Assessment of Glial Scar, Tissue Sparing, Behavioral Recovery and Axonal Regeneration following Acute Transplantation of Genetically Modified Human Umbilical Cord Blood Cells in a Rat Model of Spinal Cord Contusion

Yana O Mukhamedshina, Ekaterina E Garanina, Galina A Masgutova, Luisa R Galieva, Elvira R Sanatova, Yurii A Chelyshev, Albert A Rizvanov, Yana O Mukhamedshina, Ekaterina E Garanina, Galina A Masgutova, Luisa R Galieva, Elvira R Sanatova, Yurii A Chelyshev, Albert A Rizvanov

Abstract

Objective and methods: This study investigated the potential for protective effects of human umbilical cord blood mononuclear cells (UCB-MCs) genetically modified with the VEGF and GNDF genes on contusion spinal cord injury (SCI) in rats. An adenoviral vector was constructed for targeted delivery of VEGF and GDNF to UCB-MCs. Using a rat contusion SCI model we examined the efficacy of the construct on tissue sparing, glial scar severity, the extent of axonal regeneration, recovery of motor function, and analyzed the expression of the recombinant genes VEGF and GNDF in vitro and in vivo.

Results: Transplantation of UCB-MCs transduced with adenoviral vectors expressing VEGF and GDNF at the site of SCI induced tissue sparing, behavioral recovery and axonal regeneration comparing to the other constructs tested. The adenovirus encoding VEGF and GDNF for transduction of UCB-MCs was shown to be an effective and stable vehicle for these cells in vivo following the transplantation into the contused spinal cord.

Conclusion: Our results show that a gene delivery using UCB-MCs-expressing VEGF and GNDF genes improved both structural and functional parameters after SCI. Further histological and behavioral studies, especially at later time points, in animals with SCI after transplantation of genetically modified UCB-MCs (overexpressing VEGF and GDNF genes) will provide additional insight into therapeutic potential of such cells.

Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Fig 1. VEGF, GDNF and EGFP mRNA…
Fig 1. VEGF, GDNF and EGFP mRNA expression in vitro.
VEGF (A), GDNF (B) and EGFP (C) mRNA expression on day 5 after transduction of UCB-MCs concurrently with adenoviral vectors Ad5-VEGF/Ad5-GDNF and Ad5-EGFP, respectively. The levels of VEGF, GDNF and EGFP mRNA expression in UCB-MCs was considered 100%. Differences were statistically significant between UCB-MCs and the experimental groups in all cases (**–P

Fig 2. VEGF, GDNF and EGFP mRNA…

Fig 2. VEGF, GDNF and EGFP mRNA expression in vivo.

VEGF, GDNF (A) and EGFP…

Fig 2. VEGF, GDNF and EGFP mRNA expression in vivo.
VEGF, GDNF (A) and EGFP (B) mRNA expression on day 30 after SCI or Sham followed by injection of transduced UCB-MCs. The levels of VEGF, GDNF and EGFP mRNA expression in SCI group was considered 100%. Differences were statistically significant between SCI-group and other experimental groups (*–P

Fig 3. Tissue analysis in experimental groups.

Fig 3. Tissue analysis in experimental groups.

Injured spinal cord 30 days after SCI without…

Fig 3. Tissue analysis in experimental groups.
Injured spinal cord 30 days after SCI without therapy (A), SCI with direct injection of UCB-MCs+Ad5-VEGF+Ad5-GDNF (B) and UCB-MCs+Ad5-EGFP (C), Sham operation with direct injection of UCB-MCs+Ad5-VEGF+Ad5-GDNF (D). Images are Azur-eosin cryosections. Scale bar: 750 μm. Volume of intact tissue (E) and cavity volume (F) relative to the Th8 area of intact rats remaining 30 days after SCI in experimental groups. *–P post hoc test.

Fig 4. Visualization of grafted cells at…

Fig 4. Visualization of grafted cells at day 30 after transplantation.

