Membranes Prepared from Recombinant RGD-Silk Fibroin as Substrates for Human Corneal Cells
| Autor: | Traian V. Chirila, Neil A. Richardson, Damien G. Harkin, Elham Nili, Rebecca A. Dawson, Shuko Suzuki |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Stromal cell
Corneal Stroma Pharmaceutical Science Fibroin Organic chemistry Limbus Corneae Horseradish peroxidase Permeability Article Analytical Chemistry Cornea QD241-441 Bombyx mori Drug Discovery Fluorescence microscope Animals Humans Physical and Theoretical Chemistry Cell adhesion Cells Cultured RGD peptides Cell Proliferation cell culture Microscopy Confocal Tissue Engineering Tissue Scaffolds biology Chemistry fungi Epithelium Corneal Membranes Artificial cell adhesion Bombyx biology.organism_classification Coculture Techniques Recombinant Proteins Biomechanical Phenomena Membrane Chemistry (miscellaneous) Cell culture silk fibroin biology.protein Biophysics corneal cells Molecular Medicine Fibroins Oligopeptides |
| Zdroj: | Molecules Volume 26 Issue 22 Molecules, Vol 26, Iss 6810, p 6810 (2021) |
| ISSN: | 1420-3049 |
| DOI: | 10.3390/molecules26226810 |
| Popis: | A recombinant formulation of silk fibroin containing the arginine–glycine–aspartic acid (RGD) cell-binding motif (RGD-fibroin) offers potential advantages for the cultivation of corneal cells. Thus, we investigated the growth of corneal stromal cells and epithelial cells on surfaces created from RGD-fibroin, in comparison to the naturally occurring Bombyx mori silk fibroin. The attachment of cells was compared in the presence or absence of serum over a 90 min period and analyzed by quantification of dsDNA content. Stratification of epithelial cells on freestanding membranes was examined by confocal fluorescence microscopy and optimized through use of low molecular weight poly(ethylene glycol) (PEG 300 Da) as a porogen, the enzyme horseradish peroxidase (HRP) as a crosslinking agent, and stromal cells grown on the opposing membrane surface. The RGD-fibroin reduced the tendency of stromal cell cultures to form clumps and encouraged the stratification of epithelial cells. PEG used in conjunction with HRP supported the fabrication of more permeable freestanding RGD-fibroin membranes, that provide an effective scaffold for stromal–epithelial co-cultures. Our studies encourage the use of RGD-fibroin for corneal cell culture. Further studies are required to confirm if the benefits of this formulation are due to changes in the expression of integrins, components of the extracellular matrix, or other events at the transcriptional level. |
| Databáze: | OpenAIRE |
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