Abstract
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.
Highlights
Corneal trauma and ulceration, bacterial and viral infections, and heritable conditions are major contributors to corneal blindness, which affects over ten million individuals worldwide[1, 2]
A more practical and less labor-intensive method is the use of plastic compressed (PC) gels[26]; this technique was developed by Brown et al and central to this process is to eliminate the majority of the water content of the gel
Many polymers can be produced into nanofibers via electrospinning[29,30,31], and some electrospun constructs, such as those made of poly (PLA), poly (ε-caprolactone) (PCL), and PLGA, have excellent mechanical properties that facilitate the simulation of natural tissue constructs, as well as clinical manipulation[32,33,34]
Summary
Bacterial and viral infections, and heritable conditions are major contributors to corneal blindness, which affects over ten million individuals worldwide[1, 2]. The mechanical properties of collagen gels are improved by compression, it still can’t meet the requirement of natural cornea and is too weak to be sutured or conveniently handled for clinical applications Another novel method proposed by Connon et al is the use of templates to instruct corneal stromal cells to create native extracellular collagen matrix, which had good structural, optical, mechanical, and biological properties, as a corneal tissue equivalent. The technique of electrospinning has attracted interests in fabricating biomimetic engineering functional corneal tissue due to the close structural resemblance of the constructs to native extra cellular matrix and its high surface area–to-volume ratio and good porosity, which provide support for cell adhesion and movement, proliferation and differentiation, as well as excellent mechanical properties, easy manipulation of fiber properties, great material handling, suturability for implantation, and scalable production. We studied the activities of corneal cells (HCECs and HKs) on the designed hybrid construct and its potential for corneal tissue reconstruction
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