ARAUJO, A.V.P. Pre-clinical studies of acute and repeated-dose toxicity of synthetic phosphoethanolamine. [Dissertation]. São Paulo: Faculty of Medicine, University of São Paulo; 2017.Synthetic Phosphoethanolamine (FO-S), a monoester phospholipid has an important role on cell proliferation, induction of apoptosis, in tumor cells, without, however, significantly affecting normal cells. In this study the biological behavior and effects of acute toxicity of synthetic phosphoethanolamine (FO-S) were evaluated in single dose and repeated experiments in healthy mice, contributing to the pre-clinical validation of this compound as antitumor phospholipid. Mice of both sexes received intravenous FO-S compound at doses of 50, 100, 250, 500 and 1000 mg / kg in single dose, and 50, 100 and 250 mg / kg in repeated doses. In the single dose group, animals receiving 500 and 1000 mg / kg FO-S showed signs of toxicity, such as: 33% mortality of animals; lowering in the central and autonomic nervous system; fluctuations in hematological analyzes; increased levels of TGO and TGP; decreased creatinine; bone marrow analysis showed decreased myeloid and lymphoid populations; decrease of cells in the G0 / G1 phase of the cell cycle, as well as in the S phase, and increase in the G2 / M phase; histological changes in the heart, liver and kidneys such as hyperemia, necrosis and hyalinization. In both the single dose and repeated dose groups, there was an increase in the number of reticulocytes on the 7th day after application, as a response of the reactive bone marrow, and the cellularity analysis showed positive mitochondrial electrical potential activity. In the repeated dose group, animals receiving 50 mg / kg FO-S presented in the 14th mild anemia, the 100 mg group showed an increase in the number of leukocytes and lymphocytes and the group 250 mg fluctuations in the quantitative platelet values, which returned to the normality at 14 days. Bone marrow analysis revealed increased cell count in the myeloid compartment in the 50 mg group and increased cellularity in the erythroid compartment. Expression of CD34 marrow hematopoietic precursor cell markers was shown to be increased in the 250 mg group at 14 days and a decrease in myelodystic cell markers and CD43 lymphocyte subtypes in the groups receiving 100 and 250 mg / kg at 7 and 14 days. Animals that received 250 mg / kg presented changes in the lung parenchyma. Autophagy analysis was performed by flow cytometry that revealed no negative response, such as oxidative stress, presenting normal production of autophagic vacuoles, as well as the presence of micronuclei test did not demonstrate damage to the cells by genetic changes induced by toxicity.

Objective: The aim of this study was correlation proliferative activity, markers express stem cells, and lipid peroxides of undifferentiated stem cells of human adult dental follicle (DF) following culture. Methods: For this study, we used 8 samples from DF of impacted third molars to maintain culture conditions and evaluated the growth curve, cell viability, production of lipid peroxidation, cell cycle phases, and proliferative index during 25 days of culture. Results: Cells after culture showed characteristics of fibroblast-like type following 25th day of culture. The results of lipid peroxidation showed that stem cells in culture produce 13 nmoles/ml malondialdehyde at the start of culture, increasing until the 12th day and then began a decline that lasted until the 25th day. We revealed that DFSCs presented a significantly higher percentage of cells in S + G2/M phases by the 15th day of culture compared with cells at the start of culture. Cell surface markers revealed that cell lines were negative for HLA-DR and positive for CD90, CD44, and CD105. The expression of p21 protein, involved in the regulation of the cell cycle, showed a significant increase from the 15th to 25th day of culture. Results of cell division rates show a significant increase between the 6th and 15th day of culture. Conclusions: We conclude that the culture remained stable during the 25 days of culture, presenting the markers of stem cells and markers of control, progression, and cell proliferation that there was an increased production of lipid peroxides between the 6th and 12th days; this increase is related to the increased numbers of cells that also occurs during this period. Then, there is a significantly decline in the production of lipid peroxides and the number of cells, which is accompanied by an increase in cell unviability.