CoChat
Sign Up
Cited by 132

The DOI matches its Crossref record; 7 of the 7 references it cites were found in a registry or an index.

Neural stem cell-derived exosome as a nano-sized carrier for BDNF delivery to a rat model of ischemic stroke

Zhu, Zhi-Han, Jia, Feng, Ahmed, Waqas, Zhang, Gui-Long, Wang, Hong, Lin, Chao-Qun, Chen, Wang-Hao, Chen, Lu-Kui·2023·Neural Regeneration Research
DOI 10.4103/1673-5374.346466PMC PMC9396474
Open this paper in CoChat — free account

Abstract

JOURNAL/nrgr/04.03/01300535-202302000-00042/figure1/v/2026-05-26T054653Z/r/image-tiff Our previous study demonstrated the potential therapeutic role of human neural stem cell-derived exosomes (hNSC-Exo) in ischemic stroke. Here, we loaded brain-derived neurotrophic factor (BDNF) into exosomes derived from NSCs to construct engineered exosomes (BDNF-hNSC-Exo) and compared their effects with those of hNSC-Exo on ischemic stroke both in vitro and in vivo . In a model of H 2 O 2 -induced oxidative stress in NSCs, BDNF-hNSC-Exo markedly enhanced cell survival. In a rat middle cerebral artery occlusion model, BDNF-hNSC-Exo not only inhibited the activation of microglia, but also promoted the differentiation of endogenous NSCs into neurons. These results suggest that BDNF can improve the function of NSC-derived exosomes in the treatment of ischemic stroke. Our research may support the clinical use of other neurotrophic factors for central nervous system diseases.

