Feasibility of bacterial cellulose membrane as biological scaffold for construction of tissue engineering corneal epithelium

Abstract

Background Corneal transplantation is a primary method for the treatment of serious corneal diseases, but its application is limited because of the shortage of corneal donor. The study on tissue engineering corneal epithelium provides a new approach to corneal transplantation, and the biological scaffold materials for tissue engineering corneal epithelium is an issue of increasing concern. Bacterial cellulose membrane has been used in medical field, but its application in tissue engineering corneal epithelium deserves more researching. Objective This study was to evaluate the biocompatibility of bacterial cellulose membrane as a biological scaffold of tissue engineering corneal epithelium. Methods Corneal epithelium was isolated from 1 month-old New Zealand White rabbit. Corneal epithelial cells were cultured using explant method and identified by detecting the CK-3 expression using immunofluorescence technique. The second generation cells were inoculated on bacterial cellulose membrane and culture plate, respectively, and the growth status of the cells were examined and compared under the optical microscope. The cell activity/toxicity test was performed by LIVE/DEAD cell staining kit at the third day after inoculation to evaluate the survival rate. The ultrastructure of the cell surface was examined under the scanning electron microscope. The study was performed in accordance with the ARVO Statement. Results Rabbit corneal epithelial cells grew well 1 week after primarily cultured with a cobblestone-like appearance and positive response for CK3 antibody. The cells on the bacterial cellulose membrane presented a round shape and regular arrangement and showed the green fluorescence for LIVE/DEAD test, with the survival rate 100%. Abundant leafy protrusion, microvilli and intercellular junction were seen under the scanning electron microscope. In addition, mitosis phase of cells and many filopodia between the cells and bacterial cellulose membrane were also exhibited. Conclusions Rabbit corneal epithelial cells can grow well in bacterial cellulose membrane. Bacterial cellulose membrane has good biocompatibility, indicating that bacterial cellulose membrane can be used as new biological material for tissue engineering corneal epithelium. Key words: [Key words] Epithelium, corneal/cytology; Tissue engineering; Tissue scaffolds/microbiology; Biocompatible materials; Cells, cultured; Rabbits; Bacterial cellulose