J Vet Sci.  2013 Mar;14(1):61-67. 10.4142/jvs.2013.14.1.61.

Effects of conditioned media from human amniotic epithelial cells on corneal alkali injuries in rabbits

• Kim TH
• Park YW
• Ahn JS
• Ahn JT
• Kim SE
• Jeong MB
• Seo MS
• Kang KS
• Seo KM

• KMID: 1482814
• DOI: http://doi.org/10.4142/jvs.2013.14.1.61

Abstract

This study was performed to evaluate the effects of conditioned media (CM) from human amniotic epithelial cells (HAECs) on the corneal wound healing process. Eighteen rabbits (36 eyes) were used and randomly assigned to three groups according treatment: CM from HAECs (group 1), vehicle alone (group 2), and saline (group 3). Corneal alkali injuries were induced with 1 N sodium hydroxide. Each reagent used for treatment evaluation was injected into the dorsal bulbar subconjunctiva and the area of the corneal epithelial defect was measured every other day. Two animals from each group were euthanized at a time on days 3, 7, and 15, and the cornea was removed for histological examination. The sum of the epithelial defect areas measured on day 0 to day 6 as well as day 0 to day 14 in group 1 was significantly smaller than those of other groups. Histological examination revealed that the group 1 corneas had less inflammatory cell infiltration and showed more intact epithelial features compared to the other groups. These results suggest that CM from HAECs promote corneal wound healing in rabbits.

Keyword

conditioned media; corneal alkali injury; human amniotic epithelial cell; rabbits

MeSH Terms

Alkalies/*toxicity
Amnion/*cytology
Animals
Cornea/*injuries
Corneal Diseases/chemically induced/therapy/*veterinary
Culture Media, Conditioned/*pharmacology
Epithelial Cells/*physiology
Humans
Male
*Rabbits
Alkalies
Culture Media, Conditioned

Figure

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  Fig. 1 Changes in the mean ± SD corneal epithelial defect area after alkali injury in rabbits. Although the epithelial defect area decreased significantly on day 2, the area increased on day 4 in all groups. The area of group 1 (—) corneas then decreased and was maintained at a low value up to day 12. Although the area of group 2 (····) decreased slowly up to day 8, it increased again on day 10 and continued to be the highest among all groups up to day 12. The area of group 3 (----) had the medium value among the groups from days 6 to 12.

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  Fig. 2 Representative eyes from each group showing the epithelial defect area (bright green staining) and corneal neovascularization (white arrows on day 14). Note that the epithelial defect areas in all groups were greatly reduced within 2 days but enlarged again on day 4. Although the corneal epithelium in group 1 was intact from day 6 to 12, corneas in the other groups were recurrently de-epithelialized.

• 

  Fig. 3 Corneal histological changes observed on each day after alkali injury in the rabbits. On day 3, intact epithelium was seen in groups 1 (A) and 2 (B) while epithelial features of dysmaturation and poor attachment were found in group 3 (C). On day 7, intact epithelium was observed in group 1 (D), blister formation was seen on the alkali wound surface of a cornea in group 2 (E; white arrow), and epithelial dysmaturation and non-adherence of a cornea was found in group 3 (F; white arrow). On day 15, intact epithelium was seen in group 1 (G) and group 2 (H). Epithelial features of dysmaturation and de-epithelialization of a cornea in group 3 (I; white arrow). Black arrows indicate inflammatory cells infiltrated into the stroma in each group. H&E stain, ×10.

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