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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

Transplanted BMSCs are integrated into the vasculature or become Schwann cells.A: Determination of neurofilament. The nerve tissues were retrieved 14 days after injury and were subjected to immunohistochemistry with antibody against neurofilament in the four treatment groups. The NF-positive axons (red) are ensheathed by GFP-positive BMSCs (green), forming myelinated nerve fibers. Nuclear marker in blue. Scale bars  =  50 µm. B: The quantitative analysis of relative density of neurofilament, *P<0.05. C: Arteriogenesis induced by GFP-positive transplanted BMSCs at 14 days post implantation. Red fluorescence signified CD31, whereas green for GFP, blue for DAPI and yellow for merged. Bars  =  20 mm.
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f06: Transplanted BMSCs are integrated into the vasculature or become Schwann cells.A: Determination of neurofilament. The nerve tissues were retrieved 14 days after injury and were subjected to immunohistochemistry with antibody against neurofilament in the four treatment groups. The NF-positive axons (red) are ensheathed by GFP-positive BMSCs (green), forming myelinated nerve fibers. Nuclear marker in blue. Scale bars  =  50 µm. B: The quantitative analysis of relative density of neurofilament, *P<0.05. C: Arteriogenesis induced by GFP-positive transplanted BMSCs at 14 days post implantation. Red fluorescence signified CD31, whereas green for GFP, blue for DAPI and yellow for merged. Bars  =  20 mm.

Mentions: EGFP-labeled BMSCs transplanted in the injured nerve were observed using fluorescent microscopy. BMSCs were detected by their expression of GFP on day 7, 14 and 28, which declined over time. We found that some implanted BMSCs were located in the nerve during recovery and expressed the Schwann cell marker S100 beta. At day 14 or 28, treatment with either BMSCs alone (335.6±34.08 relative density/mm2) or BMSCs/Netrin-1 (472.6±23.60 relative density/mm2) significantly enhanced the expression of Schwann cells as compared to non-treatment (P<0.05). Furthermore, treatment with BMSCs/Netrin-1 showed a higher expression than BMSCs alone (P<0.05). At day 7, treatment with either BMSCs alone (114.8±13.77 relative density/mm2) or BMSCs/Netrin-1 (122.8±15.50 relative density/mm2) was higher than the control group (P<0.05), but there was no difference between them (P>0.05). The number of Schwann cells induced by BMSCs in the BMSC/Netrin-1 group was higher than that in the BMSC group (Fig. 6A-B). The expressions of Schwann cells in the BMSCs and BMSC/Netrin-1 group showed significant difference among day 7, 14 and 28 (all P<0.05). We also observed some double stained cells in the surface of the injured nerve at day 14, suggesting that some transplanted BMSCs were involved in vessel formation (Fig. 6C).


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)

Transplanted BMSCs are integrated into the vasculature or become Schwann cells.A: Determination of neurofilament. The nerve tissues were retrieved 14 days after injury and were subjected to immunohistochemistry with antibody against neurofilament in the four treatment groups. The NF-positive axons (red) are ensheathed by GFP-positive BMSCs (green), forming myelinated nerve fibers. Nuclear marker in blue. Scale bars  =  50 µm. B: The quantitative analysis of relative density of neurofilament, *P<0.05. C: Arteriogenesis induced by GFP-positive transplanted BMSCs at 14 days post implantation. Red fluorescence signified CD31, whereas green for GFP, blue for DAPI and yellow for merged. Bars  =  20 mm.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4585432&req=5

f06: Transplanted BMSCs are integrated into the vasculature or become Schwann cells.A: Determination of neurofilament. The nerve tissues were retrieved 14 days after injury and were subjected to immunohistochemistry with antibody against neurofilament in the four treatment groups. The NF-positive axons (red) are ensheathed by GFP-positive BMSCs (green), forming myelinated nerve fibers. Nuclear marker in blue. Scale bars  =  50 µm. B: The quantitative analysis of relative density of neurofilament, *P<0.05. C: Arteriogenesis induced by GFP-positive transplanted BMSCs at 14 days post implantation. Red fluorescence signified CD31, whereas green for GFP, blue for DAPI and yellow for merged. Bars  =  20 mm.
Mentions: EGFP-labeled BMSCs transplanted in the injured nerve were observed using fluorescent microscopy. BMSCs were detected by their expression of GFP on day 7, 14 and 28, which declined over time. We found that some implanted BMSCs were located in the nerve during recovery and expressed the Schwann cell marker S100 beta. At day 14 or 28, treatment with either BMSCs alone (335.6±34.08 relative density/mm2) or BMSCs/Netrin-1 (472.6±23.60 relative density/mm2) significantly enhanced the expression of Schwann cells as compared to non-treatment (P<0.05). Furthermore, treatment with BMSCs/Netrin-1 showed a higher expression than BMSCs alone (P<0.05). At day 7, treatment with either BMSCs alone (114.8±13.77 relative density/mm2) or BMSCs/Netrin-1 (122.8±15.50 relative density/mm2) was higher than the control group (P<0.05), but there was no difference between them (P>0.05). The number of Schwann cells induced by BMSCs in the BMSC/Netrin-1 group was higher than that in the BMSC group (Fig. 6A-B). The expressions of Schwann cells in the BMSCs and BMSC/Netrin-1 group showed significant difference among day 7, 14 and 28 (all P<0.05). We also observed some double stained cells in the surface of the injured nerve at day 14, suggesting that some transplanted BMSCs were involved in vessel formation (Fig. 6C).

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