Coupling of Fibrin Reorganization and Fibronectin Patterning by Corneal Fibroblasts in Response to PDGF BB and TGFβ1
We previously reported that corneal fibroblasts within 3D fibrin matrices secrete, bind, and organize fibronectin into tracks that facilitate cell spreading and migration. Other cells use these fibronectin tracks as conduits, which leads to the development of an interconnected cell/fibronectin netwo...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2020-08-01
|
| Series: | Bioengineering |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2306-5354/7/3/89 |
| _version_ | 1797559739503083520 |
|---|---|
| author | Miguel Miron-Mendoza Dalia Vazquez Nerea García-Rámila Hikaru R. Ikebe W. Matthew Petroll |
| author_facet | Miguel Miron-Mendoza Dalia Vazquez Nerea García-Rámila Hikaru R. Ikebe W. Matthew Petroll |
| author_sort | Miguel Miron-Mendoza |
| collection | DOAJ |
| description | We previously reported that corneal fibroblasts within 3D fibrin matrices secrete, bind, and organize fibronectin into tracks that facilitate cell spreading and migration. Other cells use these fibronectin tracks as conduits, which leads to the development of an interconnected cell/fibronectin network. In this study, we investigate how cell-induced reorganization of fibrin correlates with fibronectin track formation in response to two growth factors present during wound healing: PDGF BB, which stimulates cell spreading and migration; and TGFβ1, which stimulates cellular contraction and myofibroblast transformation. Both PDGF BB and TGFβ1 stimulated global fibrin matrix contraction (<i>p</i> < 0.005); however, the cell and matrix patterning were different. We found that, during PDGF BB-induced cell spreading, fibronectin was organized simultaneously with the generation of tractional forces at the leading edge of pseudopodia. Over time this led to the formation of an interconnected network consisting of cells, fibronectin and compacted fibrin tracks. Following culture in TGFβ1, cells were less motile, produced significant local fibrin reorganization, and formed fewer cellular connections as compared to PDGF BB (<i>p</i> < 0.005). Although bands of compacted fibrin tracks developed in between neighboring cells, fibronectin labeling was not generally present along these tracks, and the correlation between fibrin and fibronectin labeling was significantly less than that observed in PDGF BB (<i>p</i> < 0.001). Taken together, our results show that cell-induced extracellular matrix (ECM) reorganization can occur independently from fibronectin patterning. Nonetheless, both events seem to be coordinated, as corneal fibroblasts in PDGF BB secrete and organize fibronectin as they preferentially spread along compacted fibrin tracks between cells, producing an interconnected network in which cells, fibronectin and compacted fibrin tracks are highly correlated. This mechanism of patterning could contribute to the formation of organized cellular networks that have been observed following corneal injury and refractive surgery. |
| first_indexed | 2024-03-10T17:49:35Z |
| format | Article |
| id | doaj.art-22bbfd5d9eff41bfb8b9a30c35b351ea |
| institution | Directory Open Access Journal |
| issn | 2306-5354 |
| language | English |
| last_indexed | 2024-03-10T17:49:35Z |
| publishDate | 2020-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Bioengineering |
| spelling | doaj.art-22bbfd5d9eff41bfb8b9a30c35b351ea2023-11-20T09:23:18ZengMDPI AGBioengineering2306-53542020-08-01738910.3390/bioengineering7030089Coupling of Fibrin Reorganization and Fibronectin Patterning by Corneal Fibroblasts in Response to PDGF BB and TGFβ1Miguel Miron-Mendoza0Dalia Vazquez1Nerea García-Rámila2Hikaru R. Ikebe3W. Matthew Petroll4Department of Ophthalmology, UT Southwestern Medical Center, Dallas, TX 75390, USADepartment of Ophthalmology, UT Southwestern Medical Center, Dallas, TX 75390, USADepartment of Ophthalmology, UT Southwestern Medical Center, Dallas, TX 75390, USADepartment of Ophthalmology, UT Southwestern Medical Center, Dallas, TX 75390, USADepartment of Ophthalmology, UT Southwestern Medical Center, Dallas, TX 