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Treatment of rat spinal cord injury with autologous activated Schwann cells via different grafting routes

Dexiang Ban, Shiqing Feng, Guangzhi Ning

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Abstract

Objective To explore the curative effect of autologous activated Schwann cells (AASCs) in rat spinal cord injury (SCI) via different grafting routes. Methods Saphenous nerve of the adult Wistar rat was ligated for one week to activate Schwann cells. And then AASCs were cultured, passaged, purified and identified in vitro. Sixty Wistar rats were all made into SCI model at T10 and then divided into three groups randomly. One week after injury, AASCs were transplanted into the SCI rats via three different routes [group Ⅰ: tail vein; group Ⅱ: intrathecal (subarachnoid route); group Ⅲ: intraspinal cord]. BBB score was carried out to evaluate the functional recovery of the injured rats at weekly interval postinjury. Three months later, corticospinal tract (CST) was labeled by 10% BDA. Two weeks after labeling, the rats were sacrificed and then their injured spinal cord tissues were take out to carry out 10 μm fast frozen section, which was then followed by Cy3 fluorescent probe staining, NF200 and HE staining. Results AASCs passed over 4 stable passages and expressed S-100 antigen in vitro. The difference among groups could be considered sig-nificant in BBB score on the 5th week postinjury. HE staining showed that the injured cavity was evidently smaller in group Ⅲ compared with other two groups. Statistical difference also existed among groups by com-paring the percentage of positive response areas in NF200 staining. 10%BDA stain demonstrated that more regenerated axons passed through the injured cavity in group Ⅲ when compared with other two groups, which was also confirmed by immunohistochemistry stain. Conclusion The amount of grafted AASCs can be guaranteed effectively in injured epicenter via the grafting route of intraspinal cord. AASCs promote the repair after SCI by secreting multiple kinds of neurotrophic factors and bridge injured axons for regrowth. Key words: Schwann cells; Spinal cord injuries; Nerve growth factors; Axon

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Objective To explore the curative effect of autologous activated Schwann cells (AASCs) in rat spinal cord injury (SCI) via different grafting routes. Methods Saphenous nerve of the adult Wistar rat was ligated for one week to activate Schwann cells. And then AASCs were cultured, passaged, purified and identified in vitro. Sixty Wistar rats were all made into SCI model at T10 and then divided into three groups randomly. One week after injury, AASCs were transplanted into the SCI rats via three different routes [group Ⅰ: tail vein; group Ⅱ: intrathecal (subarachnoid route); group Ⅲ: intraspinal cord]. BBB score was carried out to evaluate the functional recovery of the injured rats at weekly interval postinjury. Three months later, corticospinal tract (CST) was labeled by 10% BDA. Two weeks after labeling, the rats were sacrificed and then their injured spinal cord tissues were take out to carry out 10 μm fast frozen section, which was then followed by Cy3 fluorescent probe staining, NF200 and HE staining. Results AASCs passed over 4 stable passages and expressed S-100 antigen in vitro. The difference among groups could be considered sig-nificant in BBB score on the 5th week postinjury. HE staining showed that the injured cavity was evidently smaller in group Ⅲ compared with other two groups. Statistical difference also existed among groups by com-paring the percentage of positive response areas in NF200 staining. 10%BDA stain demonstrated that more regenerated axons passed through the injured cavity in group Ⅲ when compared with other two groups, which was also confirmed by immunohistochemistry stain. Conclusion The amount of grafted AASCs can be guaranteed effectively in injured epicenter via the grafting route of intraspinal cord. AASCs promote the repair after SCI by secreting multiple kinds of neurotrophic factors and bridge injured axons for regrowth. Key words: Schwann cells; Spinal cord injuries; Nerve growth factors; Axon

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Available abstract

Objective To explore the curative effect of autologous activated Schwann cells (AASCs) in rat spinal cord injury (SCI) via different grafting routes. Methods Saphenous nerve of the adult Wistar rat was ligated for one week to activate Schwann cells. And then AASCs were cultured, passaged, purified and identified in vitro. Sixty Wistar rats were all made into SCI model at T10 and then divided into three groups randomly. One week after injury, AASCs were transplanted into the SCI rats via three different routes [group Ⅰ: tail vein; group Ⅱ: intrathecal (subarachnoid route); group Ⅲ: intraspinal cord]. BBB score was carried out to evaluate the functional recovery of the injured rats at weekly interval postinjury. Three months later, corticospinal tract (CST) was labeled by 10% BDA. Two weeks after labeling, the rats were sacrificed and then their injured spinal cord tissues were take out to carry out 10 μm fast frozen section, which was then followed by Cy3 fluorescent probe staining, NF200 and HE staining. Results AASCs passed over 4 stable passages and expressed S-100 antigen in vitro. The difference among groups could be considered sig-nificant in BBB score on the 5th week postinjury. HE staining showed that the injured cavity was evidently smaller in group Ⅲ compared with other two groups. Statistical difference also existed among groups by com-paring the percentage of positive response areas in NF200 staining. 10%BDA stain demonstrated that more regenerated axons passed through the injured cavity in group Ⅲ when compared with other two groups, which was also confirmed by immunohistochemistry stain. Conclusion The amount of grafted AASCs can be guaranteed effectively in injured epicenter via the grafting route of intraspinal cord. AASCs promote the repair after SCI by secreting multiple kinds of neurotrophic factors and bridge injured axons for regrowth. Key words: Schwann cells; Spinal cord injuries; Nerve growth factors; Axon

Key concepts: Medicine, Spinal cord, Spinal cord injury, Staining, Immunohistochemistry, H&E stain, Stain, Pathology

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