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Influence of photobiomodulation and electrical stimulation in multipotent mesenchymal cells of adipose tissue in vitro

Grant number: 19/09329-1
Support type:Scholarships in Brazil - Doctorate (Direct)
Effective date (Start): August 01, 2019
Effective date (End): July 31, 2023
Field of knowledge:Health Sciences - Physiotherapy and Occupational Therapy
Principal Investigator:Elaine Caldeira de Oliveira Guirro
Grantee:Ana Paula Ferro
Home Institution: Faculdade de Medicina de Ribeirão Preto (FMRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil

Abstract

Cutaneous wounds in the lower limbs may arise as complications related to Sickle Cell Anemia. Undifferentiated stem cells can differentiate into various types of cells to repair damaged tissues. Photobiomodulation and electrical stimulation can promote proliferation and differentiation of stem cells. The aim of the study is to investigate the effect of different parameters of photobiomodulation with laser, LED, and electrical stimulation in Mesenchymal Stem Cells (MSCs), multipotents derived from adipose tissue and fibroblasts from individuals with and without sickle cell anemia, on proliferation and nuclear morphological changes in mesenchymal stem cells. Methods: mesenchymal stem cells derived from adipose tissue will be submitted to low intensity laser applications (660 nm and 830 nm at doses of 1 J/cm2, 4 J/cm2 and 10 J/cm2, potencies of 30 mW and 100 mW), LEDs (630 nm and 850 nm, with varying frequencies from 0 to 1500 Hz and doses of 1 J/cm2, 4 J/cm2 and 10 J/cm2). The blue stimulation of tripan will be used for evaluation of cell viability, and growth curves will be used to evaluate cell proliferation at zero, 24, 48, and 72 hours. Nuclear alterations will be assessed by staining with DAPI (4-6-diamidino-2-phenylindole) within 72 hours. For the statistical analysis, in the case of normal distribution, the ANOVA followed by Tukey test will be applied. If the data do not present a normal distribution, Kruskal-Wallis will be used, followed by the Dunn test. Data processing will be performed by SPSS software (Chicago, IL, USA), version 17.0, with the critical level set at 5% (p <0.05). (AU)