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Amniotic Membrane Extract Eye Drop Promotes Limbal Stem Cell Proliferation and Corneal Epithelium Healing
OBJECTIVE: Human amniotic membrane (HAM) is used as a supporter for limbal stem cell (LSC) expansion and corneal surgery. The aim of study is to use HAM extracts from healthy donors to enhance proliferation of LSCs in vitro and in vivo. MATERIALS AND METHODS: In this interventional experimental stud...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Royan Institute
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6099140/ https://www.ncbi.nlm.nih.gov/pubmed/30123991 http://dx.doi.org/10.22074/cellj.2019.5423 |
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author | Shayan Asl, Niloufar Nejat, Farhad Mohammadi, Parvaneh Nekoukar, Abdolhossein Hesam, Saeed Ebrahimi, Marzieh Jadidi, Khosrow |
author_facet | Shayan Asl, Niloufar Nejat, Farhad Mohammadi, Parvaneh Nekoukar, Abdolhossein Hesam, Saeed Ebrahimi, Marzieh Jadidi, Khosrow |
author_sort | Shayan Asl, Niloufar |
collection | PubMed |
description | OBJECTIVE: Human amniotic membrane (HAM) is used as a supporter for limbal stem cell (LSC) expansion and corneal surgery. The aim of study is to use HAM extracts from healthy donors to enhance proliferation of LSCs in vitro and in vivo. MATERIALS AND METHODS: In this interventional experimental study, the effective and cytotoxic doses of the amniotic membrane extract eye drops (AMEED) was assessed by adding different concentrations of AMEED (0-2.0 mg/ml) to LSC cultures for 14 days. Subsequently, the expression levels of ATP-binding cassette sub-family G member 2 (ABCG2, a putative stem cell marker), cytokeratin 3 (K3, corneal maker), K12 and K19 (corneal-conjunctival cell makers) were assessed by real-time polymerase chain reaction (PCR). In the second step, the corneal epithelium of 10 rabbits was mechanically removed, and the right eye of each rabbit was treated with 1 mg/ml AMEED [every 2 hours (group 1) or every 6 hours (group 2)]. The left eyes only received an antibiotic. The corneal healing process, conjunctival infection, degree of eyelid oedema, degree of photophobia, and discharge scores were evaluated during daily assessments. Finally, corneal tissues were biopsied for pathologic evidences. RESULTS: In comparison to the positive control [10% foetal bovine serum (FBS)], 0.1-1 mg/ml AMEED induced LSC proliferation, upregulated ABCG2, and downregulated K3. There were no remarkable differences in the expression levels of K12 and K19 (P>0.05). Interestingly, in the rabbits treated with AMEED, the epithelium healing duration decreased from 4 days in the control group to 3 days in the two AMEED groups, with lower mean degrees of eyelid oedema, chemosis, and infection compared to the control group. No pathologic abnormalities were observed in either of the AMEED groups. CONCLUSION: AMEED increases LSCs proliferation ex vivo and accelerates corneal epithelium healing in vivo without any adverse effects. It could be used as a supplement for LSC expansion in cell therapy. |
format | Online Article Text |
id | pubmed-6099140 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Royan Institute |
record_format | MEDLINE/PubMed |
spelling | pubmed-60991402019-01-01 Amniotic Membrane Extract Eye Drop Promotes Limbal Stem Cell Proliferation and Corneal Epithelium Healing Shayan Asl, Niloufar Nejat, Farhad Mohammadi, Parvaneh Nekoukar, Abdolhossein Hesam, Saeed Ebrahimi, Marzieh Jadidi, Khosrow Cell J Original Article OBJECTIVE: Human amniotic membrane (HAM) is used as a supporter for limbal stem cell (LSC) expansion and corneal surgery. The aim of study is to use HAM extracts from healthy donors to enhance proliferation of LSCs in vitro and in vivo. MATERIALS AND METHODS: In this interventional experimental study, the effective and cytotoxic doses of the amniotic membrane extract eye drops (AMEED) was assessed by adding different concentrations of AMEED (0-2.0 mg/ml) to LSC cultures for 14 days. Subsequently, the expression levels of ATP-binding cassette sub-family G member 2 (ABCG2, a putative stem cell marker), cytokeratin 3 (K3, corneal maker), K12 and K19 (corneal-conjunctival cell makers) were assessed by real-time polymerase chain reaction (PCR). In the second step, the corneal epithelium of 10 rabbits was mechanically removed, and the right eye of each rabbit was treated with 1 mg/ml AMEED [every 2 hours (group 1) or every 6 hours (group 2)]. The left eyes only received an antibiotic. The corneal healing process, conjunctival infection, degree of eyelid oedema, degree of photophobia, and discharge scores were evaluated during daily assessments. Finally, corneal tissues were biopsied for pathologic evidences. RESULTS: In comparison to the positive control [10% foetal bovine serum (FBS)], 0.1-1 mg/ml AMEED induced LSC proliferation, upregulated ABCG2, and downregulated K3. There were no remarkable differences in the expression levels of K12 and K19 (P>0.05). Interestingly, in the rabbits treated with AMEED, the epithelium healing duration decreased from 4 days in the control group to 3 days in the two AMEED groups, with lower mean degrees of eyelid oedema, chemosis, and infection compared to the control group. No pathologic abnormalities were observed in either of the AMEED groups. CONCLUSION: AMEED increases LSCs proliferation ex vivo and accelerates corneal epithelium healing in vivo without any adverse effects. It could be used as a supplement for LSC expansion in cell therapy. Royan Institute 2019 2018-08-07 /pmc/articles/PMC6099140/ /pubmed/30123991 http://dx.doi.org/10.22074/cellj.2019.5423 Text en Any use, distribution, reproduction or abstract of this publication in any medium, with the exception of commercial purposes, is permitted provided the original work is properly cited http://creativecommons.org/licenses/by/2.5/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Original Article Shayan Asl, Niloufar Nejat, Farhad Mohammadi, Parvaneh Nekoukar, Abdolhossein Hesam, Saeed Ebrahimi, Marzieh Jadidi, Khosrow Amniotic Membrane Extract Eye Drop Promotes Limbal Stem Cell Proliferation and Corneal Epithelium Healing |
title | Amniotic Membrane Extract Eye Drop Promotes Limbal Stem Cell
Proliferation and Corneal Epithelium Healing |
title_full | Amniotic Membrane Extract Eye Drop Promotes Limbal Stem Cell
Proliferation and Corneal Epithelium Healing |
title_fullStr | Amniotic Membrane Extract Eye Drop Promotes Limbal Stem Cell
Proliferation and Corneal Epithelium Healing |
title_full_unstemmed | Amniotic Membrane Extract Eye Drop Promotes Limbal Stem Cell
Proliferation and Corneal Epithelium Healing |
title_short | Amniotic Membrane Extract Eye Drop Promotes Limbal Stem Cell
Proliferation and Corneal Epithelium Healing |
title_sort | amniotic membrane extract eye drop promotes limbal stem cell
proliferation and corneal epithelium healing |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6099140/ https://www.ncbi.nlm.nih.gov/pubmed/30123991 http://dx.doi.org/10.22074/cellj.2019.5423 |
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