Cytotoxicity and Immunotoxicity Assessment of NiO Nanoparticles Using the Chlamydomonas reinhardtii

2012 ◽  
Vol 198-199 ◽  
pp. 52-55
Author(s):  
Zhao Xiang Han ◽  
Dan Dan Wu ◽  
Zhen Zhu ◽  
Yu Rong Liu

Little is known about the potential behavior and ecotoxicity of nanoparticles to aquatic organisms. The cytotoxicity and immunotoxicity of NiO-Nanoparticles and Chlamydomonas reinhardtii. It shown that the mitochondrial activity was gradually reduced significantly at the different exposure concentrations for NiO nanoparticles. NiO nanoparticles had a similar effect on membrane integrity as that observed for inhibition of cellular metabolic activity, with a significant reduction in membrane integrity. The NiO nanoparticles exhibited a significantly higher degree of BOD compared to the control group. NiO nanoparticles induced increases in MDA levels with incubation time. A significantly higher percent of cells with mitochondrial depolarization was observed when cells were exposed NiO nanoparticles. NiO nanoparticles significantly elevated ROS levels more than four fold at the highest concentration and the lysozyme activity was significantly increase at all the exposure concentrations and incubation times.

2021 ◽  
Vol 49 ◽  
Author(s):  
Marcos Antônio Celestino de Sousa Filho ◽  
Luanna Soares de Melo Evangelista ◽  
Filipe Nunes Barros ◽  
Jefferson Hallisson Lustosa da Silva ◽  
Anna Monallysa Silva de Oliveira ◽  
...  

Background: Canine sperm is a very delicate cell that is quite susceptible to oxidative stress since the cytoplasm is restricted and features little antioxidant reserves. Furthermore, the sperm membrane has some polyunsaturated fatty acids sensitive to lipid peroxidation, which makes it important to addition antioxidant substances to the diluter aiming at decreasing such stress to the sperm cell, particularly during seminal cryopreservation. Several antioxidants have been used in this process in some domestic animal’s species, however, the use of palmitic acid has been little reported in works on cryopreservation of semen of the canine species. Hence, this study aimed to assess the effect of addition antioxidants palmitic acid and vitamin E to the Tris-egg yolk diluter on the semen quality of dogs after thawing.Materials, Methods & Results: Samples were collected from the ejaculates of 4 adult dogs, apparently healthy, of the American Pit Bull Terrier breed of kennels in the city of Teresina, PI, places where the pre-freezing procedures of the dog’s semen were performed. The samples were diluted in Tris citric acid fructose (3.28 g Tris-hydroxymethyl-aminomethane, 1.78 g citric acid monohydrate and 1.25 g D-fructose), dissolved in 100 mL distilled water, and added 20% egg yolk and 6% glycerol, at the concentration of 100x106 sptz/mL. The semen samples were divided into 3 mL aliquots to form 3 experimental groups: G1 - Only Tris-egg yolk (Control group); G2 - Tris-egg yolk + 100 µM palmitic acid; and G3 - Tris-egg yolk + 116 µM vitamin E. Semen was collected weekly over a period of little over 2 months. After thawing, thermorresistance test (TTR) was carried out at 0, 30, 60, and 90 min to assess spermatics motility and vigor, in addition to analysis of integrity of plasma membrane, acrosomal membrane and mitochondrial activity of the sperm, using fluorescent probes. These assessments were performed out at the Animal Reproduction Biotechnology Laboratory (LBRA/UFPI). In the TTR, G2 and G3 didn´t exhibit significant results for spermatics motility or vigor when compared with the control group. The palmitic acid and vitamin E also had no significant effects on the parameters of acrosomal membrane integrity or mitochondrial activity. However, sperm cryopreserved with the addition of palmitic acid exhibited significant differences for plasma membrane integrity, providing greater protection to the sperm cells in G2.Discussion: The palmitic acid is one of the most saturated fatty acids in human semen, with reports of great proportions also in the seminal plasma of dogs. Its main role is to protect the plasma membrane from external damage, improving viability and fertility of the sperm after cryopreservation. Data is scarce in the literature on the composition of fatty acids in canine semen and regarding the use of palmitic acid as a seminal antioxidant in that species, which grants further studies aiming to investigate such valuable information for canine reproduction. It is concluded that addition palmitic acid at 100 µM concentration to the Tris-egg yolk diluter was able to preserve the integrity of the plasma membrane during the process of cryopreservation of canine semen.Keywords: dog, semen, antioxidants, cryopreservation.Descritores: cão, sêmen, antioxidantes, criopreservação.


