Reconstruction of Tissue-Engineered Conjunctiva Using Human Amniotic Epithelial Cells and Decellularized Porcine...

Area of Science:

• Ophthalmology
• Tissue Engineering
• Biomaterials Science

Background:

• Extensive conjunctival defects pose challenges due to limited treatment options like donor scarcity and immune rejection.
• Current therapies for conjunctival defects often result in suboptimal graft outcomes.

Purpose of the Study:

• To develop a functional tissue-engineered conjunctiva for ocular surface reconstruction.
• To evaluate the efficacy of integrating decellularized porcine conjunctival matrix (DPCM), human amniotic epithelial cells (hAECs), and a perfusion culture system.

Main Methods:

• Prepared DPCM using a phospholipase A2 decellularization method, achieving 93.7% DNA removal while preserving extracellular matrix.
• Fabricated tissue-engineered constructs by seeding hAECs in RADA16-I peptide hydrogel onto DPCM scaffolds.
• Utilized a perfusion culture system to maintain cell viability and promote cell growth and differentiation.

Main Results:

• Perfusion culture significantly enhanced hAEC density, distribution uniformity, and proliferation (Ki67+) compared to static culture.
• hAECs maintained stemness marker ABCG2 expression under perfusion.
• Constructs exhibited upregulation of conjunctival epithelial markers (CK4, CK13, Muc5AC), indicating successful differentiation into a conjunctival epithelial phenotype.

Conclusions:

• The integrated approach successfully generated a functional tissue-engineered conjunctiva.
• The developed construct promotes hAEC proliferation, stemness maintenance, and differentiation.
• This tissue-engineered conjunctiva presents a promising alternative for treating ocular surface defects.