Bioinspired hydrogel surfaces to augment corneal endothelial cell monolayer formation

Area of Science:

• Biomaterials Science
• Ophthalmology
• Cell Biology

Background:

• Corneal endothelial cells (CECs) have limited regenerative capacity, leading to vision loss.
• Corneal transplantation faces donor tissue shortages.
• Developing in vitro CECs with sufficient density is crucial for regenerative therapies.

Purpose of the Study:

• To design and fabricate polyacrylamide (PA) substrates mimicking the native corneal endothelium microenvironment.
• To investigate the effect of microfabricated topography on CEC behavior.
• To enhance in vitro development of corneal endothelial cell layers.

Main Methods:

• Preparation of PA hydrogel substrates with two distinct microfabricated topographies (small and large patterns).
• Mimicking native Descemet's membrane (DM) properties with a ~50 kPa elastic modulus and Collagen IV modification.
• Culturing bovine CECs on flat, small-patterned, and large-patterned substrates.
• Analyzing cell proliferation and monolayer density.

Main Results:

• Small patterned substrates (high hexagon density) significantly increased CEC proliferation compared to large patterned substrates (p=0.0004).
• Small patterned substrates promoted denser cell monolayers than flat or large patterned substrates (p=0.001 and p<0.0001, respectively).
• Bioinspired topography augments the formation of high-density CE monolayers.

Conclusions:

• Microfabricated, bioinspired surface topographies are effective in promoting CEC proliferation and density.
• These substrates offer a promising approach for the in vitro development of corneal endothelial cell layers.
• This technology could help overcome donor shortage limitations in corneal transplantation.