Total Antioxidant Capacity Assay Research Articles

Study of Phytochemical Content and Antioxidant Properties of Musa Balbisiana Corm Extract

Musa balbisiana colla is an important plant native to India and many other Asian countries. Parts of this plant such as seeds, fruit pulp, inflorescence, pseudo-stem, and corm have been known to possess several medicinal values. In Kokrajhar district of Assam, the decoction of corm part of the plant is traditionally used as antidiabetic medicine. The present study was aimed to investigate the phytochemicals, antioxidants, trace element, and major compounds of the corm part of Musa balbisiana. Phenolic and flavonoid contents were estimated following standard protocols. The antioxidant activity of the plant was studied by ferric reducing antioxidant power assay, total antioxidant capacity, 1,1-diphenyl-2-picryl-hydrazyl, and lipid peroxidation scavenging assay. The phytochemical study revealed that the corm extract is rich in protein, carbohydrates, phenolics, and flavonoid content. Antioxidant study revealed strong free radical scavenging property of the crude corm extract of Musa balbisiana. The elemental analysis showed highest Zn content (0.2993 ppm) followed by Ni, Cu, and Mn. Pb, Cd, and Cr were not detected in the extract. GC-MS analysis showed difluroisocyanotophosphine to be the major compound of Musa balbisiana corm extract

Two new flavonoid glycosides isolated from the fruits of Catalpa ovata

Background: Catalpa ovata, widespread in China, Japan, and Korea, is receiving greater attention due to its potent pharmacological properties. Objectives: This communication report presents the results of chemical investigation and antioxidant capacity of the fruits of C. ovata. Materials and Methods: The polar fraction from the fruits of C. ovata was isolated using various chromatographic methods and their structures were identified through nuclear magnetic resonance spectrometry and high-resolution mass spectrometry, and the data were compared with the reported literature. Total antioxidant activity was analyzed using the total antioxidant capacity assay kit with a rapid ABTS method. Results: Cavonosides A and B (1 and 2), two new flavonoid glycosides, together with 13 known phenols (3–15), were obtained from the fruits of C. ovata. The antioxidant capacity of compounds 4, 7, 10, 13, and 15 was approximately equal to that of Vitamin C at the same concentration. Conclusion: This research work resulted in two new flavonoid glycosides (1 and 2) and 13 known phenols isolated from the fruits of C. ovata; among them, compounds 4, 7–10, and 13 were newly isolated from this genus. According to our results, the antioxidant capacity of compounds 4, 7, 10, 13 and 15 was approximately same as that of Vitamin C at the same concentration. We also discuss the structure–activity relationships of the isolated compounds.

Estimation of Antioxidant Potential of Indigenous Halari and French Poitu Donkey Milk by using the Total Antioxidant Capacity and Ferric Reducing Antioxidant Power Essay

Over the last decade, research has been conducted towards the uses and properties of donkey milk that has distinct chemical composition and consequently particularly nutritional properties. It is interesting to determine antioxidant potential of indigenous Halari and French Poitu donkey milk by using the total antioxidant capacity and ferric reducing antioxidant power assay. These methods are reliable, fast and with sound methodological infrastructures. The objective of the present research was to investigate the applicability of the trolox based total antioxidant capacity (TAC) and ferric reducing ability of plasma (FRAP) assay for identification of antioxidant potential of Indian Halari donkey milk and French Poitu donkey milk. The donkey milk is quite popular for its anti-aging properties in Western and European countries as cosmetic and nutraceutical preparations. However, in Indian context no such study is conducted on Halari donkey milk. The Indian Halari donkeys are untapped resource for their dairy potential. Our results showed that the whole milk of Halari donkey is at par with French Poitu donkey milk in terms of their antioxidative potential.