Visualization of grafted cells…

Fig 4. Visualization of grafted cells at day 30 after transplantation.
Visualization of grafted cells in the SCI zone 30 days after transplantation of UCB-MCs+Ad5-EGFP (A) and UCB-MCs+Ad5-VEGF+Ad5-GDNF (B). Localization of transplanted UCB-MCs+Ad5-VEGF+Ad5-GDNF after Sham operation (C). (A) HNu+ cells (red) surviving in the spinal cord contusion area, and forming cell bridges within the traumatic centromedullary cavity. The dashed boxes indicate enlarged area of A”. Arrows show some the HNu and DAPI overlap cells which were used for analysis. (A’) The same area, labeled by dashed boxes, in subsequent serial sections was stained as negative control. (B) In UCB-MCs+Ad5-VEGF+Ad5-GDNF-treated rats the contusion area with spared tissue contained mostly HNu+ cells, which expressed VEGF and GDNF. (C) HNu+/VEGF+/GDNF+ cells arranged accordingly to the nerve fibers 30 days after Sham with injection of UCB-MCs+Ad5-VEGF+Ad5-GDNF. Nuclei are stained with DAPI (blue). Scale bar: 200 (A), 10 (A’, A”), 2,5 (B) and 20 (C) μm.

Fig 5. Glial scar formation at the…

Fig 5. Glial scar formation at the lesion site indicated by GFAP.

Visualization of glial…

Fig 5. Glial scar formation at the lesion site indicated by GFAP.
Visualization of glial scar formation at the lesion site using GFAP in experimental groups: SCI UCB-MCs+Ad5-EGFP (A), SCI UCB-MCs+Ad5-VEGF+Ad5-GDNF (B), SCI (C) and Sham UCB-MCs+Ad5-VEGF+Ad5-GDNF (D). In the UCB-MCs+Ad5-VEGF+Ad5-GDNF-treated rats after SCI we detected a central zone of tissue without GFAP immunofluorescence. Scale bar: 750 μm. (E) Western-blotting analysis of GFAP on the 30th day after SCI with direct injection of UCB-MCs+Ad5-VEGF+Ad5-GDNF (1) and UCB-MCs+Ad5-EGFP (3), SCI without therapy (2), Sham operation with direct injection of UCB-MCs+Ad5-VEGF+Ad5-GDNF (4). Staining with Abs against GFAP revealed a band at 50 kDa in the samples. β-actin was used as a loading control. At day 30, western blot analysis shows reduced GFAP expression at the injury zone after UCB-MCs+Ad5-VEGF+Ad5-GDNF injection. Positive and negative controls were performed using Western Blotting control for GFAP antibodies and protein extracts from mononuclear umbilical cord blood cells, respectively. (F) Densitometry analysis demonstrated a significant change in GFAP levels relative to β-actin expression after SCI. Differences were statistically significant between SCI and other experimental groups (*P

Fig 6. Expression of GFAP, CGRP and…

Fig 6. Expression of GFAP, CGRP and GAP43 in the lesion site of spinal cord.

Fig 6. Expression of GFAP, CGRP and GAP43 in the lesion site of spinal cord.
Visualization of the distribution of GFAP, CGRP and GAP43 at the site of spinal cord contusion lesion after SCI-only (C,D), transplantation of UCB-MCs+Ad5-EGFP (A,B) and UCB-MCs+Ad5-VEGF+Ad5-GDNF after SCI (E,F,K,M,N) and Sham (G,H). In the UCB-MCs+Ad5-EGFP group (A,B) CGRP and GAP-43 expression was located in the islet of lesion zone, surrounded by glial scar. The UCB-MCs at the injury site appeared to be very closely associated with GAP43+ axons (K). The sections I,J present negative controls. Nuclei are stained with DAPI (blue). Scale bar: 100 (A-J) and 5 (K). (D) Mean labeling intensity of GAP43 of the rats in experimental groups in the SCI center. Differences were statistically significant between SCI and other experimental groups (*P < 0.01). Differences were also statistically significant between groups with injection of UCB-MCs (**P < 0.05). One-way ANOVA.

