scholarly journals Oily formulations challenge: how to evaluate their beneficial effects in hydrophilic cell-based models?

OCL ◽  
2018 ◽  
Vol 25 (5) ◽  
pp. D508 ◽  
Author(s):  
Elodie Olivier ◽  
Annabelle L’Hermitte ◽  
Patrice Rat ◽  
Melody Dutot
Keyword(s):  
Organic Solvents ◽  
High Throughput Screening ◽  
Cellular Response ◽  
Culture Medium ◽  
Recovery Period ◽  
The European Union ◽  
Cell Models ◽  
Cell Parameters ◽  
Beneficial Effects

In the European Union, Israel and India, testing cosmetic products or their ingredients on animals is prohibited. In this context, in vitro cell models play a pivotal role in the evaluation of both safety and beneficial effects of cosmetics. Oily formulations, widely used in cosmetics, are complex to study on cell models due to their lipophilic nature that doesn’t match with hydrophilic culture medium. Organic solvents are then required to solubilize oily formulations, but they can interfere with the cellular response. To avoid the use of organic solvents, we developed a method based on cells to evaluate potential beneficial effects of oily formulations. Our method, suitable for high throughput screening, consists in: (1) incubating cells with oily formulations for a short time followed by a recovery period in culture medium and (2) studying cell parameters using robust techniques such as cytofluorometry and fluorescence resonance energy transfer (FRET). Depending on the studied cell parameter, various beneficial effects can be revealed like antioxidant, anti-inflammatory and skin regeneration. The field of cell parameters is open and can be extended to new perspectives in the development of oily formulations.

scholarly journals Biological Relevance of Extra Virgin Olive Oil Polyphenols Metabolites

Antioxidants ◽  
2018 ◽  
Vol 7 (12) ◽  
pp. 170 ◽  
Author(s):  
Gabriele Serreli ◽  
Monica Deiana
Keyword(s):  
Olive Oil ◽  
Intracellular Signaling ◽  
Cellular Response ◽  
Virgin Olive Oil ◽  
Extra Virgin Olive Oil ◽  
Main Concern ◽  
Beneficial Effects ◽  
Positive Effects

Extra virgin olive oil (EVOO) polyphenols beneficial effects have widely been debated throughout the last three decades, with greater attention to hydroxytyrosol and tyrosol, which are by far the most studied. The main concern about the evaluation of EVOO phenols activities in vitro and in vivo is that the absorption and metabolism of these compounds once ingested lead to the production of different metabolites in the human body. EVOO phenols in the ingested forms are less concentrated in human tissues than their glucuronide, sulfate and methyl metabolites; on the other hand, metabolites may undergo deconjugation before entering the cells and thus act as free forms or may be reformed inside the cells so acting as conjugated forms. In most in vitro studies the presence of methyl/sulfate/glucuronide functional groups does not seem to inhibit biological activity. Parent compounds and metabolites have been shown to reach tissue concentrations useful to exert beneficial effects others than antioxidant and scavenging properties, by modulating intracellular signaling and improving cellular response to oxidative stress and pro-inflammatory stimuli. This review aims to give an overview on the reported evidence of the positive effects exerted by the main EVOO polyphenols metabolites in comparison with the parent compounds.

scholarly journals Effect of Nanostructured Scaffold on Human Adipose-Derived Stem Cells: Outcome of In Vitro Experiments

Nanomaterials ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 1822 ◽  
Author(s):  
Marina Borgese ◽  
Ludovica Barone ◽  
Federica Rossi ◽  
Mario Raspanti ◽  
Roberto Papait ◽  
...  
Keyword(s):  
Stem Cells ◽  
Culture Medium ◽  
Quantitative Polymerase Chain Reaction ◽  
Enzyme Linked Immunosorbent Assay ◽  
Adipose Derived Stem Cells ◽  
In Vitro Experiments ◽  
Fatty Acid Binding ◽  
Beneficial Effects ◽  
Angiogenesis Process

