DOI QR코드

DOI QR Code

Comparison of cytotoxicities and wound healing effects of diquafosol tetrasodium and hyaluronic acid on human corneal epithelial cells

  • Lee, Jong Heon (Department of Ophthalmology, School of Medicine, Pusan National University) ;
  • Lee, Jong Soo (Department of Ophthalmology, School of Medicine, Pusan National University) ;
  • Kim, Sujin (Department of Ophthalmology, Gyeongsang National University Changwon Hospital, School of Medicine, Gyeongsang National University) ;
  • Lee, Ji Eun (Department of Ophthalmology, School of Medicine, Pusan National University)
  • 투고 : 2016.11.01
  • 심사 : 2016.12.01
  • 발행 : 2017.03.01

초록

This study aimed to compare the cellular toxicities of three clinically used dry eye treatments; 3% diquafosol tetrasodium and hyaluronic acid at 0.3 and 0.18%. A methyl thiazolyltetrazoiun (MTT)-based calorimetric assay was used to assess cellular proliferation and a lactate dehydrogenase (LDH) leakage assay to assess cytotoxicity, using Human corneal epithelial cells (HCECs) exposed to 3% diquafosol tetrasodium, 0.3% hyaluronic acid (HA), or 0.18% HA or 1, 6 or 24 h. Cellular morphology was evaluated by inverted phase-contrast light microscopy and electron microscopy, and wound widths were measured 24 h after confluent HCECs were scratched. Diquafosol had a significant, time-dependent, inhibitory effect on HCEC proliferation and cytotoxicity. HCECs treated with diquafosol detached more from the bottoms of dishes and damaged cells showed degenerative changes, such as, reduced numbers of microvilli, vacuole formation, and chromatin of the nuclear remnant condensed along the nuclear periphery. All significantly stimulated reepithelialization of HCECs scratched, which were less observed in diquafosol. Therefore, epithelial toxicity should be considered after long-term usage of diquafosol and in overdose cases, especially in dry eye patients with pre-existing punctated epithelial erosion.

