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Analysis of Cell Viability and Gene Expression After Continuous Ultrasound Therapy in L929 Fibroblast Cells
Larissa Dragonetti Bertin1, Regina Célia Poli-Frederico, Deise Aparecida Almeida Pires Oliveira
1From the Biological and Health Sciences Center, Pitágoras Unopar University, Londrina, Paraná, Brazil (LDB, RCP-F, PDO, FBP, AFSS); and UniEVANGÉLICA - Anápolis University Center, Anápolis, Goiás, Brazil (DAAPO, RFO).
Objective:
This study aimed to analyze cell viability and gene expression interleukin 6 and vascular endothelial growth factor after continuous ultrasound therapy of 1 and 3 MHz in L929 fibroblast cells.
Design:
The L929 cells were cultivated in 12-well plates and divided into the following five groups: Group 1 (G1), nonirradiated; G2, 0.2 W/cm-1 MHz; G3, 0.5 W/cm-1 MHz; G4, 0.2 W/cm-3 MHz; and G5, 0.5 W/cm-3 MHz. The cells were irradiated at 24 and 48 hrs. Cell viability was evaluated using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) method. The gene expression analysis was assessed using real-time polymerase chain reaction.
Results:
The G2 and G3 showed a decrease in cell viability when compared with the G1 at 48 hrs (P < 0.01). The G4 and G5 presented an increase in viability (P = 0.01 and P = 0.03, respectively) in 24 to 48 hrs. The cells irradiated at an intensity of 0.5 W/cm-3 MHz at 48 hrs showed a 0.21-fold decrease in interleukin-6 gene transcripts and a 1.67-fold increase in vascular endothelial growth factor gene transcripts.
Conclusions:
Continuous ultrasound therapy with a frequency of 3 MHz at an intensity of 0.5 W/cm stimulates cell proliferation, decreases interleukin-6 gene expression, and increases vascular endothelial growth factor gene expression in L929 fibroblast cells.
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