Effects of 660 and 940nm Low-Level Laser Irradiation on Viability and Proliferation of Stem Cells from Human Exfoliated Deciduous Teeth
Effect of LLLT on SHED
Abstract
Objectives: This study evaluated the effects of 660 and 940nm lasers on viability and proliferation of stem cells from human exfoliated deciduous teeth (SHED).
Materials and Methods: SHED were cultured and divided into six groups for laser irradiation: 660nm laser at 25mW with energy densities of 1 and 2J/cm2 for 8 and 16 seconds, and 940nm laser at 100mW with the same energy densities and durations. Two additional groups served as controls with no laser exposure. Cell viability was evaluated by the methyl thiazolyl tetrazolium (MTT) assay, and cell proliferation was measured by cell counting with a Neubauer chamber and qualitatively evaluated by 4′,6-diamidino-2-phenylindole (DAPI) staining at 24 and 72 hours post- irradiation. Data were analyzed using one-way ANOVA, followed by Tamhane’s and Tukey’s tests.
Results: Irradiation with 940nm laser enhanced cell proliferation compared to both 660nm laser and control groups at 24 and 72 hours (P<0.05), as confirmed by DAPI staining. At 24 hours, all laser-treated groups showed increased cell viability relative to the control, although the increase in the 660nm/2J/cm² group was not statistically significant (P>0.05). At 72 hours, only 940nm laser group showed significantly higher cell viability and proliferation than the control group (P<0.05).
Conclusion: These findings suggest low-level laser therapy, particularly at the wavelength of 940nm enhances the viability and proliferation of stem cell, supporting its potential as a promising tool in regenerative medicine and tissue engineering.
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15. Sylvester PW. Optimization of the tetrazolium dye (MTT) colorimetric assay for cellular growth and viability. Methods Mol Biol. 2011;716:157-68.
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19. Martin-Piedra MA, Garzon I, Oliveira AC, Alfonso-Rodriguez CA, Carriel V, Scionti G, et al. Cell viability and proliferation capability of long-term human dental pulp stem cell cultures. Cytotherapy. 2014 Feb;16(2):266-77.
20. Ligasová A, Koberna K. Quantification of fixed adherent cells using a strong enhancer of the fluorescence of DNA dyes. Sci Rep. 2019 Jun 18;9(1):8701.
21. Eduardo Fde P, Bueno DF, de Freitas PM, Marques MM, Passos-Bueno MR, Eduardo Cde P, et al. Stem cell proliferation under low intensity laser irradiation: a preliminary study. Lasers Surg Med. 2008 Aug;40(6):433-8.
22. Paschalidou M, Athanasiadou E, Arapostathis K, Kotsanos N, Koidis PT, Bakopoulou A, et al. Biological effects of low-level laser irradiation (LLLI) on stem cells from human exfoliated deciduous teeth (SHED). Clin Oral Investig. 2020 Jan;24(1):167-180.
23. Almeida-Junior LA, Marques NCT, Prado MTO, Oliveira TM, Sakai VT. Effect of single and multiple doses of low-level laser therapy on viability and proliferation of stem cells from human exfoliated deciduous teeth (SHED). Lasers Med Sci. 2019 Dec;34(9):1917-1924.
24. Souza L, Rinco U, Aguiar D, de Almeida Junior L, Silva L, Oliveira T, et al. Effect of low-level laser therapy on viability and proliferation of stem cells from exfoliated deciduous teeth under different nutritional conditions. Laser Phys. 2018 Jan 17;28(2):025603.
25. Moura-Netto C, Ferreira LS, Maranduba CM, Mello-Moura ACV, Marques MM. Low-intensity laser phototherapy enhances the proliferation of dental pulp stem cells under nutritional deficiency. Braz Oral Res. 2016 May 31;30(1):S1806-83242016000100265.
26. Soares DM, Ginani F, Henriques ÁG, Barboza CA. Effects of laser therapy on the proliferation of human periodontal ligament stem cells. Lasers Med Sci. 2015 Apr;30(3):1171-4.
