Determination of Azadirachta indica extract-induced genetic damage in human lymphocytes

Determinación del daño genético inducido por extracto de Azadirachta indica en linfocitos humanos

Currently, compounds of natural origin are considered as an alternative for pest control. A study was carried out for a biopesticide based on Neem seed as raw material (Azadirachta indica), evaluating the genotoxic effect on human lymphocytes to mitigate health risks for farmers. Azadirachta indica extraction was carried out by Soxhlet method and treatments were prepared at different concentrations (C1: 0.001 mM, C2: 0.0015 mM, C3: 0.002 mM).  Hydrogen peroxide was used as positive control and phosphate buffer (PBS) as negative control. Lymphocytes were obtained from the peripheral blood sample of a healthy 25-year-old individual. Subsequently, alkaline single-cell gel electrophoresis (SCGE) or Comet Test Assay was performed with the sandwich technique for cell protection and fixation. Samples were stained with Gelred for observation with 40x objective with Optika Italy B-510 series microscope and images were captured and evaluated in Comet ScoreTM software. The %DNA of the tail was the parameter taken into account for the measurement, obtaining as a result a higher damage or degradation in treatments C2 and C3, concluding that the concentration of Azadirachta indica is directly proportional to the damage of human lymphocytes.