Sygitowicz Grazyna, Zapolska-Downar Danuta, Paluch Magdalena, Stawarski Tomasz, Sieradzki Edmund, Sitkiewicz Dariusz
Department of Biochemistry and Clinical Chemistry, Medical University of Warsaw, Banacha St. 1, 02-097 Warszawa, Poland.
Acta Pol Pharm. 2011 Sep-Oct;68(5):745-51.
In the recent years a significant development of investigations with regard to bioavailability of ocular drugs has been noticed. The corneal epithelial barrier is the main pathway for ocular penetration of topically applied ophthalmic drugs into the anterior chamber. To work out an in vitro model of bovine corneal epithelial primary cultures and exercise it for permeability research with lipophilic and hydrophilic markers, permeability coefficients estimation of the 6-carboxyfluorescein and rhodamin B was made. The corneal epithelial cultures of the 3th or 4th passage were chosen for layered culture with inserts based on the liquid-liquid interface (for the first week) and the air-liquid interface (for the two following weeks). On the 7th, 12th, 18th, 21st experiment day TER values, and on the 21st day drug permeability coefficients, were determined. The mean TER values of the 7th, 12th, 18th, 21st day of corneal epithelial culture were: 122.14, 155.14, 198.43 and; 247.43 Wcm2, respectively. The mean values of permeability coefficients on the 21st day of culture for 6-carboxyfluorescein and rhodamin B were 3.87 +/-0.10 x 10(-6)cm/s and 3.65 +/- 0.06 x 10(-6)cm/s, respectively. We state that the in vitro bovine corneal epithelial primary culture model is useful for ocular studies.
近年来，人们注意到有关眼用药物生物利用度的研究有了显著进展。角膜上皮屏障是局部应用的眼科药物进入前房的主要途径。为了建立牛角膜上皮原代培养的体外模型，并用于亲脂性和亲水性标记物的渗透性研究，对6-羧基荧光素和罗丹明B的渗透系数进行了估算。选择第3或第4代的角膜上皮培养物，基于液-液界面（第一周）和气-液界面（接下来的两周）用插入物进行分层培养。在实验的第7、12、18、21天测定跨上皮电阻（TER）值，并在第21天测定药物渗透系数。角膜上皮培养第7、12、18、21天的平均TER值分别为：122.14、155.14、198.43和247.43Ω·cm²。培养第21天6-羧基荧光素和罗丹明B的渗透系数平均值分别为3.87±0.10×10⁻⁶cm/s和3.65±0.06×10⁻⁶cm/s。我们认为体外牛角膜上皮原代培养模型对眼部研究是有用的。
