Fish Scale-Derived Scaffolds for Culturing Human Corneal Endothelial Cells
Purpose. To investigate the biocompatibility of fish scale-derived scaffolds (FSS) with primary human corneal endothelial cells (HCEnCs). Methods. HCEnCs were isolated from 30 donor corneas in a donor-matched study and plated in precoated Lab-Tek slides (n=15) and FSS (n=15). Cell morphology, prolif...
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Wiley
2018-01-01
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| Series: | Stem Cells International |
| Online Access: | http://dx.doi.org/10.1155/2018/8146834 |
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| author | Mohit Parekh Bert Van den Bogerd Nadia Zakaria Diego Ponzin Stefano Ferrari |
| author_facet | Mohit Parekh Bert Van den Bogerd Nadia Zakaria Diego Ponzin Stefano Ferrari |
| author_sort | Mohit Parekh |
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| description | Purpose. To investigate the biocompatibility of fish scale-derived scaffolds (FSS) with primary human corneal endothelial cells (HCEnCs). Methods. HCEnCs were isolated from 30 donor corneas in a donor-matched study and plated in precoated Lab-Tek slides (n=15) and FSS (n=15). Cell morphology, proliferation/migration, and glucose uptake were studied (n=30). Hoechst, ethidium homodimer, and calcein AM (HEC) staining was performed to determine viability and toxicity (n=6). The cell surface area was calculated based on calcein AM staining. HCEnCs were stained for ZO-1 (n=6) to detect tight junctions and to measure cell morphology; Ki-67 (n=6) to measure proliferating cells; and vinculin to quantify focal adhesions (n=6). The formation of de novo extracellular matrix was analyzed using histology (n=6). Results. HCEnCs attach and grow faster on Lab-Tek slides compared to the undulating topography of the FSS. At day 11, HCEnCs on Lab-Tek slide grew 100% confluent, while FSS was only 65% confluent (p=0.0883), with no significant difference in glucose uptake between the two (p=0.5181) (2.2 μg/mL in Lab-Tek versus 2.05 μg/mL in FSS). HEC staining showed no toxicity. The surface area of the cells in Lab-Tek was 409.1 μm2 compared to 452.2 μm2 on FSS, which was not significant (p=0.5325). ZO-1 showed the presence of tight junctions in both conditions; however, hexagonality was higher (74% in Lab-Tek versus 45% in FSS; p=0.0006) with significantly less polymorphic cells on Lab-Tek slides (8% in Lab-Tek versus 16% in FSS; p=0.0041). Proliferative cells were detected in both conditions (4.6% in Lab-Tek versus 4.2% in FSS; p=0.5922). Vinculin expression was marginally higher in HCEnCs cultured on Lab-Tek (234 versus 199 focal adhesions; p=0.0507). Histological analysis did not show the formation of a basement membrane. Conclusions. HCEnCs cultured on precoated FSS form a monolayer, displaying correct morphology, cytocompatibility, and absence of toxicity. FSS needs further modification in terms of structure and surface chemistry before considering it as a potential carrier for cultured HCEnCs. |
| format | Article |
| id | doaj-art-2d7323f225fe40d881edfc9d8d185efb |
| institution | Kabale University |
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| language | English |
| publishDate | 2018-01-01 |
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| spelling | doaj-art-2d7323f225fe40d881edfc9d8d185efb2025-02-03T01:03:14ZengWileyStem Cells International1687-966X1687-96782018-01-01201810.1155/2018/81468348146834Fish Scale-Derived Scaffolds for Culturing Human Corneal Endothelial CellsMohit Parekh0Bert Van den Bogerd1Nadia Zakaria2Diego Ponzin3Stefano Ferrari4International Center for Ocular Physiopathology, Fondazione Banca degli Occhi del Veneto Onlus, Venice, ItalyOphthalmology, Visual Optics and Visual Rehabilitation, Translational Neurosciences, Faculty of Medicine, University of Antwerp, Antwerp, BelgiumOphthalmology, Visual Optics and Visual Rehabilitation, Translational Neurosciences, Faculty of Medicine, University of Antwerp, Antwerp, BelgiumInternational Center for Ocular Physiopathology, Fondazione Banca degli Occhi del Veneto Onlus, Venice, ItalyInternational Center for Ocular Physiopathology, Fondazione Banca degli Occhi del Veneto Onlus, Venice, ItalyPurpose. To investigate the biocompatibility of fish scale-derived scaffolds (FSS) with primary human corneal endothelial cells (HCEnCs). Methods. HCEnCs were isolated from 30 donor corneas in a donor-matched study and plated in precoated Lab-Tek slides (n=15) and FSS (n=15). Cell morphology, proliferation/migration, and glucose uptake were studied (n=30). Hoechst, ethidium homodimer, and calcein AM (HEC) staining was performed to determine viability and toxicity (n=6). The cell surface area was calculated based on calcein AM staining. HCEnCs were stained for ZO-1 (n=6) to detect tight junctions and to measure cell morphology; Ki-67 (n=6) to measure proliferating cells; and vinculin to quantify focal adhesions (n=6). The formation of de novo extracellular matrix was analyzed using histology (n=6). Results. HCEnCs attach and grow faster on Lab-Tek slides compared to the undulating topography of the FSS. At day 11, HCEnCs on Lab-Tek slide grew 100% confluent, while FSS was only 65% confluent (p=0.0883), with no significant difference in glucose uptake between the two (p=0.5181) (2.2 μg/mL in Lab-Tek versus 2.05 μg/mL in FSS). HEC staining showed no toxicity. The surface area of the cells in Lab-Tek was 409.1 μm2 compared to 452.2 μm2 on FSS, which was not significant (p=0.5325). ZO-1 showed the presence of tight junctions in both conditions; however, hexagonality was higher (74% in Lab-Tek versus 45% in FSS; p=0.0006) with significantly less polymorphic cells on Lab-Tek slides (8% in Lab-Tek versus 16% in FSS; p=0.0041). Proliferative cells were detected in both conditions (4.6% in Lab-Tek versus 4.2% in FSS; p=0.5922). Vinculin expression was marginally higher in HCEnCs cultured on Lab-Tek (234 versus 199 focal adhesions; p=0.0507). Histological analysis did not show the formation of a basement membrane. Conclusions. HCEnCs cultured on precoated FSS form a monolayer, displaying correct morphology, cytocompatibility, and absence of toxicity. FSS needs further modification in terms of structure and surface chemistry before considering it as a potential carrier for cultured HCEnCs.http://dx.doi.org/10.1155/2018/8146834 |
| spellingShingle | Mohit Parekh Bert Van den Bogerd Nadia Zakaria Diego Ponzin Stefano Ferrari Fish Scale-Derived Scaffolds for Culturing Human Corneal Endothelial Cells Stem Cells International |
| title | Fish Scale-Derived Scaffolds for Culturing Human Corneal Endothelial Cells |
| title_full | Fish Scale-Derived Scaffolds for Culturing Human Corneal Endothelial Cells |
| title_fullStr | Fish Scale-Derived Scaffolds for Culturing Human Corneal Endothelial Cells |
| title_full_unstemmed | Fish Scale-Derived Scaffolds for Culturing Human Corneal Endothelial Cells |
| title_short | Fish Scale-Derived Scaffolds for Culturing Human Corneal Endothelial Cells |
| title_sort | fish scale derived scaffolds for culturing human corneal endothelial cells |
| url | http://dx.doi.org/10.1155/2018/8146834 |
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