Membranes Prepared from Recombinant RGD-Silk Fibroin as Substrates for Human Corneal Cells

Molecules

卷:26

Shuko Suzuki¹  Traian V. Chirila¹  Elham Nili²  Damien G. Harkin²  Rebecca A. Dawson²  Neil A. Richardson² 

[1] Queensland Eye Institute, South Brisbane, QLD 4101, Australia;

[2] School of Biomedical Sciences, Queensland University of Technology, Brisbane, QLD 4000, Australia;

关键词: silk fibroin;    RGD peptides;    corneal cells;    cell adhesion;    cell culture;   

DOI  :  10.3390/molecules26226810

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown