Positive effects of bFGF modified rat amniotic epithelial cells transplantation on transected rat optic nerve.
OBJECTIVE
Effective 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.


METHODS
A 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.


RESULTS
The 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.


CONCLUSIONS
AECs 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.
Department of Anatomy, The Second Military Medical University, Shanghai, P. R. China; Department of Orthopaedics, Changzheng Hospital, The Second Military Medical University, Shanghai, P. R. China; People's Liberation Army Clinical Center for Spinal Cord Injury, Kunming General Hospital of Chengdu Military Command, Kunming, P. R. China.
Jia-Xin
JX
Xie
People's Liberation Army Clinical Center for Spinal Cord Injury, Kunming General Hospital of Chengdu Military Command, Kunming, P. R. China.
Yu
Y
Feng
Department of Anatomy, The Second Military Medical University, Shanghai, P. R. China.
Jian-Min
JM
Yuan
Department of Neurobiology, The Second Military Medical University, Shanghai, P. R. China.
Zhen-Dong
ZD
You
Department of Anatomy, The Second Military Medical University, Shanghai, P. R. China.
Hai-Yan
HY
Lin
Department of Neurobiology, The Second Military Medical University, Shanghai, P. R. China.
Chang-Lin
CL
Lu
Department of Anatomy, The Second Military Medical University, Shanghai, P. R. China.
Jia-Jun
JJ
Xu
D000650:Amnion / Q000166:cytology; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002452:Cell Count; D002454:Cell Differentiation; D002470:Cell Survival; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism / Q000637:transplantation*; D016222:Fibroblast Growth Factor 2 / Q000235:genetics* / Q000378:metabolism; D019922:GAP-43 Protein / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D008297:Male; D009416:Nerve Regeneration / Q000502:physiology*; D009900:Optic Nerve / Q000378:metabolism* / Q000473:pathology / Q000601:surgery; D020221:Optic Nerve Injuries / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012165:Retinal Ganglion Cells / Q000166:cytology / Q000378:metabolism; D014161:Transduction, Genetic; D019076:Transgenes
D015415:Biomarkers; D019922:GAP-43 Protein; D016222:Fibroblast Growth Factor 2