Human stem cell based corneal tissue mimicking structures using laser-assisted 3D bioprinting and functional bioinks

Area of Science:

• Regenerative Medicine
• Biomaterials Science
• Ophthalmology

Background:

• High demand for native-like 3D corneal structures for transplantation.
• Limitations of current methods in replicating corneal complexity.
• Need for advanced tissue engineering techniques.

Purpose of the Study:

• To develop and optimize 3D bioprinting methods for corneal tissue fabrication.
• To utilize human stem cells for creating cornea-mimicking structures.
• To evaluate the viability, proliferation, and protein expression of bioprinted corneal tissues.

Main Methods:

• Laser-assisted bioprinting (LaBP) using human embryonic stem cell-derived limbal epithelial stem cells (hESC-LESCs) and human adipose tissue-derived stem cells (hASCs).
• Development of functional bioinks based on recombinant human laminin and collagen I.
• Bioprinting of stratified corneal epithelium, lamellar corneal stroma, and composite structures.
• Evaluation of microstructure, cell viability, proliferation, and protein markers (Ki67, p63α, p40, CK3, CK15, collagen I, VWF).
• In vitro implantation of bioprinted stromal constructs into porcine corneal organ cultures.

Main Results:

• Successful bioprinting of stratified epithelium and layered stroma using hESC-LESCs and hASCs.
• Maintained high cell viability and proliferation post-printing.
• Demonstrated expression of key corneal markers (CK3, p63α, p40) in printed epithelium.
• hASCs in stroma organized horizontally and expressed collagen I.
• Bioprinted stroma integrated with porcine corneal tissue, showing cell migration.

Conclusions:

• 3D laser-assisted bioprinting is feasible for creating human stem cell-based corneal tissues.
• The fabricated tissues mimic key structural and cellular features of native cornea.
• This approach holds significant potential for corneal regenerative medicine and transplantation.