This work is addressed to provide, by in vitro experiments, results on the repercussion that a nanostructured scaffold could have on viability, differentiation and secretion of bioactive factors of human adipose-derived stem cells (hASCs) when used in association to promote angiogenesis, a crucial condition to favour tissue regeneration. To achieve this aim, we evaluated cell viability and morphology by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay and microscopy analysis, respectively. We also investigated the expression of some of those genes involved in angiogenesis and differentiation processes utilizing quantitative polymerase chain reaction (qPCR), whereas the amounts of Vascular Endothelial Growth Factor A, Interleukin 6 and Fatty Acid-Binding Protein 4 secreted in the culture medium, were quantified by enzyme-linked immunosorbent assay (ELISA). Results suggested that, in the presence of the scaffold, cell proliferation and the exocytosis of factors involved in the angiogenesis process are reduced; by contrast, the expression of those genes involved in hASC differentiation appeared enhanced. To guarantee cell survival, the construct dimensions are, generally, smaller than clinically required. Furthermore, being the paracrine event the primary mechanism exerting the beneficial effects on injured tissues, the use of conditioned culture medium instead of cells may be convenient.

High insulin concentrations promote the in vitro growth and viability of canine preantral follicles

2013 ◽  
Vol 25 (6) ◽  
pp. 927 ◽  
Author(s):  
Michelle K. B. Serafim ◽  
Gerlane M. Silva ◽  
Ana B. G. Duarte ◽  
V. R. Araújo ◽  
T. F. P. Silva ◽  
...  
Keyword(s):  
Culture Medium ◽  
Formation Rate ◽  
Minimum Essential Medium ◽  
Preantral Follicles ◽  
Beneficial Effects ◽  
Culture Viability ◽  
High Insulin ◽  
High Concentrations ◽  
High Insulin Concentration

To determine whether the effects of different concentrations of insulin on the development of canine preantral follicles in vitro were associated or not with FSH, secondary follicles were isolated and cultured. In Experiment 1, follicles were cultured in the following media: modified minimum essential medium (CtrlMEM) alone; CtrlMEM plus 5 ng mL–1 insulin (Ins5ng); CtrlMEM plus 10 ng mL–1 insulin (Ins10ng); and CtrlMEM plus 10 μg mL–1 insulin. In Experiment 2, follicles were cultured in the same media but in the presence of sequential FSH (i.e. CtrlFSH, Ins5ngF, Ins10ngF and 10μgF, respectively). Increasing concentrations of FSH (100, 500 and 1000 ng mL–1) were added sequentially to the culture medium on Days 0, 6 and 12 of culture. Viability were assessed at the end of culture and follicular diameter and the antrum formation rate at four time points (Days 0, 6, 12 and 18). In Experiment 1, the high insulin concentration significantly increased follicular viability (P < 0.05). In contrast, in Experiment 2, viability was not affected by the inclusion of insulin. In addition, viability was significantly better in follicles cultured in CtrlFSH (P < 0.05). The diameter of follicles in the high-insulin group in Experiment 1 and high-insulin plus FSH group in Experiment 2 was superior to other groups tested. In experiment 2, the Ins10μg and Ins10μgF groups exhibited significantly higher antrum formation rates than the other groups. In conclusion, in the absence of FSH, high concentrations of insulin have beneficial effects on follicular viability. However, to promote the growth of canine preantral follicles in vitro, it is recommended that a combination of insulin and FSH be added to the medium.

scholarly journals Comparison of in Vitro Models for the Prediction of Compound Absorption across the Human Intestinal Mucosa

2004 ◽  
Vol 9 (7) ◽  
pp. 598-606 ◽  
Author(s):  
Silvia Miret ◽  
Leo Abrahamse ◽  
Els M. de Groene
Keyword(s):  
Cell Differentiation ◽  
High Throughput ◽  
High Throughput Screening ◽  
In Vitro Models ◽  
Assay System ◽  
In Vitro Assays ◽  
Cell Models ◽  
Artificial Membrane ◽  
Permeability Assay

Several in vitro assays have been developed to evaluate the gastrointestinal absorption of compounds. Our aim was to compare 3 of these methods: 1) the bio-mimetic artificial membrane permeability assay (BAMPA) method, which offers a high-throughput, noncellular approach to the measurement of passive transport; 2) the traditional Caco-2 cell assay, the use of which as a high-throughput tool is limited by the long cell differentiation time (21 days); and 3) The BioCoat™ high-throughput screening Caco-2 Assay System, which reduces Caco-2 cell differentiation to 3 days. The transport of known compounds (such as cephalexin, propranolol, or chlorothiazide) was studied at pH 7.4 and 6.5 in BAMPA and both Caco-2 cell models. Permeability data obtained was correlated to known values of human absorption. Best correlations ( r = 0.9) were obtained at pH 6.5 for BAMPA and at pH 7.4 for the Caco-2 cells grown for 21 days. The Caco-2 BioCoat™ HTS Caco-2 Assay System does not seem to be adequate for the prediction of absorption. The overall results indicate that BAMPA and the 21-day Caco-2 system can be complementary for an accurate prediction of human intestinal absorption.

