Inflammation, vascularization and goblet cell differences in LSCD: Validating animal models of corneal alkali burns

Abhinav Reddy Kethiri, Enoch Raju, Kiran Kumar Bokara, Dilip Kumar Mishra, Sayan Basu, Ch Mohan Rao, Virender Singh Sangwan, Vivek Singh

Research output: Contribution to journalArticleScientificpeer-review


Limbal stem cell deficiency (LSCD) is one of the serious cause of visual impairment and blindness with loss of corneal clarity and vascularization. Factors such as ocular burns (acids, lime, thermal), genetic disorders or infections results in the loss of limbal stem cells leading to LSCD. Reliable animal models of LSCD are useful for understanding the pathophysiology and developing novel therapeutic approaches. The purpose of the present study was to validate small and large animal models of LSCD by immunohistochemcal, clinical and histopathological comparison with human. The animal models of LSCD were created by topical administration of sodium hydroxide on the ocular surface of C57BL/6 mice (m, n = 12) and New Zealand white rabbits (r, n = 12) as per the standard existing protocol. Human corneal specimens (h, n = 12) were obtained from tissue bank who had chemical burn-induced LSCD. All samples were either paraffin embedded or frozen in cryogenic medium and the sections were processed for Hematoxylin-Eosin and Periodic Acid-Schiff staining to analyse the morphology and histopathological features of the corneal surface such as vascularization, inflammation, presence of goblet cells, epithelial hyperplasia and keratinization. Immunofluorescence was performed to distinguish between corneal (CK3+), conjunctival (CK19+) and epidermal (CK10+) epithelial phenotype. Histological analysis of corneal specimens from the three groups showed the presence of goblet cells (h:83%, m:50%, r:50%, p = 0.014), epithelial hypertrophy (h:92%, m:50%, r:66.6%, p = 0.04), epithelial hyperplasia (h:50%, m:17%, r:17%, p = 0.18), intra epithelial edema (h:42%, m:33%, r:100%, p = 0.02), stromal inflammation (h:100%, m:67%, r:67%, p = 0.01) and stromal vascularization (h:100%, m:50%, r:67%), in varying proportions. Immunostaining showed presence of total LSCD (CK19 + and/or CK10+, CK3-) in 92% of human and 50% of animal specimens. While partial LSCD (CK19 + and/or CK10+, CK3+) was seen in 8% of human and 50% of animal specimens. Our study shows the significant differences in the extent of vascularization, inflammation, epithelial thickness and goblet cell formation in mice and rabbit models of LSCD when compared to post-chemical burn LSCD in human corneas. In both mice and rabbit models complete LSCD developed in only 50% of cases and this important fact needs to be considered when working with animal models of LSCD.

Original languageEnglish
Article number107665
JournalExperimental Eye Research
Publication statusPublished - Aug 2019
Externally publishedYes
Publication typeA1 Journal article-refereed


  • Animals
  • Burns, Chemical/metabolism
  • Corneal Diseases/metabolism
  • Corneal Neovascularization/metabolism
  • Disease Models, Animal
  • Epithelial Cells/metabolism
  • Epithelium, Corneal
  • Eye Burns/chemically induced
  • Female
  • Fluorescent Antibody Technique, Indirect
  • Goblet Cells/metabolism
  • Humans
  • Immunophenotyping
  • Inflammation/metabolism
  • Keratin-19/metabolism
  • Keratin-3/metabolism
  • Keratitis/metabolism
  • Limbus Corneae/metabolism
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mucins/metabolism
  • Rabbits
  • Sodium Hydroxide/toxicity


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