Standardization of a protocol to irradiate cell cultures with low level laser
Low level laser periodontal and cell culture fibroblasts osteoblasts pre- osteoclasts orthodontic movement
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Introduction: It is impossible to apply orthodontic therapeutic laser during the orthodontic treatment and to select different times of exposure, frequency or intensity for every single cell. The whole area gets irradiated in seconds according to the laser type . This could have a proliferation effect in some cells and a harmful effect in others cells. So, it´s very important to asses the effects of a laser application on the cells involved in dental movement, to verify the effect and reaction after laser therapy.
Objective: To create and standarized a clinical protocol to irradiate culture cells involved in dental movement (Human fibroblasts, osteoblasts and pre-osteoclasts in order to understand the basics of it´s clinical effect.
Materials and methods: Gingival and periodontal fibroblasts, normal human osteoblasts (NHOst) and human osteoclast progenitor cells in culture were irradiated using a low energy level laser (As-GaAl), operated at 36.73mW, 832.79nm nm wavelength 3.75 J/cm2 and 32.40 seconds per well.
Conclusions: The cytotoxicity tests showed that irradiation is safe protocol for the four groups of cell cultures.
Domínguez, Ángela, & Criollo Gómez, W. (2017). Standardization of a protocol to irradiate cell cultures with low level laser. Revista Estomatología, 20(2), 29–35. https://doi.org/10.25100/re.v20i2.5739
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2. Meikle MC. The tissue, cellular, and molecular regulation of orthodontic tooth movement: 100 years after Carl Sandstedt. Eur J Orthod 2006; 28:221-40.
3. Vinod Krishnan, Ze’ev Davidovitch. Cellular, molecular, and tissue-level reactions to orthodontic force American Tabla 5. Promedios de citotoxicidad (%) Horas Láser Triton 6 0,22 6,6 24 0,543 48,37 Volumen 20 Nº 2 2012 34 Journal of Orthodontics & Dentofacial Orthopedics 2006; 129(4):469.
4. Masella RS, Meister M.Current concepts in the biology of orthodontic tooth movement.Am J Orthod Dentofacial Orthop 2006; 129(4):458-68.
5. Saito S, Shimizu N. Stimulatory effects of low-power laser irradiation on bone regeneration in midpalatal suture during expansion in the rat. Am J Orthod Dentofac Orthop 1997; 111:525-32.
6. Kawasaki K, Shimizu N. Effects of low-energy laser irradiation on bone remodeling during experimental tooth movement in rats. Lasers Surg Med 2000; 26:282-91.
7. King Gj, Keeling SD,Wronski TJ. Histomorphometric study of alveolar bone turnover in orthodontic tooth movement. Bone 1991; 12:401-9.
8. King GJ, Latta L, Rutenberg J, Ossi A, Keeling SD.Alveolar bone turnover and tooth movement in male rats after removal of orthodontic appliances. Am J Orthod Dentofacial Orthop 1997; 111(3):266-75.
9. Habib FA, Gama SK, Ramalho LM, Cangussú MC, Santos Neto FP, Lacerda JA, Araújo TM, Pinheiro AL. Laserinduced alveolar bone changes during orthodontic movement: a histological study on rodents. Photomed Laser Surg 2010; 28(6):823-30.
10. Yamaguchi M, Hayashi M, Fujita S, Yoshida T, Utsunomiya T, Yamamoto H, et al. Low-energy laser irradiation facilitates the velocity of tooth movement and the expressions of matrix metalloproteinase-9, cathepsin K, and alpha (v) beta (3) integrin in rats. The European Journal of Orthodontics. Eur J Orthod 2010; 32(2):131-9.
11. Abello M, Valbuena D. Efecto del láser blando (ir) en la velocidad de alineación del tratamiento ortodóncico. Tesis de postgrado Fundación C.I.E.O Bogotá, Colombia 1996.
12. Cruz DR, Kohara EK, Ribeiro MS, Wetter NU.Effects of low-intensity laser therapy on the orthodontic movement velocity of human teeth: a preliminary study.Lasers in Surgery and Medicine.2004; 35:117–120
13. Limpanichkul W, Godfrey K, Srisuk N, Rattanayatikul C. Effects of low-level laser therapy on the rate of orthodontic tooth movement. Orthodontics and Craniofacial Research. 2006; 9(1):38-43.