Thymoquinone promotes mouse mesenchymal stem cells migration in vitro and induces their immunogenicity in vivo

Mesenchymal stem cells (MSCs) have unique potentials, including migration and immunomodulation. Identification of the factors that enhance these activities can improve clinical applications of MSCs. This study aimed to investigate total antioxidant capacity (TAC) and migration potential of mouse MSCs exposed to thymoquinone (TQ) in vitro, and to examine the effect of TQ-treated MSCs on the expression of mouse immune cell markers. The results of total antioxidant capacity and wound healing assays showed that TQ increased the rate of MSCs TAC and migration in a dose- and time-dependent manner. The maximum TAC and migration were detected at 600 and 250 ng/ml of TQ, respectively. Functionally, the real-time PCR data analysis indicated that TQ induced c-Met and Cxcr4 expression and therefore, there may be a correlation between upregulation of these genes and increased MSCs migration. TQ also enhanced the up and down regulating impact of MSCs on Rorγt and Plzf expression and the expression of Tcf4 in mouse immune cells, respectively. Overall, this study declares that TQ increases the TAC of MSCs. It also proposes that TQ may, through activation of c-MET and CXCR4 signalling pathways, promote MSCs migration. TQ may also augment MSCs immunogenicity through its influence on the expression of genes involved in commitment of mouse immune system cells in vivo.

N-Doped Carbon As Peroxidase-Like Nanozymes for Total Antioxidant Capacity Assay.

N-doping strategy has been explored to enhance the activity of carbon nanozymes because the reconstructed electronic structures in N-doped carbon nanozymes are advantageous for the catalytic process. However, carbon nanozymes with high N content are still difficult to obtain due to the instability of N element under high calcination temperatures. To address this challenge, here we proposed a new N-doping strategy to fabricate highly active and specific peroxidase-like carbon nanozymes by using a high N-containing polymer (i.e., polyethylenimine (PEI)) as the N source and a natural clay mineral (i.e., montmorillonite (MMT)) as a template, respectively. We showed that the assembly of MMT with PEI protected N loss under high calcination temperatures and thus retained more catalytically active N sites. The mechanism study showed that the hydroxyl radical could be the key intermediate involved in the peroxidase-like catalysis. We then used the optimized carbon nanozyme with high and specific peroxidase-like activity (i.e., CP600-6) to detect H2O2, glucose, and ascorbic acid. Moreover, we successfully determined the total antioxidant capacity (TAC) in real samples including four commercial beverages, fresh orange juice, and three kinds of vitamin C tablets. The current study not only provides a new strategy for fabricating peroxidase-like nanozymes but also develops a facile TAC assay for future use in evaluation of antioxidant food quality and oxidative stress in healthcare.

Gallic Acid-Dextran Conjugate: Green Synthesis of a Novel Antioxidant Molecule.

A novel derivative of dextran, dextran–gallic acid (Dex–Gal), obtained from simple conjugation with gallic acid, was synthesized by an efficient free radical-mediated method. To verify the synthesis of Dex–Gal, 1H-nuclear magnetic resonance (1H-NMR), Fourier transform infrared (FTIR) spectrometry, and high-performance size-exclusion chromatography (HPSEC) were employed. The results revealed the conjugation of gallic acid with the 15.5 kDa dextran from Leuconostoc mesenteroides. Dex–Gal had a molecular weight of 11.2 kDa, indicating that the conjugation reaction was accompanied by a minor degradation of Dex–Gal. In addition, Dex–Gal contained 36.8 ± 1.4 mg gallic acid per gram dextran. These molecules were also evaluated as antioxidants using total antioxidant capacity (TAC), reducing power, ferric chelation, and superoxide radical-scavenging assays. Both polysaccharides had no ferric chelation activity. In addition, Dex–Gal was more efficient as an antioxidant agent in TAC (13 times) and was more efficient than dextran in superoxide radical-scavenging (60 times) and reducing power (90 times) assays. These data demonstrate that Dex–Gal is a natural-compound-based antioxidant with potential applications in the pharmaceutical, cosmetic, and food industries.

