Transplantation of autologous bone marrow mesenchymal stem cells for the treatment of corneal endothelium damages in rabbits

Abstract

To observe the differentiation and function of autologous bone marrow mesenchymal stem cells (MSCs) after been transplanted as a substitute of corneal endothelial cells (CECs) in vivo. Autologous MSCs in the first passage were grown in vitro on glutaraldehyde-fixed gelatin membranes and then the membranes were glued by alpha-cyanoacrylate alkyl to 7.00 mm autologous rabbits' corneal buttons whose endothelium had been mechanically removed previously. Then the buttons were sutured in place. Transplanted cultured CECs or empty gelatin membranes without any cells were used as the controls. The rabbits were observed by slit microscope and confocal microscopy at different intervals postoperatively. IOP and corneal thickness were measured by Perkin's tonometer and ultrasonic pachymeter, respectively. Twenty-one rabbits' right eyes were transplanted with MSCs, other 21 rabbits' right eyes were transplanted with CECs and another 17 rabbits' right eyes were transplanted with no-cells carriers. Two weeks after the operation, all corneal buttons were edema and began to be opaque. The corneal button began to clear and thin 8 weeks after the operation in buttons with transplantation of MSCs and 4 weeks in buttons with transplantation of CECs. The cells density of grafts decreased with time, in week 12 after the operation, the average cell density of MSCs and CECs transplantation buttons was (2105 +/- 677) cells/mm(2) and (2023 +/- 330) cells/mm(2), respectively. Scanning electronmicroscopy showed that the transplanted MSCs were differentiated into single layer irregular-shaped cells with similarity to polygonal-shaped cells. These cells had abundant microvilli but wide intercellular space. Autologous MSCs transplanted into the CECs defected area could differentiate to CECs-like cells and possess some functions of CECs.