Limits...
Conserved dopamine neurotrophic factor-transduced mesenchymal stem cells promote axon regeneration and functional recovery of injured sciatic nerve.

Liu Y, Nie L, Zhao H, Zhang W, Zhang YQ, Wang SS, Cheng L - PLoS ONE (2014)

Bottom Line: Light and electron microscopy confirmed successful regeneration of the sciatic nerve.The greater numbers of HRP-labeled neuron cell bodies and increased sciatic nerve index values (SFI) in the CDNF-MSCs group suggest that CDNF exerts neuroprotective effects in vivo.Collectively, these findings indicate that CDNF gene therapy delivered by MSCs is capable of promoting nerve regeneration and functional recovery, likely because of the significant neuroprotective and neurotrophic effects of CDNF and the superior environment offered by MSCs and collagen tubes.

View Article: PubMed Central - PubMed

Affiliation: Department of Spine Surgery, Qilu Hospital of Shandong University, Jinan, China; Shandong University Qilu Hospital Research Center for Cell Therapy, Key Laboratory of Cardiovascular Remodeling and Function Research, Qilu Hospital of Shandong University, Jinan, China.

ABSTRACT
Peripheral nerve injury (PNI) is a common disease that often results in axonal degeneration and the loss of neurons, ultimately leading to limited nerve regeneration and severe functional impairment. Currently, there are no effective treatments for PNI. In the present study, we transduced conserved dopamine neurotrophic factor (CDNF) into mesenchymal stem cells (MSCs) in collagen tubes to investigate their regenerative effects on rat peripheral nerves in an in vivo transection model. Scanning electron microscopy of the collagen tubes demonstrated their ability to be resorbed in vivo. We observed notable overexpression of the CDNF protein in the distal sciatic nerve after application of CDNF-MSCs. Quantitative analysis of neurofilament 200 (NF200) and S100 immunohistochemistry showed significant enhancement of axonal and Schwann cell regeneration in the group receiving CDNF-MSCs (CDNF-MSCs group) compared with the control groups. Myelination thickness, axon diameter and the axon-to fiber diameter ratio (G-ratio) were significantly higher in the CDNF-MSCs group at 8 and 12 weeks after nerve transection surgery. After surgery, the sciatic functional index, target muscle weight, wet weight ratio of gastrocnemius muscle and horseradish peroxidase (HRP) tracing demonstrated functional recovery. Light and electron microscopy confirmed successful regeneration of the sciatic nerve. The greater numbers of HRP-labeled neuron cell bodies and increased sciatic nerve index values (SFI) in the CDNF-MSCs group suggest that CDNF exerts neuroprotective effects in vivo. We also observed higher target muscle weights and a significant improvement in muscle atrophism in the CDNF-MSCs group. Collectively, these findings indicate that CDNF gene therapy delivered by MSCs is capable of promoting nerve regeneration and functional recovery, likely because of the significant neuroprotective and neurotrophic effects of CDNF and the superior environment offered by MSCs and collagen tubes.

Show MeSH

Related in: MedlinePlus

Evaluation of amyotrophy.(A and B) Masson’s collagen staining of sections of the transverse gastrocnemius muscle at 8 and 12 weeks after surgery. Scale bar = 50 µm (C) Quantitative analysis of the percentages of muscle fibers in each group. (D) Wet weight analysis of the gastrocnemius muscle at 4, 8 and 12 weeks after surgery. (E) Statistical analysis of the wet weight ratio of the gastrocnemius muscle. n = 5, *p<0.05, **p<0.01.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4208796&req=5

pone-0110993-g007: Evaluation of amyotrophy.(A and B) Masson’s collagen staining of sections of the transverse gastrocnemius muscle at 8 and 12 weeks after surgery. Scale bar = 50 µm (C) Quantitative analysis of the percentages of muscle fibers in each group. (D) Wet weight analysis of the gastrocnemius muscle at 4, 8 and 12 weeks after surgery. (E) Statistical analysis of the wet weight ratio of the gastrocnemius muscle. n = 5, *p<0.05, **p<0.01.

Mentions: Masson’s collagen staining method directly showed the morphological changes in the gastrocnemius muscle at 8 and 12 weeks. The percentage of muscle fibers in the CDNF-MSCs group was significantly larger than those in the No repair, MSCs, LV-MSCs and naive groups at 8 and 12 weeks (Fig. 7C). The percentage of muscle fibers between the MSCs and LV-MSCs groups did not show obvious differences at 8 and 12 weeks after surgery (n = 5, P>0.05) (Fig. 7C). We also weighed the gastrocnemius muscles at 4, 8, and 12 weeks to assess muscle innervation recovery. At 8 and 12 weeks after surgery, the wet weights of the gastrocnemius in the CDNF-MSCs group were significantly higher than those in the No repair, LV-MSCs, MSCs, and DMEM groups, suggesting that treatment with CDNF-MSCs led to greater innervation of the gastrocnemius muscle (Fig. 7D). The wet weight ratio of gastrocnemius muscle in the CDNF-MSCs group was higher than superior to those in the No repair, LV-MSCs, MSCs, and DMEM groups (<0.05), but the difference between the LV-MSCs and MSCs groups was not statistically significant (>0.05). The wet weight of gastrocnemius muscle, the percentage of muscle fibers and the wet weight ratio of gastrocnemius muscle of all repair groups were better than those in the No repair group (<0.05).