The in vitro cytotoxicity, genotoxicity and oxidative damage potential of dapagliflozin, on cultured human blood cells

Abstract Objectives Dapagliflozin (DAPA), is a potent SGLT-2 inhibitor for the treatment of patients with type 2 diabetes. DAPA has a good clinical and biological tolerance profile. However little information is available on its potential effects on cultured human blood cells. The evaluation of the in vitro cytotoxicity, genotoxicity potential and antioxidant/oxidant activity of DAPA in primary human whole blood cell cultures was aimed in this study. Materials and methods Cell viability was measured by the MTT [3,(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] and lactate dehydrogenase (LDH) leakage assays. The antioxidant/oxidant activity was determined by measuring the total antioxidant capacity (TAC) and total oxidative stress (TOS) levels. To assess the genotoxicity of DAPA, chromosomal aberration (CA) frequencies were determined. Results MTT and LDH release assay exhibited that exposure to different doses of DAPA did not changed significantly the proliferation of cells. The results of TAC and TOS assays were showed that TAC level was elevated while TOS level did not altered in DAPA-treated cells. Moreover, any increase in the frequency of CA did not found on cultures blood cells. Conclusion These data indicate that DAPA has not cytotoxic and genotoxic potential in cultured human blood cells, also, induces the increasing antioxidant activity.

Action of trichostatin A on Alzheimer's disease-like pathological changes in SH-SY5Y neuroblastoma cells.

The histone deacetylase inhibitor, trichostatin A, is used to treat Alzheimer's disease and can improve learning and memory but its underlying mechanism of action is unknown. To determine whether the therapeutic effect of trichostatin A on Alzheimer's disease is associated with the nuclear factor erythroid 2-related factor 2 (Nrf2) and Kelch-like epichlorohydrin-related protein-1 (Keap1) signaling pathway, amyloid β-peptide 25-35 (Aβ25-35) was used to induce Alzheimer's disease-like pathological changes in SH-SY5Y neuroblastoma cells. Cells were then treated with trichostatin A. The effects of trichostatin A on the expression of Keap1 and Nrf2 were detected by real-time quantitative polymerase chain reaction, western blot assays and immunofluorescence. Total antioxidant capacity and autophagy activity were evaluated by total antioxidant capacity assay kit and light chain 3-I/II levels, respectively. We found that trichostatin A increased cell viability and Nrf2 expression, and decreased Keap1 expression in SH-SY5Y cells. Furthermore, trichostatin A increased the expression of Nrf2-related target genes, such as superoxide dismutase, NAD(P)H quinone dehydrogenase 1 and glutathione S-transferase, thereby increasing the total antioxidant capacity of SH-SY5Y cells and inhibiting amyloid β-peptide-induced autophagy. Knockdown of Keap1 in SH-SY5Y cells further increased trichostatin A-induced Nrf2 expression. These results indicate that the therapeutic effect of trichostatin A on Alzheimer's disease is associated with the Keap1-Nrf2 pathway. The mechanism for this action may be that trichostatin A increases cell viability and the antioxidant capacity of SH-SY5Y cells by alleviating Keap1-mediated inhibition Nrf2 signaling, thereby alleviating amyloid β-peptide-induced cell damage.

In vitro Assessment of Antimicrobial, Antioxidant and Anticancer Activities of Some Marine Macroalgae

