Detection of Limbal Stem Cells Adhered to Melt Electrospun Silk Fibroin and Gelatin-Modified Polylactic Acid Scaffolds
Limbal stem cells (LSCs) are of paramount importance in corneal epithelial tissue repair. The cornea becomes opaque in case of limbal stem cell deficiency (LSCD), which may cause serious damage to the ocular visual function. There are many techniques to restore damaged epithelium, one of which is th...
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MDPI AG
2023-02-01
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| Series: | Polymers |
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| Online Access: | https://www.mdpi.com/2073-4360/15/3/777 |
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| author | Emilija Zdraveva Krešo Bendelja Luka Bočkor Tamara Dolenec Budimir Mijović |
| author_facet | Emilija Zdraveva Krešo Bendelja Luka Bočkor Tamara Dolenec Budimir Mijović |
| author_sort | Emilija Zdraveva |
| collection | DOAJ |
| description | Limbal stem cells (LSCs) are of paramount importance in corneal epithelial tissue repair. The cornea becomes opaque in case of limbal stem cell deficiency (LSCD), which may cause serious damage to the ocular visual function. There are many techniques to restore damaged epithelium, one of which is the transplantation of healthy cultured LSCs, usually onto a human amniotic membrane or onto bio-based engineered scaffolds in recent years. In this study, melt electrospun polylactic acid (PLA) was modified by silk fibroin or gelatin and further cultured with LSCs originating from three different donors. In terms of physicochemical properties, both modifications slightly increased PLA scaffold porosity (with a significantly larger pore area for the PLA/gelatin) and improved the scaffolds’ swelling percentage, as well as their biodegradation rate. In terms of the scaffold application function, the aim was to detect/visualize whether LSCs adhered to the scaffolds and to further determine cell viability (total number), as well as to observe p63 and CK3 expressions in the LSCs. LSCs were attached to the surface of microfibers, showing flattened conformations or 3D spheres in the formation of colonies or agglomerations, respectively. All scaffolds showed the ability to bind the cells onto the surface of individual microfibers (PLA and PLA/gelatin), or in between the microfibers (PLA/silk fibroin), with the latter showing the most intense red fluorescence of the stained cells. All scaffolds proved to be biocompatible, while the PLA/silk fibroin scaffolds showed the highest 98% viability of 2.9 × 10<sup>6</sup> LSCs, with more than 98% of p63 and less than 20% of CK3 expressions in the LSCs, thus confirming the support of their growth, proliferation and corneal epithelial differentiation. The results show the potential of these bio-engineered scaffolds to be used as an alternative clinical approach. |
| first_indexed | 2024-03-11T09:28:53Z |
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| institution | Directory Open Access Journal |
| issn | 2073-4360 |
| language | English |
| last_indexed | 2024-03-11T09:28:53Z |
| publishDate | 2023-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Polymers |
| spelling | doaj.art-dc2cfeb1334c446d904d605f73581fc22023-11-16T17:50:27ZengMDPI AGPolymers2073-43602023-02-0115377710.3390/polym15030777Detection of Limbal Stem Cells Adhered to Melt Electrospun Silk Fibroin and Gelatin-Modified Polylactic Acid ScaffoldsEmilija Zdraveva0Krešo Bendelja1Luka Bočkor2Tamara Dolenec3Budimir Mijović4Department of Fundamental Natural and Engineering Sciences, Faculty of Textile Technology, University of Zagreb, 10000 Zagreb, CroatiaCenter for Research and Knowledge Transfer in Biotechnology, University of Zagreb, 10000 Zagreb, CroatiaCenter for Applied Bioanthropology, Institute for Anthropological Research, 10000 Zagreb, CroatiaDepartment of Transfusion and Regenerative Medicine, Sestre Milosrdnice University Hospital Center, 10000 Zagreb, CroatiaDepartment of Fundamental Natural and Engineering Sciences, Faculty of Textile Technology, University of Zagreb, 10000 Zagreb, CroatiaLimbal stem cells (LSCs) are of paramount importance in corneal epithelial tissue repair. The cornea becomes opaque in case of limbal stem cell deficiency (LSCD), which may cause serious damage to the ocular visual function. There are many techniques to restore damaged epithelium, one of which is the transplantation of healthy cultured LSCs, usually onto a human amniotic membrane or onto bio-based engineered scaffolds in recent years. In this study, melt electrospun polylactic acid (PLA) was modified by silk fibroin or gelatin and further cultured with LSCs originating from three different donors. In terms of physicochemical properties, both modifications slightly increased PLA scaffold porosity (with a significantly larger pore area for the PLA/gelatin) and improved the scaffolds’ swelling percentage, as well as their biodegradation rate. In terms of the scaffold application function, the aim was to detect/visualize whether LSCs adhered to the scaffolds and to further determine cell viability (total number), as well as to observe p63 and CK3 expressions in the LSCs. LSCs were attached to the surface of microfibers, showing flattened conformations or 3D spheres in the formation of colonies or agglomerations, respectively. All scaffolds showed the ability to bind the cells onto the surface of individual microfibers (PLA and PLA/gelatin), or in between the microfibers (PLA/silk fibroin), with the latter showing the most intense red fluorescence of the stained cells. All scaffolds proved to be biocompatible, while the PLA/silk fibroin scaffolds showed the highest 98% viability of 2.9 × 10<sup>6</sup> LSCs, with more than 98% of p63 and less than 20% of CK3 expressions in the LSCs, thus confirming the support of their growth, proliferation and corneal epithelial differentiation. The results show the potential of these bio-engineered scaffolds to be used as an alternative clinical approach.https://www.mdpi.com/2073-4360/15/3/777melt electrospinningPLA/silk fibroinPLA/gelatinLSCsnumber of viable cellsimmunocytochemistry |
| spellingShingle | Emilija Zdraveva Krešo Bendelja Luka Bočkor Tamara Dolenec Budimir Mijović Detection of Limbal Stem Cells Adhered to Melt Electrospun Silk Fibroin and Gelatin-Modified Polylactic Acid Scaffolds Polymers melt electrospinning PLA/silk fibroin PLA/gelatin LSCs number of viable cells immunocytochemistry |
| title | Detection of Limbal Stem Cells Adhered to Melt Electrospun Silk Fibroin and Gelatin-Modified Polylactic Acid Scaffolds |
| title_full | Detection of Limbal Stem Cells Adhered to Melt Electrospun Silk Fibroin and Gelatin-Modified Polylactic Acid Scaffolds |
| title_fullStr | Detection of Limbal Stem Cells Adhered to Melt Electrospun Silk Fibroin and Gelatin-Modified Polylactic Acid Scaffolds |
| title_full_unstemmed | Detection of Limbal Stem Cells Adhered to Melt Electrospun Silk Fibroin and Gelatin-Modified Polylactic Acid Scaffolds |
| title_short | Detection of Limbal Stem Cells Adhered to Melt Electrospun Silk Fibroin and Gelatin-Modified Polylactic Acid Scaffolds |
| title_sort | detection of limbal stem cells adhered to melt electrospun silk fibroin and gelatin modified polylactic acid scaffolds |
| topic | melt electrospinning PLA/silk fibroin PLA/gelatin LSCs number of viable cells immunocytochemistry |
| url | https://www.mdpi.com/2073-4360/15/3/777 |
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