Tissue Engineering Using Polylactic-Co-Glycolic Acid Biodegradable Polymers as a Scaffold With Rabbit Autologous Adipose-Derived Stromal Cell in Treatment of Rabbit Corneal Stroma Defect

Abstract

Purpose: : In this study, we determined whether a polylactic-co-glycolic acid (PGLA) scaffold bearing the autologous adipose derived stem cells insert could repair the defect of rabbit corneal stroma without compromising tissue transparency.

Methods: : Adipose derived stem cells were isolated from rabbits and expanded by tissue culture. After reaching confluence, the fourth passage of cells were harvested and transfected by adenovirus carrier with Green fluorescent protein (GFP). And then the cells were mixed with PLGA meterial to form cell-scaffold constructs. After 1 week of co-culturing, they were implanted into the rabbit corneal stroma whose anterior part were removed about 200-250um thick. Before the transplantation, the adherent rate of cell to scoffold were detected. Slit lamp and confocal corneal scanning microscope were used to investigated the cornea. Ten weeks after implantation, transmission electron microscopy and histological evaluation were performed on stromal tissue. GFP expression in transplanted cornea was monitored at the laser confocal scanning microscope to determine cellular origin in the reconstructed stroma. Insertion of PLGA scaffold alone and autologous corneal stroma transplantation served as controls.

Results: : The cells attached and proliferated on the scoffolds and maintained high viability. After stromal implantation, implants gradually became transparent without corneal neovasculization over an 10-week period. During this time stromal histology was gradually restored. GFP-labeled adipose derived stem cells were subsistent in the regions bearing inserted scaffolds, suggesting that they were derived from the implants.

Conclusions: : We conclude that PLGA is a favorable scaffold for adipose derived stem cells, allowing ex vivo proliferation, and the intrastromal implantation of PLGA scaffold implants bearing autologous rabbit adipose derived stem cell can repair the defect of corneal stromal tissue reconstruction because, over an 10-week period, the implants become progressively more transparent. The tremendous applications of such tissue produced in vitro for experimental as well as clinical purposes may be considered.

Keywords: cornea: stroma and keratocytes • wound healing • transplantation