2020 ◽  
Vol 41 (1) ◽  
pp. 167 ◽  
Author(s):  
Breno Fernandes Barreto Sampaio ◽  
Bruno Gomes Nogueira ◽  
Maria Inês Lenz Souza ◽  
Eliane Vianna da Costa-e-Silva ◽  
Carmem Estefânia Serra Neto Zúccari

Plasma membrane composition has impact on phase transition from liquid crystal to gel state of cooled sperm cell. The incorporation of polyunsaturated fatty acids increases its fluidity and can contribute to sperm motility. The aim of this study was to compare the effect of adding docosahexaenoic acid (DHA) and ?-tocopherol (?-Toh) to the cooling extender, singly or combined, to the equine sperm parameters, submitted to cooling, up to 72 hours. Two ejaculates of ten stallions collected with artificial vagina were used, and evaluated for motility, plasma membrane integrity, chromatin fragmentation, mitochondrial activity and lipid peroxidation, according to the following treatments: C; DHA; ?-Toh; DHA/?-Toh; EtOH 100: and EtOH 140 (corresponding to control; 10 ng mL-1 of DHA; 2 mM of ?-Toh; : 10 ng mL-1 of DHA + 2 mM of ?-Toh; 100 µL of ethanol and 140 µL of ethanol respectively). DHA treatment showed higher motility (68.2 ± 12.3; p < 0.05) when compared to control (62.1 ± 16.2), DHA/?-Toh (61.3 ± 12.7) and EtOH (58.1 ± 8.6) groups. In lipid peroxidation assay, the control group showed 2,506.2 ± 796.4 ng of MDA 108 spermatozoa-1, being significantly higher (p < 0.05) than the groups treated with DHA (2,036.0 ± 687.0), ?-Toh (1,890.8 ± 749.5) and DHA/?-Toh (1,821.1 ± 627.2). In conclusion, ?-Toh was effective in diminishing lipid peroxidation of equine sperm subjected to cooling, and DHA improved sperm motility and, in spite of being a polyunsaturated fatty acid with high susceptibility to peroxidation, reduced lipid peroxidation.


Animals ◽  
2019 ◽  
Vol 9 (10) ◽  
pp. 810 ◽  
Author(s):  
Wen ◽  
Li ◽  
Feng ◽  
Du ◽  
Ren ◽  
...  

Grape seed procyanidin extract (GSPE) has been shown to possess antioxidative effects. This experiment was designed to study the effect of GSPE during the liquid storage of goat semen. Semen samples were collected from six sexually mature goats. The samples were treated with different concentrations of GSPE (10, 30, 50, and 70 mg/L) in basic diluent and stored at 4 °C for 120 h; samples without GSPE were used as the control group. The results showed that sperm motility, acrosome membrane integrity, mitochondrial activity, plasma membrane integrity, total antioxidative capacity (T-AOC), catalase (CAT) activity, and superoxide dismutase (SOD) activity in the treatment groups were significantly higher than in the control group, whereas malondialdehyde (MDA) content was lower than in the control group (p < 0.05). In the treatment group, sperm quality in the 30 mg/L GSPE group was significantly higher than the other groups (p < 0.05). Furthermore, artificial insemination (AI) results showed that litter sizes were higher in the 30 mg/L GSPE group than in the control group (p < 0.05). In summary, this experiment showed that adding GSPE to the basic diluent improved sperm quality and that 30 mg/L of GSPE was the most suitable concentration for the liquid preservation of goat semen at 4 °C.


2016 ◽  
Vol 27 (5) ◽  
pp. 492-496 ◽  
Author(s):  
Fabiano Palmeira Gonçalves ◽  
◽  
Gutemberg Alves ◽  
Vladi Oliveira Guimarães Júnior ◽  
Marco Antônio Gallito ◽  
...  

Abstract Bis-acryl resins are used for temporary dental restorations and have shown advantages over other materials. The aim of this work was to evaluate the in vitro cytotoxicity of two bis-acryl composite resins (Protemp 4 and Luxatemp Star), obtained at 1, 7 and 40 days after mixing the resin components, using a standardized assay employing human primary cells closely related to oral tissues. Human gingival fibroblast cell cultures were exposed for 24 h to either bis-acryl composite resins, polystyrene beads (negative control) and latex (positive control) extracts obtained after incubation by the different periods, at 37 °C under 5% CO2. Cell viability was evaluated using a multiparametric procedure involving sequential assessment (using the same cells) of mitochondrial activity (XTT assay), membrane integrity (neutral red test) and total cell density (crystal violet dye exclusion test). The cells exposed to the resin extracts showed cell viability indexes exceeding 75% after 24 h. Even when cells were exposed to extracts prepared with longer conditioning times, the bis-acryl composite resins showed no significant cytotoxic effects (p>0.05), compared to the control group or in relation to the first 24 h of contact with the products. There were no differences among the results obtained for the bis-acryl composite resins evaluated 24 h, 7 days and 40 days after mixing. It may be concluded that the bis-acryl resins Protemp 4 and Luxatemp Star were cytocompatible with human gingival fibroblasts, suggesting that both materials are suitable for use in contact with human tissues.


