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Intrathecal delivery of mesenchymal stromal cells protects the structure of altered perineuronal nets in SOD1 rats and amends the course of ALS.

Forostyak S, Homola A, Turnovcova K, Svitil P, Jendelova P, Sykova E - Stem Cells (2014)

Bottom Line: We measured the concentration of cytokines (IL-1α, MCP-1, TNF-α, GM-CSF, IL-4, and IFN-γ) in the rats' cerebrospinal fluid and found significantly higher concentrations of IL-1α and MCP-1.Our results show that PNN and cytokine homeostasis are altered in the SOD1 rat model of ALS.We also show that the administration of human MSCs is a safe procedure that delays the loss of motor function and increases the overall survival of symptomatic ALS animals, by remodeling the recipients' pattern of gene expression and having neuroprotective and immunomodulatory effects.

View Article: PubMed Central - PubMed

Affiliation: Institute of Experimental Medicine, Academy of Science of the Czech Republic, Prague, Czech Republic; Department of Neuroscience, 2nd Medical Faculty, Charles University, Prague, Czech Republic.

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WFA staining visualizes the perineuronal nets (PNNs) around motoneurons (SMI 32) in the ventral horns of wild-type (A–D), end-stage SOD1 mesenchymal stromal cell-treated (E–H) and symptomatic SOD1 sham-treated (I–L) rats. A further quantification of WFA staining intensity from the ventral horns (interrupted line) of MSC-treated SOD1 rats (violet columns) showed that these animals had a significantly better preserved PNNs in both the cervical (M) and lumbar (N) levels of the spinal cord, when compared with sham-treated SOD1 animals (green columns) and age-matched WT littermates (empty columns). Both groups of SOD1 rats had attenuated PNNs; however, this difference was not significant at the cervical level, where the MSCs were delivered, whereas at the lumbar level all SOD1 rats had a significantly weaker intensity of PNN staining (M, N). Error bars indicate SEM. Significance at: *, p ≤ .05; **, p ≤ .01; ***, p ≤ .001 (scale bars = 50 µm). Abbreviations: BMSC, bone marrow mesenchymal stromal cells; DMEM, Dulbecco's modified Eagle's medium; SOD1, superoxide dismutase 1; WFA, Wisteria floribunda agglutinin; WT, wild type.
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fig02: WFA staining visualizes the perineuronal nets (PNNs) around motoneurons (SMI 32) in the ventral horns of wild-type (A–D), end-stage SOD1 mesenchymal stromal cell-treated (E–H) and symptomatic SOD1 sham-treated (I–L) rats. A further quantification of WFA staining intensity from the ventral horns (interrupted line) of MSC-treated SOD1 rats (violet columns) showed that these animals had a significantly better preserved PNNs in both the cervical (M) and lumbar (N) levels of the spinal cord, when compared with sham-treated SOD1 animals (green columns) and age-matched WT littermates (empty columns). Both groups of SOD1 rats had attenuated PNNs; however, this difference was not significant at the cervical level, where the MSCs were delivered, whereas at the lumbar level all SOD1 rats had a significantly weaker intensity of PNN staining (M, N). Error bars indicate SEM. Significance at: *, p ≤ .05; **, p ≤ .01; ***, p ≤ .001 (scale bars = 50 µm). Abbreviations: BMSC, bone marrow mesenchymal stromal cells; DMEM, Dulbecco's modified Eagle's medium; SOD1, superoxide dismutase 1; WFA, Wisteria floribunda agglutinin; WT, wild type.

Mentions: By evaluating samples stained with immunohistochemistry (IHC), we found that WT animals have normal PNNs around MNs (Fig. 2A–2D), whereas all transgenic (Tg) rats at the end-stage of the disease had disorganized and attenuated PNN structures (Fig. 2E–2L). Interestingly, MSC-treated SOD1 rats (Fig. 2E–2H), despite having attenuated PNNs when compared with WT animals, displayed a significantly reduced deterioration of their PNN structures when compared with vehicle-treated rats (Fig. 2I–2L). Quantification of Wisteria floribunda agglutinin (WFA) staining intensity showed that all SOD1 rats had a lower fluorescent intensity at the cervical (Fig. 2M) and lumbar (Fig. 2N) levels when compared with WT age-matched littermates (p = .108 and p = .012, respectively). The delivery of MSCs into symptomatic rats significantly preserved their PNNs at the cervical (Fig. 2M) and lumbar levels of the ventral horns (Fig. 2N) when compared with vehicle-injected SOD1 rats (p = .000008 and p = .00021, respectively). We found a significant difference in WFA staining intensity between the cervical and lumbar levels within the cell-treated SOD1 group (p = .026) and no changes in the WT and vehicle-injected groups, suggesting that proximity to the site of cell application is essential to achieve the best effects implied.


