Vascular Endothelial Growth Factor Expression Patterns in non- Human Papillomavirus – Related Pterygia: An Experimental Study on Cell Spot Arrays Digital Analysis

Abstract

Purpose

The role of angiogenic factors –such as vascular endothelial growth factor (VEGF) – in the development and progression of pterygia lesions remains under investigation. In the current study, we analyzed VEGF protein expression in a series of pterygia and normal conjunctiva epithelia.

Methods

Using a liquid-based cytology assay, thirty (n = 30) cell specimens were obtained by applying a smooth scraping on conjunctiva epithelia and fixed accordingly. None of them had a history of Human Papillomavirus (HPV) infection. Similarly, the same process was applied also in normal conjunctiva epithelia (n = 10; control group). We constructed five (n = 5) slides each containing eight (n = 8) cell spots. An immunocytochemistry (ICC) assay was implemented. Digital image analysis was also performed for evaluating objectively the corresponding immunostaining intensity levels.

Results

All the examined pterygia cell samples over-expressed the marker. High staining intensity levels were detected in 15/30 (50%), whereas the rest 15/30 (50%) demonstrated moderate expression. Overall VEGF expression was statistically significantly higher in pterygia compared to normal conjunctiva epithelia (p=.0001). Concerning the other parameters, VEGF protein expression did not associate with the gender of the patients (p = 0.518), the presence of a recurrent lesion (p = 0.311), the anatomical location (p = 0.191) or with their morphology (p = 0.316). Interestingly, the recurrent lesions demonstrated the highest levels of VEGF expression.

Conclusions

VEGF overexpression is a frequent event in pterygia playing a potentially central molecular role in the progression of the lesion. Cell spot array analysis -based on liquid cytology- seems to be an innovative, easy-to-use technique for analyzing a broad variety of molecules in multiple specimens on the same slide by applying different ICC assays.

Keywords:

• Pterygia
• cornea
• ocular
• VEGF
• cytology
• image analysis

Author contributions

SM, MA, ET (Writing, analysis); OM, MP, VT (analysis, critical review); CG (proofreading).

Disclosure statement

No potential conflict of interest was reported by the author(s).

Data availability statement

Data is available on request due to privacy/ethical restrictions.