P0550ISCHEMIC PRECONDITIONING ATTENUATES RENAL ISCHEMIA-REPERFUSION INJURY THROUGH ACTIVATING THE NRF2/HO-1 PATHWAY ENHANCED AUTOPHAGY

2020 ◽  
Vol 35 (Supplement_3) ◽  
Author(s):  
Jeong Kye Hwang ◽  
HyunSu Choi ◽  
Mi-hyeong Kim
Keyword(s):  
Ischemia Reperfusion ◽  
Critical Role ◽  
Recovery Period ◽  
Kidney Injury ◽  
Mineral Oil ◽  
Renal Ischemia ◽  
Bafilomycin A1 ◽  
Beneficial Effects

Abstract Background and Aims Ischemic preconditioning (IPC), which is a brief and nonlethal episode of ischemia, confers protection against subsequent ischemia-reperfusion (I/R) through the up-regulation of endogenous protective mechanisms. Recent studies have reported that the activation of autophagy is associated with IPC. However, the underlying mechanisms still need to be determined. In this study, we investigated the role of IPC in renal I/R injury and demonstrated that IPC could ameliorate renal I/R injury by activating the Nrf2/HO-1 pathway and induction of autophagy. Method In order to gain insights into IPC–induced alterations at the cellular level, an in vitro model for IPC was designed using the human proximal tubule epithelial cell line HK-2. In vitro I/R was induced by layering mineral oil (for 60 min) over a thinfilm of media covering the cells followed by 120 min of reperfusion in normal media. IPC was performed by a 10-min period of incubation of cells under mineral oil followed by a 30-min recovery period prior to the 120 min reperfusion. To further determine the role of autophagy in IPC-induced renoprotection, the cells were treated with bafilomycin A1 (200 nM), prior to I/R injury. In a renal I/R injury model, mice were subjected to 30 min of renal ischemia followed by 24 h of reperfusion. IPC was produced by 5 min of ischemia followed by 10 min of reperfusion prior to sustained ischemia. Results We found that IPC significantly attenuated I/R-induced kidney injury and suppressed oxidative stress and inflammatory responses. IPC also increased LC3-II and decreased SQSTM/p62 and cleaved caspase-3 during renal I/R injury, as well as increased the number of intracellular double-membrane vesicles in injured renal cells. IPC-induced renal protection was efficiently attenuated by pretreatment with Bafilomycin A1 (0.3 mg/kg, IP). Pretreatment with IPC also dynamically affected the expression of Nrf2/HO-1 signaling components. The beneficial effects of IPC were abolished by Brusatol (2 mg/kg, IP), potent inhibitors of Nrf2. Moreover, the Nrf2 agonist t-BHQ (50 mg/kg, IP) showed similar activity as IPC. Conclusion In summary, the present study shows that Nrf2 plays a critical role in regulating the HO-1 production induced by I/R. Also, upregulation of Nrf2 expression might represent one of the major mechanisms whereby IPC confers protection against I/R-induced kidney injury. The beneficial effects of IPC could be attributed to its antioxidative stress and anti-inflammatory properties

scholarly journals Growth regulators, sucrose and potassium in the growth and biochemical activity of Curcuma longa L. micropropagated

2020 ◽  
Vol 1 (1) ◽  
pp. 18
Author(s):  
Meire Pereira de Souza Ferrari ◽  
Mayara Dos Santos Queiroz ◽  
Matheus Marquezini de Andrade ◽  
Jéssica Rezende Trettel ◽  
Hélida Mara Magalhães
Keyword(s):  
Antioxidant Activity ◽  
Growth Regulators ◽  
Culture Medium ◽  
Sucrose Concentration ◽  
Curcuma Longa ◽  
Naphthalene Acetic Acid ◽  
Biochemical Activity ◽  
Beneficial Effects ◽  
In Vitro Shoots