14. Youssef M, Ashkar S, Hamade E, Gutknecht N, Lampert F, Mir M. The effect of low-level laser therapy during orthodontic movement: a preliminary study. Lasers Med Sci 2008; 23(1):27-33
15. Dominguez A and Velasquez S. Acceleration effect of orthodontic movement by application of low-intensity laser. J Oral Laser Applications 2010 (10): 99-105.
16. Sousa MV, Scanavini MA, Sannomiya EK, Velasco LG, Angelieri F. Influence olowlevel laser on the speed of orthodontic movement. Photomed Laser Surg. 2011; 29(3):191-6.
17. Lim HM, Lew KK, Tay DK. A clinical investigation of the low level laser therapy in reducing orthodontic postadjustment pain. Am J Or thod Dentof a c i a l Orthop.1995; 108:614-622.
18. Harazaki M, Takahashi H, Ito A, Isshiki Y .Soft laser irradiation induced pain reduction in orthodontic treatment. Bull Tokyo Dent Coll.1998; 39:95.101.
19. Tortamano A, Lenzi D, Haddad A, Bottino M, Dominguez G, Vigorito J. Low-level laser therapy for pain caused by placement of the first orthodontic archwire: A randomized clinical trial. American Journal of Orthodontics and Dentofacial Orthopedics 2009; 136(5):662-7.
20. Xiaoting L, Yin T, Yangxi C. Interventions for pain during fixed orthodontic appliance therapy. A systematic review. Angle Orthod 2010; 80(5):925-32.
21. Domínguez A, Velásquez SA. Effect of Low-Level Laser Therapy on Pain Following Activation of Orthodontic Final Archwires: A Randomized Controlled Clinical Trial. Photomedicine and Laser Surgery. January 2013; 31(1):36-40.
22. Almeida-Lopes L, Rigau J, Zângaro RA, Guidugli-Neto J, Jaeger MM. Comparison of the low level laser therapy effects on cultured human gingival fibroblasts proliferation using different irradiance and same fluence. Lasers Surg Med. 2001; 29(2):179-84.
23. Pereira AN. Effect of low-power laser irradiation on cell growth and procollagen synthesis of cultured fibroblasts. LasersSurg Med.2002: 31(4):263-7.
24. Kreisler M. Effect of low-level GaAlAs laser irradiation on the proliferation of human periodontal ligament fibroblasts: an in vitro study. J Clin Peridontol. 2003; 30(4):353-8.
25. Choi EJ, Yim JY, Koo KT, Seol YJ, Lee YM, Ku Y, Rhyu IC, Chung CP, Kim TI. Biological effects of a semiconductor diode laser on human periodontal ligament fibroblasts. J Periodontal Implant Sci. 2010; 40(3):105-10.
26. Saygun I, Karacay S, Serdar M, Ural AU. Effects of laser irradiation on the release of basic fibroblast growth factor (bFGF), insulin like growth factor-1 (IGF-1), and receptor of IGF-1(IGFBP3) from gingival fibroblasts. Lasers Med Sci. 2008; 23(2):211-5.
27. Ozawa Y, Shimizu N, Kariya G, Abiko Y. Low-energy laser irradiation stimulates bone nodule formation at early stages of cell culture in rat calvarial cells. Bone 1998; 22:347-354.
28. Coombe AR, Darendeliler N. The effects of low level laser irradiation on osteoblastic cells. Clin Orthod Res. 2001; 4:3-14.
29. Fujihara NA, Hiraki KR, Marques MM. Irradiation at 780nm increases proliferation rate of osteoblasts independently of dexamethasone presence. Lasers Surg Med 2006; 38:332-336.
30. Soleimani M, Abbasnia E, Fathi M, Sahraei H, Fathi Y, Kaka G. The effects of lowlevel laser irradiation on differentiation and proliferation of human bone marrow mesenchymal stem cells into neurons and osteoblasts an in vitro study. Lasers Med Sci 2012; 27(2):423-30.
31. Aihara N, Yamaguchi M, Kasai K. Low– energy irradiation stimulates formation of osteoclast-like cells via RANK expression in Vitro. Lasers Med Sci 2006; 21(1): 24-33.
32. Timberlake GT. An inexpensive, automated instrument for laser irradiation of cultived cells. Photomed Laser Surg 2004; 22(3):233-9.
