Is the Photosuturing Agent, Rose Bengal, a Mutagen?

Abstract

Rose Bengal (RB) is a potential photosuturing agent that may improve standard dermatologic surgical closure techniques. However, RB produces reactive oxygen species with photoactivation and its photomutagenic potential must be considered in clinical application. We investigated cytotoxicity, mutagenicity, and singlet oxygen (SO) production of RB on epithelial Chinese hamster ovary cell line. Cells were exposed to RB concentrations: 0.1%, 0.01%, 0.001%, 0.0001%, 0.00001%; irradiated for 400 s using a high-intensity visible wavelength lamp or maintained in the dark. Cell viability was assessed by XTT assay, mutagenicity by HPRT gene mutation assay, and SO production by Sensor Green reagent. RB > 0.001% was significantly cytotoxic. Viabilities were uninfluenced by ≤0.0001% RB controls, or 30-min incubation. 49% of irradiated cells died after 24-h in 0.0001% RB. At ≥0.001% RB, >90% of cells died. Irradiating 0.00001% - 0.001% RB increased SO; levels dropped significantly between 0.01% - 0.1%. Controls exhibited negligible SO production. HPRT suggested that RB was not mutagenic (0.0001%, 0.00001%); SO induction increased between 0.00001% - 0.001%, with reduced production at higher concentrations. Pilot studies suggested irradiated 0.0001% RB is mutagenic in vitro; current data suggest RB is not photomutagenic. The contribution of RB’s cytotoxicity on observed clinical improvement of scars and mutagenic potential remains unclear, necessitating further study.

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L. Hunter-Ellul, S. Wang, J. Wickliffe and M. Wilkerson, "Is the Photosuturing Agent, Rose Bengal, a Mutagen?," Journal of Cosmetics, Dermatological Sciences and Applications, Vol. 4 No. 1, 2014, pp. 24-30. doi: 10.4236/jcdsa.2014.41004.

