Introduction: Since photobiomodulation therapy (PBMT) favors in vitro mesenchymal stem cell (MSC) preconditioning before MSC transplantation, increasing the proliferation of these cells without molecular injuries by conserving their characteristics, in the present in vitro study we analyzed the effect of PBMT on the proliferation and osteogenic differentiation of human umbilical cord mesenchymal stem cells (hUCMSCs). Methods: Irradiation with an InGaAIP Laser (660 nm, 10 mW, 2.5 J/cm 2 , 0.08 cm 2 spot size, and 10 s) was carried out. The cells were divided into four groups: CONTROL [cells grown in Dulbecco's Modified Eagle Medium (DMEM)], OSTEO (cells grown in an osteogenic medium); PBMT (cells grown in DMEM+PBMT), and OSTEO+PBMT (cells grown in an osteogenic medium plus PBMT). The cell proliferation curve was obtained over periods of 24, 48 and 72 hours using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. Osteogenic differentiation was analyzed by the formation of calcium nodules over periods of 7, 14 and 21 days. Morphometric analysis was performed to quantify the total area of nodular calcification. Results: The highest cell proliferation and cell differentiation occurred in the OSTEO+PBMT group, followed by the PBMT, OSTEO and CONTROL groups respectively, at the observed times (P < 0.05). Conclusion:PBMT enhanced the osteogenic proliferation and the differentiation of hUCMSCs during the periods tested, without causing damage to the cells and preserving their specific characteristics, a fact that may represent an innovative pretreatment in the application of stem cells.
Aims: To investigate the effects of the lectin from Punica granatum sarcotesta (PgTeL) on growth, viability, cell structure, biofilm formation and chitinase activity of Listeria monocytogenes. In addition, the effect of PgTeL on the adhesion and invasion of human cells (HeLa) was determined. Methods and Results: PgTeL showed bacteriostatic and bactericidal effects on the strains L. monocytogenes N53-1 and EGD-e, causing morphometric alterations, cell aggregation, strong deformation and cell disruption. PgTeL inhibited biofilm formation by EGD-e and N53-1 and also interfered with the adhesion and invasion processes of EGD-e and N53-1 in HeLa cells. Finally, the chitinase activity of L. monocytogenes EGD-e was reduced in the presence of PgTeL, which can be involved in the inhibition of adhesion process. Conclusion:PgTeL is an antibacterial agent against L. monocytogenes, inhibiting growth and promoting cell death, as well as impairing biofilm formation and bacterial adhesion and invasion into human cells. Significance and Impact of the Study: The results stimulate future investigations on the potential of PgTeL for protection of contamination in food products.
Lectins (carbohydrate-binding proteins) are able to distinguish different patterns of glycosylation on cell surfaces. This study investigated the effects of lectins from Alpinia purpurata inflorescence (ApuL) and Schinus terebinthifolia leaf (SteLL) on the viability of human leukemia cells (K562, chronic myeloid leukemia; JURKAT, acute lymphoblastic leukemia) and mesenchymal stem cells (MSCs) from human umbilical cords. In addition, possible immunomodulatory effects of ApuL and SteLL on MSCs were assessed by determining cytokine levels in cultures. ApuL reduced the viability of JURKAT cells (IC50: 12.5 μg/mL), inducing both apoptosis and necrosis. For K562 cells, ApuL at 50 µg/mL caused a decrease in viability, but of only 8.8%. Conversely, SteLL exerted a cytotoxic effect on K562 (IC50: 6.0 μg/mL), inducing apoptosis, while it was not cytotoxic to JURKAT. ApuL and SteLL (0.19–100 μg/mL) did not decrease MSCs viability. Treatment with ApuL strongly suppressed (99.5% reduction) the release of IL-6 by MSCs. SteLL also reduced the levels of this cytokine in culture supernatant. In conclusion, ApuL and SteLL showed potential to reduce the viability of leukemia cells, as well as immunomodulatory effect on MSCs without being toxic to them. These biological properties can be explored biomedically and biotechnologically in the future.
