Korean J Ophthalmol.  2008 Jun;22(2):123-129. 10.3341/kjo.2008.22.2.123.

Short-term Efficacy of Topical Immunosuppressive Agents on the Survival of Cultivated Allo-Conjunctival Equivalents

• Shin YJ
• Kim MK
• Oh JY
• Wee WR
• Lee JH
• Ko JH
• Lee HJ
• Lee JL
• Min BM
• Sohn YS

• KMID: 1099007
• DOI: http://doi.org/10.3341/kjo.2008.22.2.123

Abstract

PURPOSE: To investigate the short-term efficacy of topical immunosuppressive agents on the survival of cultivated allo-conjunctival equivalents. METHODS: Twenty-five eyes of New Zealand white rabbits were included. Temporal conjunctivae were trephined to a diameter of 7.5 mm, and then cultured allo-conjunctival epithelial cells on amniotic membrane were transplanted onto them. Various immunosuppressants including steroid, cyclosporine, and rapamycin were applied topically four times a day for a week. Epithelial defects and graft edema were graded daily. Numbers of inflammatory cells were measured in H&E. PKH26 and cytokeratin 4 and 7 were immunostained. RESULTS: Earlier epithelialization was observed in 1% steroid-treated eyes and defects persisted significantly in 0.5% CsA applied eyes. In histology, PKH26 positive cells considered as donor cells were only found in 1% steroid or 0.01% rapamycin applied eyes. 1% steroid- or 0.01% rapamycin-applied eyes both showed positive staining for keratin-4 and -7. Inflammatory cells were less found in 1% steroid or 0.01% rapamycin treated eyes. CONCLUSIONS: Topical steroid or rapamycin can help to suppress acute inflammation and enhance the acute survival of transplanted conjunctival cells.

Keyword

Cultivated allo-conjunctival equivalents; Steroid; Topical immunosuppressants; Rapamycin

MeSH Terms

Administration, Topical
Animals
Cell Count
*Cell Transplantation
Cells, Cultured
Conjunctiva/*cytology
Cyclosporine/pharmacology
Epithelial Cells/metabolism/*transplantation
Female
Fluorescent Antibody Technique, Indirect
Graft Survival/*drug effects
Immunosuppressive Agents/*pharmacology
Keratin-4/metabolism
Keratin-7/metabolism
Male
Organic Chemicals/metabolism
Prednisone/pharmacology
Rabbits
Sirolimus/pharmacology
Transplantation, Homologous

Figure

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  Fig. 1 Temporal conjunctivae was trephined to a diameter of 7.5 mm, and cultivated conjunctival equivalent or an allo-conjunctival graft was transplanted.

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  Fig. 2 Comparison of epithelialization (A) and graft edema (B) between untreated and treated eyes.

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  Fig. 3 Number of inflammatory cells in 1% steroid, 0.5% cyclosporine, or 0.01% rapamycin applied eyes in the cultivated conjunctival equivalent transplanted group. Histogram data represent means±SD. *p<0.05.

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  Fig. 4 PKH26 positive cells as transplanted donor cells in untreated (A), and in 0.5% cyclosporine (B), 1% steroid (C), 0.01% rapamycin (D) treated eyes in the cultivated conjunctival equivalent transplanted group. Hoechst staining in steroid (E) or 0.01% rapamycin (F) treated eyes in the cultivated conjunctival equivalent transplanted group. Magnification; ×200.

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  Fig. 5 Positive staining for keratin-4 and -7 in 1% steroid (A and B, respectively) and 0.01% rapamycin-applied eyes (C and D, respectively). Magnification; ×200.

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