Fig 7. BBB locomotor scores of rats…

Fig 7. BBB locomotor scores of rats after SCI or Sham in experimental group.

BBB…

Fig 7. BBB locomotor scores of rats after SCI or Sham in experimental group.
BBB locomotor scores of rats obtained for the SCI (A, red line), SCI UCB-MCs+Ad5-EGFP (B, red line), SCI UCB-MCs+Ad5-VEGF+Ad5-GDNF (B, black line) and Sham UCB-MCs+Ad5-VEGF+Ad5-GDNF (A, black line) groups. Statistically significant differences were detected between Sham UCB-MCs+Ad5-VEGF+Ad5-GDNF group and other groups for all days (P
All figures (7)
Similar articles
References
    1. Kuh SU, Cho YE, Yoon DH, Kim KN, Ha Y. Functional recovery after human umbilical cord blood cells transplantation with brain-derived neutrophic factor into the spinal cord injured rat. Acta Neurochirurgica (Wien). 2005;147(9): 985–992. - PubMed
    1. Yan HB, Zhang ZM, Jin DD, Wang XJ, Lu KW. The repair of acute spinal cord injury in rats by olfactory ensheathing cells graft modified by glia cell line–derived neurotrophic factor gene in combination with the injection of monoclonal antibody IN–1. Zhonghua Wai Ke Za Zhi. 2009;47(23): 1817–1820. - PubMed
    1. Kim HM, Hwang DH, Lee JE, Kim SU, Kim BG. Ex vivo VEGF delivery by neural stem cells enhances proliferation of glial progenitors, angiogenesis, and tissue sparing after spinal cord injury. PLoS One. 2009;4(3): 1–10. - PMC - PubMed
    1. Lin WP, Chen XW, Zhang LQ, Wu CY, Huang ZD, Lin JH.Effect of neuroglobin genetically modified bone marrow mesenchymal stem cells transplantation on spinal cord injury in rabbits. PLoS One. 2013;8(5): 1–9. - PMC - PubMed
    1. Jones LL, Oudega M, Bunge MB, Tuszynski MH. Neurotrophic factors, cellular bridges and gene therapy for spinal cord injury. J Physiol. 2001;533(1): 83–89. - PMC - PubMed
Show all 28 references
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Grant support
The study was supported by grants 15-04-07527 (A.A. Rizvanov) and 14-04-31246 (Y.O. Mukhamedshina) from Russian Foundation for Basic Research. Y.O. Mukhamedshina was supported by a Presidential Grant for government support of young scientists (PhD) from the Russian Federation (MK-4020.2015.7). This work was performed in accordance with Program of Competitive Growth of Kazan Federal University and a subsidy allocated to Kazan Federal University for the state assignment in the sphere of scientific activities. Some of the experiments were conducted using equipment at the Interdisciplinary Center for Collective Use of Kazan Federal University supported by Ministry of Education of Russia (ID RFMEFI59414X0003), Interdisciplinary Center for Analytical Microscopy, and Pharmaceutical Research and Education Center, Kazan (Volga Region) Federal University, Kazan, Russia.
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Fig 2. VEGF, GDNF and EGFP mRNA…
Fig 2. VEGF, GDNF and EGFP mRNA expression in vivo.
VEGF, GDNF (A) and EGFP (B) mRNA expression on day 30 after SCI or Sham followed by injection of transduced UCB-MCs. The levels of VEGF, GDNF and EGFP mRNA expression in SCI group was considered 100%. Differences were statistically significant between SCI-group and other experimental groups (*–P

Fig 3. Tissue analysis in experimental groups.

Fig 3. Tissue analysis in experimental groups.

