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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.


Related in: MedlinePlus

Transplantation of NBE-MSC-MVs and SBE-MSC-MVs improves neurological outcomes.(A) Analysis of body weight changes after injection of MSC-MVs, NBE-MSC-MVs, or SBE-MSC-MVs. Loss of body weight was significantly attenuated in rat groups treated with NBE-MSC-MVs (P < 0.011, P < 0.02) or SBE-MSC-MVs (P < 0.025, P < 0.05) compared with a group treated with PBS at day 3 and 7, respectively. (B) Torso-twisting test (left and right side). Score change from PBS group for right side was significant for NBE-MSC-MVs (P < 0.04) at day 3, and for both NBE-MSC-MVs (P < 0.025) and SBE-MSC-MVs (P < 0.03) at day 7. (C) Open field test (line cross score). Score change from PBS group was significant for NBE-MSC-MVs (P < 0.025) and SBE-MSC-MVs (P < 0.03) at day 3. (D) Beam balance test. All injection groups (MSC-MVs, NBE-MSC-MVs, and SBE-MSC-MVs) showed significant improvement compared with the PBS group at day 3 (P < 0.01, P < 0.001, P < 0.001, respectively) and day 7 (P < 0.012, P < 0.0001, P < 0.04, respectively). (E) Prehensile traction test. The score was significantly increased in animals treated with MSC-MVs (P < 0.0002) and NBE-MSC-MVs (P < 0.05) compared with that of the PBS control group at day 3, and in NBE-MSC-MVs (P < 0.005) at day 7. (F) Modified neurological severity scores (mNSS) indicate that animals treated with NBE-MSC-MVs and SBE-MSC-MVs displayed significant functional enhancement at day 3 (P < 0.0001 and P < 0.002, respectively) compared with the PBS control group, but animals treated with MSC-MVs did not. At 7 days after injection, animals treated with MSC-MVs, NBE-MSC-MVs, and SBE-MSC-MVs showed significant functional enhancement compared with that of the PBS group (P < 0.002, P < 0.0001, and P < 0.01, respectively). *P < 0.05; **P < 0.01, ***P < 0.001; n = 10 per group; Day −1, time of pMCAO surgery; Day 0, time of MV injection; Day 3 and 7, 3 and 7 days after MV injection, respectively. Sample sizes are n = 5 for sham-operated, n = 10 for NBE-MSC-MV and SBE-MSC-MV treatment groups, and n = 7 for MSC-MV treatment group.
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f1: Transplantation of NBE-MSC-MVs and SBE-MSC-MVs improves neurological outcomes.(A) Analysis of body weight changes after injection of MSC-MVs, NBE-MSC-MVs, or SBE-MSC-MVs. Loss of body weight was significantly attenuated in rat groups treated with NBE-MSC-MVs (P < 0.011, P < 0.02) or SBE-MSC-MVs (P < 0.025, P < 0.05) compared with a group treated with PBS at day 3 and 7, respectively. (B) Torso-twisting test (left and right side). Score change from PBS group for right side was significant for NBE-MSC-MVs (P < 0.04) at day 3, and for both NBE-MSC-MVs (P < 0.025) and SBE-MSC-MVs (P < 0.03) at day 7. (C) Open field test (line cross score). Score change from PBS group was significant for NBE-MSC-MVs (P < 0.025) and SBE-MSC-MVs (P < 0.03) at day 3. (D) Beam balance test. All injection groups (MSC-MVs, NBE-MSC-MVs, and SBE-MSC-MVs) showed significant improvement compared with the PBS group at day 3 (P < 0.01, P < 0.001, P < 0.001, respectively) and day 7 (P < 0.012, P < 0.0001, P < 0.04, respectively). (E) Prehensile traction test. The score was significantly increased in animals treated with MSC-MVs (P < 0.0002) and NBE-MSC-MVs (P < 0.05) compared with that of the PBS control group at day 3, and in NBE-MSC-MVs (P < 0.005) at day 7. (F) Modified neurological severity scores (mNSS) indicate that animals treated with NBE-MSC-MVs and SBE-MSC-MVs displayed significant functional enhancement at day 3 (P < 0.0001 and P < 0.002, respectively) compared with the PBS control group, but animals treated with MSC-MVs did not. At 7 days after injection, animals treated with MSC-MVs, NBE-MSC-MVs, and SBE-MSC-MVs showed significant functional enhancement compared with that of the PBS group (P < 0.002, P < 0.0001, and P < 0.01, respectively). *P < 0.05; **P < 0.01, ***P < 0.001; n = 10 per group; Day −1, time of pMCAO surgery; Day 0, time of MV injection; Day 3 and 7, 3 and 7 days after MV injection, respectively. Sample sizes are n = 5 for sham-operated, n = 10 for NBE-MSC-MV and SBE-MSC-MV treatment groups, and n = 7 for MSC-MV treatment group.

