAC Colorimetry

Abstract

The classical Fenton system composed of Fe(II) and H₂O₂ uses harsh oxidative conditions and cannot realistically simulate physiological oxidations which are less severe. Here, reactive oxygen species (ROS) were generated with a combination of CoSO₄ and H₂O₂ to provide milder conditions. DNA was used as a biologically meaningful probe for monitoring the oxidative conversion. Oxidative hazard on DNA was accomplished in ammonia/ammonium chloride buffer at 37?°C, and the Fenton reaction was stopped with trichloroacetic acid (TCA). A suitable aliquot of this solution was added to cupric ion reducing antioxidant capacity (CUPRAC) reaction mixture, and the absorbance at 450?nm was recorded. The oxidized species derived from DNA were CUPRAC-reactive while intact DNA was not. The protective effects of antioxidants (AOxs), known to have radical scavenging effects, were tested; green tea and a synthetic fetal bovine serum (FBS) were also successfully used as real ROS scavengers. Although the classical iron-based Fenton procedure applied in ethanol medium generated CUPRAC-responsive products, the proposed system was perfectly ethanol-tolerant, enabling the CUPRAC measurement of DNA oxidation products against an unaffected reagent blank. The protective effects of phenolic antioxidants, perfectly solubilized in ethanol, could also be measured.     

On the Use of Global Kinetics Models for the Investigation of Energy Deposition and Chemistry in RF Argon–Oxygen Plasmas Working in the Torr Regime

A global plasma model is used to investigate the chemistry and energy deposition in 13.56 MHz radio-frequency capacitively coupled oxygen–argon discharges under conditions usually used for the deposition of tin oxide films. These models are based on the solution of a stationary electron Boltzmann equation coupled to species balance equations including the vibrational kinetics equations of O₂. The results obtained showed that vibrational non equilibrium of O₂-molecule is not significant. The dissociation degree of O₂ was found to be around a few percents and the discharge was often moderately electronegative even for small O₂ contents in the feed gas. The ionization and energy dissipation mechanisms are mainly governed by the collisional processes involving O₂ for an oxygen feed gas composition greater than 20%. Results also showed that the predicted densities of the charged species and the electronegative character of the discharge are strongly linked to the assumption made on the homogeneity of the power deposition. On the contrary, the predicted density of O-atom is not sensitive to this assumption.     

Cold Plasma Systems and Their Application in Surface Treatments for Medicine

In this paper, a review of cold plasma setups and the physical and chemical processes leading to the generation of active species is presented. The emphasis is given to the interaction of cold plasmas with materials used in medical applications, especially medical implants as well as live cells. An overview of the different kinds of plasmas and techniques used for generation of active species, which significantly alter the surface properties of biomaterials is presented. The elemental processes responsible for the observed changes in the physio-chemical properties of surfaces when exposed to plasma are described. Examples of ongoing research in the field are given to illustrate the state-of-the-art at the more conceptual level.     

冷等离子体作用下CH_4-CO_2转化制合成气

在常温常压条件下 ,利用电晕放电 ,使 CH4 - CO2 混合气转化生成合成气 .结果表明 ,该过程中 CH4 和 CO2 的转化率与反应体系能量密度、原料气配比和流速等有关 .在 0 .1MPa气压 ,能量密度为 10 50 k J/ mol(反应体系温度低于 50 0 K) ,n( CH4 )∶ n( CO2 ) =1∶ 2条件下 CH4 和 CO2 的转化率分别超过 60 %和 50 % ,超出了热力学平衡转化率的限制 .通过调配原料的配比 ,可以得到不同 n( H2 ) / n( CO)比值的产物 .对该体系的反应机理进行了探讨 .     

