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Neuroprotective Potential of Mesenchymal Stem Cell-Based Therapy in Acute Stages of TNBS-Induced Colitis in Guinea-Pigs.

Robinson AM, Miller S, Payne N, Boyd R, Sakkal S, Nurgali K - PLoS ONE (2015)

Bottom Line: MSC and CM treatments prevented body weight loss, reduced infiltration of leukocytes into the colon wall and the myenteric plexus, facilitated repair of damaged tissue and nerve fibers, averted myenteric neuronal loss, as well as changes in neuronal subpopulations.The neuroprotective effects of MSC and CM treatments were observed as early as 24 hours after induction of inflammation even though the inflammatory reaction at the level of the myenteric ganglia had not completely subsided.The neuroprotective efficacy of MSC-based therapies can be exerted independently to their anti-inflammatory effects.

View Article: PubMed Central - PubMed

Affiliation: Centre for Chronic Diseases, College of Health and Biomedicine, Victoria University, Melbourne, Australia.

ABSTRACT

Background & aims: The therapeutic benefits of mesenchymal stem cells (MSCs), such as homing ability, multipotent differentiation capacity and secretion of soluble bioactive factors which exert neuroprotective, anti-inflammatory and immunomodulatory properties, have been attributed to attenuation of autoimmune, inflammatory and neurodegenerative disorders. In this study, we aimed to determine the earliest time point at which locally administered MSC-based therapies avert enteric neuronal loss and damage associated with intestinal inflammation in the guinea-pig model of colitis.

Methods: At 3 hours after induction of colitis by 2,4,6-trinitrobenzene-sulfonate (TNBS), guinea-pigs received either human bone marrow-derived MSCs, conditioned medium (CM), or unconditioned medium by enema into the colon. Colon tissues were collected 6, 24 and 72 hours after administration of TNBS. Effects on body weight, gross morphological damage, immune cell infiltration and myenteric neurons were evaluated. RT-PCR, flow cytometry and antibody array kit were used to identify neurotrophic and neuroprotective factors released by MSCs.

Results: MSC and CM treatments prevented body weight loss, reduced infiltration of leukocytes into the colon wall and the myenteric plexus, facilitated repair of damaged tissue and nerve fibers, averted myenteric neuronal loss, as well as changes in neuronal subpopulations. The neuroprotective effects of MSC and CM treatments were observed as early as 24 hours after induction of inflammation even though the inflammatory reaction at the level of the myenteric ganglia had not completely subsided. Substantial number of neurotrophic and neuroprotective factors released by MSCs was identified in their secretome.

Conclusion: MSC-based therapies applied at the acute stages of TNBS-induced colitis start exerting their neuroprotective effects towards enteric neurons by 24 hours post treatment. The neuroprotective efficacy of MSC-based therapies can be exerted independently to their anti-inflammatory effects.

No MeSH data available.


Related in: MedlinePlus

Effects of MSC and CM treatments on nNOS-IR myenteric neurons.No changes in nNOS-IR neurons were observed at 6 hours in any group (A-E). At 24 and 72 hours, an increase in nNOS-IR neurons was observed in sections from TNBS-only (BI-BII) and UCM-treated (EI-EII) animals compared to sham (AI-AII), but not in MSC and CM-treated groups (CI-CII, DI-DII). n = 4/group/time point. Scale bars = 50μm.
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pone.0139023.g010: Effects of MSC and CM treatments on nNOS-IR myenteric neurons.No changes in nNOS-IR neurons were observed at 6 hours in any group (A-E). At 24 and 72 hours, an increase in nNOS-IR neurons was observed in sections from TNBS-only (BI-BII) and UCM-treated (EI-EII) animals compared to sham (AI-AII), but not in MSC and CM-treated groups (CI-CII, DI-DII). n = 4/group/time point. Scale bars = 50μm.

