Abstract
Purpose: :
To reconstruct a tissue-engineered cornea in vitro with rabbit bone marrow-derived mesenchymal stem cells (MSCs) and xenogenic decullarized cornea scaffold (XDCS), and explore the possibility of being a cornea substitute in rabbit lamellar transplantation.
Methods: :
Animal protocols were in accordance with ARVO guidelines and approved by the institutional animal care and use committee. The XDCS were generated from dog corneas. The MSCs were isolated from adult rabbit bone marrow and cultured for at least 3 passages to use. The MSCs were evaluated by flow-cytometry and immuno-fluorescence methods with a spectrum of markers as shown in
Results: :
The rabbits (n=18) were divided into 4 groups by different cornea grafts in lamellar transplantation, 1) XDCS group: xenogenic decellularized corneal scaffolds (n=5); 2) MSCs groups: the tissue engineered cornea made up with XDCS and MSCs (n=5); 3) the fresh dog cornea group (n=4); and 4) the rabbit allografts group (n=4). The clinical changes of the cornea were examined by a slit lamp microscope and a Heidelberg Retina Tomograph II Rostock Cornea Module in vivo. The corneal transparency, inflammation and neovascularization level were evaluated. The cornea sections were labeled immunohistologically by anti-K3, Vimentin, and CD29 antiserum at 6-month- postoperation, respectively.Results: The MSCs from rabbit bone marrow had been identified as positive for CD90, CD29, and CD44, and negative for CD34, CD38, CD71, CD45, CD117, CD14, CD86, and CD80. The MSCs can be induced to differentiate into lipid tissue by a routine protocol and into K3-positive cornea epithelial cells on the XDCS for 14 days. After a rabbit lamellar transplantation, no neovascularization was observed on the cornea graft in MSCs group 6 months after the transplantation while the new vessels grew on the cornea graft in XDSC group only 1 month post-operation. Confocal microscopy showed the regeneration of cornea nerve fiber into the decellularized cornea graft in XDCS groups, but not in the MSCs group.
Keywords: transplantation • cornea: basic science • neovascularization