Interação de matriz polimérica de polietilenoglicol (PEG) com corante artificial de alaranjado de acridina e natural de açafrão nos parâmetros de viabilidade e atividade fagocítica

• Autor(a) principal: Silva, Luciana do Carmo Paulino
• Data de Publicação: 2020
• Tipo de documento: Dissertação
• Idioma: por
• Título da fonte: Repositório Institucional da UFMT
• Texto Completo: http://ri.ufmt.br/handle/1/5303
• topic: Biomaterial; Curcuma longa; EPEC; Leucócitos; Polímeros; CNPQ::CIENCIAS EXATAS E DA TERRA; Biomaterial; Curcuma longa; EPEC; Leukocytes; Polymers

Leukocytes are cells present in the blood, produced in the bone marrow, with the body's defense function. They can divide into polymorphonuclear cells (PMNs) and mononuclear cells (MNs), and this second group is widely used in immunological investigations, increasing in number in cases of infections. In this sense, for a long time in investigations in phagocytic processes, acridine orange is used as a dye with high sensitivity, staining live and dead cells differently. The acridine orange staining method is already well described in the literature and its use is quick to quantify due to its toxicity. In Brazil, the tuber of the turmeric plant is popularly known as saffron, and its rhizome produces a yellow powder, widely used in medicine, as a spice and natural color. In this sense, a new cell staining method was tested, using acridine orange in different concentrations and also saffron as a natural cell dye. In order to prolong this fluorescence process, polyethylene glycol (PEG) was used, which has the capacity to form porous structures, improving the release speed and showing the ability to adsorb organic compounds that improve biological functions. For this study, 39 peripheral blood samples (8mL) were collected from clinically healthy individuals in tubes with EDTA. Peripheral blood mononuclear cell populations were separated by density gradient using Ficoll-Paque, after being centrifuged at 1600rpm for 40 minutes, the phagocyte ring removed, washed twice in PBS, the cells counted and adjusted. Saffron and acridine orange (AA adsorbed or not with PEG) were used in concentrations of 100ng / mL and 100pg / mL, at 0 hours, 24 hours and 168 hours. The assay was carried out with five groups: the group with only cells, cells + PEG, cells + acridine orange, cells + saffron and cells + PEG + orange acridine. In the rheological analysis, the same groups were tested, noting that both the viscosity and the flow were not altered in the presence of PEG, AA and saffron. In cell viability, all groups were homogeneous, with a viability index greater than 90%. In the phagocytosis analysis it was observed for the concentrations of ng / mL without PEG (33.74 ± 0.50) and with PEG (36.98 ± 0.27) these did not differ (p <0.05). However, when we analyzed the groups on the pg / mL scale (p <0.0001), we noticed a difference between the groups with PEG (41.28 ± 0.41) and without the presence of the polymer (50.11 ± 0.56). Analyzing the phagocytosis of the Cells + EPEC group (17.6 ± 2.30) with the group containing cells, turmeric and EPEC (48.8 ± 6.97), this showed high phagocytosis rates compared to the control group (cells + EPEC). For the analysis of the microbicidal index, the same groups analyzed in phagocytosis were used, where there was no significant difference between the groups analyzed with p> 0.05. In the anion and SOD analyzes, there is also no significant difference, only the SOD of the group with saffron shows a difference (p <0.05). These results demonstrate the polyethylene glycol (PEG) adsorbed to the acridine orange dye in the pg / mL concentration and showed a modified release for up to 168 hours and do not interfere with cellular processes and that the saffron of the earth fulfills the role of cellular dye, but interferes in some activities biological factors, such as phagocytosis and SOD, emphasizing the importance of further studies for its use.