33. Domínguez A, Clarkson A and López R. An in vitro study of the reaction of 35 Revista Estomatología y Salud periodontal and gingival fibroblasts to low- level laser irradiation: A pilot study. J Oral Laser Applications 2008; 8:235-244.
34. Dominguez A, Castro P, Morales M. An In Vitro Study of the Reaction of Human Osteoblasts to Low-level Laser Irradiation. Journal of Oral Laser Applications 2009; 9(1):21-28
35. Domínguez A, Bayona G, and Casas A. In vitro response of Human Pre-osteoclasts to low intensity Laser irradiation. Journal of research in Biology 2012; 2 (8):1-9.
2. Meikle MC. The tissue, cellular, and molecular regulation of orthodontic tooth movement: 100 years after Carl Sandstedt. Eur J Orthod 2006; 28:221-40.
3. Vinod Krishnan, Ze’ev Davidovitch. Cellular, molecular, and tissue-level reactions to orthodontic force American Tabla 5. Promedios de citotoxicidad (%) Horas Láser Triton 6 0,22 6,6 24 0,543 48,37 Volumen 20 Nº 2 2012 34 Journal of Orthodontics & Dentofacial Orthopedics 2006; 129(4):469.
4. Masella RS, Meister M.Current concepts in the biology of orthodontic tooth movement.Am J Orthod Dentofacial Orthop 2006; 129(4):458-68.
5. Saito S, Shimizu N. Stimulatory effects of low-power laser irradiation on bone regeneration in midpalatal suture during expansion in the rat. Am J Orthod Dentofac Orthop 1997; 111:525-32.
6. Kawasaki K, Shimizu N. Effects of low-energy laser irradiation on bone remodeling during experimental tooth movement in rats. Lasers Surg Med 2000; 26:282-91.
7. King Gj, Keeling SD,Wronski TJ. Histomorphometric study of alveolar bone turnover in orthodontic tooth movement. Bone 1991; 12:401-9.
8. King GJ, Latta L, Rutenberg J, Ossi A, Keeling SD.Alveolar bone turnover and tooth movement in male rats after removal of orthodontic appliances. Am J Orthod Dentofacial Orthop 1997; 111(3):266-75.
9. Habib FA, Gama SK, Ramalho LM, Cangussú MC, Santos Neto FP, Lacerda JA, Araújo TM, Pinheiro AL. Laserinduced alveolar bone changes during orthodontic movement: a histological study on rodents. Photomed Laser Surg 2010; 28(6):823-30.
10. Yamaguchi M, Hayashi M, Fujita S, Yoshida T, Utsunomiya T, Yamamoto H, et al. Low-energy laser irradiation facilitates the velocity of tooth movement and the expressions of matrix metalloproteinase-9, cathepsin K, and alpha (v) beta (3) integrin in rats. The European Journal of Orthodontics. Eur J Orthod 2010; 32(2):131-9.
11. Abello M, Valbuena D. Efecto del láser blando (ir) en la velocidad de alineación del tratamiento ortodóncico. Tesis de postgrado Fundación C.I.E.O Bogotá, Colombia 1996.
12. Cruz DR, Kohara EK, Ribeiro MS, Wetter NU.Effects of low-intensity laser therapy on the orthodontic movement velocity of human teeth: a preliminary study.Lasers in Surgery and Medicine.2004; 35:117–120
13. Limpanichkul W, Godfrey K, Srisuk N, Rattanayatikul C. Effects of low-level laser therapy on the rate of orthodontic tooth movement. Orthodontics and Craniofacial Research. 2006; 9(1):38-43.
14. Youssef M, Ashkar S, Hamade E, Gutknecht N, Lampert F, Mir M. The effect of low-level laser therapy during orthodontic movement: a preliminary study. Lasers Med Sci 2008; 23(1):27-33
15. Dominguez A and Velasquez S. Acceleration effect of orthodontic movement by application of low-intensity laser. J Oral Laser Applications 2010 (10): 99-105.
16. Sousa MV, Scanavini MA, Sannomiya EK, Velasco LG, Angelieri F. Influence olowlevel laser on the speed of orthodontic movement. Photomed Laser Surg. 2011; 29(3):191-6.
17. Lim HM, Lew KK, Tay DK. A clinical investigation of the low level laser therapy in reducing orthodontic postadjustment pain. Am J Or thod Dentof a c i a l Orthop.1995; 108:614-622.