Injured spinal cord 30 days after SCI without…

Fig 3. Tissue analysis in experimental groups.
Injured spinal cord 30 days after SCI without therapy (A), SCI with direct injection of UCB-MCs+Ad5-VEGF+Ad5-GDNF (B) and UCB-MCs+Ad5-EGFP (C), Sham operation with direct injection of UCB-MCs+Ad5-VEGF+Ad5-GDNF (D). Images are Azur-eosin cryosections. Scale bar: 750 μm. Volume of intact tissue (E) and cavity volume (F) relative to the Th8 area of intact rats remaining 30 days after SCI in experimental groups. *–P post hoc test.

Fig 4. Visualization of grafted cells at…

Fig 4. Visualization of grafted cells at day 30 after transplantation.

Visualization of grafted cells…

Fig 4. Visualization of grafted cells at day 30 after transplantation.
Visualization of grafted cells in the SCI zone 30 days after transplantation of UCB-MCs+Ad5-EGFP (A) and UCB-MCs+Ad5-VEGF+Ad5-GDNF (B). Localization of transplanted UCB-MCs+Ad5-VEGF+Ad5-GDNF after Sham operation (C). (A) HNu+ cells (red) surviving in the spinal cord contusion area, and forming cell bridges within the traumatic centromedullary cavity. The dashed boxes indicate enlarged area of A”. Arrows show some the HNu and DAPI overlap cells which were used for analysis. (A’) The same area, labeled by dashed boxes, in subsequent serial sections was stained as negative control. (B) In UCB-MCs+Ad5-VEGF+Ad5-GDNF-treated rats the contusion area with spared tissue contained mostly HNu+ cells, which expressed VEGF and GDNF. (C) HNu+/VEGF+/GDNF+ cells arranged accordingly to the nerve fibers 30 days after Sham with injection of UCB-MCs+Ad5-VEGF+Ad5-GDNF. Nuclei are stained with DAPI (blue). Scale bar: 200 (A), 10 (A’, A”), 2,5 (B) and 20 (C) μm.

Fig 5. Glial scar formation at the…

Fig 5. Glial scar formation at the lesion site indicated by GFAP.

Visualization of glial…

Fig 5. Glial scar formation at the lesion site indicated by GFAP.
Visualization of glial scar formation at the lesion site using GFAP in experimental groups: SCI UCB-MCs+Ad5-EGFP (A), SCI UCB-MCs+Ad5-VEGF+Ad5-GDNF (B), SCI (C) and Sham UCB-MCs+Ad5-VEGF+Ad5-GDNF (D). In the UCB-MCs+Ad5-VEGF+Ad5-GDNF-treated rats after SCI we detected a central zone of tissue without GFAP immunofluorescence. Scale bar: 750 μm. (E) Western-blotting analysis of GFAP on the 30th day after SCI with direct injection of UCB-MCs+Ad5-VEGF+Ad5-GDNF (1) and UCB-MCs+Ad5-EGFP (3), SCI without therapy (2), Sham operation with direct injection of UCB-MCs+Ad5-VEGF+Ad5-GDNF (4). Staining with Abs against GFAP revealed a band at 50 kDa in the samples. β-actin was used as a loading control. At day 30, western blot analysis shows reduced GFAP expression at the injury zone after UCB-MCs+Ad5-VEGF+Ad5-GDNF injection. Positive and negative controls were performed using Western Blotting control for GFAP antibodies and protein extracts from mononuclear umbilical cord blood cells, respectively. (F) Densitometry analysis demonstrated a significant change in GFAP levels relative to β-actin expression after SCI. Differences were statistically significant between SCI and other experimental groups (*P

Fig 6. Expression of GFAP, CGRP and…

Fig 6. Expression of GFAP, CGRP and GAP43 in the lesion site of spinal cord.

