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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 administration increases neurogenesis and reduces reactive astrogliosis at the ischemic boundary in the rat brain.(A) Compared with PBS control treatment, the number of DCX-positive cells at the ischemic boundary significantly increased immediately after NBE-MSC-MV treatment. *P < 0.05, **P < 0.01, ***P < 0.001, n = 5 per group. (B) Representative micrographs of α-SMA-positive vessels in ipsilateral hippocampal vessels at day 7. Compared with PBS control treatment, the number of α-SMA-positive vessels increased in the ipsilateral hippocampal area after NBE-MSC-MV treatment. Data are presented as mean numbers of α-SMA-positive vessels per group. *P < 0.05, n = 5 per group. (C) The number of GFAP-positive cells significantly decreased in response to NBE-MSC-MV transplantation compared with that of PBS control injection at day 7. Data are presented as percent of GFAP-positive area/field. *P < 0.05, n = 5 per group. Scale bar = 200 μm. Values are indicated as means ± SD.
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f3: NBE-MSC-MV administration increases neurogenesis and reduces reactive astrogliosis at the ischemic boundary in the rat brain.(A) Compared with PBS control treatment, the number of DCX-positive cells at the ischemic boundary significantly increased immediately after NBE-MSC-MV treatment. *P < 0.05, **P < 0.01, ***P < 0.001, n = 5 per group. (B) Representative micrographs of α-SMA-positive vessels in ipsilateral hippocampal vessels at day 7. Compared with PBS control treatment, the number of α-SMA-positive vessels increased in the ipsilateral hippocampal area after NBE-MSC-MV treatment. Data are presented as mean numbers of α-SMA-positive vessels per group. *P < 0.05, n = 5 per group. (C) The number of GFAP-positive cells significantly decreased in response to NBE-MSC-MV transplantation compared with that of PBS control injection at day 7. Data are presented as percent of GFAP-positive area/field. *P < 0.05, n = 5 per group. Scale bar = 200 μm. Values are indicated as means ± SD.

Mentions: Previous studies show that ischemic stroke promotes spontaneous neurogenesis in the subventricular zone (SVZ) and subgranular zone of the hippocampus and induces migration of neuroblasts to the ischemic boundary2021. To examine the effect of MSC-MVs treated with brain extract on the proliferation of neural progenitor cells and immature neurons in the ischemic region, sagittal sections of brain tissue were stained for doublecortin (DCX), a marker of migrating neuroblast progenitor cells, 7 days after MV injection. The therapeutic efficacy and key signalling-protein profiles did not significantly differ between NBE-MSC-MVs and SBE-MSC-MVs (see Fig. 1). Therefore, we used NBE-MSC-MVs to perform immunohistochemical analyses and investigate the underlying mechanisms of brain tissue regeneration. The number of DCX-positive migrating neuroblasts in the lateral ventricle area and hippocampal region of stroke-affected hemispheres significantly increased after treatment with NBE-MSC-MVs compared to that of PBS (Fig. 3A), suggesting that NBE-MSC-MVs potentially promote neurogenesis by neural stem cells in the hippocampus and SVZ, and these cells migrate toward the ischemic boundary after stroke. By contrast, only a few DCX-positive cells confined to the SVZ were detected contralateral to the ischemic lesion in untreated rats (data not shown).


Microvesicles from brain-extract — treated mesenchymal stem cells improve neurological functions in a rat model of ischemic stroke
NBE-MSC-MV administration increases neurogenesis and reduces reactive astrogliosis at the ischemic boundary in the rat brain.(A) Compared with PBS control treatment, the number of DCX-positive cells at the ischemic boundary significantly increased immediately after NBE-MSC-MV treatment. *P < 0.05, **P < 0.01, ***P < 0.001, n = 5 per group. (B) Representative micrographs of α-SMA-positive vessels in ipsilateral hippocampal vessels at day 7. Compared with PBS control treatment, the number of α-SMA-positive vessels increased in the ipsilateral hippocampal area after NBE-MSC-MV treatment. Data are presented as mean numbers of α-SMA-positive vessels per group. *P < 0.05, n = 5 per group. (C) The number of GFAP-positive cells significantly decreased in response to NBE-MSC-MV transplantation compared with that of PBS control injection at day 7. Data are presented as percent of GFAP-positive area/field. *P < 0.05, n = 5 per group. Scale bar = 200 μm. Values are indicated as means ± SD.
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getmorefigures.php?uid=PMC5016792&req=5

f3: NBE-MSC-MV administration increases neurogenesis and reduces reactive astrogliosis at the ischemic boundary in the rat brain.(A) Compared with PBS control treatment, the number of DCX-positive cells at the ischemic boundary significantly increased immediately after NBE-MSC-MV treatment. *P < 0.05, **P < 0.01, ***P < 0.001, n = 5 per group. (B) Representative micrographs of α-SMA-positive vessels in ipsilateral hippocampal vessels at day 7. Compared with PBS control treatment, the number of α-SMA-positive vessels increased in the ipsilateral hippocampal area after NBE-MSC-MV treatment. Data are presented as mean numbers of α-SMA-positive vessels per group. *P < 0.05, n = 5 per group. (C) The number of GFAP-positive cells significantly decreased in response to NBE-MSC-MV transplantation compared with that of PBS control injection at day 7. Data are presented as percent of GFAP-positive area/field. *P < 0.05, n = 5 per group. Scale bar = 200 μm. Values are indicated as means ± SD.
Mentions: Previous studies show that ischemic stroke promotes spontaneous neurogenesis in the subventricular zone (SVZ) and subgranular zone of the hippocampus and induces migration of neuroblasts to the ischemic boundary2021. To examine the effect of MSC-MVs treated with brain extract on the proliferation of neural progenitor cells and immature neurons in the ischemic region, sagittal sections of brain tissue were stained for doublecortin (DCX), a marker of migrating neuroblast progenitor cells, 7 days after MV injection. The therapeutic efficacy and key signalling-protein profiles did not significantly differ between NBE-MSC-MVs and SBE-MSC-MVs (see Fig. 1). Therefore, we used NBE-MSC-MVs to perform immunohistochemical analyses and investigate the underlying mechanisms of brain tissue regeneration. The number of DCX-positive migrating neuroblasts in the lateral ventricle area and hippocampal region of stroke-affected hemispheres significantly increased after treatment with NBE-MSC-MVs compared to that of PBS (Fig. 3A), suggesting that NBE-MSC-MVs potentially promote neurogenesis by neural stem cells in the hippocampus and SVZ, and these cells migrate toward the ischemic boundary after stroke. By contrast, only a few DCX-positive cells confined to the SVZ were detected contralateral to the ischemic lesion in untreated rats (data not shown).

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