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Microvesicles from brain-extract — treated mesenchymal stem cells improve neurological functions in a rat model of ischemic stroke

View Article: PubMed Central - PubMed

ABSTRACT

Transplantation of mesenchymal stem cells (MSCs) was reported to improve functional outcomes in a rat model of ischemic stroke, and subsequent studies suggest that MSC-derived microvesicles (MVs) can replace the beneficial effects of MSCs. Here, we evaluated three different MSC-derived MVs, including MVs from untreated MSCs (MSC-MVs), MVs from MSCs treated with normal rat brain extract (NBE-MSC-MVs), and MVs from MSCs treated with stroke-injured rat brain extract (SBE-MSC-MVs), and tested their effects on ischemic brain injury induced by permanent middle cerebral artery occlusion (pMCAO) in rats. NBE-MSC-MVs and SBE-MSC-MVs had significantly greater efficacy than MSC-MVs for ameliorating ischemic brain injury with improved functional recovery. We found similar profiles of key signalling proteins in NBE-MSC-MVs and SBE-MSC-MVs, which account for their similar therapeutic efficacies. Immunohistochemical analyses suggest that brain-extract—treated MSC-MVs reduce inflammation, enhance angiogenesis, and increase endogenous neurogenesis in the rat brain. We performed mass spectrometry proteomic analyses and found that the total proteomes of brain-extract—treated MSC-MVs are highly enriched for known vesicular proteins. Notably, MSC-MV proteins upregulated by brain extracts tend to be modular for tissue repair pathways. We suggest that MSC-MV proteins stimulated by the brain microenvironment are paracrine effectors that enhance MSC therapy for stroke injury.

No MeSH data available.


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NBE-MSC-MV and SBE-MSC-MV injection reduces infarct size in a rat stroke model.(A) Schematic diagram depicting the experimental design of this study. Numerals in the black bar represent the days of experiment. Day −1, time of pMCAO surgery; Day 0, time of MV injection; Day 3 and 7, 3 and 7 days after MV injection. Downward arrows indicate the days for behavioural tests and weight measurement. Upward arrows indicate the days for treatments and brain tissue sampling. (B) Representative TTC-stained brain sections from rats treated with PBS (control), MSC-MVs, NBE-MSC-MVs, and SBE-MSC-MVs show ischemic lesions on day 7 after MV treatment. (C) Bar graph depicting infarct size in all animals. The infarct area in the ipsilateral hemisphere is expressed as a percentage of the contralateral hemisphere area. Values are indicated as means ± SEM. **P < 0.01 compared with controls. Sample size n = 5 for sham control, n = 10 for NBE-MSC-MV and SBE-MSC-MV treatment groups, and n = 7 for MSC-MV treatment group.
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f2: NBE-MSC-MV and SBE-MSC-MV injection reduces infarct size in a rat stroke model.(A) Schematic diagram depicting the experimental design of this study. Numerals in the black bar represent the days of experiment. Day −1, time of pMCAO surgery; Day 0, time of MV injection; Day 3 and 7, 3 and 7 days after MV injection. Downward arrows indicate the days for behavioural tests and weight measurement. Upward arrows indicate the days for treatments and brain tissue sampling. (B) Representative TTC-stained brain sections from rats treated with PBS (control), MSC-MVs, NBE-MSC-MVs, and SBE-MSC-MVs show ischemic lesions on day 7 after MV treatment. (C) Bar graph depicting infarct size in all animals. The infarct area in the ipsilateral hemisphere is expressed as a percentage of the contralateral hemisphere area. Values are indicated as means ± SEM. **P < 0.01 compared with controls. Sample size n = 5 for sham control, n = 10 for NBE-MSC-MV and SBE-MSC-MV treatment groups, and n = 7 for MSC-MV treatment group.

Mentions: Brain infarction area (the ratio of the damaged ipsilateral area to the contralateral hemisphere area) was measured via 2,3,5,-Triphenyl tetrazolium chloride (TTC) staining of the brain on post-treatment day 7. Infarct lesions primarily appeared in the cortex and striatum. The infarction area was demonstrably larger in ischemic brains treated with PBS than in those treated with NBE-MSC-MVs and SBE-MSC-MVs (Fig. 2). The infarction area of PBS-treated (control) rats was 59.19 ± 3.95%. By contrast, the infarction area was significantly reduced in rats treated with NBE-MSC-MVs (42.31 ± 2.80%) or SBE-MSC-MVs (43.00 ± 2.98%) (P < 0.01). A rat group treated with MSC-MVs showed a tendency of reduction in infarction area (45.70 ± 7.95%), but this reduction was not statistically significant for the given sample size. There was no significant difference in lesion volume between groups treated with NBE-MSC-MVs or SBE-MSC-MVs, indicating that there is no significant difference between normal and stroke-injured brain extracts for MV stimulation. These combined results suggest that both normal and injured brain extracts can enhance the therapeutic effects of MSC-MVs by reducing infarction area, and extract from injured brains does not confer greater efficacy than extract from normal brains in promoting the therapeutic effect of MSC-MVs.