THE PRESENT study evaluated ethanol, methanol, ethyl acetate, hexane, chloroform and acetone extracts of five green and red seaweed species from Abu-Qir bay, Alexandria, Egypt for their antimicrobial, antioxidant activities and cytotoxicity against four cell lines. Chloroform extracts of Ulva lactuca and Ulva fasciata exhibited the highest inhibition zones against the tested pathogenic bacteria (Klebsiella pneumoniae and Proteus mirabilis) and fungi (Aspergillus flavus, Aspergillus fumingatus and Aspergillus niger) as measured by disc diffusion method. The extracts of U. lactuca and U. fasciata showed the highest antioxidant activity (IC50 6.32±0.29mg/ml and 6.61±0.27mg/ml, respectively), using DPPH (2, 2- diphenyl-1- picrylhydrazyl) scavenging method and total antioxidant capacity assay (2.13 and 1.51mg ascorbic acid equivalent/ gram dry weight, respectively). Cytotoxicity test using MTT assay showed that U. lactuca extract had strong activity against MCF-7 and Hela cell lines (IC50 10.83±1.0, 12.43±1.3μg/ml, respectively), while U. fasciata had strong activity against PC3 and HePG2 cell lines (IC50 12.99±1.2, 16.75±1.5μg/ml, respectively). As analyzed by Ultraviolet spectra, Fourier Transform Infrared spectra and Gas Chromatography Mass Spectroscopy after purification by column chromatography, the most active antimicrobial fractions in U. lactuca and U. fasciata extracts include an aromatic compound that have different active group (-C=O, phenyl ring and –OH); its molecular weight of Di-isooctyl Phthalate equals 390.56g/mol and Butylated Hydroxytoluene equals 220.356g/mol, respectively. The studied seaweeds have distinct active metabolites being a promising source of antimicrobial and antiproliferative compounds that can be used effectively in pharmaceutical drug industry.

Evaluation of Phytochemicals, Antioxidants, Trace Elements in Kigelia africana Fruit Extracts and Chemical Profiling Analysis Using UHPLC-qTOF-MS2 Spectrometry.

The study aimed at evaluating the phytochemical composition, antioxidant potentials and the levels of trace elements in the fruit extract of Kigelia africana obtained by different extraction solvents in order to ascertain its numerous pharmacological activities and identify the different chemical compounds responsible for these activities. The crude extract in ethanol and four other solvent fractions (hexane, ethylacetate, butanol and aqueous) were obtained for phytochemical screening. Antioxidant potentials of K. africana fruit were investigated spectrophotometrically using hydroxyl ion scavenging (OH-) activity, metal ion chelating activity, anti-lipid peroxidation activity as well as total antioxidant capacity assays. Trace element (Mn, Zn, Cd, Ni, Cu, Pb, Cr, Co and Fe) levels were measured using a plasma-emission spectrometer that has an auto sampler AS 93-plus and coupled with Nebulizer CETAC U-6000AT+ after microwave acid digestion of the fruit extracts. Chemical identification was performed using ultra-high-pressure liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UHPLC-qTOF-MS2). Kigelia africana fruit extracts obtained showed a variety of bioactive phytochemical compounds including phenolic acids, flavonoids, saponins, tannins and glycosides. The total antioxidant capacity activities of the aqueous, butanol, ethanol, hexane and ethylacetate extracts are 15.04, 52.11, 44.95, 79.27 and 175.20mg AAE/g. Metal ion chelating activity showed significant correlation with lipid peroxidation inhibition activity at p ≤ 0.01 and with OH- scavenging activity at p ≤ 0.05. PCA analysis revealed that all the extract/fractions have higher total antioxidant activities compared to aqueous extract with hexane extract exhibiting the highest radical scavenging potential. HCA showed similarities with three well-defined clusters and PLS regression was used to predict total antioxidant activity. High sensitivity by low values of limits of detection and quantification was observed ranging from 0.021 to 0.085mg/ml and 0.063 to 0.258mg/ml for Zn and Fe respectively. Ethylacetate extract had high concentration of Fe (0.5656mg/kg). For the standardization of the K. africana fruit extract, 244 chemical compounds were identified by measuring m/z values with threshold override of 100,000 and analysing mass spectrometer fragmentation behaviour while 16 of these were confirmed. Kigelia africana fruit extract is a good source of antioxidant and possess maximum accepted concentration of trace elements according to European legislation (1881/2006/EC). The metabolites identified exhibited numerous pharmacological activities. The method and results suggest the applicability for commercial use of this K. africana fruit in the treatment of oxidative-related diseases. Graphical abstract The phytochemical, antioxidant and trace element composition of crude ethanol extract, hexane, butanol, aqueous and ethylacetate extracts of Kigelia africana fruit were determined. The fruit extracts were found to possess good antioxidant activity, maximum acceptable amount of essential trace elements as well as the presence of bioactive phytochemicals. K. africana fruit would be an ideal candidate in improving human health and thus the management of oxidative-related diseases such as diabetes, by involving in the antioxidant defense system against free radical generation.