Fig 6. Expression of GFAP, CGRP and GAP43 in the lesion site of spinal cord.
Visualization of the distribution of GFAP, CGRP and GAP43 at the site of spinal cord contusion lesion after SCI-only (C,D), transplantation of UCB-MCs+Ad5-EGFP (A,B) and UCB-MCs+Ad5-VEGF+Ad5-GDNF after SCI (E,F,K,M,N) and Sham (G,H). In the UCB-MCs+Ad5-EGFP group (A,B) CGRP and GAP-43 expression was located in the islet of lesion zone, surrounded by glial scar. The UCB-MCs at the injury site appeared to be very closely associated with GAP43+ axons (K). The sections I,J present negative controls. Nuclei are stained with DAPI (blue). Scale bar: 100 (A-J) and 5 (K). (D) Mean labeling intensity of GAP43 of the rats in experimental groups in the SCI center. Differences were statistically significant between SCI and other experimental groups (*P < 0.01). Differences were also statistically significant between groups with injection of UCB-MCs (**P < 0.05). One-way ANOVA.

Fig 7. BBB locomotor scores of rats…

Fig 7. BBB locomotor scores of rats after SCI or Sham in experimental group.

BBB…

Fig 7. BBB locomotor scores of rats after SCI or Sham in experimental group.
BBB locomotor scores of rats obtained for the SCI (A, red line), SCI UCB-MCs+Ad5-EGFP (B, red line), SCI UCB-MCs+Ad5-VEGF+Ad5-GDNF (B, black line) and Sham UCB-MCs+Ad5-VEGF+Ad5-GDNF (A, black line) groups. Statistically significant differences were detected between Sham UCB-MCs+Ad5-VEGF+Ad5-GDNF group and other groups for all days (P
All figures (7)
Similar articles
References
    1. Kuh SU, Cho YE, Yoon DH, Kim KN, Ha Y. Functional recovery after human umbilical cord blood cells transplantation with brain-derived neutrophic factor into the spinal cord injured rat. Acta Neurochirurgica (Wien). 2005;147(9): 985–992. - PubMed
    1. Yan HB, Zhang ZM, Jin DD, Wang XJ, Lu KW. The repair of acute spinal cord injury in rats by olfactory ensheathing cells graft modified by glia cell line–derived neurotrophic factor gene in combination with the injection of monoclonal antibody IN–1. Zhonghua Wai Ke Za Zhi. 2009;47(23): 1817–1820. - PubMed
    1. Kim HM, Hwang DH, Lee JE, Kim SU, Kim BG. Ex vivo VEGF delivery by neural stem cells enhances proliferation of glial progenitors, angiogenesis, and tissue sparing after spinal cord injury. PLoS One. 2009;4(3): 1–10. - PMC - PubMed
    1. Lin WP, Chen XW, Zhang LQ, Wu CY, Huang ZD, Lin JH.Effect of neuroglobin genetically modified bone marrow mesenchymal stem cells transplantation on spinal cord injury in rabbits. PLoS One. 2013;8(5): 1–9. - PMC - PubMed
    1. Jones LL, Oudega M, Bunge MB, Tuszynski MH. Neurotrophic factors, cellular bridges and gene therapy for spinal cord injury. J Physiol. 2001;533(1): 83–89. - PMC - PubMed
Show all 28 references
Publication types
MeSH terms
Substances
Related information
Grant support
The study was supported by grants 15-04-07527 (A.A. Rizvanov) and 14-04-31246 (Y.O. Mukhamedshina) from Russian Foundation for Basic Research. Y.O. Mukhamedshina was supported by a Presidential Grant for government support of young scientists (PhD) from the Russian Federation (MK-4020.2015.7). This work was performed in accordance with Program of Competitive Growth of Kazan Federal University and a subsidy allocated to Kazan Federal University for the state assignment in the sphere of scientific activities. Some of the experiments were conducted using equipment at the Interdisciplinary Center for Collective Use of Kazan Federal University supported by Ministry of Education of Russia (ID RFMEFI59414X0003), Interdisciplinary Center for Analytical Microscopy, and Pharmaceutical Research and Education Center, Kazan (Volga Region) Federal University, Kazan, Russia.