27. Huang YY, Chen AC, Carroll JD, Hamblin MR. Biphasic dose response in low level light therapy. Dose Response. 2009 Sep 1;7(4):358-83.
28. Pandeshwar P, Roa MD, Das R, Shastry SP, Kaul R, Srinivasreddy MB. Photobiomodulation in oral medicine: a review. J Investig Clin Dent. 2016 May;7(2):114-26.
29. Tunér J, Jenkins PA. Parameter Reproducibility in Photobiomodulation. Photomed Laser Surg. 2016 Mar;34(3):91-2.
2. Marques NP, Lopes CS, Marques NCT, Cosme-Silva L, Oliveira TM, Duque C, et al. A preliminary comparison between the effects of red and infrared laser irradiation on viability and proliferation of SHED. Lasers Med Sci. 2019 Apr;34(3):465-471.
3. Bakhtiar H, Mazidi S A, Mohammadi Asl S, Ellini MR, Moshiri A, Nekoofar MH, et al. The role of stem cell therapy in regeneration of dentine-pulp complex: a systematic review. Prog Biomater. 2018 Dec;7(4):249-268.
4. Tocci A, Forte L. Mesenchymal stem cell: use and perspectives. Hematol J. 2003;4(2):92-6.
5. Martinez Saez D, Sasaki RT, Neves AD, da Silva MC. Stem Cells from Human Exfoliated Deciduous Teeth: A Growing Literature. Cells Tissues Organs. 2016;202(5-6):269-280.
6. Sanz AR, Carrión FS, Chaparro AP. Mesenchymal stem cells from the oral cavity and their potential value in tissue engineering. Periodontol 2000. 2015 Feb;67(1):251-67.
7. Borzabadi-Farahani A. Effect of low-level laser irradiation on proliferation of human dental mesenchymal stem cells; a systemic review. J Photochem Photobiol B. 2016 Sep;162:577-582.
8. Xie F, He J, Chen Y, Hu Z, Qin M, Hui T. Multi-lineage differentiation and clinical application of stem cells from exfoliated deciduous teeth. Hum Cell. 2020 Apr;33(2):295-302.
9. Rosa V, Dubey N, Islam I, Min KS, Nör JE. Pluripotency of Stem Cells from Human Exfoliated Deciduous Teeth for Tissue Engineering. Stem Cells Int. 2016;2016:5957806..
10. Ginani F, Soares DM, de Oliveira Rocha HA, de Souza LB, Barboza CAG. Low-level laser irradiation induces in vitro proliferation of stem cells from human exfoliated deciduous teeth. Lasers Med Sci. 2018 Jan;33(1):95-102.
11. Zaccara IM, Ginani F, Mota-Filho HG, Henriques ÁC, Barboza CA. Effect of low-level laser irradiation on proliferation and viability of human dental pulp stem cells. Lasers Med Sci. 2015 Dec;30(9):2259-64.
12. Marques NCT, Neto NL, Prado MTO, Vitor LLR, Oliveira RC, Sakai VT, et al. Effects of PBM in different energy densities and irradiance on maintaining cell viability and proliferation of pulp fibroblasts from human primary teeth. Lasers Med Sci. 2017 Sep;32(7):1621-1628.
13. Fernandes AP, Junqueira Mde A, Marques NC, Machado MA, Santos CF, Oliveira TM, et al. Effects of low-level laser therapy on stem cells from human exfoliated deciduous teeth. J Appl Oral Sci. 2016 Jul-Aug;24(4):332-7
14. Navaei-Nigjeh M, Moloudizargari M, Baeeri M, Gholami M, Lotfibakhshaiesh N, Soleimani M et al. Reduction of marginal mass required for successful islet transplantation in a diabetic rat model using adipose tissue-derived mesenchymal stromal cells. Cytotherapy. 2018 Sep;20(9):1124-1142.
15. Sylvester PW. Optimization of the tetrazolium dye (MTT) colorimetric assay for cellular growth and viability. Methods Mol Biol. 2011;716:157-68.
