겔형태 인공누액이 각막실질세포 및 결막상피세포에 미치는 영향
Effects of Gel Type Artificial Tears on Human Corneal Keratocytes and Conjunctival Epithelial Cells
2
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
목적: 겔형태 인공누액이 인체각막실질세포 및 결막상피세포에 미치는 영향을 비교하고자 하였다. 대상과 방법:인체 각막실질세포 및 결막상피세포를 배양하여 Soothe®, Systane® 농도별로 접촉 후MTT 분석법으로 세포 생존율, 젖산탈수소효소 (lactate dehydrogenase, LDH) 분석법으로 세포손상정도를 측정하였다. 세포자멸사 유무 확인 위해 형광현미경 및 유세포분석을 시행하고 전자현미경상 세포형태학적변화를 관찰하였다. 결과:Soothe®와 Systane®에서 약물농도가 높을수록, 접촉시간이 길수록 각결막세포의 증식억제력이 증가되었다. 각결막세포 모두에서 Soothe®는 노출시간과 농도에 비례하여 높은 LDH가 측정되나 Systane® 은 안정적이었다. 두 약제 모두 형광현미경상 세포사소견을 보였고 유세포분석상 세포자멸사를 보였다. Soothe® 에 비해 Systane®에서 세포형태학적 손상이 경미하였다. 결론:Soothe®와 Systane® 의 각막실질세포 및 결막상피세포 증식억제는 세포자멸사와 연관 있으며, 약물의 농도 및 노출시간에 비례하여 심하게 나타났고, Systane®이 Soothe®보다 각결막세포에 미치는 손상이 적었다.
Abstract
Purpose : To evaluate the biological effects and cytotoxicity of gel type artificial tears on the human corneal keratocytes and conjunctival cells in vitro. Methods : Human corneal keratocytes and conjunctival epithelial cells were exposed of Soothe® and Systane® at variable concentration respectively. Evaluations were conducted with MTT-based calorimetric assay for measuring the metabolic activity and the lactate dehydrogenase (LDH) assay for cellular damage. Apoptotic response was examined by fluorescent microscope, flow cytometric analysis and cellular morphologic finding was evaluated with transmission electron microscope. Results : The inhibitory effect of the corneal keratocytes and conjunctival cells proliferation increased at higher concentrations and longer exposure times of Soothe® and Systane®. The LDH titers increased after Soothe® was exposed, but Systane® showed no significant difference. Soothe® and Systane® showed fluorescence, which represent apoptic cells. In flow cytometry, the maximal apoptotic response developed in both artificial tears. Systane® showed less edema, cytoplasimc and nuclear degeneration of cells than that of Soothe®. Conclusions : The apoptotic response of Soothe® and Systane® is associated with the inhibitory effect of human corneal keratocytes and conjunctival epithelial cells proliferation. For inhibition of cellular proliferation of human corneal keratocytes and conjunctival epithelial cells, Systane® may be less severe at higher concentration and longer exposure time than Soothe®.
참고문헌 (26)
- Lemp MA, "Report of the National Eye Institute", Industry workshop on Clinical Trials in Dry Eyes, 1995, pp.21, 221~232
- Perry HD, "Dry eye diagnosis and management in 2004", Curr Opin Ophthalmol, 15, pp.299~304 (2004)
- Smith RE, "The tear film complex pathogenesis and emerging therapies for dry eyes", Cornea, 24, pp.1~7 (2005)
- Balaram M, "Efficacy and tolerability outcomes after punctual occlusion with silicone plugs in dry eye syndrome", Am J Ophthalmol, 131, pp.30~36 (2001)
- Isreb MA, "Correlation of lipid layer thickness measurements with fluorescein tear film break-up time and Schirmer`s test", Eye, 17, pp.79~83 (2003)
- Greiner JV, "Treatment of dry eye with a metastable formulation containing lipids and dineric interfacial molecules", Presented at the American Academy of Ophthalmology Annual Meeting, ~ (2004)
- Korb DR, "The effect of two novel lubricant eye drops on tear film lipid layer thickness in subjects with dry eye symptoms", Optom Vis Sci, 82, pp.594~601 (2005)
- Petricek I, "Hydroxypropyl-guar gellable lubricant eye drops for dry eye treatment", Expert Opin Pharmacother, 9, pp.1431~1436 (2008)
- Christensen MT, "Clinical evaluation of an HP-guar gellable lubricant eye drop for the relief of dryness of the eye", Curr Eye Res, 28, pp.55~62 (2004)
- Ubels JL, "Pre-clinical investigation of the efficacy of an artificial tear solution containing hydroxypropyl-guar as a gelling agent", Curr Eye Res, 28, pp.437~444 (2004)
- Schein OD, "Prevalence of dry eye among the elderly", Am J Ophthalmol, 124, pp.723~729 (1997)
- 윤경철, "안구건조증 환자에서 제대혈청 점안액 치료 후 눈물막 및 안구표면의 변화", 대한안과학회지, 46(2), pp.7~7 (2005), 대한안과학회
- 허환, "안구건조증 환자에서 국소 테스토스테론 젤의 치료효과", 대한안과학회지, 47(8), pp.1259~1265 (2006), 대한안과학회
- Trees GR, "Effect of artificial tear solutions and saline on tear film evaporation", Optom Vis Sci, 67, pp.886~890 (1990)
- Korb DR, "The effect of two novel lubricant eye drops on tear film lipid layer thickness in subjects with dry eye symptoms", Optom Vis Sci, 82, pp.594~601 (2005)
- Hartstein I, "An open-label evaluation of HP-Guar gellable lubricant eye drops for the improvement of dry eye signs and symptoms in a moderate dry eye adult population", Curr Med Res Opin, 21, pp.225~260 (2005)
- Sall KN, "An evaluation of the efficacy of a cyclosporine-based dry eye therapy when used with marked artificial tears as supportive therapy in dry eye", Eye Contact Lens, 32, pp.21~26 (2006)
- Gifford P, "A clinical evaluation of Systane", Contact Lens Anterior Eye, 29, pp.31~40 (2006)
- Adams J, "Morphologic and physiologic effects of artificial tear formulations on corneal epithelial derived cells", Cornea, 11, pp.234~241 (1992)
- Berdy GJ, "Preservative-free artificial tear preparations Assessment of corneal epithelial toxic effects", Arch Ophthalmol, 110, pp.528~532 (1992)
- Diebold Y, "Carbomer-versus cellulose-based artificial-tear formulations: morphologic and toxicologic effects on a corneal cell line", Cornea, 17, pp.433~440 (1998)
- Kasper K, "Toxicity of a new moistening agent and preservative in vitro", Ophthalmologe, 105, pp.557~562 (2008)
- Alder R, "Mechanism of photoreceptor death in retinal degeneration. From the cell biology of the 1990s to the ophthalmology or 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)
- Muller G, "Biocompatibility of index of antiseptic agents by parallel assessment of antimicrobial activity and cellular cytotoxicity", J Antimicrob Chemother, 61, pp.1281~1287 (2008)
- Meloni M, "Occludin gene expression as an early in vitro sign for mild eye irritation assessment", Toxicol In Vitro, 24, pp.276~285 (2010)