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Fig 3. Tissue analysis in experimental groups.
Fig 3. Tissue analysis in experimental groups.
Injured spinal cord 30 days after SCI without therapy (A), SCI with direct injection of UCB-MCs+Ad5-VEGF+Ad5-GDNF (B) and UCB-MCs+Ad5-EGFP (C), Sham operation with direct injection of UCB-MCs+Ad5-VEGF+Ad5-GDNF (D). Images are Azur-eosin cryosections. Scale bar: 750 μm. Volume of intact tissue (E) and cavity volume (F) relative to the Th8 area of intact rats remaining 30 days after SCI in experimental groups. *–P post hoc test.
Fig 4. Visualization of grafted cells at…
Fig 4. Visualization of grafted cells at day 30 after transplantation.
Visualization of grafted cells in the SCI zone 30 days after transplantation of UCB-MCs+Ad5-EGFP (A) and UCB-MCs+Ad5-VEGF+Ad5-GDNF (B). Localization of transplanted UCB-MCs+Ad5-VEGF+Ad5-GDNF after Sham operation (C). (A) HNu+ cells (red) surviving in the spinal cord contusion area, and forming cell bridges within the traumatic centromedullary cavity. The dashed boxes indicate enlarged area of A”. Arrows show some the HNu and DAPI overlap cells which were used for analysis. (A’) The same area, labeled by dashed boxes, in subsequent serial sections was stained as negative control. (B) In UCB-MCs+Ad5-VEGF+Ad5-GDNF-treated rats the contusion area with spared tissue contained mostly HNu+ cells, which expressed VEGF and GDNF. (C) HNu+/VEGF+/GDNF+ cells arranged accordingly to the nerve fibers 30 days after Sham with injection of UCB-MCs+Ad5-VEGF+Ad5-GDNF. Nuclei are stained with DAPI (blue). Scale bar: 200 (A), 10 (A’, A”), 2,5 (B) and 20 (C) μm.
Fig 5. Glial scar formation at the…
Fig 5. Glial scar formation at the lesion site indicated by GFAP.
Visualization of glial scar formation at the lesion site using GFAP in experimental groups: SCI UCB-MCs+Ad5-EGFP (A), SCI UCB-MCs+Ad5-VEGF+Ad5-GDNF (B), SCI (C) and Sham UCB-MCs+Ad5-VEGF+Ad5-GDNF (D). In the UCB-MCs+Ad5-VEGF+Ad5-GDNF-treated rats after SCI we detected a central zone of tissue without GFAP immunofluorescence. Scale bar: 750 μm. (E) Western-blotting analysis of GFAP on the 30th day after SCI with direct injection of UCB-MCs+Ad5-VEGF+Ad5-GDNF (1) and UCB-MCs+Ad5-EGFP (3), SCI without therapy (2), Sham operation with direct injection of UCB-MCs+Ad5-VEGF+Ad5-GDNF (4). Staining with Abs against GFAP revealed a band at 50 kDa in the samples. β-actin was used as a loading control. At day 30, western blot analysis shows reduced GFAP expression at the injury zone after UCB-MCs+Ad5-VEGF+Ad5-GDNF injection. Positive and negative controls were performed using Western Blotting control for GFAP antibodies and protein extracts from mononuclear umbilical cord blood cells, respectively. (F) Densitometry analysis demonstrated a significant change in GFAP levels relative to β-actin expression after SCI. Differences were statistically significant between SCI and other experimental groups (*P

Fig 6. Expression of GFAP, CGRP and…

Fig 6. Expression of GFAP, CGRP and GAP43 in the lesion site of spinal cord.