키워드

참고문헌

  1. Uchino M, Nishiwaki Y, Michikawa T, Shirakawa K, Kuwahara E, Yamada M, Dogru M, Schaumberg DA, Kawakita T, Takebayashi T, Tsubota K. Prevalence and risk factors of dry eye disease in Japan: Koumi study. Ophthalmology. 2011;118:2361-2367. https://doi.org/10.1016/j.ophtha.2011.05.029
  2. Lee JE, Kim NM, Yang JW, Kim SJ, Lee JS, Lee JE. A randomised controlled trial comparing a thermal massager with artificial teardrops for the treatment of dry eye. Br J Ophthalmol. 2014;98:46-51. https://doi.org/10.1136/bjophthalmol-2013-303742
  3. Gomes JA, Amankwah R, Powell-Richards A, Dua HS. Sodium hyaluronate (hyaluronic acid) promotes migration of human corneal epithelial cells in vitro. Br J Ophthalmol. 2004;88:821-825. https://doi.org/10.1136/bjo.2003.027573
  4. Yamaguchi M, Tsubota K, Watanabe H, Ohashi Y. The safety and efficacy of long-term treatment with 3% diquafosol ophthalmic solution for dry eye. Atarashii Ganka (J Eye). 2012;29:527-535.
  5. Takaoka-Shichijo Y, Sakamoto A, Nakamura M. Effect of diquafosol tetrasodium on MUC5AC secretion by rabbit conjunctival tissues. Atarashii Ganka (J Eye). 2011;28:261-265.
  6. Li Y, Kuang K, Yerxa B, Wen Q, Rosskothen H, Fischbarg J. Rabbit conjunctival epithelium transports fluid, and P2Y2(2) receptor agonists stimulate $ Cl^{-}$ and fluid secretion. Am J Physiol Cell Physiol. 2001;281:C595-602. https://doi.org/10.1152/ajpcell.2001.281.2.C595
  7. Lee JE, Sun Y, Gjorstrup P, Pearlman E. Inhibition of corneal inflammation by the resolvin E1. Invest Ophthalmol Vis Sci. 2015;56:2728-2736. https://doi.org/10.1167/iovs.14-15982
  8. Arita R, Suehiro J, Haraguchi T, Maeda S, Maeda K, Tokoro H, Amano S. Topical diquafosol for patients with obstructive meibomian gland dysfunction. Br J Ophthalmol. 2013;97:725-729. https://doi.org/10.1136/bjophthalmol-2012-302668
  9. Lee JS, Lee SU, Che CY, Lee JE. Comparison of cytotoxicity and wound healing effect of carboxymethylcellulose and hyaluronic acid on human corneal epithelial cells. Int J Ophthalmol. 2015;8:215-221.
  10. Lee JS, Lee JE, Kim N, Oum BS. Comparison of the conjunctival toxicity of topical ocular antiallergic agents. J Ocul Pharmacol Ther. 2008;24:557-562. https://doi.org/10.1089/jop.2008.0071
  11. Park YS. Physiology of body fluid. In: Kang DH, editor. Physiology. 5th ed. Seoul: Sin-Kwang; 2000. p.585-606.
  12. Garrett Q, Khandekar N, Shih S, Flanagan JL, Simmons P, Vehige J, Willcox MD. Betaine stabilizes cell volume and protects against apoptosis in human corneal epithelial cells under hyperosmotic stress. Exp Eye Res. 2013;108:33-41. https://doi.org/10.1016/j.exer.2012.12.001
  13. Kim K. Acid-base balance. In: Sung H, Kim KH, editors. Physiology. 3rd ed. Seoul: Eui-hak; 2006. p.326-327.
  14. Ayaki M, Iwasawa A, Yaguchi S, Koide R. In vitro assessment of the cytotoxicity of anti-allergic eye drops using 5 cultured corneal and conjunctival cell lines. J Oleo Sci. 2011;60:139-144. https://doi.org/10.5650/jos.60.139
  15. Pauly A, Brasnu E, Riancho L, Brignole-Baudouin F, Baudouin C. Multiple endpoint analysis of BAC-preserved and unpreserved antiallergic eye drops on a 3D-reconstituted corneal epithelial model. Mol Vis. 2011;17:745-755.
  16. Brasnu E, Brignole-Baudouin F, Riancho L, Guenoun JM, Warnet JM, Baudouin C. In vitro effects of preservative-free tafluprost and preserved latanoprost, travoprost, and bimatoprost in a conjunctival epithelial cell line. Curr Eye Res. 2008;33:303-312. https://doi.org/10.1080/02713680801971857
  17. De Saint Jean M, Debbasch C, Brignole F, Rat P, Warnet JM, Baudouin C. Toxicity of preserved and unpreserved antiglaucoma topical drugs in an in vitro model of conjunctival cells. Curr Eye Res. 2000;20:85-94. https://doi.org/10.1076/0271-3683(200002)2021-DFT085
  18. Baudouin C, Labbe A, Liang H, Pauly A, Brignole-Baudouin F. Preservatives in eyedrops: the good, the bad and the ugly. Prog Retin Eye Res. 2010;29:312-334. https://doi.org/10.1016/j.preteyeres.2010.03.001
  19. Champeau EJ, Edelhauser HF. The effect of ophthalmic preservatives on the ocular surface: conjunctival and corneal uptake and distribution of Benzalkonium chloride and chlorhexidine digluconate. In: Holly FJ, editor. The preocular tear film in health, disease and contact lens wear. Lubbock, TX: Dry eye Institute, Inc.; 1986.
  20. Dutot M, Pouzaud F, Larosche I, Brignole-Baudouin F, Warnet JM, Rat P. Fluoroquinolone eye drop-induced cytotoxicity: role of preservative in P2X7 cell death receptor activation and apoptosis. Invest Ophthalmol Vis Sci . 2006;47:2812-2819. https://doi.org/10.1167/iovs.06-0224
  21. Feng MM, Baryla J, Liu H, Laurie GW, McKown RL, Ashki N, Bhayana D, Hutnik CM. Cytoprotective effect of lacritin on human corneal epithelial cells exposed to benzalkonium chloride in vitro. Curr Eye Res . 2014;39:604-610. https://doi.org/10.3109/02713683.2013.859275
  22. Guzman-Aranguez A, Calvo P, Ropero I, Pintor J. In vitro effects of preserved and unpreserved anti-allergic drugs on human corneal epithelial cells. J Ocul Pharmacol Ther. 2014;30:790-798. https://doi.org/10.1089/jop.2014.0030
  23. Byun YS, Yoo YS, Kwon JY, Joo JS, Lim SA, Whang WJ, Mok JW, Choi JS, Joo CK. Diquafosol promotes corneal epithelial healing via intracellular calcium-mediated ERK activation. Exp Eye Res. 2016;143:89-97. https://doi.org/10.1016/j.exer.2015.10.013
  24. Mohamed YH, Uematsu M, Ueki R, Inoue D, Fujikawa A, Sasaki H, Kitaoka T. Acute Corneal Toxicity of Diquas. Pharmacology. 2016;98:56-61. https://doi.org/10.1159/000445691
  25. Lazarus HM, Imperia PS, Botti RE, Mack RJ, Lass JH. An in vitro method which assesses corneal epithelial toxicity due to antineoplastic, preservative and antimicrobial agents. Lens Eye Toxic Res. 1989;6:59-85.
  26. Tripathi BJ, Tripathi RC. Cytotoxic effects of benzalkonium chloride and chlorobutanol on human corneal epithelial cells in vitro. Lens Eye Toxic Res. 1989;6:395-403.

피인용 문헌

  1. Hialid®/Hyalein® (sodium hyaluronate) ophthalmic solution in dry eye syndrome: a profile of its use vol.34, pp.9, 2017, https://doi.org/10.1007/s40267-018-0540-0