75390, USAWe previously reported that corneal fibroblasts within 3D fibrin matrices secrete, bind, and organize fibronectin into tracks that facilitate cell spreading and migration. Other cells use these fibronectin tracks as conduits, which leads to the development of an interconnected cell/fibronectin network. In this study, we investigate how cell-induced reorganization of fibrin correlates with fibronectin track formation in response to two growth factors present during wound healing: PDGF BB, which stimulates cell spreading and migration; and TGFβ1, which stimulates cellular contraction and myofibroblast transformation. Both PDGF BB and TGFβ1 stimulated global fibrin matrix contraction (<i>p</i> < 0.005); however, the cell and matrix patterning were different. We found that, during PDGF BB-induced cell spreading, fibronectin was organized simultaneously with the generation of tractional forces at the leading edge of pseudopodia. Over time this led to the formation of an interconnected network consisting of cells, fibronectin and compacted fibrin tracks. Following culture in TGFβ1, cells were less motile, produced significant local fibrin reorganization, and formed fewer cellular connections as compared to PDGF BB (<i>p</i> < 0.005). Although bands of compacted fibrin tracks developed in between neighboring cells, fibronectin labeling was not generally present along these tracks, and the correlation between fibrin and fibronectin labeling was significantly less than that observed in PDGF BB (<i>p</i> < 0.001). Taken together, our results show that cell-induced extracellular matrix (ECM) reorganization can occur independently from fibronectin patterning. Nonetheless, both events seem to be coordinated, as corneal fibroblasts in PDGF BB secrete and organize fibronectin as they preferentially spread along compacted fibrin tracks between cells, producing an interconnected network in which cells, fibronectin and compacted fibrin tracks are highly correlated. This mechanism of patterning could contribute to the formation of organized cellular networks that have been observed following corneal injury and refractive surgery.https://www.mdpi.com/2306-5354/7/3/89fibronectincorneal fibroblastsfibrin3-D matricescollective cell migration |
| spellingShingle | Miguel Miron-Mendoza Dalia Vazquez Nerea García-Rámila Hikaru R. Ikebe W. Matthew Petroll Coupling of Fibrin Reorganization and Fibronectin Patterning by Corneal Fibroblasts in Response to PDGF BB and TGFβ1 Bioengineering fibronectin corneal fibroblasts fibrin 3-D matrices collective cell migration |
| title | Coupling of Fibrin Reorganization and Fibronectin Patterning by Corneal Fibroblasts in Response to PDGF BB and TGFβ1 |
| title_full | Coupling of Fibrin Reorganization and Fibronectin Patterning by Corneal Fibroblasts in Response to PDGF BB and TGFβ1 |
| title_fullStr | Coupling of Fibrin Reorganization and Fibronectin Patterning by Corneal Fibroblasts in Response to PDGF BB and TGFβ1 |
| title_full_unstemmed | Coupling of Fibrin Reorganization and Fibronectin Patterning by Corneal Fibroblasts in Response to PDGF BB and TGFβ1 |
| title_short | Coupling of Fibrin Reorganization and Fibronectin Patterning by Corneal Fibroblasts in Response to PDGF BB and TGFβ1 |
| title_sort | coupling of fibrin reorganization and fibronectin patterning by corneal fibroblasts in response to pdgf bb and tgfβ1 |
| topic | fibronectin corneal fibroblasts fibrin 3-D matrices collective cell migration |
| url | https://www.mdpi.com/2306-5354/7/3/89 |
| work_keys_str_mv | AT miguelmironmendoza couplingoffibrinreorganizationandfibronectinpatterningbycornealfibroblastsinresponsetopdgfbbandtgfb1 AT daliavazquez couplingoffibrinreorganizationandfibronectinpatterningbycornealfibroblastsinresponsetopdgfbbandtgfb1 AT nereagarciaramila couplingoffibrinreorganizationandfibronectinpatterningbycornealfibroblastsinresponsetopdgfbbandtgfb1 AT hikarurikebe couplingoffibrinreorganizationandfibronectinpatterningbycornealfibroblastsinresponsetopdgfbbandtgfb1 AT wmatthewpetroll couplingoffibrinreorganizationandfibronectinpatterningbycornealfibroblastsinresponsetopdgfbbandtgfb1 |