Safety and feasibility of intrastromal injection of cultivated human corneal stromal keratocytes as cell-based therapy for corneal opacities
Purpose: To evaluate the safety and feasibility of intrastromal injection of human corneal stromal keratocytes (CSKs) and its therapeutic effect on a rodent early corneal opacity model. Methods: Twelve research-grade donor corneas were used in primary culture to generate quiescent CSKs and activated...
| Main Authors: | , , , , , , , , , , |
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| Other Authors: | |
| Format: | Journal Article |
| Language: | English |
| Published: |
2019
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| Subjects: | |
| Online Access: | https://hdl.handle.net/10356/84634 http://hdl.handle.net/10220/49158 |
| _version_ | 1826121298008342528 |
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| author | Yam, Gary Hin-Fai Fuest, Matthias Nur Zahirah M. Yusoff Mehta, Jodhbir Singh Goh, Tze-Wei Bandeira, Francisco Setiawan, Melina Seah, Xin-Yi Lwin, Nyein-Chan Stanzel, Tisha P. Ong, Hon-Shing |
| author2 | School of Materials Science & Engineering |
| author_facet | School of Materials Science & Engineering Yam, Gary Hin-Fai Fuest, Matthias Nur Zahirah M. Yusoff Mehta, Jodhbir Singh Goh, Tze-Wei Bandeira, Francisco Setiawan, Melina Seah, Xin-Yi Lwin, Nyein-Chan Stanzel, Tisha P. Ong, Hon-Shing |
| author_sort | Yam, Gary Hin-Fai |
| collection | NTU |
| description | Purpose: To evaluate the safety and feasibility of intrastromal injection of human corneal stromal keratocytes (CSKs) and its therapeutic effect on a rodent early corneal opacity model. Methods: Twelve research-grade donor corneas were used in primary culture to generate quiescent CSKs and activated stromal fibroblasts (SFs). Single and repeated intrastromal injections of 2 to 4 × 104 cells to rat normal corneas (n = 52) or corneas with early opacities induced by irregular phototherapeutic keratectomy (n = 16) were performed, followed by weekly examination of corneal response under slit-lamp biomicroscopy and in vivo confocal microscopy with evaluation of haze level and stromal reflectivity, and corneal thickness using anterior segment optical coherence tomography (AS-OCT). Time-lapse tracing of Molday ION–labelled cells was conducted using Spectralis OCT and label intensity was measured. Corneas were collected at time intervals for marker expression by immunofluorescence, cell viability, and apoptosis assays. Results: Injected CSKs showed proper marker expression with negligible SF-related features and inflammation, hence maintaining corneal clarity and stability. The time-dependent loss of injected cells was recovered by repeated injection, achieving an extended expression of human proteoglycans inside rat stroma. In the early corneal opacity model, intrastromal CSK injection reduced stromal reflectivity and thickness, resulting in recovery of corneal clarity, whereas noninjected corneas were thicker and had haze progression. Conclusions: We demonstrated the safety, feasibility, and therapeutic efficacy of intrastromal CSK injection. The cultivated CSKs can be a reliable cell source for potential cell-based therapy for corneal opacities. |
| first_indexed | 2024-10-01T05:30:23Z |
| format | Journal Article |
| id | ntu-10356/84634 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T05:30:23Z |
| publishDate | 2019 |
| record_format | dspace |
| spelling | ntu-10356/846342023-07-14T15:51:14Z Safety and feasibility of intrastromal injection of cultivated human corneal stromal keratocytes as cell-based therapy for corneal opacities Yam, Gary Hin-Fai Fuest, Matthias Nur Zahirah M. Yusoff Mehta, Jodhbir Singh Goh, Tze-Wei Bandeira, Francisco Setiawan, Melina Seah, Xin-Yi Lwin, Nyein-Chan Stanzel, Tisha P. Ong, Hon-Shing School of Materials Science & Engineering Science::Medicine Intrastromal Injection Corneal Opacities Purpose: To evaluate the safety and feasibility of intrastromal injection of human corneal stromal keratocytes (CSKs) and its therapeutic