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Netrin-1 overexpression in bone marrow mesenchymal stem cells promotes functional recovery in a rat model of peripheral nerve injury.

Ke X, Li Q, Xu L, Zhang Y, Li D, Ma J, Mao X - J Biomed Res (2015)

Bottom Line: The results showed that BMSCs infected by Ad5-Netrin-1-EGFP significantly improved the function of the sciatic nerve, and led to increased expression of BDNF and NGF (P<0.05).Moreover, 28 days after injury, more Schwann cells were found in BMSCs infected by Ad5-Netrin-1-EGFP compared to control BMSCs.In conclusion, transplantation of BMSCs that produce netrin-1 improved the function of the sciatic nerve after injury.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, Affiliated Hospital of Jiangsu University , Zhenjiang, Jiangsu 212001 , China .

ABSTRACT
Transplantation of bone marrow mesenchymal stem cells (BMSCs) has been developed as a new method of treating diseases of the peripheral nervous system. While netrin-1 is a critical molecule for axonal path finding and nerve growth, it may also affect vascular network formation. Here, we investigated the effect of transplanting BMSCs that produce netrin-1 in a rat model of sciatic nerve crush injury. We introduced a sciatic nerve crush injury, and then injected 1×10(6) BMSCs infected by a recombinant adenovirus expressing netrin-1 Ad5-Netrin-1-EGFP or culture medium into the injured part in the next day. At day 7, 14 and 28 after injection, we measured motor nerve conduction and detected mRNA expressions of netrin-1 receptors UNC5B and Deleted in Colorectal Cancer (DCC), and neurotrophic factors brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) by real-time PCR. We also detected protein expressions of BDNF and NGF by Western blotting assays and examined BMSCs that incorporated into myelin and vascellum. The results showed that BMSCs infected by Ad5-Netrin-1-EGFP significantly improved the function of the sciatic nerve, and led to increased expression of BDNF and NGF (P<0.05). Moreover, 28 days after injury, more Schwann cells were found in BMSCs infected by Ad5-Netrin-1-EGFP compared to control BMSCs. In conclusion, transplantation of BMSCs that produce netrin-1 improved the function of the sciatic nerve after injury. This method may be a new treatment of nerve injury.

No MeSH data available.


Related in: MedlinePlus

Netrin-1 and GFP are expressed in transfected BMSCs.A: Netrin-1 protein expression in BMSCs after Ad5-EGFP-Netrin-1 successful transfection. Netrin-1 is at 67 kDa.β-actin acted as the internal control. Lane 1: Ad5-EGFP; Lane 2: Ad5-Netrin-1-EGFP (50 particles/cell); Lane 3: Ad5-Netrin-1-EGFP (100 particles/cell). B: BMSCs infected with Ad5-EGFP-Netrin-1 at 24h, EGFP is expressed in most BMSCs (Scale bars  =  50 µm).
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f01: Netrin-1 and GFP are expressed in transfected BMSCs.A: Netrin-1 protein expression in BMSCs after Ad5-EGFP-Netrin-1 successful transfection. Netrin-1 is at 67 kDa.β-actin acted as the internal control. Lane 1: Ad5-EGFP; Lane 2: Ad5-Netrin-1-EGFP (50 particles/cell); Lane 3: Ad5-Netrin-1-EGFP (100 particles/cell). B: BMSCs infected with Ad5-EGFP-Netrin-1 at 24h, EGFP is expressed in most BMSCs (Scale bars  =  50 µm).

Mentions: To determine whether Ad5-EGFP-Netrin-1 sustained netrin-1 expression, we examined netrin-1 expression in BMSCs infected with Ad5-Netrin-1-EGFP at different multiplicities of infection. After infection of BMSCs with Ad5-Netrin-1-EGFP, netrin-1 was detected by Western blotting assay. EGFP-expression of BMSCs was examined by fluorescence microscopy (Fig. 1A-B).