Mentions: Changes in inhibitory and excitatory myenteric neurons underlie inflammation-induced colonic dysmotility (16–18); therefore we investigated the effects of MSC-based therapies on these neurons. Inhibitory neurons were identified by using nNOS immunoreactivity in wholemount LMMP preparations of the distal colon (Fig 10). The number and proportion of nNOS-IR neurons were comparable between all groups at the 6 hour time point (sham: 54±2 cells/area, 20±1%, TNBS: 57±3 cells/area, 22±1%, MSC: 57±2 cells/area, 22±2%, CM: 56±2 cells/area, 21±2%, UCM: 57±3 cells/area, 22±3%, Figs 10A–10E and 11). At 24 hours, the quantity of nNOS-IR myenteric neurons increased in the colon in TNBS-only (65±1 cells/area) and UCM-treated (66±3 cells/area) guinea-pigs compared to sham (53±1 cells/area), MSC (53±2 cells/area) and CM-treated (55±2 cells/area) animals (P<0.001 for all; Figs 10AI–10EI and 11A). The proportion of nNOS-IR neurons to the total number of neurons 24 hours post induction of colitis was greater in TNBS-only (31.9±1.8%) and UCM-treated (31.7±1.2%) guinea-pigs compared to sham (20±0.5%), MSC (21±1%), and CM-treated (22±1%) animals (P<0.001 for all; Fig 11B). At 72 hours, the number of nNOS-IR neurons in preparations of the distal colon from TNBS-only (65±3 cells/area) and UCM-treated (64±2 cells/area) guinea-pigs was higher compared to sham (53±1 cells/area), MSC (54±2 cells/area) and CM-treated (54±2 cells/area) animals (P<0.05 for all; Figs 10AII–10EII and 11A). The proportion of nNOS-IR neurons at 72 hours was greater in colon preparations from TNBS-only (32±2%) and UCM-treated (30±0.5%) guinea-pigs compared to sham (20±0.5%), MSC (21±1%), and CM-treated (21±1%) animals (P<0.001 for all; Fig 11B).


Neuroprotective Potential of Mesenchymal Stem Cell-Based Therapy in Acute Stages of TNBS-Induced Colitis in Guinea-Pigs.

Robinson AM, Miller S, Payne N, Boyd R, Sakkal S, Nurgali K - PLoS ONE (2015)

Effects of MSC and CM treatments on nNOS-IR myenteric neurons.No changes in nNOS-IR neurons were observed at 6 hours in any group (A-E). At 24 and 72 hours, an increase in nNOS-IR neurons was observed in sections from TNBS-only (BI-BII) and UCM-treated (EI-EII) animals compared to sham (AI-AII), but not in MSC and CM-treated groups (CI-CII, DI-DII). n = 4/group/time point. Scale bars = 50μm.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4580595&req=5