近十年烧伤感染创面细菌培养及药物敏感试验

为分析烧伤感染创面细菌及药物敏感的变化情况,采集了1993-2004年贵阳中医学院-附院烧伤病人的创面分泌物作细菌培养及体外药物敏感试验,总结了十年来五个不同时期烧伤感染创面细菌培养及药物敏感试验的结果.结果表明,397例病人培养出433株细菌,其中革兰氏阴性杆菌79.2%（铜绿脓假单胞杆菌38%,埃希氏副大肠杆菌34%）,革兰氏阳性珠菌20.8%（金色黄色葡萄球菌20%）;细菌对所用的抗生素耐药性都很强,耐药性80%～100%的抗生素所占比例从1993-1994年的58.3%增至2003-2004的80%.结论：烧伤感染创面分泌物培养出的细菌革兰氏阴性杆菌多于革兰氏阳性球菌,且G杆菌的总检出率逐年升高,主要致病菌是铜绿脓假单胞杆菌,埃希氏副大肠杆菌,金色黄色葡萄球菌;无论是革兰氏阴性杆菌还是革兰氏阳性球菌对所用的抗生素耐药性都很强,且耐药率逐年升高.     

A Novel Fluorescent Probe for Hydrogen Peroxide and Its Application in Bio-Imaging

Hydrogen peroxide (H₂O₂) plays an important role in the human body and monitoring its level is meaningful due to the relationship between its level and diseases. A fluorescent sensor (CMB) based on coumarin was designed and its ability for detecting hydrogen peroxide by fluorescence signals was also studied. The CMB showed an approximate 25-fold fluorescence enhancement after adding H₂O₂ due to the interaction between the CMB and H₂O₂ and had the potential for detecting physiological H₂O₂. It also showed good biocompatibility and permeability, allowing it to penetrate cell membranes and zebrafish tissues, thus it can perform fluorescence imaging of H₂O₂ in living cells and zebrafish. This probe is a promising tool for monitoring the level of H₂O₂ in related physiological and pathological research.     

Modeling Alleviative Effects of Ca,Mg, and K on Cu-Induced Oxidative Stress in Grapevine Roots Grown Hydroponically

The aim of this study was to determine the pattern of alleviation effects of calcium (Ca), magnesium (Mg), and potassium (K) on copper (Cu)-induced oxidative toxicity in grapevine roots. Root growth, Cu and cation accumulation, reactive oxygen species (ROS) production, and antioxidant activities were examined in grapevine roots grown in nutrient solutions. The experimental setting was divided into three sets; each set contained a check (Hoagland solution only) and four treatments of simultaneous exposure to 15 μM Cu with four cation levels (i.e., Ca set: 0.5, 2.5, 5, and 10 mM Ca; Mg set: 0.2, 2, 4, and 8 mM Mg; K set: 0.6, 2.4, 4.8, and 9.6 mM K). A damage assessment model (DAM)-based approach was then developed to construct the dose-effect relationship between cation levels and the alleviation effects on Cu-induced oxidative stress. Model parameterization was performed by fitting the model to the experimental data using a nonlinear regression estimation. All data were analyzed by a one-way analysis of variance (ANOVA), followed by multiple comparisons using the least significant difference (LSD) test. The results showed that significant inhibitory effects on the elongation of roots occurred in grapevine roots treated with 15 μM Cu. The addition of Ca and Mg significantly mitigated phytotoxicity in root growth, whereas no significant effect of K treatment on root growth was found. With respect to oxidative stress, ROS and malondialdehyde (MDA) contents, as well as antioxidant (superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX)) activities, were stimulated in the roots after exposure to 15 μM Cu for three days. Moreover, H₂O₂ levels decreased significantly as Ca, Mg, and K concentrations increased, indicating that the coexistence of these cations effectively alleviated Cu-induced oxidative stress; however, alleviative effects were not observed in the assessment of the MDA content and antioxidant enzyme activities. Based on the DAM, an exponential decay equation was developed and successfully applied to characterize the alleviative effects of Ca, Mg, and K on the H₂O₂ content induced by Cu in the roots. In addition, compared with Mg and K, Ca was the most effective cation in the alleviation of Cu-induced ROS. Based on the results, it could be concluded that Cu inhibited root growth and Ca and Mg absorption in grapevines, and stimulated the production of ROS, lipid peroxidation, and antioxidant enzymes. Furthermore, the alleviation effects of cations on Cu-induced ROS were well described by the DAM-based approach developed in the present study.