Fig 6. Expression of GFAP, CGRP and GAP43 in the lesion site of spinal cord.
Visualization of the distribution of GFAP, CGRP and GAP43 at the site of spinal cord contusion lesion after SCI-only (C,D), transplantation of UCB-MCs+Ad5-EGFP (A,B) and UCB-MCs+Ad5-VEGF+Ad5-GDNF after SCI (E,F,K,M,N) and Sham (G,H). In the UCB-MCs+Ad5-EGFP group (A,B) CGRP and GAP-43 expression was located in the islet of lesion zone, surrounded by glial scar. The UCB-MCs at the injury site appeared to be very closely associated with GAP43+ axons (K). The sections I,J present negative controls. Nuclei are stained with DAPI (blue). Scale bar: 100 (A-J) and 5 (K). (D) Mean labeling intensity of GAP43 of the rats in experimental groups in the SCI center. Differences were statistically significant between SCI and other experimental groups (*P < 0.01). Differences were also statistically significant between groups with injection of UCB-MCs (**P < 0.05). One-way ANOVA.

Fig 7. BBB locomotor scores of rats…

Fig 7. BBB locomotor scores of rats after SCI or Sham in experimental group.

BBB…

Fig 7. BBB locomotor scores of rats after SCI or Sham in experimental group.
BBB locomotor scores of rats obtained for the SCI (A, red line), SCI UCB-MCs+Ad5-EGFP (B, red line), SCI UCB-MCs+Ad5-VEGF+Ad5-GDNF (B, black line) and Sham UCB-MCs+Ad5-VEGF+Ad5-GDNF (A, black line) groups. Statistically significant differences were detected between Sham UCB-MCs+Ad5-VEGF+Ad5-GDNF group and other groups for all days (P
All figures (7)
Similar articles
References
    1. Kuh SU, Cho YE, Yoon DH, Kim KN, Ha Y. Functional recovery after human umbilical cord blood cells transplantation with brain-derived neutrophic factor into the spinal cord injured rat. Acta Neurochirurgica (Wien). 2005;147(9): 985–992. - PubMed
    1. Yan HB, Zhang ZM, Jin DD, Wang XJ, Lu KW. The repair of acute spinal cord injury in rats by olfactory ensheathing cells graft modified by glia cell line–derived neurotrophic factor gene in combination with the injection of monoclonal antibody IN–1. Zhonghua Wai Ke Za Zhi. 2009;47(23): 1817–1820. - PubMed
    1. Kim HM, Hwang DH, Lee JE, Kim SU, Kim BG. Ex vivo VEGF delivery by neural stem cells enhances proliferation of glial progenitors, angiogenesis, and tissue sparing after spinal cord injury. PLoS One. 2009;4(3): 1–10. - PMC - PubMed
    1. Lin WP, Chen XW, Zhang LQ, Wu CY, Huang ZD, Lin JH.Effect of neuroglobin genetically modified bone marrow mesenchymal stem cells transplantation on spinal cord injury in rabbits. PLoS One. 2013;8(5): 1–9. - PMC - PubMed
    1. Jones LL, Oudega M, Bunge MB, Tuszynski MH. Neurotrophic factors, cellular bridges and gene therapy for spinal cord injury. J Physiol. 2001;533(1): 83–89. - PMC - PubMed
Show all 28 references
Publication types
MeSH terms
Substances
Related information
Grant support
The study was supported by grants 15-04-07527 (A.A. Rizvanov) and 14-04-31246 (Y.O. Mukhamedshina) from Russian Foundation for Basic Research. Y.O. Mukhamedshina was supported by a Presidential Grant for government support of young scientists (PhD) from the Russian Federation (MK-4020.2015.7). This work was performed in accordance with Program of Competitive Growth of Kazan Federal University and a subsidy allocated to Kazan Federal University for the state assignment in the sphere of scientific activities. Some of the experiments were conducted using equipment at the Interdisciplinary Center for Collective Use of Kazan Federal University supported by Ministry of Education of Russia (ID RFMEFI59414X0003), Interdisciplinary Center for Analytical Microscopy, and Pharmaceutical Research and Education Center, Kazan (Volga Region) Federal University, Kazan, Russia.
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Cite
Copy Download .nbib
Format: AMA APA MLA NLM
Fig 6. Expression of GFAP, CGRP and…
Fig 6. Expression of GFAP, CGRP and GAP43 in the lesion site of spinal cord.
Visualization of the distribution of GFAP, CGRP and GAP43 at the site of spinal cord contusion lesion after SCI-only (C,D), transplantation of UCB-MCs+Ad5-EGFP (A,B) and UCB-MCs+Ad5-VEGF+Ad5-GDNF after SCI (E,F,K,M,N) and Sham (G,H). In the UCB-MCs+Ad5-EGFP group (A,B) CGRP and GAP-43 expression was located in the islet of lesion zone, surrounded by glial scar. The UCB-MCs at the injury site appeared to be very closely associated with GAP43+ axons (K). The sections I,J present negative controls. Nuclei are stained with DAPI (blue). Scale bar: 100 (A-J) and 5 (K). (D) Mean labeling intensity of GAP43 of the rats in experimental groups in the SCI center. Differences were statistically significant between SCI and other experimental groups (*P < 0.01). Differences were also statistically significant between groups with injection of UCB-MCs (**P < 0.05). One-way ANOVA.
Fig 7. BBB locomotor scores of rats…
Fig 7. BBB locomotor scores of rats after SCI or Sham in experimental group.
BBB locomotor scores of rats obtained for the SCI (A, red line), SCI UCB-MCs+Ad5-EGFP (B, red line), SCI UCB-MCs+Ad5-VEGF+Ad5-GDNF (B, black line) and Sham UCB-MCs+Ad5-VEGF+Ad5-GDNF (A, black line) groups. Statistically significant differences were detected between Sham UCB-MCs+Ad5-VEGF+Ad5-GDNF group and other groups for all days (P
All figures (7)