Intrathecal delivery of mesenchymal stromal cells protects the structure of altered perineuronal nets in SOD1 rats and amends the course of ALS.

Forostyak S, Homola A, Turnovcova K, Svitil P, Jendelova P, Sykova E - Stem Cells (2014)

WFA staining visualizes the perineuronal nets (PNNs) around motoneurons (SMI 32) in the ventral horns of wild-type (A–D), end-stage SOD1 mesenchymal stromal cell-treated (E–H) and symptomatic SOD1 sham-treated (I–L) rats. A further quantification of WFA staining intensity from the ventral horns (interrupted line) of MSC-treated SOD1 rats (violet columns) showed that these animals had a significantly better preserved PNNs in both the cervical (M) and lumbar (N) levels of the spinal cord, when compared with sham-treated SOD1 animals (green columns) and age-matched WT littermates (empty columns). Both groups of SOD1 rats had attenuated PNNs; however, this difference was not significant at the cervical level, where the MSCs were delivered, whereas at the lumbar level all SOD1 rats had a significantly weaker intensity of PNN staining (M, N). Error bars indicate SEM. Significance at: *, p ≤ .05; **, p ≤ .01; ***, p ≤ .001 (scale bars = 50 µm). Abbreviations: BMSC, bone marrow mesenchymal stromal cells; DMEM, Dulbecco's modified Eagle's medium; SOD1, superoxide dismutase 1; WFA, Wisteria floribunda agglutinin; WT, wild type.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4321196&req=5

fig02: WFA staining visualizes the perineuronal nets (PNNs) around motoneurons (SMI 32) in the ventral horns of wild-type (A–D), end-stage SOD1 mesenchymal stromal cell-treated (E–H) and symptomatic SOD1 sham-treated (I–L) rats. A further quantification of WFA staining intensity from the ventral horns (interrupted line) of MSC-treated SOD1 rats (violet columns) showed that these animals had a significantly better preserved PNNs in both the cervical (M) and lumbar (N) levels of the spinal cord, when compared with sham-treated SOD1 animals (green columns) and age-matched WT littermates (empty columns). Both groups of SOD1 rats had attenuated PNNs; however, this difference was not significant at the cervical level, where the MSCs were delivered, whereas at the lumbar level all SOD1 rats had a significantly weaker intensity of PNN staining (M, N). Error bars indicate SEM. Significance at: *, p ≤ .05; **, p ≤ .01; ***, p ≤ .001 (scale bars = 50 µm). Abbreviations: BMSC, bone marrow mesenchymal stromal cells; DMEM, Dulbecco's modified Eagle's medium; SOD1, superoxide dismutase 1; WFA, Wisteria floribunda agglutinin; WT, wild type.
Mentions: By evaluating samples stained with immunohistochemistry (IHC), we found that WT animals have normal PNNs around MNs (Fig. 2A–2D), whereas all transgenic (Tg) rats at the end-stage of the disease had disorganized and attenuated PNN structures (Fig. 2E–2L). Interestingly, MSC-treated SOD1 rats (Fig. 2E–2H), despite having attenuated PNNs when compared with WT animals, displayed a significantly reduced deterioration of their PNN structures when compared with vehicle-treated rats (Fig. 2I–2L). Quantification of Wisteria floribunda agglutinin (WFA) staining intensity showed that all SOD1 rats had a lower fluorescent intensity at the cervical (Fig. 2M) and lumbar (Fig. 2N) levels when compared with WT age-matched littermates (p = .108 and p = .012, respectively). The delivery of MSCs into symptomatic rats significantly preserved their PNNs at the cervical (Fig. 2M) and lumbar levels of the ventral horns (Fig. 2N) when compared with vehicle-injected SOD1 rats (p = .000008 and p = .00021, respectively). We found a significant difference in WFA staining intensity between the cervical and lumbar levels within the cell-treated SOD1 group (p = .026) and no changes in the WT and vehicle-injected groups, suggesting that proximity to the site of cell application is essential to achieve the best effects implied.

Bottom Line: We measured the concentration of cytokines (IL-1α, MCP-1, TNF-α, GM-CSF, IL-4, and IFN-γ) in the rats' cerebrospinal fluid and found significantly higher concentrations of IL-1α and MCP-1.Our results show that PNN and cytokine homeostasis are altered in the SOD1 rat model of ALS.We also show that the administration of human MSCs is a safe procedure that delays the loss of motor function and increases the overall survival of symptomatic ALS animals, by remodeling the recipients' pattern of gene expression and having neuroprotective and immunomodulatory effects.

View Article: PubMed Central - PubMed

Affiliation: Institute of Experimental Medicine, Academy of Science of the Czech Republic, Prague, Czech Republic; Department of Neuroscience, 2nd Medical Faculty, Charles University, Prague, Czech Republic.

Show MeSH
Related in: MedlinePlus