Veterinary Research Forum
Vet Res Forum

DNA damage in dental pulp mesenchymal stem cells: An in vitro study

Document Type : Original Article

Authors

Jaime Sardá Aramburú Junior 1
Tiago Luis Eilers Treichel 1
Saulo Tadeu Lemos Pinto Filho 1
Sergio Alexandre Gerhke 2
Alencar Kolinski Machado 3
Francine Carla Cadoná 3
Ivana Beatrice Mânica da Cruz 3
Ney Luis Pippi 1

1 Graduate Program in Veterinary Medicine, Federal University of Santa Maria, Santa Maria, Brazil

2 Biotecnos Research Center, Santa Maria, Rio Grande do Sul, Brazil; Catholic University of Uruguay, Montevideo, Uruguay

3 Morphology Department, Federal University of Santa Maria, Santa Maria, Brazil

https://doi.org/10.30466/vrf.2018.33083
Abstract
The aim of this study was to evaluate the potential use of a DNA comet assay, DNA fragmentation fluorimetric assay and reactive oxygen species levels as potential biomarkers of genome conditions of dental pulp stem cells (DPSCs) isolated from dog canine teeth. Mesenchymal stem cells were isolated from the dental pulp collected from dog teeth. The results obtained suggest the ideal moment for clinical application of cellular therapy for this type of cell. The cell culture was maintained with Dulbecco’s modified Eagle’s medium supplemented with 10.00% fetal bovine serum for eight passages. During each passage, cell proliferation, oxidative stress and level of DNA fragmentation were assessed by3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) assay, testing 2,7 dichlorodihydro-fluorescein-diacetate and PicoGreen®, respectively. There were important differences among the first three DPSC passages compared to passages 4–8 and a large number of nuclei with some levels of DNA damage (30.00 to 40.00% in initial DPSC passages and > 50.00% in late passages), indicating in vitro DPSC genomic fragility. Within the limitations of this study, the results suggest these relatively simple and inexpensive approaches - comet and DNA fragmentation assays - could help sort stem cells with less DNA damage for use in research or therapies.

Keywords

Dental pulp
DNA damage
Genotoxicity
Mesenchymal stem cell
Oxidative stress

Subjects

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Veterinary Research Forum
Volume 9, Issue 4
Autumn 2018
Pages 293-299

  • Receive Date 12 November 2018

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