Impact of Chronic Hepatitis B Virus Infection On Semen Parameters of Fertile Men

Aim: To investigate the effect of chronic hepatitis B virus (CHB) infection on semen parameters of males with proven fertility. Material & methods: 61 fertile men with CHB infection and a control group of 42 HBV-negative fertile men were assessed for serum and semen HBV viral loads by quantitative real-time PCR, complete semen analysis, hypo-osmotic swelling test, seminal total peroxide, total antioxidant capacity assay and sperm DNA integrity evaluation. Results: CHB men demonstrated significantly lower sperm motility, sperm viability and sperm DNA integrity, seminal total antioxidant capacity assay and significantly increased seminal total peroxide and oxidative stress index compared with controls. CHB cases with positive semen HBV DNA showed a significant increase in seminal total peroxide and oxidative stress index compared with CHB cases with negative semen HBV DNA. Conclusion: CHB has a negative burden on seminal parameters in fertile men.

Comparative Evaluation of Antioxidant Potential in Thermally Processed, Underutilized Food Grains of the Himalayan Region

Aims: To determine the bioactive components and total antioxidant capacity (TAC) in selected underutilized crops of the Himalayan region viz. Barnyard millet, Grain amaranth, Rice bean, Black soybean and Horsegram.
 Study Design: Experimental design (Lab experiment).
 Place and Duration of Study: Department of Foods and Nutrition, College of Home Science, G. B. Pant University of Agriculture and Technology, Pantnagar, Uttarakhand, in the year 2016-18.
 Methodology: We applied different processing techniques (covered pan cooking and pressure cooking) in the underutilized crops and analyzed the total phenol, total flavonoids and total antioxidant capacity (Ferric reducing antioxidant power (FRAP) assay and (2, 2-Diphenyl-1-picrylhydrazyl) (DPPH) by using standard methods.
 Results: It was found that black soybean had highest phenolic content after both thermal treatments (3233.76 mg GAE/100 g for pan cooked and 1883.11 mg GAE/100 g for pressure cooked samples) and TAC by both FRAP (6423.76 mg TE/100 g for pan-cooked and 4415.58 mg TE/100 g for pressure cooked) and DPPH (536.41 and 453.98 mg TE/100g for pan and pressure cooked samples, respectively) method. Among raw samples, rice bean contained the highest flavonoid content and TAC by FRAP assay. In contrast, raw grain Amaranth showed the lowest phenolic content. Further, pressure cooking was found to be better for barnyard millet, while in pulses, pan cooking yielded the best results (in terms of increased value/lower losses).
 Conclusion: It can be concluded that among pulses pan cooked black soybean was found to have a good store of bioactive compounds as compared to rice bean and horse gram. The pressure cooking method was found to be suitable for millet like a barnyard.

Antioxidant Effects and Suppression of Nitric Oxide Production in LPS-stimulated Macrophages by Fractions of Vaccinium leschenaultii Wight

ABSTRACTSolvent fractions from leaf and bark extracts of Vaccinium leschenaultii were screened for their antioxidant, cell viability, and nitric oxide (NO) inhibitory potential. Antioxidant activities were determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH), total antioxidant activity (TAC) and Oxygen Radical Antioxidant Capacity (ORAC) assays. Sulforhodamine B assay was used to estimate the cell viability of RAW 264.7 cells. Griess reagent was used to measure the NO production, which was induced by lipopolysaccharide (LPS) on RAW 264.7 cells. Higher antioxidant activity was observed in the fraction of V. leschenaultii bark ethyl acetate: methanol extract (VBEM, 60/40, v/v) in DPPH (IC50 = 20.80 µg mL−1), TAC (14.90 mmol uric acid equivalents/mg extract) and ORAC (454.58 µmol trolox equivalents/mg extract) assays compared to the leaf extracts. Similarly, VBEM (60/40) fraction also showed 83.87% of RAW 264.7 cell viability. V. leschenaultii leaf ethyl acetate: methanol extract (VLCE, 70/30, v/v) inhibited the NO production (90.06%, IC50 = 1.61 µg mL−1) and showed dose-dependent activity without inhibiting the macrophages. V. leschenaultii leaf and bark fractions from the methanol extract inhibited the free radicals including NO by modulating the macrophage-mediated inflammatory functions in the in vitro system.