effect on a rodent early corneal opacity model. Methods: Twelve research-grade donor corneas were used in primary culture to generate quiescent CSKs and activated stromal fibroblasts (SFs). Single and repeated intrastromal injections of 2 to 4 × 104 cells to rat normal corneas (n = 52) or corneas with early opacities induced by irregular phototherapeutic keratectomy (n = 16) were performed, followed by weekly examination of corneal response under slit-lamp biomicroscopy and in vivo confocal microscopy with evaluation of haze level and stromal reflectivity, and corneal thickness using anterior segment optical coherence tomography (AS-OCT). Time-lapse tracing of Molday ION–labelled cells was conducted using Spectralis OCT and label intensity was measured. Corneas were collected at time intervals for marker expression by immunofluorescence, cell viability, and apoptosis assays. Results: Injected CSKs showed proper marker expression with negligible SF-related features and inflammation, hence maintaining corneal clarity and stability. The time-dependent loss of injected cells was recovered by repeated injection, achieving an extended expression of human proteoglycans inside rat stroma. In the early corneal opacity model, intrastromal CSK injection reduced stromal reflectivity and thickness, resulting in recovery of corneal clarity, whereas noninjected corneas were thicker and had haze progression. Conclusions: We demonstrated the safety, feasibility, and therapeutic efficacy of intrastromal CSK injection. The cultivated CSKs can be a reliable cell source for potential cell-based therapy for corneal opacities. NRF (Natl Research Foundation, S’pore) NMRC (Natl Medical Research Council, S’pore) MOH (Min. of Health, S’pore) Published version 2019-07-05T07:09:19Z 2019-12-06T15:48:44Z 2019-07-05T07:09:19Z 2019-12-06T15:48:44Z 2018 Journal Article Yam, G. H.-F., Fuest, M., Nur Zahirah M. Yusoff., Goh, T.-W., Bandeira, F., Setiawan, M., . . . Mehta, J. S. (2018). Safety and feasibility of intrastromal injection of cultivated human corneal stromal keratocytes as cell-based therapy for corneal opacities. Investigative Opthalmology & Visual Science, 59(8), 3340-3354. doi:10.1167/iovs.17-23575 https://hdl.handle.net/10356/84634 http://hdl.handle.net/10220/49158 10.1167/iovs.17-23575 en Investigative Opthalmology & Visual Science © 2018 The Author(s) (Published by ARVO). This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License. 15 p. application/pdf |
| spellingShingle | Science::Medicine Intrastromal Injection Corneal Opacities Yam, Gary Hin-Fai Fuest, Matthias Nur Zahirah M. Yusoff Mehta, Jodhbir Singh Goh, Tze-Wei Bandeira, Francisco Setiawan, Melina Seah, Xin-Yi Lwin, Nyein-Chan Stanzel, Tisha P. Ong, Hon-Shing Safety and feasibility of intrastromal injection of cultivated human corneal stromal keratocytes as cell-based therapy for corneal opacities |
| title | Safety and feasibility of intrastromal injection of cultivated human corneal stromal keratocytes as cell-based therapy for corneal opacities |
| title_full | Safety and feasibility of intrastromal injection of cultivated human corneal stromal keratocytes as cell-based therapy for corneal opacities |
| title_fullStr | Safety and feasibility of intrastromal injection of cultivated human corneal stromal keratocytes as cell-based therapy for corneal opacities |
| title_full_unstemmed | Safety and feasibility of intrastromal injection of cultivated human corneal stromal keratocytes as cell-based therapy for corneal opacities |
| title_short | Safety and feasibility of intrastromal injection of cultivated human corneal stromal keratocytes as cell-based therapy for corneal opacities |
| title_sort | safety and feasibility of intrastromal injection of cultivated human corneal stromal keratocytes as cell based therapy for corneal opacities |
| topic | Science::Medicine Intrastromal Injection Corneal Opacities |
| url | https://hdl.handle.net/10356/84634 http://hdl.handle.net/10220/49158 |
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