Curcuma longa L. is a plant widely used for its pharmacological and medicinal properties, however, does not have a conclusive micropropagation protocol.  The objective was to evaluate how growth regulators, sucrose and potassium influence the growth and biochemical activity of Curcuma longa seedlings grown in vitro.  Shoots were inoculated in culture medium supplemented with 6-benzylaminpurine - BAP (8.88 and 17.76 μM), Kinetin - KIN (0.92 and 2.16 μM) and naphthalene acetic acid - NAA (2.16 and 7.20 μM), potassium iodide KI (25 and 50 μM) and sucrose (30 and 60 gL-1). Growth regulators at the lowest concentration and 30 gL-1 sucrose promoted increases in growth of C. longa. Antioxidant activity was high in all treatments. The activity of the enzymes catalase and ascorbato peroxidase were increased in the two treatments that contained the highest concentrations of growth regulators. It is concluded that the addition of growth regulators such as cytokinins and auxins are fundamental to increase the number of leaves and growth of shoots in C. longa. Since the best concentrations are in the range of 4.44 to 8.88 μM of (BAP) and 2.16 of NAA. The addition of supplemental potassium in the culture medium is not necessary, and the beneficial effects of doubling the usual sucrose concentration are nullified if other constituents of the medium are altered. The antioxidant activity and enzymes had their activity altered, especially in treatments that did not contain growth regulators.

139 OXIDATIVE STRESS INDUCED CHANGES OF GLUTATHIONE CONCENTRATION IN IN VITRO-PRODUCED BOVINE BLASTOCYSTS

2007 ◽  
Vol 19 (1) ◽  
pp. 187 ◽  
Author(s):  
M. Aksoy ◽  
C. Wrenzycki ◽  
D. Herrmann ◽  
H. Niemann
Keyword(s):  
Oxidative Stress ◽  
De Novo ◽  
Critical Role ◽  
Recovery Period ◽  
Transcript Abundance ◽  
Culture Period ◽  
In Vitro Production ◽  
Tertiary Butyl ◽  
Beneficial Effects

Glutathione (GSH) plays a critical role in the elimination of reactive oxygen species (ROS) by acting as substrate for GSH peroxidase (GPX) which results in the oxidation of GSH. Glutathione reductase (GSR) regenerates GSH from oxidized glutathione (GSSG). In bovine oocytes, GSH concentration increases and is thought to have beneficial effects on embryo development. The goal of the present study was to analyze the GSH content in bovine blastocysts during blastocoel expansion and to determine the effects of tertiary-butyl hydroperoxide (tBH) in causing oxidative stress. It is metabolized by GPX with conversion of GSH to GSSG. Blastocysts were generated with standard protocols of in vitro production (IVP) using SOF medium supplemented with FAF-BSA and were collected at Days 7/8 of culture (Day 0 = IVF). Diameters of blastocysts were measured and embryos were allocated to 4 groups: early blastocysts (ebla, less than 135 �m), blastocysts (bla, 135–180 �m), expanded blastocysts (exbla, greater than 180 �m), and hatched blastocysts (hbla). Pools of the different categories (3 replicates, 36 embryos in total) were frozen at -80�C until assayed for GSH by HPLC. Total cell numbers were determined using the Hoechst 33342 stain. Blastocysts from the 4 groups were cultured together for 30 min in tBH (20 �M) containing SOF medium to induce oxidative stress. This was followed by another culture period for 1 or 8 h. Samples of blastocysts were frozen at -80�C at 0 h, 30 min, 1 h, and 8 h to assess GSH levels (Cereser et al. 2001 J. Chromatogr. B. Biomed. Sci. Appl. 752, 123–132) and transcript abundance. Semi-quantitative real-time RT-PCR was used to determine the relative abundance (RA) of GPX and GSR transcripts. Differences between groups were tested using ANOVA followed by a Tukey or Student&apos;s test (P &le; 0.05). The grade of expansion significantly increased GSH concentration in bovine blastocysts. Expanded blastocysts (0.38 &plusmn; 0.04 ng) and hatched blastocysts (0.46 &plusmn; 0.05 ng) had a higher GSH content than early blastocysts (0.21 &plusmn; 0.01 ng) and blastocysts (0.27 &plusmn; 0.02 ng). However, GSH concentration remained constant in blastomeres during expansion and hatching as the number of cells increased from 54.9 &plusmn; 3.5 (ebla) to 152.3 &plusmn; 4.7 (hbla). After induction of oxidative stress, blastocysts showed a significant decrease in GSH content after a 1-h recovery period, followed by an increase in their GSH content after a period of 8 h (0 h: 0.26 &plusmn; 0.01 ng; 30 min: 0.25 &plusmn; 0.01 ng; 1 h: 0.20 &plusmn; 0.01 ng; 8 h: 0.32 &plusmn; 0.01 ng, respectively). This was accompanied by a significant increase of the RA of GSR transcripts in blastocysts collected after the 8-h recovery period compared to embryos at the other time points of the treatment. GPX mRNA abundance remained stable during the entire culture period. In the present study, GSH concentration was determined in bovine embryos during blastocoel expansion for the first time. The increased GSH content might to some extent be due to de novo GSH synthesis. GSSG is reduced to GSH most likely by GSR. Results show that bovine IVP embryos may serve as a new model for the study of oxidative stress.

