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. 2020 Dec 19;63:103161. doi: 10.1016/j.ebiom.2020.103161

Fig. 1.

Fig. 1

IP administration of hPMSC protects against ischemic injury in the MCAO stroke model.

(a) The area of infarction (‘white’ area) in each brain slice was visualized using TTC staining and measured using Image-J (NIH). Unstained (‘white’) regions in brain slices were combined to generate a composite tissue injury score for each brain. Each point represents one animal (sham (n = 7), MCAO (n = 6), MCAO+hPMSC (n = 8)). Compared to sham, there was a significant increase in infarct size in MCAO mice (⁎⁎⁎⁎p<0.0001, one-way ANOVA). By comparison infarct size in hPMSC-treated MCAO mice was significantly reduced (⁎⁎⁎⁎p<0.0001, Student's t-test).

(b) Total cerebral perfusion measured using Laser Speckle imaging. ‘Perfusion’ reflects total cerebral flow signal measured in selected tissue areas. Measurements are expressed as perfusion units (PU), using a fixed scale (arbitrary units). Significant differences were observed between cerebral perfusion in MCAO (n = 6) and sham (n = 6) mice (⁎⁎⁎⁎p<0.0001, one-way ANOVA). hPMSC-treated MCAO (n = 7) mice showed significant preservation of perfusion compared to non-treated post-MCAO group (⁎⁎⁎⁎P<0.0001, Student's t-test).

(c, d) Cerebral perfusion in each pair of ipsilateral (c) and contralateral (d) brain hemispheres were normalized to the average sham total CBF as the reference point.

(c) A large (83%) decrease in cerebral perfusion of the ipsilateral side of MCAO brain was observed compared to the sham operated group (⁎⁎⁎⁎p<0.0001, one-way ANOVA);

(d) A significant decrease (65%) in cerebral perfusion of the contralateral hemisphere of MCAO brain was observed compared to the sham group (⁎⁎⁎⁎p<0.0001, one-way ANOVA). hPMSC treatment significantly preserved blood flow normal distribution between the hemispheres (far right bars; c, d); significant differences determined by Student's t-test, ⁎⁎⁎⁎p<0.0001 and ⁎⁎⁎⁎p<0.0001 for comparison of hPMSC-treated MCAO mice to untreated MCAO ipsilateral (c) / contralateral (d) hemispheres, respectively.

(e) Neurological scores 24 h post-reperfusion. Significant differences in neurological scores were seen between untreated MCAO (n = 8) and hPMSC-treated MCAO (n = 8) to sham (n = 8) group (⁎⁎⁎⁎p<0.0001, one-way ANOVA). There was a significant improvement in neurological score in hPMSC-treated MCAO mice compared to untreated MCAO (⁎⁎⁎⁎p<0.0001, Student's t-test).

(f) Neuronal degeneration in MCAO with hPMSC therapy. Neurons (black arrows), degenerating neurons (black arrows head), glial cells (red arrows head), and spongiform regions (red arrows) were visualized using Nissl staining of brain sections in untreated and hPMSC-treated MCAO groups.

Two-tailed Student's t-test analysis revealed significant differences in proportions of (g) neurons (*p = 0.02), (h) degenerating neurons (*p = 0.04), and (i) glial cell (*p = 0.02) numbers in hPMSC-treated MCAO brain sections compared to untreated MCAO sections. In all graphs, data represent means ± SEM. (j) Representative images after immunohistochemistry staining for Iba-1.

(h) Quantification of numbers of Iba-1+ microglia in the border between striatum and cortex (average of 3 different fields) of ipsilateral hemisphere of MCAO and MCAO+hPMSC groups (NS, p = 0.5, Student's t-test, n = 4 per group).

(i) Microglial activation in ipsilateral hemispheres of MCAO and MCAO+hPMSC groups was determined by cell body length (⁎⁎p = 0.002, Student's t-test). (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)