References

    1. Kuh SU, Cho YE, Yoon DH, Kim KN, Ha Y. Functional recovery after human umbilical cord blood cells transplantation with brain-derived neutrophic factor into the spinal cord injured rat. Acta Neurochirurgica (Wien). 2005;147(9): 985–992.
    1. Yan HB, Zhang ZM, Jin DD, Wang XJ, Lu KW. The repair of acute spinal cord injury in rats by olfactory ensheathing cells graft modified by glia cell line–derived neurotrophic factor gene in combination with the injection of monoclonal antibody IN–1. Zhonghua Wai Ke Za Zhi. 2009;47(23): 1817–1820.
    1. Kim HM, Hwang DH, Lee JE, Kim SU, Kim BG. Ex vivo VEGF delivery by neural stem cells enhances proliferation of glial progenitors, angiogenesis, and tissue sparing after spinal cord injury. PLoS One. 2009;4(3): 1–10.
    1. Lin WP, Chen XW, Zhang LQ, Wu CY, Huang ZD, Lin JH.Effect of neuroglobin genetically modified bone marrow mesenchymal stem cells transplantation on spinal cord injury in rabbits. PLoS One. 2013;8(5): 1–9.
    1. Jones LL, Oudega M, Bunge MB, Tuszynski MH. Neurotrophic factors, cellular bridges and gene therapy for spinal cord injury. J Physiol. 2001;533(1): 83–89.
    1. Gluckman E, Locatelli F. Umbilical cord blood transplants. Opin Hematol. 2000;7(6): 353–357.
    1. Weiss M, Troyer D. Stem cells in the umbilical cord. Stem Cell Rev. 2007;2: 155–162.
    1. Park SI, Lim JY, Jeong CH, Kim SM, Jun JA, Jeun SS, et al. Human umbilical cord blood-derived mesenchymal stem cell therapy promotes functional recovery of contused rat spinal cord through enhancement of endogenous cell proliferation and oligogenesis. J Biomed Biotechnol. 2012: 1–8.
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    1. Mukhamedshina YO, Shaymardanova GF, Garanina EE, Salafutdinov II, Rizvanov AA, Islamov RR, et al. Adenoviral vector carrying glial cell-derived neurotrophic factor for direct gene therapy or human umbilical cord blood cell-mediated therapy of spinal cord injury in rat. Spinal Cord. 2015. September 29 10.1038/sc.2015.161 [Epub ahead of print]
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