Abstract

PurposeEffective therapy for visual loss caused by optic nerve injury or diseases has not been achieved even though the optic nerve has the regeneration potential after injury. This study was designed to modify amniotic epithelial cells (AECs) with basic fibroblast growth factor (bFGF) gene, preliminarily investigating its effect on transected optic nerve.MethodsA human bFGF gene segment was delivered into rat AECs (AECs/hbFGF) by lentiviral vector, and the gene expression was examined by RT-PCR and ELISA. The AECs/hbFGF and untransfected rat AECs were transplanted into the transected site of the rat optic nerve. At 28 days post transplantation, the survival and migration of the transplanted cells was observed by tracking labeled cells; meanwhile retinal ganglion cells (RGCs) were observed and counted by employing biotin dextran amine (BDA) and Nissl staining. Furthermore, the expression of growth associated protein 43 (GAP-43) within the injury site was examined with immunohistochemical staining.ResultsThe AECs/hbFGF was proven to express bFGF gene and secrete bFGF peptide. Both AECs/hbFGF and AECs could survive and migrate after transplantation. RGCs counting implicated that RGCs numbers of the cell transplantation groups were significantly higher than that of the control group, and the AECs/hbFGF group was significantly higher than that of the AECs group. Moreover GAP-43 integral optical density value in the control group was significantly lower than that of the cell transplantation groups, and the value in the AECs/hbFGF group was significantly higher than that of the AECs group.ConclusionsAECs modified with bFGF could reduce RGCs loss and promote expression of GAP-43 in the rat optic nerve transected model, facilitating the process of neural restoration following injury.

Highlights

  • The optic nerve derives from the embryonic optic stalks in the optic vesicles located in the diencephalon; the microenvironment within the optic nerve is the same as within the central nervous system (CNS), which contains astrocytes, oligodendrocytes, microglias

  • retinal ganglion cells (RGCs) counting implicated that RGCs numbers of the cell transplantation groups were significantly higher than that of the control group, and the amniotic epithelial cells (AECs)/human bFGF (hbFGF) group was significantly higher than that of the AECs group

  • AECs modified with Basic fibroblast growth factor (bFGF) could reduce RGCs loss and promote expression of growth associated protein 43 (GAP-43) in the rat optic nerve transected model, facilitating the process of neural restoration following injury

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Summary

Introduction

The optic nerve derives from the embryonic optic stalks in the optic vesicles located in the diencephalon; the microenvironment within the optic nerve is the same as within the central nervous system (CNS), which contains astrocytes, oligodendrocytes, microglias. Axons of the RGCs form optic nerve, once injury happens the axons will ordinarily fail to regrow through the lesion site while the RGCs will die gradually, leading to permanent vision loss [1]. The failure of the optic nerve to regenerate after injury or in neurodegenerative diseases remains a major clinical and scientific problem. The injured RGCs axons were proven to be able to regenerate in certain extent, which made it possible for improving the regenerative potential of the optic nerve [2, 3]. Several animal studies have indicated that the application of bFGF could possiblely promote the survival of RGCs, and stimulates regrowth of the axons within the injured optic nerve [5, 6]

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