Overview

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.4103/1673-5374.346466

Cites

7 of 7 references verified0 citation errors · 0 not checkable

Statistics

Figures

Figure 1 The features of hNSCs and engineered exosomes (Exo). (A) Immunofluorescence showed the expression of Nestin (red), Sox2 (green), MOG (red), Tuj1 (red), and GFAP (green). Nuclei were stained by DAPI (blue). Scale bars: 100 μm in the MOG panel; 50 μm in the Tuj1 panel; 20 μm in Nestin, Sox2 and GFAP panels. (B) hNSC-Exo and BDNF-hNSC-Exo showed cup-shaped exosome morphology under TEM. Scale bars: 200 μm. (C) NTA analysis indicated a similar size range for hNSC-Exo and BDNF-hNSC-Exo. (D) Western blot analysis of exosomal marker proteins including CD81, HSP70, and TSG101 and calnexin (as a negative control). (E) Representative micrographs of exosome uptake in NSCs incubated with PKH67-labeled exosomes (green) for 48 hours. Nuclei were stained by DAPI (blue). Scale bars: 20 μm. BDNF: Brain-derived neurotrophic factor; GFAP: glial fibrillary acidic protein; hNSC: human neural stem cell; HSP70: heat shock protein 70; MOG: myelin oligodendrocyte glycoprotein; TSG101: tumor susceptibility 101; Tuj1: β-tubulin III.
Figure 1 The features of hNSCs and engineered exosomes (Exo). (A) Immunofluorescence showed the expression of Nestin (red), Sox2 (green), MOG (red), Tuj1 (red), and GFAP (green). Nuclei were stained by DAPI (blue). Scale bars: 100 μm in the MOG panel; 50 μm in the Tuj1 panel; 20 μm in Nestin, Sox2 and GFAP panels. (B) hNSC-Exo and BDNF-hNSC-Exo showed cup-shaped exosome morphology under TEM. Scale bars: 200 μm. (C) NTA analysis indicated a similar size range for hNSC-Exo and BDNF-hNSC-Exo. (D) Western blot analysis of exosomal marker proteins including CD81, HSP70, and TSG101 and calnexin (as a negative control). (E) Representative micrographs of exosome uptake in NSCs incubated with PKH67-labeled exosomes (green) for 48 hours. Nuclei were stained by DAPI (blue). Scale bars: 20 μm. BDNF: Brain-derived neurotrophic factor; GFAP: glial fibrillary acidic protein; hNSC: human neural stem cell; HSP70: heat shock protein 70; MOG: myelin oligodendrocyte glycoprotein; TSG101: tumor susceptibility 101; Tuj1: β-tubulin III.CC BY-NC-SA · PubMed Central
Figure 2 BDNF-hNSC-Exo promotes the survival and differentiation of neural stem cells in vitro. (A) The expression level of BDNF was measured by western blotting after the addition of BDNF-hNSC-Exo to cultured neural stem cells. Data were normalized to β-actin expression. (B) The differentiation efficiency of BDNF-hNSC-Exo was evaluated using quantitative reverse transcription-polymerase chain reaction. Data were normalized to GAPDH mRNA expression. Cells were treated with a 500 μM solution H2O2 for 5 hours. (C) Western blot analysis of Tuj1 in neural stem cells. Cells were treated with a 500 μM solution H2O2 for 5 hours. Data were normalized to GAPDH expression. (D) Immunofluorescence showed that the ratio of Tuj1-positive cells (red) in the BDNF-hNSC-Exo group was higher than that in the control group. The percentage of Tuj1-positive cells was obtained by counting cells in three visual fields. Cells were treated with a 500 μM solution H2O2 for 5 hours. Nuclei were stained by DAPI (blue). Scale bars: 50 μm. (E) BDNF-hNSC-Exo remarkably increased cell survival in the H2O2 stress model (CCK8 assay). Cells were treated with a 500 μM solution H2O2 for 5 hours. (F) TUNEL-positive cells. The proportion of TUNEL-positive cells in the BDNF-hNSC-Exo group was remarkably lower than that in the PBS group. Cells were treated with a 500 μM solution H2O2 for 5 hours. Nuclei were stained by DAPI (blue). Scale bars: 50 μm. (G) Western blotting showed that the expressions of apoptosis-related proteins cleaved-caspase-3 and Bax were significantly decreased in the BDNF-hNSC-Exo group compared with the other two groups. Cells were treated with a 500 μM solution H2O2 for 5 hours. Data were normalized to GAPDH expression. All experiments were conducted in triplicate. All data are presented as mean ± SEM. *P < 0.05 (one-way analysis of variance followed by Tukey’s post hoc test). Bax: Bcl2 associated X, apoptosis regulator; BDNF: brain-derived neurotrophic factor; Exo: exosomes; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; hNSC: human neural stem cell; Tuj1: β-tubulin III.
Figure 2 BDNF-hNSC-Exo promotes the survival and differentiation of neural stem cells in vitro. (A) The expression level of BDNF was measured by western blotting after the addition of BDNF-hNSC-Exo to cultured neural stem cells. Data were normalized to β-actin expression. (B) The differentiation efficiency of BDNF-hNSC-Exo was evaluated using quantitative reverse transcription-polymerase chain reaction. Data were normalized to GAPDH mRNA expression. Cells were treated with a 500 μM solution H2O2 for 5 hours. (C) Western blot analysis of Tuj1 in neural stem cells. Cells were treated with a 500 μM solution H2O2 for 5 hours. Data were normalized to GAPDH expression. (D) Immunofluorescence showed that the ratio of Tuj1-positive cells (red) in the BDNF-hNSC-Exo group was higher than that in the control group. The percentage of Tuj1-positive cells was obtained by counting cells in three visual fields. Cells were treated with a 500 μM solution H2O2 for 5 hours. Nuclei were stained by DAPI (blue). Scale bars: 50 μm. (E) BDNF-hNSC-Exo remarkably increased cell survival in the H2O2 stress model (CCK8 assay). Cells were treated with a 500 μM solution H2O2 for 5 hours. (F) TUNEL-positive cells. The proportion of TUNEL-positive cells in the