The Neurotoxicity of Organic Solvents Studied Using Synaptosomes and Neural Cell Cultures

1994 ◽  
Vol 22 (3) ◽  
pp. 175-179
Author(s):  
Leila Naskali ◽  
Maria Engelke ◽  
Hanna Tähti
Keyword(s):  
Organic Solvents ◽  
Cell Types ◽  
Cell Models ◽  
Cns Depressant ◽  
Optimal Functioning ◽  
Neurotoxic Effects ◽  
Lipid Protein Interaction ◽  
Solvent Molecules ◽  
Integral Proteins

The most common acute neurotoxic effect of organic solvents is their central nervous system (CNS) depressant effect. The molecular mechanism underlying this effect is not known. The purpose of our studies has been to evaluate the adverse effects of organic solvents on the CNS in vitro. Synaptosomal membranes, whole brain reaggregate and astrocyte cultures were studied. Our results suggest that cell membrane integral proteins are targets for solvent impact, but that there are differences among various cell types. In addition to lipophilicity, the structure of solvent molecules seems to be important when considering CNS toxicity. Organic solvents increase the fluidity of the membranes, which may disturb the lipid-protein interaction and the optimal functioning of the enzyme. However, direct effects of solvents on proteins cannot be excluded. In vitro cell models can be used in methods designed to predict acute neurotoxic effects of foreign compounds, and in studies of neurotoxic mechanisms.

scholarly journals Discovery of Novel Antigiardiasis Drug Candidates

2014 ◽  
Vol 58 (12) ◽  
pp. 7303-7311 ◽  
Author(s):  
Liudmila Kulakova ◽  
Andrey Galkin ◽  
Catherine Z. Chen ◽  
Noel Southall ◽  
Juan J. Marugan ◽  
...  
Keyword(s):  
High Throughput Screening ◽  
Standard Care ◽  
Parasitic Infection ◽  
The United States ◽  
The European Union ◽  
Current Standard ◽  
Content Type ◽  
Drug Candidates

ABSTRACTGiardiasis is a severe intestinal parasitic disease caused byGiardia lamblia, which inflicts many people in poor regions and is the most common parasitic infection in the United States. Current standard care drugs are associated with undesirable side effects, treatment failures, and an increasing incidence of drug resistance. As follow-up to a high-throughput screening of an approved drug library, which identified compounds lethal toG. lambliatrophozoites, we have determined the minimum lethal concentrations of 28 drugs and advanced 10 of them toin vivostudies in mice. The results were compared to treatment with the standard care drug, metronidazole, in order to identify drugs with equal or better anti-Giardiaactivities. Three drugs, fumagillin, carbadox, and tioxidazole, were identified. These compounds were also potent against metronidazole-resistant humanG. lambliaisolates (assemblages A and B), as determined inin vitroassays. Of these three compounds, fumagillin is currently an orphan drug used within the European Union to treat microsporidiosis in immunocompromised individuals, whereas carbadox and tioxidazole are used in veterinary medicine. A dose-dependent study of fumagillin in a giardiasis mouse model revealed that the effective dose of fumagillin was ∼100-fold lower than the metronidazole dose. Therefore, fumagillin may be advanced to further studies as an alternative treatment for giardiasis when metronidazole fails.

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