Fenpropathrin induces testicular damage, apoptosis, and genomic DNA damage in adult rats: Protective role of camel milk

Fenpropathrin (FNP) is a member of the synthetic pyrethroids. Herein, the present study was conducted to investigate, for the first time, the potentially harmful effects of FNP on the reproductive system of male rats. In addition, the prophylactic or concurrent influence of camel milk (CM) was assessed. Adult male rats were divided into five groups; control, vehicle control (oil), CM (2ml/rat/day), FNP (15mg/kg bwt/60 days), CM/FNP (prophylaxis) and FNP /CM (co-treated) groups. Sperm morphology, count, serum testosterone (TES), luteinizing hormone (LH) and follicle-stimulating hormone (FSH), thiobarbituric acid reactive substances (TBARS), total antioxidant capacity (TAC), superoxide dismutase (SOD), testicular enzymes, and comet assay analysis were estimated. In addition, histopathology, the ultrastructure of testicular tissue and apoptosis were evaluated. Reduced body weight and gonadosomatic index were observed in the FNP exposed group. TES, LH, FSH were markedly declined following FNP treatment. SOD and TAC concentrations were reduced while PC and TBARS were significantly elevated in FNP group indicating oxidative stress. Furthermore, FNP induced DNA damage and apoptosis in the testis which was evidenced histopathologically and by electron microscope examination. CM significantly counteracted FNP reprotoxic effects, particularly at the prophylactic routine (CM/FNP) than the co-exposure (FNP/CM) one. Conclusively, these findings verified that CM could be a potential candidate therapy against FNP reprotoxic impacts.

Studies on antioxidant, hepatoprotective, and vasculoprotective potential of Viola odorata and Wrightia tinctoria.

We have reported the antidyslipidemic, antihypertensive, and Ca++ channel blocking activities of Viola odorata (VO) and Wrightia tinctoria (WT). This study extends our understanding of their therapeutic potential by exploring the effects on biomarkers of hepatic and vascular dysfunction together with phytochemical standardization and antioxidant potential. Total phenolic compounds, total flavonoids content, and proanthocyanins of the methanolic extracts were identified using HPLC. Antioxidant capacity was measured using the in vitro assays. Two studies of 6-week duration were conducted on a high-fat diet rat model to test the leaves and seed extracts of VO and WT (300 and 600mg/kg) for their effect on biomarkers for hepatic and vascular dysfunction. The HPLC analysis showed high contents of total phenolic compounds, total flavonoids content, and proanthocyanins along with distinctive phenolic composition. Both extracts exhibited significant antioxidant potential in 1,1-diphenyl-2-picrylhydrazyl, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid), fluorescence recovery after photobleaching, and total antioxidant capacity (TAC) assays, comparable with synthetic standard antioxidants. The in vivo studies indicated a significant reduction in the high-fat-diet-induced rise in serum uric acid, phosphorus, aspartate aminotransferase, alanine aminotransferase, and gamma-glutamyl transferase. This study indicates the potential of VO and WT to protect from vascular and hepatic damage and an antioxidant effect, thus making these herbs strong candidates for managing cardiometabolic disorders.