2011 ◽  
Vol 14 (1) ◽  
pp. 21-27 ◽  
Author(s):  
A. Dziekońska ◽  
J. Strzeżek

Boar variability affects sperm metabolism activity in liquid stored semen at 5°CMetabolic activity of boar spermatozoa, liquid stored for three days at 5°C, was measured using bioluminescence for ATP content, fluorescent assay (JC fluorochrome) of mitochondrial activity and oxygen consumption. Sperm motility and plasma membrane integrity (PMI) were simultaneously analyzed. Apart from the statistically significant effect (P < 0.001) of semen storage time, the importance of the individual source of the ejaculate for the analyzed parameters of metabolic efficiency of spermatozoa was shown. This phenomenon was manifested in the interaction of the individual source of the ejaculate with spermatozoa motility, integrity of their membranes and metabolic activity with the passing time of semen preservation. Recorded results indicate that the individual factor may have a significant influence on the technological usefulness of boar spermatozoa for liquid storage. Quality analyses conducted on boar semen stored at 5°C may be used for pre-selection of boars producing sperm with an enhanced tolerance to cold shock.


Zygote ◽  
2018 ◽  
Vol 27 (1) ◽  
pp. 17-24 ◽  
Author(s):  
Andressa Dalmazzo ◽  
Daniel de Souza Ramos Angrimani ◽  
João Diego A. Losano ◽  
Carolina C. Rocha ◽  
Carlos A. B. Sobrinho ◽  
...  

SummaryThe aim of this study was to compare different concentrations of soy lecithin (LEC0.01%, LEC0.05% and LEC0.1%) with egg yolk (Control) in cooling extenders during the storage of semen at 5ºC for 5 days. Twelve dogs (n = 12) were selected, and semen was cooled and assessed after 2, 24, 48, 72, 96 or 120 h. At each time point, sperm were analyzed for kinetic patterns (using computer-assisted sperm analysis), mitochondrial activity (3′3- diaminobenzidine assay), lipid peroxidation (TBARS assay), DNA fragmentation (SCSA®) and plasma and acrosome membrane integrity (eosin/nigrosin and fast green/rose Bengal stains, respectively). The Control group (1814.4 ± 197.2) presented the highest rates of lipid peroxidation at 120 h. Conversely, progressive motility (42.8 ± 4%), linearity (45.4 ± 1%), and VAP (88 ± 3%) were higher in the Control group. In addition, there was lower mitochondrial activity in the Control group at 72 h. Therefore, our data show that lecithin used at these concentrations was not able to maintain sperm viability at as high qualities as would egg yolk. Moreover, the decrease in high mitochondrial activity and the persistence of sperm motility may indicate a compensatory mechanism in canine spermatozoa (i.e., glycolytic pathway). Furthermore, these higher lipid peroxidation indexes could indicate the necessity for future therapy using extenders and antioxidants over a long cooling time for dog sperm.


2018 ◽  
Vol 29 (5) ◽  
pp. 507-512
Author(s):  
Miriam Zaccaro Scelza ◽  
Isleine Portal Caldas ◽  
Jhony Medeiros de Mattos ◽  
Felipe Oliveira ◽  
Waldimir Carvalho ◽  
...  

Abstract This study aimed to compare the cytotoxicity of the Vita AC12, Lava Ultimate, Vita Enamic and InSync indirect restorative materials. Extracts of each material were prepared by incubation for 1, 7 and 40 days, with daily washing. Human gingival fibroblasts were exposed to the extracts, and cell viability was evaluated by sequential assessment of mitochondrial activity (XTT), membrane integrity (NRU) and cell density (CVDE). Extracts of polystyrene beads and latex fragments were used as negative and positive controls, respectively. Differences between groups and experimental times were evaluated by analysis of variance. At the 24 h extraction, significant differences between the control and both Vita AC-12 and InSync were observed in the XTT assay (p<0.05), and between the control and both Enamic and Lava Ultimate, in the CVDE assay (p<0.05). AC12, Lava Ultimate, and InSync presented significantly lower cell viability than Enamic and the control group, in the NRU assay (p<0.05). The Vita Enamic and Lava Ultimate hybrid ceramic-like materials presented better biocompatibility at the 24 h extraction time point than the AC12 and InSync ceramic materials. However, a simulation of the removal of toxic components by biological fluids, conducted by using longer extraction times and daily washing, led to the absence of cytotoxicity in all the tested restorative materials. These findings can be viewed as positive for the clinical indication of these restorative materials, considering their contact with adjacent soft tissues for extended periods of time.