BDNF-hNSC-Exo group was remarkably lower than that in the PBS group. Cells were treated with a 500 μM solution H2O2 for 5 hours. Nuclei were stained by DAPI (blue). Scale bars: 50 μm. (G) Western blotting showed that the expressions of apoptosis-related proteins cleaved-caspase-3 and Bax were significantly decreased in the BDNF-hNSC-Exo group compared with the other two groups. Cells were treated with a 500 μM solution H2O2 for 5 hours. Data were normalized to GAPDH expression. All experiments were conducted in triplicate. All data are presented as mean ± SEM. *P < 0.05 (one-way analysis of variance followed by Tukey’s post hoc test). Bax: Bcl2 associated X, apoptosis regulator; BDNF: brain-derived neurotrophic factor; Exo: exosomes; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; hNSC: human neural stem cell; Tuj1: β-tubulin III.CC BY-NC-SA · PubMed Central
Figure 3 BDNF-hNSC-Exo improves neurological deficits and brain damage in rats following cerebral ischemia. (A) Experimental design. (B) BDNF-hNSC-Exo increases the expression of BDNF in the infarct areas. Data were normalized to GAPDH expression. (C–E) Evaluation of the behavioral function of MCAO rats at 1, 3, 7, 14, and 28 days after BDNF-hNSC-Exo treatment by rotarod, postural reflex, and mNSS tests. n = 7–10 rats/group. (F) TTC staining showed that BDNF-hNSC-Exo significantly reduced the volume of cerebral infarction. White indicates infact region. n = 3 rats/group. All data are mean ± SEM. *P < 0.05 (one-way analysis of variance followed by Tukey’s post hoc test). BDNF: Brain-derived neurotrophic factor; Exo: exosomes; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; hNSC: human neural stem cell; MCAO: middle cerebral artery occlusion; mNSS: modified neurological severity score; PBS: phosphate-buffered saline; TTC: 2,3,5-triphenyltetrazolium chloride.
Figure 3 BDNF-hNSC-Exo improves neurological deficits and brain damage in rats following cerebral ischemia. (A) Experimental design. (B) BDNF-hNSC-Exo increases the expression of BDNF in the infarct areas. Data were normalized to GAPDH expression. (C–E) Evaluation of the behavioral function of MCAO rats at 1, 3, 7, 14, and 28 days after BDNF-hNSC-Exo treatment by rotarod, postural reflex, and mNSS tests. n = 7–10 rats/group. (F) TTC staining showed that BDNF-hNSC-Exo significantly reduced the volume of cerebral infarction. White indicates infact region. n = 3 rats/group. All data are mean ± SEM. *P < 0.05 (one-way analysis of variance followed by Tukey’s post hoc test). BDNF: Brain-derived neurotrophic factor; Exo: exosomes; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; hNSC: human neural stem cell; MCAO: middle cerebral artery occlusion; mNSS: modified neurological severity score; PBS: phosphate-buffered saline; TTC: 2,3,5-triphenyltetrazolium chloride.CC BY-NC-SA · PubMed Central
Figure 4 BDNF-hNSC-Exo inhibit neuroinflammation and promote neurogenesis in the peri-infarct zone of rats. (A) Immunofluorescence double-labeling demonstrated that more functional neurons in the peri-infarct area were generated in the BDNF-hNSC-Exo and hNSC-Exo groups than in the PBS group. BrdU (red) co-localized with Tuj1 (green). Nuclei were stained by DAPI (blue). Scale bar: 20 μm. n = 5 rats/group. (B) Double immunofluorescence staining showed that in the BDNF-hNSC-Exo group, the proportion of BrdU/GFAP double-positive cells in the peri-infarct area was lower than that in the hNSC-Exo group and the PBS group on day 28 after treatment. BrdU (red) co-localized with GFAP (green). Nuclei were stained by DAPI (blue). Scale bar: 20 μm. n = 5 rats/group. (C) Immunofluorescence showed that BDNF-hNSC-Exo significantly reduced the expression of Iba1 (red), indicating reduced neuroinflammation. Nuclei were stained by DAPI (blue). Scale bar: 20 μm. n = 5 rats/group. All data are shown as the mean ± SEM. *P < 0.05 (one-way analysis of variance followed by Tukey’s post hoc test). BDNF: Brain-derived neurotrophic factor; BrdU: bromodeoxyuridine; Exo: exosomes; GFAP: glial fibrillary acidic protein; hNSC: human neural stem cell; Iba1: induction of brown adipocytes 1; MCAO: middle cerebral artery occlusion; PBS: phosphate-buffered saline; Tuj1: β-tubulin.
Figure 4 BDNF-hNSC-Exo inhibit neuroinflammation and promote neurogenesis in the peri-infarct zone of rats. (A) Immunofluorescence double-labeling demonstrated that more functional neurons in the peri-infarct area were generated in the BDNF-hNSC-Exo and hNSC-Exo groups than in the PBS group. BrdU (red) co-localized with Tuj1 (green). Nuclei were stained by DAPI (blue). Scale bar: 20 μm. n = 5 rats/group. (B) Double immunofluorescence staining showed that in the BDNF-hNSC-Exo group, the proportion of BrdU/GFAP double-positive cells in the peri-infarct area was lower than that in the hNSC-Exo group and the PBS group on day 28 after treatment. BrdU (red) co-localized with GFAP (green). Nuclei were stained by DAPI (blue). Scale bar: 20 μm. n = 5 rats/group. (C) Immunofluorescence showed that BDNF-hNSC-Exo significantly reduced the expression of Iba1 (red), indicating reduced neuroinflammation. Nuclei were stained by DAPI (blue). Scale bar: 20 μm. n = 5 rats/group. All data are shown as the mean ± SEM. *P < 0.05 (one-way analysis of variance followed by Tukey’s post hoc test). BDNF: Brain-derived neurotrophic factor; BrdU: bromodeoxyuridine; Exo: exosomes; GFAP: glial fibrillary acidic protein; hNSC: human neural stem cell; Iba1: induction of brown adipocytes 1; MCAO: middle cerebral artery occlusion; PBS: phosphate-buffered saline; Tuj1: β-tubulin.CC BY-NC-SA · PubMed Central