Mentions: In response to tissue injury, tissue repair and regeneration signals may be delivered to and sensed by resident stem cells. These stem cells then generate MVs, which are paracrine mediators that can reprogram the fate of other local stem cells19. Systemic or local administration of MSCs provides functional benefits in a number of animal stroke models. We hypothesised that MVs from MSCs significantly contribute to the observed benefits of MSC administration, and signalling molecules secreted from a damaged microenvironment could enhance the therapeutic effect of MSC-MVs. To test these hypotheses, we analysed the therapeutic effect of NBE-MSC-MVs, SBE-MSC-MVs, and untreated MSC-MVs. To examine the therapeutic effects of these three MV preparations, we induced a pMCAO model of the ischemic brain in rats. A single injection of MVs (0.2 mg/kg rat body weight) of NBE-MSC-MVs, SBE-MSC-MVs, and MSC-MVs was administered 48 h after the induction of pMCAO via the right common carotid artery. After induction of ischemic stroke, body weight change was observed in all the rats. Loss of body weight was significantly attenuated in groups treated with NBE-MSC-MVs or SBE-MSC-MVs compared with that of a control group treated with phosphate buffered saline (PBS) on days 3 and 7, but was not significantly different than that of rats treated with MSC-MVs (Fig. 1A).


Microvesicles from brain-extract — treated mesenchymal stem cells improve neurological functions in a rat model of ischemic stroke
Transplantation of NBE-MSC-MVs and SBE-MSC-MVs improves neurological outcomes.(A) Analysis of body weight changes after injection of MSC-MVs, NBE-MSC-MVs, or SBE-MSC-MVs. Loss of body weight was significantly attenuated in rat groups treated with NBE-MSC-MVs (P < 0.011, P < 0.02) or SBE-MSC-MVs (P < 0.025, P < 0.05) compared with a group treated with PBS at day 3 and 7, respectively. (B) Torso-twisting test (left and right side). Score change from PBS group for right side was significant for NBE-MSC-MVs (P < 0.04) at day 3, and for both NBE-MSC-MVs (P < 0.025) and SBE-MSC-MVs (P < 0.03) at day 7. (C) Open field test (line cross score). Score change from PBS group was significant for NBE-MSC-MVs (P < 0.025) and SBE-MSC-MVs (P < 0.03) at day 3. (D) Beam balance test. All injection groups (MSC-MVs, NBE-MSC-MVs, and SBE-MSC-MVs) showed significant improvement compared with the PBS group at day 3 (P < 0.01, P < 0.001, P < 0.001, respectively) and day 7 (P < 0.012, P < 0.0001, P < 0.04, respectively). (E) Prehensile traction test. The score was significantly increased in animals treated with MSC-MVs (P < 0.0002) and NBE-MSC-MVs (P < 0.05) compared with that of the PBS control group at day 3, and in NBE-MSC-MVs (P < 0.005) at day 7. (F) Modified neurological severity scores (mNSS) indicate that animals treated with NBE-MSC-MVs and SBE-MSC-MVs displayed significant functional enhancement at day 3 (P < 0.0001 and P < 0.002, respectively) compared with the PBS control group, but animals treated with MSC-MVs did not. At 7 days after injection, animals treated with MSC-MVs, NBE-MSC-MVs, and SBE-MSC-MVs showed significant functional enhancement compared with that of the PBS group (P < 0.002, P < 0.0001, and P < 0.01, respectively). *P < 0.05; **P < 0.01, ***P < 0.001; n = 10 per group; Day −1, time of pMCAO surgery; Day 0, time of MV injection; Day 3 and 7, 3 and 7 days after MV injection, respectively. Sample sizes are n = 5 for sham-operated, n = 10 for NBE-MSC-MV and SBE-MSC-MV treatment groups, and n = 7 for MSC-MV treatment group.
