Tissue-engineered cornea constructed with compressed collagen and laser-perforated electrospun mat

While Plastic Compressed (PC) collagen technique is often used to fabricate bioengineered constructs, PC collagen gels are too weak to be sutured or conveniently handled for clinical applications. To overcome this limitation, electrospun poly (lactic-co-glycolide) (PLGA) mats, which have excellent biocompatibility and mechanical properties, were combined with PC collagen to fabricate sandwich-like hybrid constructs. By laser-perforating holes with different sizes and spacings in the electrospun mats to regulate the mechanical properties and light transmittance of the hybrid constructs, we produced hybrid constructs with properties very suitable to apply in corneal tissue engineering. The maximum tensile stress of the optimal hybrid construct was 3.42 +/- 0.22 MPa. The light transmittance of the hybrid construct after perforation was approximately 15-fold higher than before, and light transmittance increased gradually with increasing time. After immersing into PBS for 7 days, the transmittance of the optimal construct changed from 63 +/- 2.17% to 72 +/- 1.8% under 500 nm wavelength. The live/dead staining, cell proliferation assay and immunohistochemistry study of human corneal epithelial cells (HCECs) and human keratocytes (HKs) cultured on the optimal hybrid construct both demonstrated that the cells adhered, proliferated, and maintained their phenotype well on the material. In addition, after culturing for 2 weeks, the HCECs could form stratified layers. Thus, our designed construct is suitable for the construction of engineered corneal tissue.