무방부제 인공누액이 인체각막상피세포에 미치는 영향
Effect of Preservative-free Artificial Eye Drop on Human Corneal Epithelial Cell in vitro
1
조회
0
다운로드
1
인용
0
참고문헌
키워드
초록
목적: 임상에서 사용되는 무방부제 인공누액들이 각막상피세포에 미치는 영향을 알아보고자 하였다. 대상과 방법: 인체각막상피를 일차 배양하여 KynexⓇ, Hyalein Mini 0.3%Ⓡ, Refresh PlusⓇ와 접촉시킨 후 MTT 및 lactate dehydrogenase (LDH) assay를 이용하여 세포증식력 및 독성을 비교하였으며, 약제의 구성성분을 분석하고, 유세포 분석을 통하여 세포자멸사를 확인하였다. 결과: MTT 및 세포이주, 창상회복에서 세 약제 모두 대조군과 큰 차이를 보이지 않았다. LDH 역가는 노출시간이 길어질수록 증가하는 양상을 보였으나, 세 약제 모두 대조군에 비해 의미 있는 차이를 보이지 않았고, 유세포 분석에서는 대조군과 세 약제 모두에서 24시간째 세포자멸사와 함께 세포괴사의 소견을 보였다. 세 약제의 주성분을 제외한 전해질, pH, 삼투압 등의 차이는 없었다. 결론: 임상에서 흔히 사용되는 일회용 인공누액제인 KynexⓇ, Hyalein Mini 0.3%Ⓡ, Refresh PlusⓇ의 경우 각막상피세포에 미치는 영향이 미미하여 안전하게 사용할 수 있을 것으로 생각된다.
Abstract
Purpose: To investigate the biologic effects of preservative-free artificial tear drops on cultured human corneal epithelial cells in vitro. Methods: Efficacies of the preservative-free artificial tear drops-KynexⓇ, Hyalein Mini 0.3%Ⓡ, and Refresh PlusⓇ-were evaluated using the MTT assay. Cell damage was determined using the lactate dehydrogenase (LDH) assay. Cellular proliferation was determined using a migration and wound-healing assay. The ingredients of the drugs were analyzed. Apoptotic response was evaluated with flow cytometric analysis. Results: Metabolic activity of the corneal epithelial cells showed similar activity to that of the control. Cellular migration and proliferation also were not significantly different between the preservative-free artificial tear drop groups and the control. The LDH titers tended to increase for up to 24 hours after exposure to the preservative-free artificial tear drops, but there was no significant difference in LDH titers between the control groups and the artificial tear drop-treated groups. Apoptosis and necrosis were observed using flow cytometry at 24 hours in all groups. The electrolyte levels, pHs and osmolarities of the three drugs were not significantly different. Conclusions: The clinically available preservative-free artificial tear drops KynexⓇ, Hyalein Mini 0.3%Ⓡ, and Refresh PlusⓇ had no significant toxic effects on corneal epithelial cells and thus can be used safely.
참고문헌 (23)
- Ayaki M, "Cytotoxicity of ophthalmic solutions with and without preservatives to human corneal endothelial cells, epithelial cells and conjuncival epithelial cells", Clin Experiment Ophthalmol, 36, pp.553~559 (2008)
- Tonjum AM, "Effects of benzalkonium chloride upon the corneal epithelium studies with scanning electron microscopy", Acta Ophthalmol, 53, pp.358~366 (1975)
- Burstein NL, "Coneal cytotoxictiy of topically applied drugs, vehicles and preservatives", Surv Opthalmol, 25, pp.15~30 (1980)
- Burstein NL, "Preservative cytotoxic threshold for benzalkonium chloride and chlorhexidine digluconate in cat and rabbit corneas", Invest Opthlamol Vis Sci, 19, pp.308~313 (1980)
- Collin HB, "The effect of ophthalmic preservatives on the healing rate of the rabbit corneal epithelium after keratectomy", Am J Opthalmol Physiol Opt, 59, pp.215~222 (1982)
- Mosmann TJ, "Rapid colorimetric assay for cellular growth and sruvival: Application to proliferation and cytotoxic assays", J Immunolo Methods, 65, pp.55~63 (1983)
- Garrett Q, "Carboxymethylcellulose binds to human corneal epithelial cells and is a modulator of corneal epithelial wound healing", Invest Ophthalmol Vis Sci, 48, pp.1559~1567 (2007)
- Berdy GJ, "Preservative-free artificial tear preparations; assessment of corneal epithelial toxic effects", Arch Opthalmol, 110, pp.528~532 (1992)
- Johnson ME, "Effectiveness of sodium hyaluronate eyedrops in the treatment of dry eye", Graefes Arch Clin Exp Opthalmol, 244, pp.109~112 (2006)
- Miyauchi S, "The effect of sodium hyaluronate on the migration of rabbit corneal epithelium", J Ocul Pharmacol, 6, pp.91~99 (1990)
- Nishida T, "Hyaluronate stimulate corneal epithelial migration", Exp Eye Res, 53, pp.753~758 (1991)
- Gomes JA, "Sodium hyaluronate (hyaluronic acid) promotes migration of human corneal epithelial cells in vitro", Br J Opthalmol, 88, pp.821~825 (2004)
- Chung JH, "Experimental corneal alkali wound healing", Acta Ophthalmol Suppl, 187, pp.1~35 (1988)
- Chung JH, "Hyaluronate in healing of corneal alkali wound in the rabbit", Exp Eye Res, 48, pp.569~576 (1989)
- Garrett Q, "Carboxymethyl cellulose stimulate rabbit corneal epithelial wound healing", Curr Eye Res, 33, pp.567~573 (2008)
- Garrett Q, "Carboxymethylcellulose binds to human corneal epithelial cells and is a modulator of corneal epithelial wound healing", Invest Opthalmol Vis Sci, 48, pp.1559~1567 (2007)
- Ahee JA, "Decreased incidence of epithelial defects during laser in situ keratomileusis using intraoperative nonpreserved carboxymethylcellulose sodium 0.5% solution", J Cataract Refract Surg, 28, pp.1651~1654 (2002)
- Adler R, "Mechanism of photoreceptor death in retinal degeneration. From the cell biology of the 1990s to the ophthalmology of the 21st century", Arch Ophthalmol, 114, pp.79~83 (1996)
- Thompson CB, "Apoptosis in the pathogenesis and treatment of disease", Science, 267, pp.1456~1462 (1995)
- Wee WR, "Effect of artificial tears on cultured keratocytes in vitro", Cornea, 109, pp.860~863 (1995)
- Park YS, "Physiology of body fluid. In Physiology, 5th ed", pp.5~ (2000), Sin-Kwang publishing & printing
- Freshney RI, "The culture environment: Substrate, gas phase, medium, and temparature, 2nd ed", pp.70~71 (1987), Wiley-Liss
- Kim K, "Acid-base balance", pp.326~327 (1996), Eui-hak publishing & printing