Conserved dopamine neurotrophic factor-transduced mesenchymal stem cells promote axon regeneration and functional recovery of injured sciatic nerve.

Liu Y, Nie L, Zhao H, Zhang W, Zhang YQ, Wang SS, Cheng L - PLoS ONE (2014)

Evaluation of amyotrophy.(A and B) Masson’s collagen staining of sections of the transverse gastrocnemius muscle at 8 and 12 weeks after surgery. Scale bar = 50 µm (C) Quantitative analysis of the percentages of muscle fibers in each group. (D) Wet weight analysis of the gastrocnemius muscle at 4, 8 and 12 weeks after surgery. (E) Statistical analysis of the wet weight ratio of the gastrocnemius muscle. n = 5, *p<0.05, **p<0.01.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4208796&req=5

pone-0110993-g007: Evaluation of amyotrophy.(A and B) Masson’s collagen staining of sections of the transverse gastrocnemius muscle at 8 and 12 weeks after surgery. Scale bar = 50 µm (C) Quantitative analysis of the percentages of muscle fibers in each group. (D) Wet weight analysis of the gastrocnemius muscle at 4, 8 and 12 weeks after surgery. (E) Statistical analysis of the wet weight ratio of the gastrocnemius muscle. n = 5, *p<0.05, **p<0.01.
Mentions: Masson’s collagen staining method directly showed the morphological changes in the gastrocnemius muscle at 8 and 12 weeks. The percentage of muscle fibers in the CDNF-MSCs group was significantly larger than those in the No repair, MSCs, LV-MSCs and naive groups at 8 and 12 weeks (Fig. 7C). The percentage of muscle fibers between the MSCs and LV-MSCs groups did not show obvious differences at 8 and 12 weeks after surgery (n = 5, P>0.05) (Fig. 7C). We also weighed the gastrocnemius muscles at 4, 8, and 12 weeks to assess muscle innervation recovery. At 8 and 12 weeks after surgery, the wet weights of the gastrocnemius in the CDNF-MSCs group were significantly higher than those in the No repair, LV-MSCs, MSCs, and DMEM groups, suggesting that treatment with CDNF-MSCs led to greater innervation of the gastrocnemius muscle (Fig. 7D). The wet weight ratio of gastrocnemius muscle in the CDNF-MSCs group was higher than superior to those in the No repair, LV-MSCs, MSCs, and DMEM groups (<0.05), but the difference between the LV-MSCs and MSCs groups was not statistically significant (>0.05). The wet weight of gastrocnemius muscle, the percentage of muscle fibers and the wet weight ratio of gastrocnemius muscle of all repair groups were better than those in the No repair group (<0.05).

Bottom Line: Light and electron microscopy confirmed successful regeneration of the sciatic nerve.The greater numbers of HRP-labeled neuron cell bodies and increased sciatic nerve index values (SFI) in the CDNF-MSCs group suggest that CDNF exerts neuroprotective effects in vivo.Collectively, these findings indicate that CDNF gene therapy delivered by MSCs is capable of promoting nerve regeneration and functional recovery, likely because of the significant neuroprotective and neurotrophic effects of CDNF and the superior environment offered by MSCs and collagen tubes.

View Article: PubMed Central - PubMed

Affiliation: Department of Spine Surgery, Qilu Hospital of Shandong University, Jinan, China; Shandong University Qilu Hospital Research Center for Cell Therapy, Key Laboratory of Cardiovascular Remodeling and Function Research, Qilu Hospital of Shandong University, Jinan, China.

ABSTRACT
Peripheral nerve injury (PNI) is a common disease that often results in axonal degeneration and the loss of neurons, ultimately leading to limited nerve regeneration and severe functional impairment. Currently, there are no effective treatments for PNI. In the present study, we transduced conserved dopamine neurotrophic factor (CDNF) into mesenchymal stem cells (MSCs) in collagen tubes to investigate their regenerative effects on rat peripheral nerves in an in vivo transection model. Scanning electron microscopy of the collagen tubes demonstrated their ability to be resorbed in vivo. We observed notable overexpression of the CDNF protein in the distal sciatic nerve after application of CDNF-MSCs. Quantitative analysis of neurofilament 200 (NF200) and S100 immunohistochemistry showed significant enhancement of axonal and Schwann cell regeneration in the group receiving CDNF-MSCs (CDNF-MSCs group) compared with the control groups. Myelination thickness, axon diameter and the axon-to fiber diameter ratio (G-ratio) were significantly higher in the CDNF-MSCs group at 8 and 12 weeks after nerve transection surgery. After surgery, the sciatic functional index, target muscle weight, wet weight ratio of gastrocnemius muscle and horseradish peroxidase (HRP) tracing demonstrated functional recovery. Light and electron microscopy confirmed successful regeneration of the sciatic nerve. The greater numbers of HRP-labeled neuron cell bodies and increased sciatic nerve index values (SFI) in the CDNF-MSCs group suggest that CDNF exerts neuroprotective effects in vivo. We also observed higher target muscle weights and a significant improvement in muscle atrophism in the CDNF-MSCs group. Collectively, these findings indicate that CDNF gene therapy delivered by MSCs is capable of promoting nerve regeneration and functional recovery, likely because of the significant neuroprotective and neurotrophic effects of CDNF and the superior environment offered by MSCs and collagen tubes.

Show MeSH
Related in: MedlinePlus