pone.0139023.g010: Effects of MSC and CM treatments on nNOS-IR myenteric neurons.No changes in nNOS-IR neurons were observed at 6 hours in any group (A-E). At 24 and 72 hours, an increase in nNOS-IR neurons was observed in sections from TNBS-only (BI-BII) and UCM-treated (EI-EII) animals compared to sham (AI-AII), but not in MSC and CM-treated groups (CI-CII, DI-DII). n = 4/group/time point. Scale bars = 50μm.
Mentions: Changes in inhibitory and excitatory myenteric neurons underlie inflammation-induced colonic dysmotility (16–18); therefore we investigated the effects of MSC-based therapies on these neurons. Inhibitory neurons were identified by using nNOS immunoreactivity in wholemount LMMP preparations of the distal colon (Fig 10). The number and proportion of nNOS-IR neurons were comparable between all groups at the 6 hour time point (sham: 54±2 cells/area, 20±1%, TNBS: 57±3 cells/area, 22±1%, MSC: 57±2 cells/area, 22±2%, CM: 56±2 cells/area, 21±2%, UCM: 57±3 cells/area, 22±3%, Figs 10A–10E and 11). At 24 hours, the quantity of nNOS-IR myenteric neurons increased in the colon in TNBS-only (65±1 cells/area) and UCM-treated (66±3 cells/area) guinea-pigs compared to sham (53±1 cells/area), MSC (53±2 cells/area) and CM-treated (55±2 cells/area) animals (P<0.001 for all; Figs 10AI–10EI and 11A). The proportion of nNOS-IR neurons to the total number of neurons 24 hours post induction of colitis was greater in TNBS-only (31.9±1.8%) and UCM-treated (31.7±1.2%) guinea-pigs compared to sham (20±0.5%), MSC (21±1%), and CM-treated (22±1%) animals (P<0.001 for all; Fig 11B). At 72 hours, the number of nNOS-IR neurons in preparations of the distal colon from TNBS-only (65±3 cells/area) and UCM-treated (64±2 cells/area) guinea-pigs was higher compared to sham (53±1 cells/area), MSC (54±2 cells/area) and CM-treated (54±2 cells/area) animals (P<0.05 for all; Figs 10AII–10EII and 11A). The proportion of nNOS-IR neurons at 72 hours was greater in colon preparations from TNBS-only (32±2%) and UCM-treated (30±0.5%) guinea-pigs compared to sham (20±0.5%), MSC (21±1%), and CM-treated (21±1%) animals (P<0.001 for all; Fig 11B).

Bottom Line: MSC and CM treatments prevented body weight loss, reduced infiltration of leukocytes into the colon wall and the myenteric plexus, facilitated repair of damaged tissue and nerve fibers, averted myenteric neuronal loss, as well as changes in neuronal subpopulations.The neuroprotective effects of MSC and CM treatments were observed as early as 24 hours after induction of inflammation even though the inflammatory reaction at the level of the myenteric ganglia had not completely subsided.The neuroprotective efficacy of MSC-based therapies can be exerted independently to their anti-inflammatory effects.

View Article: PubMed Central - PubMed

Affiliation: Centre for Chronic Diseases, College of Health and Biomedicine, Victoria University, Melbourne, Australia.

ABSTRACT

Background & aims: The therapeutic benefits of mesenchymal stem cells (MSCs), such as homing ability, multipotent differentiation capacity and secretion of soluble bioactive factors which exert neuroprotective, anti-inflammatory and immunomodulatory properties, have been attributed to attenuation of autoimmune, inflammatory and neurodegenerative disorders. In this study, we aimed to determine the earliest time point at which locally administered MSC-based therapies avert enteric neuronal loss and damage associated with intestinal inflammation in the guinea-pig model of colitis.

Methods: At 3 hours after induction of colitis by 2,4,6-trinitrobenzene-sulfonate (TNBS), guinea-pigs received either human bone marrow-derived MSCs, conditioned medium (CM), or unconditioned medium by enema into the colon. Colon tissues were collected 6, 24 and 72 hours after administration of TNBS. Effects on body weight, gross morphological damage, immune cell infiltration and myenteric neurons were evaluated. RT-PCR, flow cytometry and antibody array kit were used to identify neurotrophic and neuroprotective factors released by MSCs.

Results: MSC and CM treatments prevented body weight loss, reduced infiltration of leukocytes into the colon wall and the myenteric plexus, facilitated repair of damaged tissue and nerve fibers, averted myenteric neuronal loss, as well as changes in neuronal subpopulations. The neuroprotective effects of MSC and CM treatments were observed as early as 24 hours after induction of inflammation even though the inflammatory reaction at the level of the myenteric ganglia had not completely subsided. Substantial number of neurotrophic and neuroprotective factors released by MSCs was identified in their secretome.

Conclusion: MSC-based therapies applied at the acute stages of TNBS-induced colitis start exerting their neuroprotective effects towards enteric neurons by 24 hours post treatment. The neuroprotective efficacy of MSC-based therapies can be exerted independently to their anti-inflammatory effects.

No MeSH data available.


Related in: MedlinePlus