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1] Y. Kamegaya, W. Farinelli, A. V. Vila Echague, et al., “Evaluation of Photochemical Tissue Bonding for Closure of Skin Incisions and Excisions,” Lasers in Surgery and Medicine, Vol. 37, No. 4, 2005, pp. 264-270. http://dx.doi.org/10.1002/lsm.20221
[2] S. Tsao, M. Yao, H. Tsao, et al., “Light-Activated Tissue Bonding for Excisional Wound Closure: A Split-Lesion Clinical Trial,” British Journal of Dermatology, Vol. 166, No. 3, 2012, pp. 555-563.
http://dx.doi.org/10.1111/j.1365-2133.2011.10710.x
[3] M. Yao, A. Yaroslavsky, F. Henry, et al., “Photo-Toxicity Is Not Associated with Photochemical Tissue Bonding of Skin,” Lasers in Surgery and Medicine, Vol. 42, No. 2, 2010, pp. 123-131.
http://dx.doi.org/10.1002/lsm.20869
[4] D. Olivier, S. Douillard, I. Lhommeau, et al., “Photodynamic Treatment of Culture Medium Containing Serum Induces Long-Lasting Toxicity in Vitro,” Radiation Research, Vol. 172, No. 4, 2009, pp. 451-462. http://dx.doi.org/10.1667/RR1646.1
[5] S. Y. Kim and J.-W. Park, “Cellular Defense against Singlet Oxygen-Induced Oxidative Damage by Cytosolic NADP+-dependent Isocitrate Dehydrogenase,” Free Radical Research, Vol. 37, No. 3, 2003, pp. 309-316. http://dx.doi.org/10.1080/1071576021000050429
[6] B. P. Chan, I. E. Kochevar and R. W. Redmond, “Enhancement of Porcine Skin Graft Adherence Using a Light-Activated Process,” Journal of Surgical Research, Vol.108, No. 1, 2002, pp. 77-84.
http://dx.doi.org/10.1006/jsre.2002.6516
[7] S. Tsao, M. Yao, F. P. Henry, et al., “Photochemical Tissue Bonding for Closure of Excision Wounds: A Firstin-Human Clinical Trial Dermatology,” Journal of Investigative Dermatology, Vol. 129, No. S37, 2009.
[8] B. P. Chan and K.-F. So, “Photochemical Cross-Linking Improves the Physicochemical Properties of Collagen Scaffolds,” Journal of Biomedical Materials Research, Vol. 75A, No. 3, 2005, pp. 689-701. http://dx.doi.org/10.1002/jbm.a.30469
[9] L. Mulroy, J. Kim, I. Wu, et al., “Photochemical Keratodesmos for Repair of Lamellar Corneal Incisions,” Investigative Ophthalmology & Visual Science, Vol. 41, 2000, pp. 3335-3340.
[10] A. Camoirano, S. De Flora and T. A. Dahl, “Genotoxicity of Volatile and Secondary Reactive Oxygen Species Generated by Photosensitization,” Environmental and Molecular Mutagenesis, Vol. 21, No. 3, 1993, pp. 219-228. http://dx.doi.org/10.1002/em.2850210304
[11] B. P. Chan, O. C. M. Chan and K.-F. So, “Effects of Photochemical Crosslinking on the Microstructure of Collagen and a Feasibility Study on Controlled Protein Release,” Acta Biomaterlialia, Vol. 4, No. 6, 2008, pp. 1627-1636. http://dx.doi.org/10.1016/j.actbio.2008.06.007
[12] I. E. Kochevar and R. W. Redmond, “Photosensitized Production of Singlet Oxygen,” Methods in Enzymology, Vol. 319, 2000, pp. 20-28. http://dx.doi.org/10.1016/S0076-6879(00)19004-4
[13] A. Webster, D. Britton, A. Apap-Bologna, et al., “A Dye-Photosensitized Reaction that Generates Stable Protein-Protein Crosslinks,” Analytical Biochemistry, Vol. 179, No. 1, 1989, pp. 154-157.
http://dx.doi.org/10.1016/0003-2697(89)90216-9
[14] P. C. C. Lee and M. A. J. Rodgers, “Laser Flash Photokinetic Studies of Rose Bengal Sensitized Photodynamic Interactions of Nucleotides and DNA,” Photochemistry and Photobiology, Vol. 45, No. 1, 1987, pp. 79-86. http://dx.doi.org/10.1111/j.1751-1097.1987.tb08407.x
[15] L.-O. Klotz, K. Briviba and H. Sies, “Singlet Oxygen Mediates the Activation of JNK by UVA Radiation in Human Skin Fibroblasts,” FEBS Letters, Vol. 408, No. 3, 1997, pp. 289-291.
http://dx.doi.org/10.1016/S0014-5793(97)00440-7
[16] M. Wiener, D. Damian and J. Thompson, “Systemic Phototoxicity Following Intralesional Rose Bengal for Subcutaneous Melanoma Metastases,” Dermatology, Vol. 216, No. 4, 2008, pp. 361-362. http://dx.doi.org/10.1159/000117707
[17] P. Bilski, B. Kukielczak and C. Chignell, “Photo-Production and Direct Spectral Detection of Singlet Molecular Oxygen (1O2) in Keratinocytes Stained with Rose Bengal,” Photochemistry and Photobiology, Vol. 68, No. 5, 1998, pp. 675-678.
[18] R. P. G. Feenstra and S. C. G. Tseng, “What Is Actually Stained by Rose Bengal?” Archives of Ophthalmology, Vol. 110, No. 7, 1992, pp. 984-993. http://dx.doi.org/10.1001/archopht.1992.0108 0190090035
[19] R. Costa de Oliveir, D. T. Ribeir, R. G. Nigro, et al., “Singlet Oxygen Induced Mutation Spectrum in Mammalian Cells,” Nucleic Acids Research, Vol. 20, No. 16, 1992, pp. 4319-4323. http://dx.doi.org/10. 1093/nar/20.16.4319
[20] R. S. Gupta and B. Singh, “Mutagenic Responses of Five Independent Genetic Loci in CHO Cells to a Variety of Mutagens: Development and Characteristics of a Mutagen Screening System Based on Selection of Multiple Drug-Resistant Markers,” Mutation Research, Vol. 94, No. 2, 1982, pp. 449-466. http://dx.doi.org/10.1016/0027-5107(82)90307-4
[21] L. Hunter, S. Wang, J. Wickliffe, et al., “Photoactivated Rose Bengal, a Possible Mutagen in Chinese Hamster Ovary Cells,” Journal of Investigative Dermatology, Vol. 130, No. S138, 2010.
[22] A. L. Tajirian and D. J. Goldberg, “A Review of Sutures and Other Skin Closure Materials,” Journal of Cosmetic and Laser Therapy, Vol. 12, No. 6, 2010, pp. 296-302. http://dx.doi.org/10.3109/147641 72.2010.538413
[23] E. P. Tierney, R. L. Moy and D. J. Kouba, “Rapid Absorbing Gut Suture versus 2-Octylethylcyanoacrylate Tissue Adhesive in the Epidermal Closure of Linear Repairs,” Journal of Drugs in Dermatology, Vol. 8, No. 2, 2009, pp. 115-119.
[24] A. D. Altman, V. M. Allen, S. A. McNeil, et al., “Pfannenstiel Incision Closure: A Review of Current Skin Closure Techniques,” Journal of Obstetrics & Gynaecology Canada, Vol. 31, No. 6, 2009, pp. 514-520.
[25] J. W. Eichenbaum, P. H. Pevsner, G. Pivawer, et al., “A Murine Photochemical Stroke Model with Histologic Correlates of Apoptotic and Nonapoptotic Mechanisms,” Journal of Pharmacological and Toxicological Methods, Vol. 47, No. 2, 2002, pp. 67-71. http://dx.doi.org/10.1016/S1056-8719(02) 00215-0
[26] S. Basu-Modak and R. Tyrrell, “Singlet Oxygen: A Primary Effector in the Ultraviolet A/Near-Visible Light Induction of the Human Heme Oxygenase Gene,” Cancer Research, Vol. 53, 1993, pp. 4505-4510.
[27] V. Au and S. A. Madison, “Effects of Singlet Oxygen on the Extracellular Matrix Protein Collagen: Oxidation of the Collagen Crosslink Histi-Dinohydroxylsinonorleucine and Histidine,” Archives of Biochemistry and Biophysics, Vol. 384, No. 1, 2000, pp. 133-142. http://dx.doi.org/10.1006/abbi. 2000.2070
[28] E. Wachter, C. Dees, J. Harkins, et al., “Topical Rose Bengal: Pre-Clinical Evaluation of Pharma-Cokinetics and Safety,” Lasers in Surgery and Medicine, Vol. 32, No. 2, 2003, pp. 101-110. http://dx.doi.org/10.1002/lsm.10138
[29] L. Cronin, M. Moffitt, D. Mawad, et al., “An in Vitro Study of the Photodynamic Effect of Rose Bengal on Trichophyton rubrum,” Journal of Biophotonics, 2012, pp. 1-9.

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