Low-level laser irradiation protects the chick embryo chorioallantoic membrane from UV cytotoxicity

Amira Hammami, Mohamed Amri, Meherzia Mokni


Low-level laser therapy or photobiomodulation is the medical use of a very low intensity light in the red to near infrared (wavelengths in the range of 630-940 nm). The present work was conducted to explore the effects of both UV and low-level laser irradiation (LLLI) on microcirculation using the in vivo model of the chick embryo chorioallantoic membrane (CAM). The effects were assessed by measuring lipid peroxidation and antioxidant enzyme activity. Cell cytotoxicity, survival and intracellular reactive oxygen species (ROS) of the CAM were also evaluated. We found that UV irradiation induced alterations of the vessels, leading to bleeding and extravasation. This effect was intensified after 60 min of exposure to UV irradiation, leading to marked edema. UVA irradiation increased cell cytotoxicity as assessed by lactate dehydrogenase (LDH) release (56.23% of control) and reduced cell viability as assessed by decreased fluorescein diacetate (FDA) fluorescence (56.23% of control). Pretreatment with LLLI prior to UV exposure protected the CAM tissue from UV-mediated cell death. This protective effect was supported by the observation of significantly inhibited lipid peroxidation (from 0.3±0.004 for UV, to 0.177±0.012 after LLLI pretreatment), ROS and O2-production, as indicated by respective dihydrorhodamine (DHR) and dihydroethidium (DHE) intensities (from 132.78% of control for UVA, to 95.90% of control for L-UV (DHR), and from 127.34% of control for UVA, to 82.03% of control for L-UV (DHE)), and by preventing the increase in oxidative activities. LLLI efficiently protected CAM cells from UV-induced oxidative stress and appeared as a safe protective pretreatment against UV irradiation.


Received: July 6, 2017; Revised: July 26, 2017; Accepted: July 27, 2017; Published online: August 28, 2017

How to cite this article: Hammami A, Amri M, Mokni M. Low-level laser irradiation protects the chick embryo chorioallantoic membrane from UV cytotoxicityc. Arch Biol Sci. 2018;70(1):119-27.


low level laser irradiation; UV irradiation; chorioallantoic membrane; cytotoxicity; vascular protection

Full Text:



Kondo S. The roles of cytokines in photoaging. J Dermatol Sci. 2000;23:S30-6.

Rittie L, Fisher GJ. UV-light-induced signal cascades and skin aging. Ageing Res Rev. 2002;1(4):705-20.

Manela-Azulay M, Bagatin E. Cosmeceuticals vitamins. Clin Dermatol. 2009;27(5):469-74.

Kert J, Rose L. Clinical laser therapy: Low level laser therapy. Copenhagen, Denmark: Medical Laser Technology Group; 1989. 240 p.

Niemz MH. Laser-tissue interactions: Fundamentals and applications. New York: Springer Verlag; 2007.

Mezghani S, Hammami A, Amri M. Low-level laser therapy: effects on human face aged skin and Cell viability of hela cells exposed to UV radiation. Arch Biol Sci. 2015;67(1):25-9.

Bergmeyer HU. New values for the molar extinction coefficients of NADH and NADPH for the use in routine laboratories. Z Klin Chem Klin Biochem. 1975;13(11):507-8. Chinese.

Draper HH, Hadley M. Malondialdehyde determination as index of lipid peroxidation. Methods Enzymol.1990;186:421-31.

Misra HP, Fridovich I. The role of superoxide anion in autoxidation of epinephrine and a simple assay for SOD. J Biol Chem.1972;247(10):3170-5.

Aebi H. Catalase in vitro. Methods Enzymol.1984;105:121-6.

Chance B, Maehly AC. Assay of catalases and peroxidases. In Methods Enzymol. 1955;2:764-75.

Ohnishi ST, Barr JK. A simple method of quantitating protein using the biuret and phenol reagent. Anal Biochem. 1978;86(1):193-200.

Stern J, Lewis WH. The colorimetric estimation of calcium in serum with ocresolphthalein complexone. Clin Chim Acta.1957;2(6):576-80.

Fraikin GY, Strakhovskaya MG, Rubin AB. The role of membrane-bound porphyrin-type compound as endogenous sensitizer in photodynamic damage to yeast plasma membranes. J Photochem Photobiol. B Biol. 1996;34:129-35.

Edwards AM, Silva E. Effect of visible light on selected enzymes, vitamins and amino acids. J Photochem Photobiol. 2001;63(1-3):126-31.

Menezes S, Coulomb B, Lebreton C, Dubertret L. Non-coherent near infrared radiation protects normal human dermal fibroblasts from solar ultraviolet toxicity. J Invest Dermatol. 1998;111(4):629-33.

Walker MD, Rumpf S, Baxter GD, Hirst DG, Lowe AS. Effect of low-intensity laser irradiation (660 nm) on a radiation-impaired wound-healing model in murine skin.Lasers Surg Med. 2000;26(1):41-47.

Basford JR. Low-level laser treatment of pain and wounds. Mayo Clin Proc.1986;61(8):671-5.

Steg PG, Gal D, Rongion AJ, De Jesus ST, Clarke RH. Effect of argon laser irradiation on rabbit aortic smooth muscle. Inser J M Cardiovasc Res. 1988;22:747-53.

Hamamichi S, Nishigori H. Establishment of a chick embryo shell-less culture system and its use to observe change in behavior caused by nicotine and substances from cigarette smoke. Toxicol Lett. 2001;119(2):95-102.

Ribatti D. The chick embryo chorioallantoic membrane (CAM) assay. Reprod Toxicol. 2017;70:97-101.