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f1: Transplantation of NBE-MSC-MVs and SBE-MSC-MVs improves neurological outcomes.(A) Analysis of body weight changes after injection of MSC-MVs, NBE-MSC-MVs, or SBE-MSC-MVs. Loss of body weight was significantly attenuated in rat groups treated with NBE-MSC-MVs (P < 0.011, P < 0.02) or SBE-MSC-MVs (P < 0.025, P < 0.05) compared with a group treated with PBS at day 3 and 7, respectively. (B) Torso-twisting test (left and right side). Score change from PBS group for right side was significant for NBE-MSC-MVs (P < 0.04) at day 3, and for both NBE-MSC-MVs (P < 0.025) and SBE-MSC-MVs (P < 0.03) at day 7. (C) Open field test (line cross score). Score change from PBS group was significant for NBE-MSC-MVs (P < 0.025) and SBE-MSC-MVs (P < 0.03) at day 3. (D) Beam balance test. All injection groups (MSC-MVs, NBE-MSC-MVs, and SBE-MSC-MVs) showed significant improvement compared with the PBS group at day 3 (P < 0.01, P < 0.001, P < 0.001, respectively) and day 7 (P < 0.012, P < 0.0001, P < 0.04, respectively). (E) Prehensile traction test. The score was significantly increased in animals treated with MSC-MVs (P < 0.0002) and NBE-MSC-MVs (P < 0.05) compared with that of the PBS control group at day 3, and in NBE-MSC-MVs (P < 0.005) at day 7. (F) Modified neurological severity scores (mNSS) indicate that animals treated with NBE-MSC-MVs and SBE-MSC-MVs displayed significant functional enhancement at day 3 (P < 0.0001 and P < 0.002, respectively) compared with the PBS control group, but animals treated with MSC-MVs did not. At 7 days after injection, animals treated with MSC-MVs, NBE-MSC-MVs, and SBE-MSC-MVs showed significant functional enhancement compared with that of the PBS group (P < 0.002, P < 0.0001, and P < 0.01, respectively). *P < 0.05; **P < 0.01, ***P < 0.001; n = 10 per group; Day −1, time of pMCAO surgery; Day 0, time of MV injection; Day 3 and 7, 3 and 7 days after MV injection, respectively. Sample sizes are n = 5 for sham-operated, n = 10 for NBE-MSC-MV and SBE-MSC-MV treatment groups, and n = 7 for MSC-MV treatment group.
Mentions: In response to tissue injury, tissue repair and regeneration signals may be delivered to and sensed by resident stem cells. These stem cells then generate MVs, which are paracrine mediators that can reprogram the fate of other local stem cells19. Systemic or local administration of MSCs provides functional benefits in a number of animal stroke models. We hypothesised that MVs from MSCs significantly contribute to the observed benefits of MSC administration, and signalling molecules secreted from a damaged microenvironment could enhance the therapeutic effect of MSC-MVs. To test these hypotheses, we analysed the therapeutic effect of NBE-MSC-MVs, SBE-MSC-MVs, and untreated MSC-MVs. To examine the therapeutic effects of these three MV preparations, we induced a pMCAO model of the ischemic brain in rats. A single injection of MVs (0.2 mg/kg rat body weight) of NBE-MSC-MVs, SBE-MSC-MVs, and MSC-MVs was administered 48 h after the induction of pMCAO via the right common carotid artery. After induction of ischemic stroke, body weight change was observed in all the rats. Loss of body weight was significantly attenuated in groups treated with NBE-MSC-MVs or SBE-MSC-MVs compared with that of a control group treated with phosphate buffered saline (PBS) on days 3 and 7, but was not significantly different than that of rats treated with MSC-MVs (Fig. 1A).

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