Ethnopharmacobotanical information reports that Parkinsonia aculeata infusion is used to control diabetes-related complications and dyslipidemia. However, few studies are reported on the safe use of this species. The aim of this study is to evaluate the acute toxicity, embryotoxicity and cytotoxicity of a polar fraction obtained from hydroethanolic extract of P. aculeata (PfrHEPA). For the acute toxicity test, we considered the Up and Down method which the guidelines are described by the Organization for Economic Cooperation and Development (OECD N°425). The animals were treated with PfrHEPA (2000 mg/kg) or with distilled water (10 ml/kg) by gavage and observed from Day 1 to14. For embryotoxicity assay, zebrafish embryos were exposed to PfrHEPA (100 mg/L) and toxicity parameters were observed during four consecutive days. The cytotoxicity of PfrHEPA (5, 10, 25, 50, 75 and 100 μg/ml, respectively) was performed on normal cell lines (mesenchymal stem cells, African green monkey renal cells and mouse pre-adipocytes 3 T3-L1 using the MTT salt reduction assay. In the acute toxicity test, no mortality was observed in mice treated with PfrHEPA (2000 mg/kg), as well as behavioral changes, histopathological abnormalities and hematological and biochemical variables. In the embryotoxicity test, no abnormal changes related to the toxicological parameters were observed in the period of 96 h. Regarding the cytotoxicity assay, PfrHEPA showed no cytotoxic effect on the normal cell lines tested, with an IC50 value > 100 μg/ml. These results suggest the safe use of P. aculeata, however, more trials are needed for PfrHEPA to be presented as new safe therapeutic proposal for the control of metabolic disorders.
Introdução: A geometria fractal tem a capacidade de descrever a forma irregular ou fragmentada de objetos considerados complexos, portanto, pode ser uma ferramenta útil para análise computadorizada de imagens médicas. Neste contexto, a análise fractal vem sendo aplicada na caracterização de células de diferentes tecidos humanos, inclusive as células-tronco mesenquimais (CTMs). Objetivos: Utilizar a análise fractal para avaliar in vitro a diferenciação adipogênica das células-tronco mesenquimais irradiadas. Métodos: Essa pesquisa tem aprovação do Comitê de Ética e Pesquisa em Seres Humanos – CCS/UFPE (2.259.272). As CTMs foram obtidas do cordão umbilical, sendo cultivadas em meio DMEM baixa glicose, suplementado com soro fetal bovino, fator de crescimento e antibióticos e mantidas na estufa a 37 ºC, 80% umidade e 5% CO2 por 21 dias. Após este tempo, as CTMs foram separadas em dois grupos de estudo: o NR (não irradiado) e o IR (irradiado). O grupo IR foi irradiado, in vitro, com dose de 3 Gy a partir de uma fonte de cobalto- 60 (Gammacell 2020 Excel) com taxa de dose de 2,657 kGy/h. Nos grupos NR e IR foi induzido o processo de diferenciação adipogênica durante 21 dias, quando foram capturadas 20 fotomicrografias das células, sendo 10 do grupo NR e 10 do grupo IR. As imagens foram processadas e o contorno da membrana celular foi definido por meio do software de imagem GIMP e a análise fractal foi realizada por meio do software ImageJ (NIH, EUA) com adição do puglin FracLac, que possibilita o cálculo da dimensão fractal. A análise estatística foi realizada pelo teste das médias com representação para o teste normal, assumindo dois grupos independentes com confiança de 95%. Resultados: A média da dimensão fractal das células do grupo NR foi de 1,14 ± 0,05 (mediana 1,13) e a do grupo IR foi de 1,12 ± 0,02 (mediana 1,12). Não foi observada diferença significativa dos dados de dimensão fractal entre os grupos, mas observou-se uma maior dispersão dos dados no grupo NR em relação ou grupo IR. É provável que haja uma relação com a alteração da estrutura do citoesqueleto celular após a irradiação, com a desorganização da rede de actina e a irregularidade dos componentes internos celulares(1, 2, 3). Conclusões: A análise fractal mostrou-se ser um método computacional eficaz para caracterizar as CTMs após a irradiação e diferenciação celular.
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