Navegando por Palavras-chave "Cell Culture"

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    Comprovação de claim de ativos cosméticos in vitro utilizando modelo tridimensional de cultura celular em adipócitos
    (Universidade Federal de São Paulo (UNIFESP), 2020-03-23) Ueoka, Andreza Rodrigues [UNIFESP]; Lopes, Patricia Santos [UNIFESP]; Universidade Federal de São Paulo
    In vitro safety and efficacy assesment models of cosmetic actives are increasingly popular as there is a ban on the use of animals in 39 countries and some Brazilian states. In this context, a new in vitro model of three-dimensional coculture of adipose and endothelial cells was proposed for the in vitro safety assessment and proof of efficacy for improvement skin appearence of cellulite and aged skin. The safety trials of the assets Centella asiatica, horse chestnut, witch hazel, caffeine, Blend and Ecoblend were performed by cytotoxicity assays in three different types of cells strains, phototoxicity, genotoxicity with flow cytometry method and cytotoxicity in three-dimensional coculture of adipose and endothelial cells and the efficacy assay was performed by the Oil Red O method in single-layer cells and three-dimensional coculture. The results showed that the active Centella asiatica, Blend and caffeine in the concentration 2.5% were more cytotoxic in endothelial cells than in Balb/c 3T3 cells, since they provided a decreasement of about 10%, 98% and 20%, respectively. In the cytotoxicity assay, the active Centella asiatica, witch hazel, Blend and caffeine at the concentration of 2.5% were more cytotoxic in 3T3-L1 cells than in Balb/c 3T3 cells, since they led to a decreasement of about 8%, 24%, 99% and 20%, respectively. None of the evaluated actives presented phototoxicity and genotoxicity. The three-dimensional coculture of adipose and endothelial cells presented greater cellular viability when compared to single-layer cells at the same concentration. In the efficacy test, the Centella asiatica, horse chestnut, witch hazel, caffeine and Blend in the highest test concentration caused a reduction in the amount of intracellular lipids around 40%, 12%, 27%, 35% and 30%, respectively. It was possible to conclude that the proposed model of three-dimensional co-culture for in vitro evaluation of safety and efficacy of cosmetic assets was effective and advantageous in relation to the 2D model, since it presents better representation of adipose tissue with concentrations closer to cell use and organization more similar to the in vivo system.
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    Influência Do Meio De Cultivo Condicionado De Células-Tronco Mesenquimais No Desenvolvimento Embrionário Murino Até O Estágio De Blastocisto
    (Universidade Federal de São Paulo (UNIFESP), 2018-05-22) Silveira, Larissa Berioni Rodrigues Da [UNIFESP]; Turco, Edson Guimaraes Lo [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)
    Objective: Evaluate The Influence Of The Conditioned Culture Medium From Endometrial Mesenchymal Stem Cells In The Embryonic Cell Culture Method: 544 Murine Embryos Were Divided In 3 Groups: Control (Regular Embryonic Culture Medium); Supplemented (Embryonic Culture Medium Supplemented With Conditioned Imdm Medium After Endometrial Mesenchymal Stem Cell Culture); And Negative Control (Regular Embryonic Culture Medium Supplemented With Imdm Medium). Embryonic Development Was Evaluated In Each Group And Culture Media Were Collected For Metabolomic Analysis By Mass Spectrometry. The Data Were Normalized By Z-Score And Submitted To The Pca And Pls-Da Tests Using The Software Metaboanalyst 3.0, To Identify The Molecular Markers. Results: In The Comparison Between The Three Groups, The Control Group Presented A Higher Blastocyst Rate On The Sixth Day Of Embryo Culture In Relation To The Supplemented And Negative Control Groups (0,3 ± 0,160; 0,01±0,01; 0,00±0,00, Respectively) , While The Rate Of Degenerate Was High
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    Mecanismo de ação molecular envolvido na captação de Ca2+ de compostos presentes na Cecropia glaziouii Sneth (procianidinas, flavanóis e flavonóides) em culturas de células musculares lisas vasculares e células endoteliais
    (Universidade Federal de São Paulo (UNIFESP), 2020-04-07) Dermondes, Gabriella Trettel [UNIFESP]; Landman, Maria Teresa Riggio De Lima [UNIFESP]; http://lattes.cnpq.br/8687378770717503; http://lattes.cnpq.br/0174008735696387; Universidade Federal de São Paulo
    The vast majority of drugs used in therapy derived from active products taken from nature. Cecropia glaziouii Sneth is a tree typical of tropical and subtropical regions of Latin America, and is widely used in folk medicine. Previous studies from our group demonstrated that the aqueous extract (EA) of Cecropia glaziouii Sneth and its butanolic fraction (FBut) have hypotensive and antihypertensive activities which were related to relaxation of vascular smooth muscle by in vitro experiments. The same activitieswere obtained after oral chronic administration of EA and FbutBased on these data, this work aims to analyze the molecular mechanisms of action of compounds present in C. glaziouii Sneth in calcium mobilization by cultured vascular smooth muscle and endothelial cells. The mechanisms of action of the following major Cecropia compounds were analyzed in calcium mobilization, by spectrophotofluorimetry: isovitexine, isoorientine, catechin, epicatechin, procyanidin B2, procyanidin B3 and orientin. The results were obtained, by comparison with agonists (KCl, ACh, Ang II and BK) and antagonists (verapamil, atropine, losartan and HOE140) with known mechanisms of action. It was verified that Cecropia compounds influence the mobilization of intracellular calcium in both cell types. In endothelial cells, all compounds tested, except isoorientin, act similarly to verapamil, blocking L-type calcium channels; and the compounds isovitexin, epicatechin and procyanidin B3 have a mixed effect, also acting through the muscarinic pathway via activation of M3 metabotropic receptors. All compounds, except isoorientin, act in the pathway involving AT1 receptors, since their activity were blocked by losartan, while isovitexin, isoorientin, epicatechin and procyanidin B3 present an effect on calcium mobilization via NO pathway, since their action was blocked by the antagonist HOE 140. In vascular smooth muscle cells, the observed effect suggests that the all the tested compounds act via L-type calcium channels. In the case of isovitexin and isoorientin this effect was associated with an action involving AT1 receptors. In conclusion the tested compounds present in Cecropia glaziouii Sneth, tested in cultured endothelial and vascular smooth muscle cells, mostly present a mixed mechanism of action on intracellular calcium mobilization, acting in more than one pathway involved in justifying the popular antihypertensive use of the plant.