Pretreatment of nucleus pulposus mesenchymal stem cells with appropriate concentration of H2O2 enhances their ability to treat intervertebral disc degeneration View Full Text


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Article Info

DATE

2022-07-26

AUTHORS

Yu-yao Zhang, Zhi-lei Hu, Yu-han Qi, Hai-yin Li, Xian Chang, Xiao-xin Gao, Chen-hao Liu, Yue-yang Li, Jin-hui Lou, Yu Zhai, Chang-qing Li

ABSTRACT

BackgroundNucleus pulposus mesenchymal stem cells (NPMSCs) transplantation is a promising treatment for intervertebral disc degeneration (IVDD). However, the transplanted NPMSCs exhibited weak cell proliferation, high cell apoptosis, and a low ability to resist the harsh microenvironment of the degenerated intervertebral disc. There is an urgent need to explore feasible methods to enhance the therapeutic efficacy of NPMSCs transplantation.ObjectiveTo identify the optimal concentration for NPMSCs pretreatment with hydrogen peroxide (H2O2) and explore the therapeutic efficacy of NPMSCs transplantation using H2O2 pretreatment in IVDD.MethodsRat NPMSCs were pretreated with different concentrations (range from 25 to 300 μM) of H2O2. The proliferation, reactive oxygen species (ROS) level, and apoptosis of NPMSCs were detected by cell counting kit-8 (CCK-8) assay, 5-ethynyl-2′-deoxyuridine (EdU) staining, and flow cytometry in vitro. The underlying signalling pathways were explored utilizing Western blotting. A rat needle puncture-stimulated IVDD model was established. X-ray, histological staining, and a multimode small animal live imaging system were used to evaluate the therapeutic effect of H2O2-pretreated NPMSCs in vivo.ResultsNPMSCs pretreated with 75 μM H2O2 demonstrated the strongest elevated cell proliferation by inhibiting the Hippo pathway (P < 0.01). Meanwhile, 75 μM H2O2-pretreated NPMSCs exhibited significantly enhanced antioxidative stress ability (P < 0.01), which is related to downregulated Brd4 and Keap1 and upregulated Nrf2. NPMSCs pretreated with 75 μM H2O2 also exhibited distinctly decreased apoptosis (P < 0.01). In vivo experiments verified that 75 μM H2O2-pretreated NPMSCs-transplanted rats exhibited an enhanced disc height index (DHI% = 90.00 ± 4.55, P < 0.01) and better histological morphology (histological score = 13.5 ± 0.5, P < 0.01), which means 75 μM H2O2-pretreated NPMSCs can better adapt to the environment of degenerative intervertebral discs and promote the repair of IVDD.ConclusionsPretreatment with 75 μM H2O2 was the optimal concentration to improve the proliferation, antioxidative stress, and antiapoptotic ability of transplanted NPMSCs, which is expected to provide a new feasible method to improve the stem cell therapy efficacy of IVDD. More... »

PAGES

340

References to SciGraph publications

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  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1186/s13287-022-03031-7

    DOI

    http://dx.doi.org/10.1186/s13287-022-03031-7

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    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/35883157


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    28 schema:description BackgroundNucleus pulposus mesenchymal stem cells (NPMSCs) transplantation is a promising treatment for intervertebral disc degeneration (IVDD). However, the transplanted NPMSCs exhibited weak cell proliferation, high cell apoptosis, and a low ability to resist the harsh microenvironment of the degenerated intervertebral disc. There is an urgent need to explore feasible methods to enhance the therapeutic efficacy of NPMSCs transplantation.ObjectiveTo identify the optimal concentration for NPMSCs pretreatment with hydrogen peroxide (H2O2) and explore the therapeutic efficacy of NPMSCs transplantation using H2O2 pretreatment in IVDD.MethodsRat NPMSCs were pretreated with different concentrations (range from 25 to 300 μM) of H2O2. The proliferation, reactive oxygen species (ROS) level, and apoptosis of NPMSCs were detected by cell counting kit-8 (CCK-8) assay, 5-ethynyl-2′-deoxyuridine (EdU) staining, and flow cytometry in vitro. The underlying signalling pathways were explored utilizing Western blotting. A rat needle puncture-stimulated IVDD model was established. X-ray, histological staining, and a multimode small animal live imaging system were used to evaluate the therapeutic effect of H2O2-pretreated NPMSCs in vivo.ResultsNPMSCs pretreated with 75 μM H2O2 demonstrated the strongest elevated cell proliferation by inhibiting the Hippo pathway (P < 0.01). Meanwhile, 75 μM H2O2-pretreated NPMSCs exhibited significantly enhanced antioxidative stress ability (P < 0.01), which is related to downregulated Brd4 and Keap1 and upregulated Nrf2. NPMSCs pretreated with 75 μM H2O2 also exhibited distinctly decreased apoptosis (P < 0.01). In vivo experiments verified that 75 μM H2O2-pretreated NPMSCs-transplanted rats exhibited an enhanced disc height index (DHI% = 90.00 ± 4.55, P < 0.01) and better histological morphology (histological score = 13.5 ± 0.5, P < 0.01), which means 75 μM H2O2-pretreated NPMSCs can better adapt to the environment of degenerative intervertebral discs and promote the repair of IVDD.ConclusionsPretreatment with 75 μM H2O2 was the optimal concentration to improve the proliferation, antioxidative stress, and antiapoptotic ability of transplanted NPMSCs, which is expected to provide a new feasible method to improve the stem cell therapy efficacy of IVDD.
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    35 ConclusionsPretreatment
    36 H2O2
    37 Hippo pathway
    38 IVDD model
    39 Keap1
    40 Kit-8
    41 NPMSCs
    42 Nrf2
    43 ObjectiveTo
    44 Western blotting
    45 X-ray
    46 ability
    47 antiapoptotic ability
    48 antioxidative stress
    49 antioxidative stress ability
    50 apoptosis
    51 appropriate concentration
    52 blotting
    53 cell apoptosis
    54 cell proliferation
    55 cell therapy efficacy
    56 cell transplantation
    57 cells
    58 concentration
    59 cytometry
    60 degenerated intervertebral discs
    61 degeneration
    62 degenerative intervertebral disc
    63 different concentrations
    64 disc degeneration
    65 disc height index
    66 disk
    67 effect
    68 efficacy
    69 elevated cell proliferation
    70 environment
    71 experiments
    72 feasible method
    73 harsh microenvironment
    74 height index
    75 higher cell apoptosis
    76 histological morphology
    77 histological staining
    78 hydrogen peroxide
    79 imaging system
    80 index
    81 intervertebral disc
    82 intervertebral disc degeneration
    83 levels
    84 live imaging system
    85 low ability
    86 mesenchymal stem cell transplantation
    87 mesenchymal stem cells
    88 method
    89 microenvironment
    90 model
    91 morphology
    92 need
    93 new feasible method
    94 npMSCs
    95 nucleus
    96 optimal concentration
    97 oxygen species levels
    98 pathway
    99 peroxide
    100 pretreatment
    101 proliferation
    102 promising treatment
    103 rats
    104 reactive oxygen species levels
    105 repair
    106 signaling pathways
    107 species level
    108 staining
    109 stem cell transplantation
    110 stem cells
    111 stress
    112 stress ability
    113 system
    114 therapeutic effect
    115 therapeutic efficacy
    116 therapy efficacy
    117 transplantation
    118 treatment
    119 underlying signaling pathways
    120 upregulated Nrf2
    121 urgent need
    122 vivo
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    124 weak cell proliferation
    125 μM H2O2
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