Verification

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Citations

Cites

Research works printed but not linked: 35

  1. Ahmad, 2022, Novel therapeutic interventions for combating Parkinson's disease and prospects of Nose-to-Brain drug delivery, Biochem Pharmacol, 195, 114849, 10.1016/j.bcp.2021.114849 · DOI
  2. Arumugam, 2006, Gamma secretase-mediated Notch signaling worsens brain damage and functional outcome in ischemic stroke, Nat Med, 12, 621, 10.1038/nm1403 · DOI
  3. Belayev, 2017, Neuroprotectin D1 upregulates Iduna expression and provides protection in cellular uncompensated oxidative stress and in experimental ischemic stroke, Cell Death Differ, 24, 1091, 10.1038/cdd.2017.55 · DOI
  4. Chen, 2020, Exosome-shuttled circSHOC2 from IPASs regulates neuronal autophagy and ameliorates ischemic brain injury via the miR-7670-3p/SIRT1 axis, Mol Ther Nucleic Acids, 22, 657, 10.1016/j.omtn.2020.09.027 · DOI
  5. Fang, 2020, Small extracellular vesicles derived from human mesenchymal stromal cells prevent group 2 innate lymphoid cell-dominant allergic airway inflammation through delivery of miR-146a-5p, J Extracell Vesicles, 9, 1723260, 10.1080/20013078.2020.1723260 · DOI
  6. Feng, 2021, Emerging exosomes and exosomal miRNAs in spinal cord injury, Front Cell Dev Biol, 9, 703989, 10.3389/fcell.2021.703989 · DOI
  7. Fu, 2019, CAR exosomes derived from effector CAR-T cells have potent antitumour effects and low toxicity, Nat Commun, 10, 4355, 10.1038/s41467-019-12321-3 · DOI
  8. Goldstein, 2007, Acute ischemic stroke treatment in 2007, Circulation, 116, 1504, 10.1161/CIRCULATIONAHA.106.670885 · DOI
  9. Hamblin, 2022, Modulation of gene expression on a transcriptome-wide level following human neural stem cell transplantation in aged mouse stroke brains, Exp Neurol, 347, 113913, 10.1016/j.expneurol.2021.113913 · DOI
  10. Hao, 2021, Memantine, NMDA receptor antagonist, attenuates ox-LDL-induced inflammation and oxidative stress via activation of BDNF/TrkB signaling pathway in HUVECs, Inflammation, 44, 659, 10.1007/s10753-020-01365-z · DOI
  11. Huang, 2021, A conductive dual-network hydrogel composed of oxidized dextran and hyaluronic-hydrazide as BDNF delivery systems for potential spinal cord injury repair, Int J Biol Macromol, 167, 434, 10.1016/j.ijbiomac.2020.11.206 · DOI
  12. Hughes, 2020, Association of blood pressure lowering with incident dementia or cognitive impairment:a systematic review and meta-analysis, Jama, 323, 1934, 10.1001/jama.2020.4249 · DOI · Paper page
  13. Kim, 2020, Exosomes:cell-derived nanoplatforms for the delivery of cancer therapeutics, Int J Mol Sci, 22, 14, 10.3390/ijms22010014 · DOI
  14. Lai, 2020, Restoration of L-OPA1 alleviates acute ischemic stroke injury in rats via inhibiting neuronal apoptosis and preserving mitochondrial function, Redox Biol, 34, 101503, 10.1016/j.redox.2020.101503 · DOI
  15. Leira, 2020, Preserving stroke care during the COVID-19 pandemic:Potential issues and solutions, Neurology, 95, 124, 10.1212/WNL.0000000000009713 · DOI
  16. Li, 2010, Coexistence of quiescent and active adult stem cells in mammals, Science, 327, 542, 10.1126/science.1180794 · DOI · Paper page
  17. Li, 2021, Recovery of post-stroke cognitive and motor deficiencies by Shuxuening injection via regulating hippocampal BDNF-mediated neurotrophin/Trk signaling, Biomed Pharmacother, 141, 111828, 10.1016/j.biopha.2021.111828 · DOI
  18. Lv, 2021, Progress in clinical trials of stem cell therapy for cerebral palsy, Neural Regen Res, 16, 1377, 10.4103/1673-5374.300979 · DOI
  19. Ma, 2021, Temporal trend and attributable risk factors of stroke burden in China, 1990-2019: an analysis for the Global Burden of Disease Study 2019, Lancet Public Health, 6, e897, 10.1016/S2468-2667(21)00228-0 · DOI
