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Prolonged Inflammation and Infectious Changes in the Corneal Epithelium Are Associated with Persistent Epithelial Defect (PED)

Purpose: Failure of rapid re-epithelialization within 10–14 days after corneal injury, even with standard supportive treatment, is referred to as persistent corneal epithelial (CE) defect (PED). Though an array of genes regulates reepithelization, their mechanisms are poorly understood. We sought to...

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Autores principales: Dutta, Tanmoy, Sangwan, Jyoti, Mondal, Moumita, Vohra, Mehak, Nidhi, Vatsala, Gour, Abha, Kapur, Neha, Gupta, Nidhi, Bhowmick, Tuhin, Chandru, Arun, Mathur, Umang, Sangwan, Virender Singh, Acharya, Manisha, Tiwari, Anil
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9960897/
https://www.ncbi.nlm.nih.gov/pubmed/36839533
http://dx.doi.org/10.3390/pathogens12020261
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author Dutta, Tanmoy
Sangwan, Jyoti
Mondal, Moumita
Vohra, Mehak
Nidhi, Vatsala
Gour, Abha
Kapur, Neha
Gupta, Nidhi
Bhowmick, Tuhin
Chandru, Arun
Mathur, Umang
Sangwan, Virender Singh
Acharya, Manisha
Tiwari, Anil
author_facet Dutta, Tanmoy
Sangwan, Jyoti
Mondal, Moumita
Vohra, Mehak
Nidhi, Vatsala
Gour, Abha
Kapur, Neha
Gupta, Nidhi
Bhowmick, Tuhin
Chandru, Arun
Mathur, Umang
Sangwan, Virender Singh
Acharya, Manisha
Tiwari, Anil
author_sort Dutta, Tanmoy
collection PubMed
description Purpose: Failure of rapid re-epithelialization within 10–14 days after corneal injury, even with standard supportive treatment, is referred to as persistent corneal epithelial (CE) defect (PED). Though an array of genes regulates reepithelization, their mechanisms are poorly understood. We sought to understand the network of genes driving the re-epithelialization in PED. Method: After obtaining informed consent, patients underwent an ophthalmic examination. Epithelial scrapes and tears samples of six PED patients and six individuals (control) undergoing photorefractive keratectomy (PRK) were collected. RNA isolation and quantification were performed using either the epithelial scrape taken from PED patients or from HCLE cells treated with control tears or tears of PED patients. Quantitative real-time polymerase chain reaction (qRT-PCR) was performed to detect the expression of a few important genes in CE homeostasis, inflammation, and cell–cell communication, viz., Kruppel-like factor 4 (KLF4), GPX4, IL6, TNFα, STING, IL8, desmoglein, and E-cadherin, among others. Their expressions were normalized with their respective housekeeping genes and fold changes were recorded. KLF4 localization and MMPs activity was carried out via immunofluorescence and zymography, respectively. Results: KLF4, a transcription factor important for CE homeostasis, was upregulated in tears-treated HCLE cells and downregulated in PED patients compared to the healthy PRK group. Cell–cell communication genes were also upregulated in tears-treated cells, whereas they were downregulated in the PED tissue group. Genes involved in proinflammation (IL6, 282-fold; TNFα, 43-fold; IL8, 4.2-fold) were highly upregulated in both conditions. MMP9 activity increased upon tears treatment. Conclusions: This study suggests that tears create an acute proinflammatory milieu driving the PED disease pathology, whereas the PED patients scrapes are an indicator of the chronic stage of the disease. Interferons, pro-inflammatory genes, and their pathways are involved in PED, which can be a potential target for inducing epithelialization of the cornea.
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spelling pubmed-99608972023-02-26 Prolonged Inflammation and Infectious Changes in the Corneal Epithelium Are Associated with Persistent Epithelial Defect (PED) Dutta, Tanmoy Sangwan, Jyoti Mondal, Moumita Vohra, Mehak Nidhi, Vatsala Gour, Abha Kapur, Neha Gupta, Nidhi Bhowmick, Tuhin Chandru, Arun Mathur, Umang Sangwan, Virender Singh Acharya, Manisha Tiwari, Anil Pathogens Article Purpose: Failure of rapid re-epithelialization within 10–14 days after corneal injury, even with standard supportive treatment, is referred to as persistent corneal epithelial (CE) defect (PED). Though an array of genes regulates reepithelization, their mechanisms are poorly understood. We sought to understand the network of genes driving the re-epithelialization in PED. Method: After obtaining informed consent, patients underwent an ophthalmic examination. Epithelial scrapes and tears samples of six PED patients and six individuals (control) undergoing photorefractive keratectomy (PRK) were collected. RNA isolation and quantification were performed using either the epithelial scrape taken from PED patients or from HCLE cells treated with control tears or tears of PED patients. Quantitative real-time polymerase chain reaction (qRT-PCR) was performed to detect the expression of a few important genes in CE homeostasis, inflammation, and cell–cell communication, viz., Kruppel-like factor 4 (KLF4), GPX4, IL6, TNFα, STING, IL8, desmoglein, and E-cadherin, among others. Their expressions were normalized with their respective housekeeping genes and fold changes were recorded. KLF4 localization and MMPs activity was carried out via immunofluorescence and zymography, respectively. Results: KLF4, a transcription factor important for CE homeostasis, was upregulated in tears-treated HCLE cells and downregulated in PED patients compared to the healthy PRK group. Cell–cell communication genes were also upregulated in tears-treated cells, whereas they were downregulated in the PED tissue group. Genes involved in proinflammation (IL6, 282-fold; TNFα, 43-fold; IL8, 4.2-fold) were highly upregulated in both conditions. MMP9 activity increased upon tears treatment. Conclusions: This study suggests that tears create an acute proinflammatory milieu driving the PED disease pathology, whereas the PED patients scrapes are an indicator of the chronic stage of the disease. Interferons, pro-inflammatory genes, and their pathways are involved in PED, which can be a potential target for inducing epithelialization of the cornea. MDPI 2023-02-06 /pmc/articles/PMC9960897/ /pubmed/36839533 http://dx.doi.org/10.3390/pathogens12020261 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Dutta, Tanmoy
Sangwan, Jyoti
Mondal, Moumita
Vohra, Mehak
Nidhi, Vatsala
Gour, Abha
Kapur, Neha
Gupta, Nidhi
Bhowmick, Tuhin
Chandru, Arun
Mathur, Umang
Sangwan, Virender Singh
Acharya, Manisha
Tiwari, Anil
Prolonged Inflammation and Infectious Changes in the Corneal Epithelium Are Associated with Persistent Epithelial Defect (PED)
title Prolonged Inflammation and Infectious Changes in the Corneal Epithelium Are Associated with Persistent Epithelial Defect (PED)
title_full Prolonged Inflammation and Infectious Changes in the Corneal Epithelium Are Associated with Persistent Epithelial Defect (PED)
title_fullStr Prolonged Inflammation and Infectious Changes in the Corneal Epithelium Are Associated with Persistent Epithelial Defect (PED)
title_full_unstemmed Prolonged Inflammation and Infectious Changes in the Corneal Epithelium Are Associated with Persistent Epithelial Defect (PED)
title_short Prolonged Inflammation and Infectious Changes in the Corneal Epithelium Are Associated with Persistent Epithelial Defect (PED)
title_sort prolonged inflammation and infectious changes in the corneal epithelium are associated with persistent epithelial defect (ped)
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9960897/
https://www.ncbi.nlm.nih.gov/pubmed/36839533
http://dx.doi.org/10.3390/pathogens12020261
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