Abstract
Purpose :
To explore the feasibility of 3D bioprinting induced bone marrow mesenchymal stem cells (MSCs) transdifferentiating into corneal epithelial cells in a limbal stem cell deficiency (LSCD) model in rats.
Methods :
Rat MSCs were isolated and purified using a gradient isolation procedure. The cells were induced by a 3D bioprinting system with rat corneal stromal cells (CSCs) to mimicking the native cell composition. The induced MSCs were identified by immunofluorescence staining, flow cytometry, and scanning electron microscopy (SEM). A corneal LSCD model was produced in rat eyes by alkali injury. The eyes without any transplant served as controls. Amniotic membranes (AM), uninduced MSCs, or MSCs induced by 3D bioprinting system, were transplanted onto the cornea of the model. The therapeutic effects were evaluated by slit lamp observation, hematoxylin and eosin staining, immunohistochemistry staining, and confocal laser corneal microscopy.
Results :
Cultivated MSCs were positive for CD29, CD44, and CD90, but negative for CD34, CD45, CD133, and CK12, with typical MSCs characteristics revealed by SEM. After induced in 3D bioprinting system, the MSCs expressed positive staining for CK12 with corneal epithelial cell characteristics confirmed by SEM; the induced MSCs were unchanged on the amnion. Compared with the other three groups, the corneal opacity, fluorescence staining, and neovascularization grades were significantly decreased in the induced MSCs group, both on postoperative week four and ten.
Conclusions :
The induced MSCs can trans differentiate into corneal epithelial cells in vitro. The induced MSCs by 3D bioprinting system have remarkable effects on the treatment of corneal alkali burn and the reconstruction of the corneal surface of rats.
This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.