Rabbit conjunctival epithelial cells

P Saha, J Yang, VHL Lee. (1998). Existence of a p-glycoprotein drug efflux pump in cultured rabbit conjunctival epithelial cells. Invest Opthalmol Vis Sci 39 1221-1226. 

J. J. Yang, H. Ueda, K. Kim, and V. H. Lee. Meeting future challenges in topical ocular drug delivery Development of an air-interfaced primary culture of rabbit conjunctival epithelial cells on a permeable support for drug transport studies. J Control Release 65 1-11 (2000). 

Figure 13.3 A flowchart illustrating various steps for the preparation and maintenance of primary rabbit conjunctival epithelial cell layers, cultured under liquid-covered and air-interfaced conditions (See also color insert).

H. J. Gukasyan, V. H. Lee, K. J. Kim, and R. Kannan. Net glutathione secretion across primary cultured rabbit conjunctival epithelial cell layers. Invest Ophthalmol Vis Sci 43 1154-1161 (2002)... 

M. G. Qaddoumi, H. Ueda, J. Yang, J. Davda, V. Labhasetwar, and V. H. Lee. The characteristics and mechanisms of uptake of PLGA nanoparticles in rabbit conjunctival epithelial cell layers. Pharm Res 21 641-648 (2004)... 

S. K. Basu, I. S. Haworth, M. B. Bolger, and V. H. Lee. Proton-driven dipeptide uptake in primary cultured rabbit conjunctival epithelial cells. Invest Ophthalmol... 

Qaddoumi et al. [65] studied the uptake of PLGA nanoparticles in rabbit conjunctival epithelial cell culture. The highest uptake by cultured conjunctival cells was achieved for the smallest particles (100 nm), compared to larger 800 nm and 10 pm particles. A study of the fate of the tiny 100-nm particles following 2 h of cultured cells exposure to a 0.5 mg/mL dose showed that 6% was internalized by conjunctival epithelial cells, 1.5% was surface-bound, whereas the remainder of the dose was found in the donor medium. In an in vivo rabbit eye study [66] on the uptake of poly(hexyl cyanoacrylate) nanoparticles, 6 h postinstillation into the conjunctival sac, it was found that the fraction that was internalized by conjunctival epithelial cells was only 1% of the dose reflecting in vivo precorneal elimination... 

Because mucin and/or cilia systems of AIC cultured epithelial cells may work as a barrier for drug transport, lower Papp values are expected in cell layers cultured in AIC than in LCC methods. However, it was interesting to note that no significant differences in Rapp values were observed between the cell layers cultured with the two methods (Table 9.1). This is in contrast to solute permeabilities reported previously for cell layers cultured with LCC versus AIC methods [76, 80], For example, Yang et al. reported that Rapp of lipophilic solutes (e.g., various /3-blockers) across the primary cultured conjunctival epithelial cell layer are about threefold lower when cultured under AIC than LCC conditions, suggesting that the permeability of AIC cultured cell layers generally better reflects that of the excised tissue than LCC counterparts. Mathias et al. [76] also reported that the permeability of hydrophilic solutes across the primary rabbit tracheal epithelial cell layer cultured under AIC conditions was only half of that observed for cell layers cultured under... 

The effects of sodium chlorite on membrane components and antioxidant depletion have been studied in rabbit corneal epithelial cells, human conjunctival epithelial cells, phospholipid vesicles prepared from egg yolk and GSH in solution. Incubation of phospholipid vesicles with 3.5 mmol sodium chlorite/l for up to 2 h had no effect, whereas incubation for 48 h resulted in lipid depletion and an increase in lipid oxidation. Sodium chlorite was found to be a very potent GSH oxidizing agent at a GSH/sodium chlorite ratio of 0.5, GSH was depleted after 5 min. GSH depletion was also seen in rabbit corneal epithelial cells and human conjunctival epithelial cells incubated with 3.5 mmol sodium chlorite/l or 0.55 mmol sodium chlorite/l. At 3.5 mmol/l, sodium chlorite caused rapid loss of cell viability in the corneal cells, as assessed by trypan blue staining and loss of adherence. At 0.55 mmol/l, sodium chlorite had very little effect over the first few hours but decreased viability after 24 h. The conjunctival cells appeared to be less sensitive than the corneal cells. No oxidatively modified lipids could be detected in the cells following sodium chlorite treatment, and no effects were seen in levels of cytosolic antioxidants (Ingram et al., 2003). 

In a subsequent study, rabbit corneal epithelial and human conjunctival epithelial cells were incubated with 0, 5, 25, 50, 100, 250, 500 or 750 mg sodium chlorite/l. Depletion of cellular adenosine triphosphate (ATP) occurred at concentrations above 100 mg/l for rabbit corneal epithelial cells and above 50 mg/ I for human conjunctival epithelial cells. GSH was depleted progressively between 5 and 250 mg/l in both cell types. The authors concluded that sodium chlorite is of low toxicity to ocular cells (Ingram et al., 2004). 

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