Abstract
Purpose :
To evaluate the feasibility of silk fibroin (SF) / poly(L-lactic acid-co-ε-caprolactone) (PLCL) scaffolds for reconstructing conjunctival epithelium.
Methods :
Characteristics and mechanical properties of SF/PLCL scaffolds, such as surface microstructure, surface wettability and tensile strength were analyzed. Rabbit conjunctival epithelial cells (rCjECs) were harvested from the palpebral conjunctiva containing fornix of New Zealand white rabbits. To assess the cell–scaffold interaction, CjECs were cultured on the electrospun scaffolds. Cell morphology, phenotype, and inflammatory reaction were observed using scanning electron microscopy (SEM) and the quantitative polymerase chain reaction (qPCR). CCK-8 kit, BrdU staining and live/dead staining was used to observe cell proliferation and viability. Epithelial cell stratification was observed using hematoxylin–eosin (H&E) staining. After culture for 3 days in vitro, the cell-seeded scaffolds were implanted subcutaneously into nude mice for 1, 2 and 4 weeks, followed by evaluation of the biocompatibility of the scaffold.
Results :
Scanning electron micrographs (SEM) showed that SF/PLCL scaffolds were composed of defect-free nanofibers with smooth and homogeneous fiber morphology. Water contact angle measurement demonstrated that the scaffolds were hydrophilic. Scanning electron micrographs and in-vitro proliferation assays showed that the cells adhered and proliferated well on the scaffolds. The results of qPCR showed excellent expression of conjunctival epithelial cells gene, with reduced expression of inflammatory mediators. H&E staining showed that the engineered conjunctiva constructed by SF/PLCL scaffolds was consist of 2–4 layers epithelium. Furthermore, SF/PLCL scaffolds transplanted subcutaneously exhibited excellent biocompatibility.
Conclusions :
SF/PLCL scaffolds facilitate the formation of conjunctiva epithelium and thus may represent a promising scaffold repairing damaged or diseased conjunctival tissue.
This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.