Assessment of Corneal Stromal Remodeling and Regeneration after Photorefractive Keratectomy
Abstract This study utilizes high resolution multi-dimensional imaging to identify temporal and spatial changes in cell/extracellular matrix (ECM) patterning mediating cell migration, fibrosis, remodeling and regeneration during wound healing. Photorefractive keratectomy (PRK) was performed on rabbi...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2018-08-01
|
| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-018-30372-2 |
| _version_ | 1818996529486102528 |
|---|---|
| author | Pouriska B. Kivanany Kyle C. Grose Madhavi Tippani Shan Su W. Matthew Petroll |
| author_facet | Pouriska B. Kivanany Kyle C. Grose Madhavi Tippani Shan Su W. Matthew Petroll |
| author_sort | Pouriska B. Kivanany |
| collection | DOAJ |
| description | Abstract This study utilizes high resolution multi-dimensional imaging to identify temporal and spatial changes in cell/extracellular matrix (ECM) patterning mediating cell migration, fibrosis, remodeling and regeneration during wound healing. Photorefractive keratectomy (PRK) was performed on rabbits. In some cases, 5([4,6-dichlorotriazin-2yl]-amino)fluorescein (DTAF) was applied immediately after surgery to differentiate native vs. cell-secreted collagen. Corneas were assessed 3–180 days postoperatively using in vivo confocal microscopy, and cell/ECM patterning was evaluated in situ using multiphoton and second harmonic generation (SHG) imaging. 7 days post-PRK, migrating fibroblasts below the ablation site were co-aligned with the stromal lamellae. At day 21, randomly patterned myofibroblasts developed on top of the ablation site; whereas cells underneath were elongated, co-aligned with collagen, and lacked stress fibers. Over time, fibrotic tissue was remodeled into more transparent stromal lamellae. By day 180, stromal thickness was almost completely restored. Stromal regrowth occurred primarily below the ablation interface, and was characterized by co-localization of gaps in DTAF labeling with elongated cells and SHG collagen signaling. Punctate F-actin labeling was detected along cells co-aligned with DTAF and non-DTAF labeled collagen, suggesting cell-ECM interactions. Overall, collagen lamellae appear to provide a template for fibroblast patterning during wound healing that mediates stromal repopulation, regeneration and remodeling. |
| first_indexed | 2024-12-20T21:31:11Z |
| format | Article |
| id | doaj.art-40d118bf16d9433881159e7adc92e021 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-20T21:31:11Z |
| publishDate | 2018-08-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-40d118bf16d9433881159e7adc92e0212022-12-21T19:26:02ZengNature PortfolioScientific Reports2045-23222018-08-018111410.1038/s41598-018-30372-2Assessment of Corneal Stromal Remodeling and Regeneration after Photorefractive KeratectomyPouriska B. Kivanany0Kyle C. Grose1Madhavi Tippani2Shan Su3W. Matthew Petroll4Department of Ophthalmology, UT Southwestern Medical CenterDepartment of Ophthalmology, UT Southwestern Medical CenterDepartment of Ophthalmology, UT Southwestern Medical CenterDepartment of Ophthalmology, UT Southwestern Medical CenterDepartment of Ophthalmology, UT Southwestern Medical CenterAbstract This study utilizes high resolution multi-dimensional imaging to identify temporal and spatial changes in cell/extracellular matrix (ECM) patterning mediating cell migration, fibrosis, remodeling and regeneration during wound healing. Photorefractive keratectomy (PRK) was performed on rabbits. In some cases, 5([4,6-dichlorotriazin-2yl]-amino)fluorescein (DTAF) was applied immediately after surgery to differentiate native vs. cell-secreted collagen. Corneas were assessed 3–180 days postoperatively using in vivo confocal microscopy, and cell/ECM patterning was evaluated in situ using multiphoton and second harmonic generation (SHG) imaging. 7 days post-PRK, migrating fibroblasts below the ablation site were co-aligned with the stromal lamellae. At day 21, randomly patterned myofibroblasts developed on top of the ablation site; whereas cells underneath were elongated, co-aligned with collagen, and lacked stress fibers. Over time, fibrotic tissue was remodeled into more transparent stromal lamellae. By day 180, stromal thickness was almost completely restored. Stromal regrowth occurred primarily below the ablation interface, and was characterized by co-localization of gaps in DTAF labeling with elongated cells and SHG collagen signaling. Punctate F-actin labeling was detected along cells co-aligned with DTAF and non-DTAF labeled collagen, suggesting cell-ECM interactions. Overall, collagen lamellae appear to provide a template for fibroblast patterning during wound healing that mediates stromal repopulation, regeneration and remodeling.https://doi.org/10.1038/s41598-018-30372-2 |
| spellingShingle | Pouriska B. Kivanany Kyle C. Grose Madhavi Tippani Shan Su W. Matthew Petroll Assessment of Corneal Stromal Remodeling and Regeneration after Photorefractive Keratectomy Scientific Reports |
| title | Assessment of Corneal Stromal Remodeling and Regeneration after Photorefractive Keratectomy |
| title_full | Assessment of Corneal Stromal Remodeling and Regeneration after Photorefractive Keratectomy |
| title_fullStr | Assessment of Corneal Stromal Remodeling and Regeneration after Photorefractive Keratectomy |
| title_full_unstemmed | Assessment of Corneal Stromal Remodeling and Regeneration after Photorefractive Keratectomy |
| title_short | Assessment of Corneal Stromal Remodeling and Regeneration after Photorefractive Keratectomy |
| title_sort | assessment of corneal stromal remodeling and regeneration after photorefractive keratectomy |
| url | https://doi.org/10.1038/s41598-018-30372-2 |
| work_keys_str_mv | AT pouriskabkivanany assessmentofcornealstromalremodelingandregenerationafterphotorefractivekeratectomy AT kylecgrose assessmentofcornealstromalremodelingandregenerationafterphotorefractivekeratectomy AT madhavitippani assessmentofcornealstromalremodelingandregenerationafterphotorefractivekeratectomy AT shansu assessmentofcornealstromalremodelingandregenerationafterphotorefractivekeratectomy AT wmatthewpetroll assessmentofcornealstromalremodelingandregenerationafterphotorefractivekeratectomy |