Microvesicles from brain-extract — treated mesenchymal stem cells improve neurological functions in a rat model of ischemic stroke
NBE-MSC-MV and SBE-MSC-MV injection reduces infarct size in a rat stroke model.(A) Schematic diagram depicting the experimental design of this study. Numerals in the black bar represent the days of experiment. Day −1, time of pMCAO surgery; Day 0, time of MV injection; Day 3 and 7, 3 and 7 days after MV injection. Downward arrows indicate the days for behavioural tests and weight measurement. Upward arrows indicate the days for treatments and brain tissue sampling. (B) Representative TTC-stained brain sections from rats treated with PBS (control), MSC-MVs, NBE-MSC-MVs, and SBE-MSC-MVs show ischemic lesions on day 7 after MV treatment. (C) Bar graph depicting infarct size in all animals. The infarct area in the ipsilateral hemisphere is expressed as a percentage of the contralateral hemisphere area. Values are indicated as means ± SEM. **P < 0.01 compared with controls. Sample size n = 5 for sham control, n = 10 for NBE-MSC-MV and SBE-MSC-MV treatment groups, and n = 7 for MSC-MV treatment group.
© Copyright Policy - open-access
Related In: Results  -  Collection

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f2: NBE-MSC-MV and SBE-MSC-MV injection reduces infarct size in a rat stroke model.(A) Schematic diagram depicting the experimental design of this study. Numerals in the black bar represent the days of experiment. Day −1, time of pMCAO surgery; Day 0, time of MV injection; Day 3 and 7, 3 and 7 days after MV injection. Downward arrows indicate the days for behavioural tests and weight measurement. Upward arrows indicate the days for treatments and brain tissue sampling. (B) Representative TTC-stained brain sections from rats treated with PBS (control), MSC-MVs, NBE-MSC-MVs, and SBE-MSC-MVs show ischemic lesions on day 7 after MV treatment. (C) Bar graph depicting infarct size in all animals. The infarct area in the ipsilateral hemisphere is expressed as a percentage of the contralateral hemisphere area. Values are indicated as means ± SEM. **P < 0.01 compared with controls. Sample size n = 5 for sham control, n = 10 for NBE-MSC-MV and SBE-MSC-MV treatment groups, and n = 7 for MSC-MV treatment group.
Mentions: Brain infarction area (the ratio of the damaged ipsilateral area to the contralateral hemisphere area) was measured via 2,3,5,-Triphenyl tetrazolium chloride (TTC) staining of the brain on post-treatment day 7. Infarct lesions primarily appeared in the cortex and striatum. The infarction area was demonstrably larger in ischemic brains treated with PBS than in those treated with NBE-MSC-MVs and SBE-MSC-MVs (Fig. 2). The infarction area of PBS-treated (control) rats was 59.19 ± 3.95%. By contrast, the infarction area was significantly reduced in rats treated with NBE-MSC-MVs (42.31 ± 2.80%) or SBE-MSC-MVs (43.00 ± 2.98%) (P < 0.01). A rat group treated with MSC-MVs showed a tendency of reduction in infarction area (45.70 ± 7.95%), but this reduction was not statistically significant for the given sample size. There was no significant difference in lesion volume between groups treated with NBE-MSC-MVs or SBE-MSC-MVs, indicating that there is no significant difference between normal and stroke-injured brain extracts for MV stimulation. These combined results suggest that both normal and injured brain extracts can enhance the therapeutic effects of MSC-MVs by reducing infarction area, and extract from injured brains does not confer greater efficacy than extract from normal brains in promoting the therapeutic effect of MSC-MVs.

View Article: PubMed Central - PubMed

ABSTRACT

Transplantation of mesenchymal stem cells (MSCs) was reported to improve functional outcomes in a rat model of ischemic stroke, and subsequent studies suggest that MSC-derived microvesicles (MVs) can replace the beneficial effects of MSCs. Here, we evaluated three different MSC-derived MVs, including MVs from untreated MSCs (MSC-MVs), MVs from MSCs treated with normal rat brain extract (NBE-MSC-MVs), and MVs from MSCs treated with stroke-injured rat brain extract (SBE-MSC-MVs), and tested their effects on ischemic brain injury induced by permanent middle cerebral artery occlusion (pMCAO) in rats. NBE-MSC-MVs and SBE-MSC-MVs had significantly greater efficacy than MSC-MVs for ameliorating ischemic brain injury with improved functional recovery. We found similar profiles of key signalling proteins in NBE-MSC-MVs and SBE-MSC-MVs, which account for their similar therapeutic efficacies. Immunohistochemical analyses suggest that brain-extract&mdash;treated MSC-MVs reduce inflammation, enhance angiogenesis, and increase endogenous neurogenesis in the rat brain. We performed mass spectrometry proteomic analyses and found that the total proteomes of brain-extract&mdash;treated MSC-MVs are highly enriched for known vesicular proteins. Notably, MSC-MV proteins upregulated by brain extracts tend to be modular for tissue repair pathways. We suggest that MSC-MV proteins stimulated by the brain microenvironment are paracrine effectors that enhance MSC therapy for stroke injury.

No MeSH data available.


Related in: MedlinePlus