Denuded human amniotic membrane seeding bone marrow stromal cells as an effective composite matrix stimulates axonal outgrowth of rat neural cortical cells in vitro.
BACKGROUND
Previous studies have shown that axonal outgrowth in the damaged central nervous system is closely related to the local microenvironment. Transplantation of bone marrow stromal cells (BMSC) or BMSC with some biomaterials has been used to treat various central nervous system diseases with some success. In the current study, we investigated if BMSC on denuded human amniotic membrane (DhAM) as a composite matrix could stimulate axonal outgrowth or not.


METHODS
After completely removing the cells on the amniotic membrane with a tryptic and mechanical approach, we seeded BMSC on it. The MTS was applied to test the cytotoxicity of DhAM compared with PLGA and PLL. The morphology of the BMSC was observed by light, electronic and laser confocal microscopy. We also used four kinds of substance (PLL, DhAM, BMSC + PLL, BMSC + DhAM) to coculturing with the cortical neurons. Finally, the lengths of axons in each group were studied using the positive axon-specific marker NF-H.


RESULTS
The DhAM was devoid of cellular components and only its intact basement membrane was left. BMSC grew on the substrate and proliferated with a flat to fusiform morphology. In the MTS test, the results indicated that BMSC cultured in DhAM extract had a high survival rate (> 80%). Moreover, the cortical neural axons in the experimental group (BMSC + DhAM) were longer (287.37 +/- 12.72 microm) than in the other groups (P < 0.01).


CONCLUSIONS
This study demonstrates that the DhAM was a good carrier to support growth of BMSC and BMSC on DhAM was an effective composite matrix to support the outgrowth of the axons of rat cortical neurons in vitro. Future studies of the use of the composite matrix in disorders are planned.
Liang
Hong Sheng
HS
Department of Neurosurgery, The First Affiliated Hospital of Harbin Medical University, Harbin, 150001, China.
Liang
Peng
P
Xu
Ye
Y
Wu
Jia Ning
JN
Liang
Tao
T
Xu
Xiao Ping
XP
Liu
En Zhong
EZ
D000650:Amnion / Q000166:cytology / Q000378:metabolism*; D000818:Animals; D001485:Basement Membrane / Q000166:cytology / Q000378:metabolism*; D016026:Bone Marrow Transplantation / Q000379:methods*; D001925:Brain Damage, Chronic / Q000601:surgery; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D018920:Coculture Techniques; D005109:Extracellular Matrix / Q000637:transplantation*; D005260:Female; D020439:Growth Cones / Q000502:physiology*; D006801:Humans; D009416:Nerve Regeneration / Q000502:physiology; D009473:Neuronal Plasticity / Q000502:physiology; D051381:Rats; D020127:Recovery of Function / Q000502:physiology; D017154:Stromal Cells / Q000637:transplantation*; D023822:Tissue Engineering; D014182:Transplantation, Autologous