Corneal epithelial cell growth over tethered-protein/peptide surface-modified hydrogels.
In this study, we investigated the corneal epithelial cell growth rate and adhesion to novel hydrogels with (1) extracellular matrix proteins [fibronectin, laminin, substance P, and insulin-like growth factor-1 (IGF-1)] and (2) peptide sequences [RGD and fibronectin adhesion-promoting peptide (FAP)] tethered to their surface on poly(ethylene glycol) (PEG) chains. The growth rate to confluence of primary rabbit cornea epithelial cells was compared for plain polymethacrylic acid-co-hydroxyethyl methacrylate (PHEMA/MAA) hydrogels, PHEMA/MAA hydrogels coated with extracellular matrix proteins or peptides, and PHEMA/MAA hydrogels with tethered extracellular matrix proteins or peptides on the surface. The development of focal adhesions by the epithelial cells grown on the surfaces was determined by F-actin staining. Little to no epithelial cell growth occurred on the plain hydrogel surfaces throughout the 15-day culture period. Of the coated hydrogels, only the fibronectin-coated surfaces showed a significant increase in cell growth compared to plain hydrogels (p < 0.009). However, even these surfaces reached a maximum of only 20% confluence. Laminin, fibronectin adhesion-promoting peptide (FAP), and fibronectin/laminin (1:1) tether-modified hydrogels all achieved 100% confluence by the end of the culture period, although the rates at which confluence was reached differed. F-actin staining showed that focal adhesions were formed for the laminin, FAP, and fibronectin/laminin tether-modified surfaces. The results support the hypothesis that tethering certain extracellular matrix proteins and/or peptides to the hydrogel surface enhances epithelial cell growth and adhesion, compared with that seen for protein-coated or plain hydrogel surfaces.
LSU Eye Center, LSU Health Sciences Center, 2020 Gravier Street, Suite B, New Orleans, Louisiana 70112, USA. jjacob@lsuhsc.edu
Jean T
JT
Jacob
James R
JR
Rochefort
Jingjing
J
Bi
Bryan M
BM
Gebhardt
D000818:Animals; D002448:Cell Adhesion; D002455:Cell Division; D002478:Cells, Cultured; D003315:Cornea / Q000166:cytology*; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism; D005109:Extracellular Matrix / Q000737:chemistry*; D022001:Focal Adhesions; D020100:Hydrogels / Q000737:chemistry*; D007633:Keratins / Q000378:metabolism; D010455:Peptides / Q000737:chemistry*; D011817:Rabbits
D020100:Hydrogels; D010455:Peptides; D007633:Keratins