Mowbray M, McLintock S, Weerakoon R, Lomatschinsky N, Jones S, Rossi AG, Weller RB. Enzyme-independent NO stores in human skin: quantification and influence of UV radiation. J Invest Dermatol. 2009;129(4):834-42.

Stryer L. Biochemistry. 4th ed. New York: W H Freeman and Company; 1995. 732 p.

Kowaluk EA, Seth P, Fung HL. Metabolic activation of sodium nitroprusside to nitric oxide in vascular smooth muscle. J Pharmacol Exp Ther. 1992;262(3):916-22.

Keeble JD, Moore PK. Pharmacology and potential therapeutic applications of nitric oxide–releasing non-steroidal anti-inflammatory and related nitric oxide-donating drugs. Br J Pharmacol. 2002;137(3):295-310.

Schwentker A, Vodovotz Y, Weller R, Billiar TR. Nitric oxide and wound repair: role of cytokines? Nitric Oxide. 2002;7(1):1-10.

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;69(3):161-7.

Dube A, Bock C, Bauer E, Kohli R,Gupta PK. He-Ne laser irradiation protects B-lymphoblasts from UVA-induced DNA damage. Radiation Environ Biophys. 2001;40(1):77-82.

Glubisz AW, Fraczek J, Moncznik JS, Friedlein G, Mikolajczyk T, Sarna T, Pryjma J. Effect of UVA and 8-Methoxypsoralen, 4, 6, 4’-trimethylangelicin or chlorpromazine on apoptosis of lymphocytes and their recognition by monocytes. J Physio Pharmacol. 2010;61(1):107-14.

Sahu KH, Mohanty SK, Gupta PK. He-Ne laser (632.8 nm) pre-irradiation gives protection against DNA damage induced by a near-infrared trapping beam. J Biophotonics. 2009;2(3):140-4.

Cao Z, Lindsa G,IsaacsNW. Mitochondrial peroxiredoxins.SubCell Biochem. 2007;44: 295-315.

Graves JA, Metukuri M, Scott D, Rothermund K, Prochownik EV. Regulation of reactive oxygen species homeostasis by peroxiredoxins and c-Myc. J Biol Chem. 2009;284(10):6520-9.

Zhao H, Yi X, Hu Z, Hu M, Chen S, Dong X. RNAi-mediated knockdown of catalase causes cell cycle arrest in SL-1 cells and results in low survival rate of Spodopteralitura (Fabricius). PLoS One. 2013;8(3):e59527.

Li ZH, Zlabek V, Velisek J, Grabic R, Machova J, Kolarova J, Li P, Randak T. Acute toxicity of carbamazepine to juvenile rainbow trout (Oncorhynchus mykiss): effects on antioxidant responses, hematological parameters and hepatic EROD. Ecotoxicol Environ Saf. 2011;74(3):319-27.

Shaw BJ, Al-Bairuty G, Handy RD. Effects of waterborne copper nanoparticles and copper sulphate on rainbow trout, (Oncorhynchus mykiss): physiology and accumulation. Aquat Toxicol. 2012;116-117:90-101.

Foyer CH, Descourvières P, Kunert KJ. Protection against oxygen radicals: An important defence mechanism studied in transgenic plants. Plant Cell Environ. 1994;17:507-23.

John S, Kale M, Rathore N, Bhatnagar D. Protective effect of vitamin E in dimethoate and malathion induced oxidative stress in rat erythrocytes. J Nutr Biochem. 2001;12(9):500-4.

Krishnan N, Kodrík D. Antioxidant enzymes in Spodopteralittoralis (Boisduval): are they enhanced to protect gut tissues during oxidative stress. J Insect Physiol.2006;52(1):11-20.

Karthi S, Sankari R, Shivakumar MS. Ultraviolet-B light induced oxidative stress: effects on the antioxidant response of Spodopteralitura. J Photoch Photobiol B. 2014;135:1-6.

Lesser MP. Oxidative stress in marine environments: biochemistry and physiological ecology. Annu Rev Physiol. 2006;68:253-78.

Lubart R, Eichler M, Lavi R, Friedman H, Shainberg A. Low-energy laser irradiation promotes cellular redox activity. Photomed Laser Surg. 2005;23(1):3-9.

Lavi R, Shainberg A, Friedmann H, Shneyvays V, Rickover O, Eichler M, Kaplan D, Lubart R. Low energy visible light induces reactive oxygen species generation and stimulates an increase of intracellular calcium concentration in cardiac cells. J Biol Chem. 2003;278(42):40917-22.

Kokoska ER, Wolff AB, Smith GS, Miller TA. Epidermal growth factor-induced cytoprotection in human intestinal cells involves intracellular calcium signaling. J Surg Res. 2000;88(2):97-103.

Krizaj D, Copenhagen DR. Calcium regulation in photoreceptors. Front Biosci. 2002;7:d2023-44.

Levonen AL, Landar A, Ramachandran A, Ceaser EK, Dickinson DA, Zanoni G, Morrow JD, Darley-Usmar VM. Cellular mechanisms of redox cell signalling: role of cysteine modification in controlling antioxidant defences in response to electrophilic lipid oxidation products. Biochem J. 2004;378(Pt 2):373-82.

Bello RI, Alcaín FJ, Gómez-Díaz C, López-Lluch G, Navas P, Villalba JM. Hydrogen peroxide- and cell-density-regulated expression of NADH-cytochrome b5 reductase in HeLa cells. J Bioenerg Biomembr. 2003;35(2):169-79.


  • There are currently no refbacks.


Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.