Effect of Helium-Neon Laser and Sodium Hypochlorite on Calf Thymus Double-Stranded Deoxyribonucleic Acid Molecule: An in Vitro Experimental Study

Document Type : Original Paper


1 Department of Pharmacology and Toxicology, College of Pharmacy, Hawler Medical University, ErbilIraq

2 Department of Physiology, Medical Physics, College of Medicine, Diyala University


Introduction: Low-energy helium-neon (He-Ne) laser beam lightis used in combination with sodium hypochlorite (Na2HOCl3) for clinical purposes. Regarding this, the present study aimed to investigate the effect of He-Ne laser (632.8 nm) and sodium hypochlorite on the calf thymus double-stranded deoxyribonucleic acid (ctdsDNA) molecule. 
Materials and Methods: For the purpose of the study, ctdsDNA solutions (30µg/ml) were exposed to He-Ne laser (632.8 nm) light in the absence and presence of different concentrations of sodium hypochlorite for up to 60 sec. The levels of nucleic acids released as uncontaminated and contaminated proteins were considered as the markers of DNA damage in terms of hypochromasia (i.e., DNA strand breakage) and hyperchromasia (i.e., of DNA strands separation).
 Results: The mean concentration of nucleic acids insignificantly (P > 0.05) decreased after exposure to laser light irradiation (hypochromic effect). Furthermore, laser irradiation insignificantly and inconsistency protected the ctdsDNA molecules from the effect of sodium hypochlorite.  Sodium hypochlorite at concentrations of 1 and 3 mmol reduced the levels of the nucleic acids released from contaminated protein by 29.2% and 78.3% of the pre-incubated levels (hyporchromasia effect). The He-Ne laser (632.8 nm) irradiation induced hypochromic effect on the uncontaminated and contaminated proteins, while sodium hypochlorite induced a remarkable hyperchromic effect at higher concentrations.
Conclusion: As the finding indicated, a short time He-Ne laser light (632.8 nm) irradiation exerted minor significant effect on the ctdsDNA molecule. This laser light did not interact with sodium hypochlorite as a synergistic combination against the ctdsDNA molecule.  


Main Subjects

  1. References


    1. Lan CC, Wu CS, Chiou MH, Chiang TY, Yu HS. Low-energy helium-neon laser induces melanocyte proliferation via interaction with type IV collagen: visible lightas a therapeutic option for vitiligo. Br J Dermatol. 2009 Aug;161(2):273-80. Doi: 10.1111/j.1365-2133.2009.09152.x.
    2. Reichelt J, Winter J, Meister J, Frentzen M, Kraus D. A novel blue light laser system for surgical applications in dentistry: evaluation of specific laser-tissue interactions in monolayer cultures. Clin Oral Investig. 2017 May;21(4):985-94. Doi: 10.1007/s00784-016-1864-6.
    3. Houreld NN, Abrahamse H. Effectiveness of helium neon laser irradiation on viability and cytotoxicity of diabetic-wounded fibroblast cells. Photomed Laser Surg. 2007 Dec; 25(6): 474-81. Doi: 10.1089/pho.2007.1095.
    4. Lu T, Zhang YC, Wong M, Feiveson A, Gaza R, Stoffle N, et al. Detection of DNA damage by space radiation in human fibroblasts flown on the International Space Station. Life Sci Space Res (Amst). 2017 Feb; 12: 24-31. DOI: 10.1016/j.lssr.2016.12.004.
    5.  Hawkins DH, Abrahamse H. The role of laser fluence in cell viability, proliferation, and membrane integrity of wounded human skin fibroblasts following helium-neon laser irradiation. Lasers Surg Med. 2006 Jan; 38(1): 74-83. DOI: 10.1002/lsm.20271.
    6. Fahimipour F, Houshmand B., Alemi P, Asnaashari M, Tafti MA., Akhoundikharanagh F, et al. The effect of He-Ne and Ga-Al-As lasers on the healing of oral mucosa in diabetic mice. J Photochem Photobiol B. 2016 Jun; 159: 149-54. DOI: 10.1016/j.jphotobiol.2016.03.020.
    7. Kohli R, Gupta, PK. Irradiance dependence of the He-Ne laser-induced protection against UVC radiation in E. coli strains. J Photochem Photobiol B. 2003 Mar; 69(3): 161-7. DOI: 10.1016/S1011-1344(03)00018-6.
    8. Zaichkina SI, Rozanova O.M, Diukina, AR, Simonova NB, Romanchenko SP, Sorokina SS, et al. Influence of low-dose-rate red and near-infrared radiations on the level of reactive oxygen species, the genetic apparatus and the tumor growth in mice in vivo. Biofizika. 2013 Sep-Oct; 58(5): 897-903. DOI: 10.1134/S0006350913050199.
    9. Novoselova EG, Glushkova OV, Khrenov MO, Chernenkov DA, Lunin SM, Novoselova TV, et al. Protective effect of low-power laser radiation in acute toxic stress. Biofizika. 2007 Jan-Feb; 52(1): 137-40.
    10. Al-Nimer MS, Al-Deen SM, Abdul Lateef Z. Rofecoxib prevents ctdsDNA against damage induced by copper sulfate and ultraviolet B radiation in vitro study. J Basic Clin Pharm. 2010 Dec; 2(1): 21-5.
    11. Layne E. Spectrophotometric and turbidimetric methods for measuring proteins. Methods Enzymol.1957; 10:  447-55. DOI: 10.1016/S0076-6879(57)03413-8.
    12. Scopes RK. Measurement of protein by spectrophotometry at 205 nm. Anal Biochem. 1974 May; 59(1): 277-82. DOI: 10.1016/0003-2697(74)90034-7.
    13. Stoscheck CM. Quantitation of protein. Methods Enzymol. 1990; 182: 50-68.
    14. Al-Nimer MS, Mshatat SF, Abdulla HI. Saliva α-synuclein and a high extinction coefficient protein: A novel approach in assessment biomarkers of Parkinson's disease. N Am J Med Sci. 2014 Dec; 6(12): 633-7. DOI: 10.4103/1947-2714.147980.
    15. Evans DH, Abrahamse H. Efficacy of three different laser wavelengths for in vitro wound healing. Photodermatol Photoimmunol Photomed. 2008 Aug; 24(4): 199-210. . DOI: 10.1111/j.1600-0781.2008.00362.x.
    16. Li Y, Gao L, Han R. A combination of He-Ne laser irradiation and exogenous NO application efficiently protect wheat seedling from oxidative stress caused by elevated UV-B stress. Environ Sci Pollut Res Int. 2016 Dec; 23(23): 23675-82. DOI: 10.1007/s11356-016-7567-3.
    17. Masuda M, Suzuki T, Friesen MD, Ravanat JL, Cadet J, Pignatelli B, et al. Chlorination of guanosine and other nucleosides by hypochlorous acid and myeloperoxidase of activated human neutrophils: Catalysis by nicotine and trimethylamine. J Biol Chem. 2001 Nov 2; 276(44): 40486-96. DOI: 10.1074/jbc.M102700200.
    18. Badouard C, Masuda M, Nishino H, Cadet J, Favier A, Ravanat JL. Detection of chlorinated DNA and RNA nucleosides by HPLC coupled to tandem mass spectrometry as potential biomarkers of inflammation. J Chromatogr B Analyt Technol Biomed Life Sci. 2005 Nov 15; 827(1): 26-31.
    19. Cadet J, Wagner, JR. DNA base damage by reactive oxygen species, oxidizing agents, and UV radiation. Cold Spring Harb Perspect Biol. 2013 Feb 1; 5(2). DOI: 10.1101/cshperspect.a012559.
    20. Hawkins CL, Davies MJ. Hypochlorite-induced damage to DNA, RNA, and polynucleotides: formation of chloramines and nitrogen-centered radicals. Chem Res Toxicol. 2002 Jan; 15(1): 83-92.
    21. Botton G, Pires CW, Cadoná FC, Machado AK, Azzolin VF, Cruz IB, et al. Toxicity of irrigating solutions and pharmacological associations used in pulpectomy of primary teeth. Int Endod J. 2016 Aug; 49(8): 746-54. DOI: 10.1111/iej.12509.
    22. Swanson M J, Baribault ME, Israel JN, Bae NS. Telomere protein RAP1 levels are affected by cellular aging and oxidative stress. Biomed Rep. 2016 Aug; 5(2): 181-7. Doi: 10.3892/br.2016.707.
    23. Gómez S, Bravo P, Morales R, Romero A, Oyarzún A. Resin penetration in artificial enamel carious lesions after using sodium hypochlorite as a deproteinization agent. J Clin Pediatr Dent. 2014 Fall; 39(1): 51-6. DOI: 10.17796/jcpd.39.1.e72570275387527r.
    24. Lee HJ, Cho MJ, Chang SK. Ratiometric Signaling of Hypochlorite by the Oxidative Cleavage of Sulfonhydrazide-Based Rhodamine-Dansyl Dyad. Inorg Chem. 2015 Sep 8; 54(17): 8644-9. DOI: 10.1021/acs.inorgchem.5b01284.
    25. Xhevdet A, Stubljar D, Kriznar I, Jukic T, Skvarc M, Veranic P, Ihan A.  The disinfecting efficacy of root canals with laser photodynamic therapy. J Lasers Med Sci. 2014 Winter; 5(1): 19-26.