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

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...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Elham Nili, Damien G. Harkin, Rebecca A. Dawson, Neil A. Richardson, Shuko Suzuki, Traian V. Chirila
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	silk fibroin RGD peptides corneal cells cell adhesion cell culture Organic chemistry QD241-441
Acceso en línea:	https://doaj.org/article/3dc00f7b878540eb8831d5ada83229c6
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:3dc00f7b878540eb8831d5ada83229c6
record_format	dspace
spelling	oai:doaj.org-article:3dc00f7b878540eb8831d5ada83229c62021-11-25T18:27:25ZMembranes Prepared from Recombinant RGD-Silk Fibroin as Substrates for Human Corneal Cells10.3390/molecules262268101420-3049https://doaj.org/article/3dc00f7b878540eb8831d5ada83229c62021-11-01T00:00:00Zhttps://www.mdpi.com/1420-3049/26/22/6810https://doaj.org/toc/1420-3049A 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 <i>Bombyx mori</i> 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.Elham NiliDamien G. HarkinRebecca A. DawsonNeil A. RichardsonShuko SuzukiTraian V. ChirilaMDPI AGarticlesilk fibroinRGD peptidescorneal cellscell adhesioncell cultureOrganic chemistryQD241-441ENMolecules, Vol 26, Iss 6810, p 6810 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	silk fibroin RGD peptides corneal cells cell adhesion cell culture Organic chemistry QD241-441
spellingShingle	silk fibroin RGD peptides corneal cells cell adhesion cell culture Organic chemistry QD241-441 Elham Nili Damien G. Harkin Rebecca A. Dawson Neil A. Richardson Shuko Suzuki Traian V. Chirila Membranes Prepared from Recombinant RGD-Silk Fibroin as Substrates for Human Corneal Cells
description	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 <i>Bombyx mori</i> 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.
format	article
author	Elham Nili Damien G. Harkin Rebecca A. Dawson Neil A. Richardson Shuko Suzuki Traian V. Chirila
author_facet	Elham Nili Damien G. Harkin Rebecca A. Dawson Neil A. Richardson Shuko Suzuki Traian V. Chirila
author_sort	Elham Nili
title	Membranes Prepared from Recombinant RGD-Silk Fibroin as Substrates for Human Corneal Cells
title_short	Membranes Prepared from Recombinant RGD-Silk Fibroin as Substrates for Human Corneal Cells
title_full	Membranes Prepared from Recombinant RGD-Silk Fibroin as Substrates for Human Corneal Cells
title_fullStr	Membranes Prepared from Recombinant RGD-Silk Fibroin as Substrates for Human Corneal Cells
title_full_unstemmed	Membranes Prepared from Recombinant RGD-Silk Fibroin as Substrates for Human Corneal Cells
title_sort	membranes prepared from recombinant rgd-silk fibroin as substrates for human corneal cells
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/3dc00f7b878540eb8831d5ada83229c6
work_keys_str_mv	AT elhamnili membranespreparedfromrecombinantrgdsilkfibroinassubstratesforhumancornealcells AT damiengharkin membranespreparedfromrecombinantrgdsilkfibroinassubstratesforhumancornealcells AT rebeccaadawson membranespreparedfromrecombinantrgdsilkfibroinassubstratesforhumancornealcells AT neilarichardson membranespreparedfromrecombinantrgdsilkfibroinassubstratesforhumancornealcells AT shukosuzuki membranespreparedfromrecombinantrgdsilkfibroinassubstratesforhumancornealcells AT traianvchirila membranespreparedfromrecombinantrgdsilkfibroinassubstratesforhumancornealcells
_version_	1718411127839260672

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

Ejemplares similares