Netrin-1 overexpression in bone marrow mesenchymal stem cells promotes functional recovery in a rat model of peripheral nerve injury.

Ke X, Li Q, Xu L, Zhang Y, Li D, Ma J, Mao X - J Biomed Res (2015)

Netrin-1 and GFP are expressed in transfected BMSCs.A: Netrin-1 protein expression in BMSCs after Ad5-EGFP-Netrin-1 successful transfection. Netrin-1 is at 67 kDa.β-actin acted as the internal control. Lane 1: Ad5-EGFP; Lane 2: Ad5-Netrin-1-EGFP (50 particles/cell); Lane 3: Ad5-Netrin-1-EGFP (100 particles/cell). B: BMSCs infected with Ad5-EGFP-Netrin-1 at 24h, EGFP is expressed in most BMSCs (Scale bars  =  50 µm).
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Related In: Results  -  Collection

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f01: Netrin-1 and GFP are expressed in transfected BMSCs.A: Netrin-1 protein expression in BMSCs after Ad5-EGFP-Netrin-1 successful transfection. Netrin-1 is at 67 kDa.β-actin acted as the internal control. Lane 1: Ad5-EGFP; Lane 2: Ad5-Netrin-1-EGFP (50 particles/cell); Lane 3: Ad5-Netrin-1-EGFP (100 particles/cell). B: BMSCs infected with Ad5-EGFP-Netrin-1 at 24h, EGFP is expressed in most BMSCs (Scale bars  =  50 µm).
Mentions: To determine whether Ad5-EGFP-Netrin-1 sustained netrin-1 expression, we examined netrin-1 expression in BMSCs infected with Ad5-Netrin-1-EGFP at different multiplicities of infection. After infection of BMSCs with Ad5-Netrin-1-EGFP, netrin-1 was detected by Western blotting assay. EGFP-expression of BMSCs was examined by fluorescence microscopy (Fig. 1A-B).

Bottom Line: The results showed that BMSCs infected by Ad5-Netrin-1-EGFP significantly improved the function of the sciatic nerve, and led to increased expression of BDNF and NGF (P<0.05).Moreover, 28 days after injury, more Schwann cells were found in BMSCs infected by Ad5-Netrin-1-EGFP compared to control BMSCs.In conclusion, transplantation of BMSCs that produce netrin-1 improved the function of the sciatic nerve after injury.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, Affiliated Hospital of Jiangsu University , Zhenjiang, Jiangsu 212001 , China .

ABSTRACT
Transplantation of bone marrow mesenchymal stem cells (BMSCs) has been developed as a new method of treating diseases of the peripheral nervous system. While netrin-1 is a critical molecule for axonal path finding and nerve growth, it may also affect vascular network formation. Here, we investigated the effect of transplanting BMSCs that produce netrin-1 in a rat model of sciatic nerve crush injury. We introduced a sciatic nerve crush injury, and then injected 1×10(6) BMSCs infected by a recombinant adenovirus expressing netrin-1 Ad5-Netrin-1-EGFP or culture medium into the injured part in the next day. At day 7, 14 and 28 after injection, we measured motor nerve conduction and detected mRNA expressions of netrin-1 receptors UNC5B and Deleted in Colorectal Cancer (DCC), and neurotrophic factors brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) by real-time PCR. We also detected protein expressions of BDNF and NGF by Western blotting assays and examined BMSCs that incorporated into myelin and vascellum. The results showed that BMSCs infected by Ad5-Netrin-1-EGFP significantly improved the function of the sciatic nerve, and led to increased expression of BDNF and NGF (P<0.05). Moreover, 28 days after injury, more Schwann cells were found in BMSCs infected by Ad5-Netrin-1-EGFP compared to control BMSCs. In conclusion, transplantation of BMSCs that produce netrin-1 improved the function of the sciatic nerve after injury. This method may be a new treatment of nerve injury.

No MeSH data available.


Related in: MedlinePlus