Deciphering the Antioxidant and Free Radical-Scavenging Capacity of Anthocleista vogelii

The empirical survey comparatively explores the antioxidant activity and the free radical-scavenging capacity of various fractions of Anthocleista vogelii leaves. The dried and pulverized leaves of A. vogelii were subjected to cold maceration for 4, 8 and 12 days using 70 % methanol and filtered. The filtrate was concentrated to a slurry crude extract by means of a rotary evaporator at 40ºC with a salvaged yield of 34.10 %. Exactly 0.23 kg of crude extract was defatted and fractionated using two chromatographic techniques in chloroform-methanol-hexane solvent system (3:21 v/v). The fractions were screened qualitatively for polyphenolic compounds. The fractions containing these bioactive compounds were quantified and further subjected to free radical-scavenging activity: DPPH assay, ferric reducing power (FRP) assay, hydrogen peroxide scavenging activity (HPA) and total antioxidant capacity (TAC) respectively. There was marked significant difference (p<0.05) in the total phenolic and flavonoid contents (TPC and TFC) among the fractions with ample amount found in n-hexane fraction. There were marked significant differences (p<0.05) in the IC 50 values among fractions for DPPH, FRP, HPA and TAC assays. Thus, the ample antioxidant activity of n-hexane fraction among other fractions was further validated by the extrapolated IC 50 values. The study inferred that A. vogelii leaves are possible sources of antioxidants with health benefits, especially for disease conditions characterized by oxidative stress. Keywords: Anthocleista vogelii , Antioxidant activity, Free radical-scavenging capacity, IC 50 value, Oxidative stress DOI : 10.7176/JNSR/9-12-09 Publication date :June 30 th 2019

The cytotoxic and oxidative effects of restorative materials in cultured human gingival fibroblasts

The aim of this study was to evaluate the cytotoxic and oxidative effects of the most commonly used dental restorative materials on human gingival fibroblast cells (HGFCs). HGFCs were obtained from healthy individuals. The tested restorative materials were a microhybrid resin based composite, a compomer resin, a glass ionomer cement, and an amalgam alloy. One hundred eight cylindirical samples, 10 mm in diameter and 2 mm in height, were prepared according to ISO 10993-12:2002 specifications (n = 9 in the tested subgroups). Freshly prepared and aged samples in artificial saliva at 37 °C (7 and 21 d) were placed into well plates and incubated. Wells without dental materials were constituted as the control group. After 72 h incubation period, cytotoxicity was determined using the neutral red (NR) assay. Oxidative alterations were assessed using total antioxidant capacity (TAC) and total oxidant status (TOS) assay kits. Data were analyzed using the ANOVA and LSD post hoc tests. All tested materials led to significant decreases in the cell viability rates (33–73%) compared to the control group. Glass ionomer and resin composite were found to be more cytotoxic than amalgam alloy and compomer. The highest TAC level was observed in glass ionomer after seven-day aging and these changes prevented an increase in TOS levels. Increases in TAC levels after seven-day aging in all groups exhibited significant differences with freshly prepared samples (p < 0.05). In all material groups, TOS levels of freshly prepared samples differed statistically and significantly from samples aged for 7 and 21 d (p < 0.05). The data obtained suggested that all the tested materials exhibited cytotoxic and pro-oxidant features. Freshly prepared samples caused higher TOS levels. However, oxidant status induced by materials decreased over time.

Chemical Composition and Biological Activities of Essential Oils from Two Wild Algerian Medicinal Plants: Mentha pulegium L. and Lavandula stoechas L.