16. Feoktistova M, Geserick P, Leverkus M. Crystal Violet Assay for Determining Viability of Cultured Cells. Cold Spring Harb Protoc. 2016 Apr 1;2016(4):pdb.prot087379.
17. Felice DL, Sun J, Liu RH. A modified methylene blue assay for accurate cell counting. Journal of Functional Foods. 2009 Jan 1;1(1):109-18.
18. Avelar-Freitas BA, Almeida VG, Pinto MC, Mourão FA, Massensini AR, Martins-Filho OA, et al. Trypan blue exclusion assay by flow cytometry. Braz J Med Biol Res. 2014 Apr;47(4):307-15.
19. Martin-Piedra MA, Garzon I, Oliveira AC, Alfonso-Rodriguez CA, Carriel V, Scionti G, et al. Cell viability and proliferation capability of long-term human dental pulp stem cell cultures. Cytotherapy. 2014 Feb;16(2):266-77.
20. Ligasová A, Koberna K. Quantification of fixed adherent cells using a strong enhancer of the fluorescence of DNA dyes. Sci Rep. 2019 Jun 18;9(1):8701.
21. Eduardo Fde P, Bueno DF, de Freitas PM, Marques MM, Passos-Bueno MR, Eduardo Cde P, et al. Stem cell proliferation under low intensity laser irradiation: a preliminary study. Lasers Surg Med. 2008 Aug;40(6):433-8.
22. Paschalidou M, Athanasiadou E, Arapostathis K, Kotsanos N, Koidis PT, Bakopoulou A, et al. Biological effects of low-level laser irradiation (LLLI) on stem cells from human exfoliated deciduous teeth (SHED). Clin Oral Investig. 2020 Jan;24(1):167-180.
23. Almeida-Junior LA, Marques NCT, Prado MTO, Oliveira TM, Sakai VT. Effect of single and multiple doses of low-level laser therapy on viability and proliferation of stem cells from human exfoliated deciduous teeth (SHED). Lasers Med Sci. 2019 Dec;34(9):1917-1924.
24. Souza L, Rinco U, Aguiar D, de Almeida Junior L, Silva L, Oliveira T, et al. Effect of low-level laser therapy on viability and proliferation of stem cells from exfoliated deciduous teeth under different nutritional conditions. Laser Phys. 2018 Jan 17;28(2):025603.
25. Moura-Netto C, Ferreira LS, Maranduba CM, Mello-Moura ACV, Marques MM. Low-intensity laser phototherapy enhances the proliferation of dental pulp stem cells under nutritional deficiency. Braz Oral Res. 2016 May 31;30(1):S1806-83242016000100265.
26. Soares DM, Ginani F, Henriques ÁG, Barboza CA. Effects of laser therapy on the proliferation of human periodontal ligament stem cells. Lasers Med Sci. 2015 Apr;30(3):1171-4.
27. Huang YY, Chen AC, Carroll JD, Hamblin MR. Biphasic dose response in low level light therapy. Dose Response. 2009 Sep 1;7(4):358-83.
28. Pandeshwar P, Roa MD, Das R, Shastry SP, Kaul R, Srinivasreddy MB. Photobiomodulation in oral medicine: a review. J Investig Clin Dent. 2016 May;7(2):114-26.
29. Tunér J, Jenkins PA. Parameter Reproducibility in Photobiomodulation. Photomed Laser Surg. 2016 Mar;34(3):91-2.
| Issue | Vol 23 (Continuously Published Article-Based) | |
| Section | Original Article | |
| Keywords | ||
| Stem Cells Low-Level Light Therapy Cell Proliferation Cell Survival | ||
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How to Cite
1.
Rahimpour S, Azizi A, Hodjat M, Vahedi M. Effects of 660 and 940nm Low-Level Laser Irradiation on Viability and Proliferation of Stem Cells from Human Exfoliated Deciduous Teeth. Front Dent. 2026;23.
