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Doctoral Thesis
DOI
https://doi.org/10.11606/T.17.2024.tde-12062024-132637
Document
Author
Full name
Ana Paula Ferro
Institute/School/College
Knowledge Area
Date of Defense
Published
Ribeirão Preto, 2024
Supervisor
Committee
Guirro, Elaine Caldeira de Oliveira (President)
Amaral, André Capaldo
Renno, Ana Claudia Muniz
Rocha, José Carlos Tatmatsu
Title in Portuguese
Influência da fotobiomodulação em células mesenquimais multipotentes de tecido adiposo in vitro e fibroblastos de pacientes portadores de anemia falciforme
Keywords in Portuguese
Célula tronco mesenquimal
Cicatrização
Fotobiomodulação
Viabilidade celular
Abstract in Portuguese
Feridas cutâneas em membros inferiores podem surgir como complicações relacionadas a anemia falciforme. Células-tronco indiferenciadas podem se diferenciar em vários tipos de células para reparar tecidos lesados. A fotobiomodulação pode promover a proliferação e diferenciação de células estaminais. O objetivo do estudo é investigar o efeito de diferentes parâmetros de fotobiomodulação com Laser e LED em células-tronco mesenquimais (CTMs), multipotentes derivadas do tecido adiposo e fibroblastos de indivíduos com anemia falciforme, sobre a proliferação e as possíveis alterações morfológicas nucleares em células tronco mesenquimais. Métodos: Células tronco mesenquimais derivadas do tecido adiposo foram submetidas a aplicações de Laser de baixa intensidade (660 nm e 830 nm; com doses de 0,5 J, 2 J e 4 J, potências de 40 mW e 100 mW) e LED (630 nm e 850 nm, com frequência variável de 0 a 1500 Hz e doses de 0,5 J, 2 J e 4 J). Após a irradiação, as células foram análisadas em 24, 48 e 72 horas com marcação de Hoechest (células viáveis) e Iodeto de Propidio (células mortas), para análise da taxa de proliferação, morte e viabilidade celular. O ensaio de cicatrização de ferida foi realizado por meio de um arranhão na camada celular com uma pipeta de 1000 µL e a análise fo realizada com o microscópio confocal e mesa motorizada (LSM 710®, Zeiss, Alemanha). Ambos os comprimentos de onda do laser e do LED foram efetivos para a proliferação celular, no entanto a viabilidade celular com o laser nas primeiras 48 horas após a irradiação, o comprimento de onda de 660 nm e potência de 100 mW mostrou resultados significativos, e com o avançar do tempo, em 72 horas, o laser de 830 nm teve aumento significativamente maior, que o laser de 660 nm. Com o LED apenas o comprimento de onda de 630 nm teve diferença significativa. Em relação ao ensaio de cicatrização, houve redução significativa dos paraêtros utilizados comparado com o grupo controle. Conclui-se que diferentes parâmetros físicos da FBM, possui respostas biológicas que interferem na taxa de proliferação, morte e consequentemente na viabilidade celular.
Title in English
Influence of photobiomodulation on multipotent mesenchymal cells of adipose tissue in vitro and fibroblasts of patients with sickle cell anemia
Keywords in English
Cell viability
Healing
Mesenchymal stem cell
Photobiomodulation
Abstract in English
Skin wounds on the lower limbs can arise as complications related to sickle cell anemia. Undifferentiated stem cells can differentiate into various cell types to repair injured tissue. Photobiomodulation can promote the proliferation and differentiation of stem cells. The objective of the study is to investigate the effect of different laser and LED photobiomodulation parameters on multipotent mesenchymal stem cells (MSCs), derived from adipose tissue and fibroblasts from individuals with sickle cell anemia, on proliferation and possible nuclear morphological changes in cells mesenchymal trunk. Methods: Mesenchymal stem cells derived from adipose tissue were subjected to low-intensity laser applications (660 nm and 830 nm; with doses of 0.5 J, 2 J and 4 J, powers of 40 mW and 100 mW) and LED (630 nm and 850 nm, with variable frequency from 0 to 1500 Hz and doses of 0.5 J, 2 J and 4 J). After irradiation, the cells were analyzed at 24, 48 and 72 hours with Hoechest staining (viable cells) and Propidium Iodide (dead cells), to analyze the rate of proliferation, death and cell viability. The wound healing assay was performed by scratching the cell layer with a 1000 µL pipette and the analysis was carried out using a confocal microscope and motorized table (LSM 710®, Zeiss, Germany). Both laser and LED wavelengths were effective for cell proliferation, however cell viability with the laser in the first 48 hours after irradiation, the wavelength of 660 nm and power of 100 mW showed significant results, and As time progressed, in 72 hours, the 830 nm laser had a significantly greater increase than the 660 nm laser. With LED, only the 630 nm wavelength had a significant difference. Regarding the healing test, there was a significant reduction in the parameters used compared to the control group. It is concluded that different physical parameters of FBM have biological responses that interfere with the rate of proliferation, death and consequently cell viability.
 
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Publishing Date
2024-07-11
 
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