The aim of the present study was to determine the chemical composition, antioxidant and antimicrobial activities of Algerian Mentha pulegium L. and Lavandula stoechas L. essential oils (EOs). The essential oils of M. pulegium and L. stoechas were obtained by hydrodistillation and their yields was 3.96 ± 0.12 and 1.46 ± 0.06 % (v/w), respectively. GC-MS analyses allowed the identification of 27 components representing 98.06 % of the total oil for M. pulegium, and 68 components representing 93.51 % of the total for L. steochas. The main components of M. pulegium EO were pulegone (81.04 %), piperitenone (13.60 %) and trans-p-mentha-8-en-3-one (1.35 %). Fenchone (50.29 %), camphor (14.02 %) and bornyl acetate (5.60 %) were the main components of L. stoechas EO. The antioxidant activity of EOs was estimated by total antioxidant capacity, DPPH and ABTS radical scavenging assays, ferric-reducing power test and β-carotene bleaching assay. Both EOs exhibited a moderate antioxidant activity compared to BHT, vitamin C and trolox. Antibacterial studies showed good activity against four Gram positive and Gram negative bacteria: Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa, Escherichia coli, as well as good inhibitory properties (MIC), suggesting that Algerian M. pulegium and L. steochas EOs can be used in food and pharmaceutical industries as antimicrobial and preservative agents.

A QCM-D sensing strategy for investigating the real-time effects of oxidative stress on the viscoelastic properties of pre-osteoblast cells

Oxidative stress results from an imbalance in the cellular redox state which can cause damage to the cell cytoskeleton and morphology, thereby affecting cell viscoelasticity. In this study, we introduce a quartz crystal microbalance with dissipation (QCM-D) sensing strategy that enables the real-time assessment of the changes in the viscoelastic properties of an MC3T3 cell monolayer in response to oxidative stress. This was successfully achieved by tracking the changes in energy dissipation (ΔD-response) caused by the interaction of H2O2 with the cell monolayer adhered to a poly-d-lysine-coated polystyrene quartz sensor. While a return to baseline values of the ΔD-response was obtained at 325 min (recovery point) after the incubation of cells with 25 μM H2O2, higher concentrations (50 μM-10 mM) exhibited no recovery. We successfully validated using scanning electron, atomic force and fluorescence microscopy that at the recovery point the cell morphology recovered from 25 μM H2O2 exposure, whereas, higher concentrations (50 μM-10 mM) resulted in the shrinkage of the cytoskeleton, alteration in cell morphology, and decrease in cell density due to apoptosis/necrosis, supported by a cell viability assay. Finally, total antioxidant capacity assay (TAC) confirmed that while cells’ metabolic state recovered from 25 μM H2O2 treatment, higher H2O2 concentrations led to a decline in the TAC. Altogether, these results showed that the viscoelastic properties of cells can be used to investigate cell recovery from oxidative stress, which may allow us to distinguish oxidative eustress from distress. Overall, we have demonstrated that QCM-D is a viable tool for measuring the effects of oxidative stress on the viscoelastic properties of MC3T3 cells.

Synthesis, crystal structures, Hirshfeld surface analysis and spectroscopic studies of two Schiff bases of anisaldehyde and their urease and acetylcholinesterase inhibitory and antioxidant properties

The growing demand of pharmaceutical industry for more effective drugs requires new molecules with promising medicinal activities. In the present work, a natural product anisaldehyde was treated with hydrazine and 3,5-dichloroaniline to synthesize their Schiff bases, ASB1 and ASB2, which were assessed for various bioactivities. ASB1 was synthesized by conventional reflux method while ASB2 was synthesized by reflux as well as by mechanochemical grinding method which gave higher yield. The bases were recrystalised, and their structures were elucidated based on XRD and spectroscopic studies. Hirshfeld surface analysis was also carried out. They showed considerable urease inhibitory activity, almost comparable with the standard thiourea. The activity of ASB1 was much higher than ASB2. Acetylcholinesterase inhibitory activity of ASB1 was also higher than that of ASB2. The antioxidant activities were determined using DPPH, ABTS radical scavenging and total antioxidant capacity (TAC) assays. The bases were very poor scavengers of DPPH radical. However, they showed considerable anti-radical activity against ABTS radical, ASB2 being more active than ASB1, while ASB1 showed higher TAC than ASB2. In conclusion, the bases appeared to have good drugability as inhibitors of urease and acetylcholinesterase enzymes. They can be easily synthesized for possible large-scale applications. The grinding method proved to be more efficient than the reflux method.