2011 ◽  
Vol 493-494 ◽  
pp. 37-42 ◽  
Author(s):  
G.V.O. Fernandes ◽  
Gutemberg Alves ◽  
A.B.R. Linhares ◽  
Marcelo Henrique Prado da Silva ◽  
J.M. Granjeiro

The pursuit for an ideal bone substitute remains the main focus of many tissue engineering researchers. Among the myriad types of grafts available, synthetic bone grafts are of special importance, because it is available in large amounts, reduce the surgical trauma and eliminate the risk of diseases’ transmission. In this context, bioactive glasses have received attention mostly due to its described biocompatibility and rapid rate of surface reactivity when compared with other materials, allowing for faster interactions with the local tissue. The addition of niobium to this material has been shown as increasing the chemical resistance of the compound and providing greater stability. However, alterations on the chemical composition of biomaterials may impact on its biocompatibility. Therefore, the aim of this study was to evaluate thein vitrobiocompatibility of bioglass-Niobium (BgNb) granules, in comparison with standard commercial bioglass (Biogran®) throughout an interesting multiparametrical approach, employing Phenol 2% and dense polystyrene beads as positive and negative controls, respectively. Extracts from each material were prepared by 24 hours incubation in culture medium (DMEM). Human primary osteoblasts were then exposed for 24 hours to each extract and cell viability was evaluated by three parameters: mitochondrial activity (XTT method), membrane integrity (neutral red dye uptake) and cell density (crystal violet dye exclusion test). BgNb extracts were highly compatible, since the levels of viable cells were similar to the control group (unexposed cells), on all parameters studied. The mean cell density on the Biogran® group was slightly lower than BgNb, even though this material was also non-cytotoxic. The excellentin vitroresponse for BgNb granules indicates the suitability of this material to future studies on its biological and physical properties when appliedin vivo.


2019 ◽  
Author(s):  
Mahdieh Mehdipour ◽  
Hossein Daghigh Kia ◽  
Abouzar Najafi

AbstractThe aim of our research was to examine the effects of crocin (0.5 (C0.5), 1 (C1) and 1.5 (C1.5) mM) and naringenin (50 (N50), 100 (N100) and 150 (N150) µM) in cryopreservation extender for freezing rooster semen. Sperm motility, viability, abnormalities, membrane integrity, mitochondrial activity, apoptosis status, lipid peroxidation (LP), GPX, SOD, TAC, the mRNA expression of pro-apoptotic (CASPASE 3) and anti-apoptotic (Bcl-2) genes, fertility and hatchability rate were investigated following freeze-thawing. C1 and N100 resulted in the higher (P < 0.05) total motility and progressive motility in comparison to the control group. C1 and N100 improved viability, membrane integrity and reduced lipid peroxidation. We found much higher values for mitochondria activity with C1 and N100 respect to the control group. The C1 and N100 showed lower percentages of early apoptosis when compared with control group. Also, C1 and N100 had higher TAC when compared with control group. The mRNA expression of BCL-2 in the C1 and N100 group were significantly higher than that of other treatments. The expression of CASPASES 3 was significantly reduced in C1 and N100 group (P < 0.05) when compared to control group. Significantly higher percentage of fertility and hatching rate were observed in C1 and N100 compared to the control group. In conclusion, crocin at 1 mM and naringenin at 100 µM seem to improve the post-thawing rooster semen quality, fertility and could protect the sperm against excessive ROS generation by reducing the pro-apoptotic (CASPASE 3) and increasing anti-apoptotic (Bcl-2) genes.


2012 ◽  
Vol 518-523 ◽  
pp. 942-945 ◽  
Author(s):  
Zhao Xiang Han ◽  
Meng Zhang ◽  
Chun Xia Lv

Abstract. Little is known about the potential behavior and ecotoxicity of nanoparticles to aquatic organisms. To carefully address this issue, we conducted a comprehensive toxicity assessment, including NiO nanoparticles accumulation and toxicity tests as well as Lipid peroxidation (LPO), MDA content, mitochondrial depolarization and Reactive oxygen radical (ROS) analysis using Gracilaria lemaneiformis as a model organism. Our results demonstrated that the concentration of NiO Nanoparticles accumulated in Gracilaria lemaneiformis increased with the extended incubation time and a considerable amount of NiO Nanoparticles were still present in the bodies of Gracilaria lemaneiformis. Exposure to NiO nanoparticles led to significant reductions of cell viability between 20 and 60% relative to controls. Elevated lipid peroxidation in Gracilaria lemaneiformis exposed to NiO nanoparticles compared to control and induced increases in MDA levels with incubation time. Significantly enhanced percent of mitochondrial depolarization in nanometer size compared to controls and NiO nanoparticles significantly elevated ROS levels more than three fold at the highest concentration. Taken together, these findings indicate that NiO nanoparticles exposure, especially for long periods of time, may exert negative impact on population of aquatic organisms and on food web dynamics in aquatic systems.


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