18. Harazaki M, Takahashi H, Ito A, Isshiki Y .Soft laser irradiation induced pain reduction in orthodontic treatment. Bull Tokyo Dent Coll.1998; 39:95.101.
19. Tortamano A, Lenzi D, Haddad A, Bottino M, Dominguez G, Vigorito J. Low-level laser therapy for pain caused by placement of the first orthodontic archwire: A randomized clinical trial. American Journal of Orthodontics and Dentofacial Orthopedics 2009; 136(5):662-7.
20. Xiaoting L, Yin T, Yangxi C. Interventions for pain during fixed orthodontic appliance therapy. A systematic review. Angle Orthod 2010; 80(5):925-32.
21. Domínguez A, Velásquez SA. Effect of Low-Level Laser Therapy on Pain Following Activation of Orthodontic Final Archwires: A Randomized Controlled Clinical Trial. Photomedicine and Laser Surgery. January 2013; 31(1):36-40.
22. Almeida-Lopes L, Rigau J, Zângaro RA, Guidugli-Neto J, Jaeger MM. Comparison of the low level laser therapy effects on cultured human gingival fibroblasts proliferation using different irradiance and same fluence. Lasers Surg Med. 2001; 29(2):179-84.
23. Pereira AN. Effect of low-power laser irradiation on cell growth and procollagen synthesis of cultured fibroblasts. LasersSurg Med.2002: 31(4):263-7.
24. Kreisler M. Effect of low-level GaAlAs laser irradiation on the proliferation of human periodontal ligament fibroblasts: an in vitro study. J Clin Peridontol. 2003; 30(4):353-8.
25. Choi EJ, Yim JY, Koo KT, Seol YJ, Lee YM, Ku Y, Rhyu IC, Chung CP, Kim TI. Biological effects of a semiconductor diode laser on human periodontal ligament fibroblasts. J Periodontal Implant Sci. 2010; 40(3):105-10.
26. Saygun I, Karacay S, Serdar M, Ural AU. Effects of laser irradiation on the release of basic fibroblast growth factor (bFGF), insulin like growth factor-1 (IGF-1), and receptor of IGF-1(IGFBP3) from gingival fibroblasts. Lasers Med Sci. 2008; 23(2):211-5.
27. Ozawa Y, Shimizu N, Kariya G, Abiko Y. Low-energy laser irradiation stimulates bone nodule formation at early stages of cell culture in rat calvarial cells. Bone 1998; 22:347-354.
28. Coombe AR, Darendeliler N. The effects of low level laser irradiation on osteoblastic cells. Clin Orthod Res. 2001; 4:3-14.
29. Fujihara NA, Hiraki KR, Marques MM. Irradiation at 780nm increases proliferation rate of osteoblasts independently of dexamethasone presence. Lasers Surg Med 2006; 38:332-336.
30. Soleimani M, Abbasnia E, Fathi M, Sahraei H, Fathi Y, Kaka G. The effects of lowlevel laser irradiation on differentiation and proliferation of human bone marrow mesenchymal stem cells into neurons and osteoblasts an in vitro study. Lasers Med Sci 2012; 27(2):423-30.
31. Aihara N, Yamaguchi M, Kasai K. Low– energy irradiation stimulates formation of osteoclast-like cells via RANK expression in Vitro. Lasers Med Sci 2006; 21(1): 24-33.
32. Timberlake GT. An inexpensive, automated instrument for laser irradiation of cultived cells. Photomed Laser Surg 2004; 22(3):233-9.
33. Domínguez A, Clarkson A and López R. An in vitro study of the reaction of 35 Revista Estomatología y Salud periodontal and gingival fibroblasts to low- level laser irradiation: A pilot study. J Oral Laser Applications 2008; 8:235-244.
34. Dominguez A, Castro P, Morales M. An In Vitro Study of the Reaction of Human Osteoblasts to Low-level Laser Irradiation. Journal of Oral Laser Applications 2009; 9(1):21-28
35. Domínguez A, Bayona G, and Casas A. In vitro response of Human Pre-osteoclasts to low intensity Laser irradiation. Journal of research in Biology 2012; 2 (8):1-9.
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- Pilar León, Angela Domínguez, Laser therapy and biochemical markers in the acceleration of orthodontic dental movement: a review of the literature , Revista Estomatología: Vol. 21 No. 2 (2013)
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