  20. Marko, 2020, Trends of r-tPA (recombinant tissue-type plasminogen activator) treatment and treatment-influencing factors in acute ischemic stroke, Stroke, 51, 1240, 10.1161/STROKEAHA.119.027921 · DOI
  21. Muheremu, 2021, Sustained delivery of neurotrophic factors to treat spinal cord injury, Transl Neurosci, 12, 494, 10.1515/tnsci-2020-0200 · DOI
  22. Song, 2013, Apoptosis signal-regulating kinase 1 (ASK1) is linked to neural stem cell differentiation after ischemic brain injury, Exp Mol Med, 45, e69, 10.1038/emm.2013.134 · DOI
  23. Tian, 2021, Targeted delivery of neural progenitor cell-derived extracellular vesicles for anti-inflammation after cerebral ischemia, Theranostics, 11, 6507, 10.7150/thno.56367 · DOI
  24. Vogel, 2018, Neural stem cell derived extracellular vesicles:Attributes and prospects for treating neurodegenerative disorders, EBioMedicine, 38, 273, 10.1016/j.ebiom.2018.11.026 · DOI
  25. Xiong, 2021, Brain-derived neurotrophic factor and its related enzymes and receptors play important roles after hypoxic-ischemic brain damage, Neural Regen Res, 16, 1453, 10.4103/1673-5374.303033 · DOI
  26. Xiong, 2016, Functions and mechanisms of microglia/macrophages in neuroinflammation and neurogenesis after stroke, Prog Neurobiol, 142, 23, 10.1016/j.pneurobio.2016.05.001 · DOI
  27. Yuan, 2017, Macrophage exosomes as natural nanocarriers for protein delivery to inflamed brain, Biomaterials, 142, 1, 10.1016/j.biomaterials.2017.07.011 · DOI · Paper page
  28. Yuan, 2021, Neural stem cell-derived exosomes regulate neural stem cell differentiation through miR-9-Hes1 axis, Front Cell Dev Biol, 9, 601600, 10.3389/fcell.2021.601600 · DOI
  29. Zhang, 2018, Neural stem cells alleviate inflammation via neutralization of IFN-γnegative effect in ischemic stroke model, J Biomed Nanotechnol, 14, 1178, 10.1166/jbn.2018.2568 · DOI
  30. Zhang, 2020, Exosomes derived from human neural stem cells stimulated by interferon gamma improve therapeutic ability in ischemic stroke model, J Adv Res, 24, 435, 10.1016/j.jare.2020.05.017 · DOI
  31. Zhang, 2016, Mesenchymal stromal cell therapy in ischemic stroke, J Neurorestoratol, 4, 79, 10.2147/JN.S119874 · DOI
  32. Zhang, 2019, Sevoflurane prevents miR-181a-induced cerebral ischemia/reperfusion injury, Chem Biol Interact, 308, 332, 10.1016/j.cbi.2019.06.008 · DOI
  33. Zhang, 2016, Neural stem cells and ischemic brain, J Stroke, 18, 267, 10.5853/jos.2016.00206 · DOI
  34. Zhang, 2016, Exosomes in stroke pathogenesis and therapy, J Clin Invest, 126, 1190, 10.1172/JCI81133 · DOI
  35. Zhong, 2020, Neural stem cell-derived exosomes facilitate spinal cord functional recovery after injury by promoting angiogenesis, Exp Biol Med (Maywood), 245, 54, 10.1177/1535370219895491 · DOI

Cited by

Sorted by citations

See all 132

Metadata

Authors
Zhu, Z.-H., Jia, F., Ahmed, W., Zhang, G.-L., Wang, H., Lin, C.-Q., Chen, W.-H., & Chen, L.-K.
Affiliations
  • Department of Neurosurgery, School of Medicine, Southeast University, Nanjing, Jiangsu Province, China
  • Department of Neurosurgery, Neuroscience Center, Cancer Center, Integrated Hospital of Traditional Chinese Medicine, Southern Medical University, Guangzhou, Guangdong Province, China
  • Department of Neurosurgery, The Second Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong Province, China
  • Department of Neurosurgery, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China
ORCID
Chen, Lu-Kui
Venue
Neural Regeneration Research
Publisher
Ovid Technologies (Wolters Kluwer Health)
Published
2022-06-02
Volume
18
Issue
2
Pages
404-409
ISSN
  • 1673-5374
  • 1876-7958
DOI
10.4103/1673-5374.346466
PMC
PMC9396474
Cited by
132
References
7
Language
en

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review