Methylprednisolone inhibits the proliferation of endogenous neural stem cells in nonhuman primates with spinal cord injury

Restricted access

OBJECTIVE

The aim of this work was to investigate the effects of methylprednisolone on the proliferation of endogenous neural stem cells (ENSCs) in nonhuman primates with spinal cord injury (SCI).

METHODS

A total of 14 healthy cynomolgus monkeys (Macaca fascicularis) (4–5 years of age) were randomly divided into 3 groups: the control group (n = 6), SCI group (n = 6), and methylprednisolone therapy group (n = 2). Only laminectomy was performed in the control animals at T-10. SCI was induced in monkeys using Allen’s weight-drop method (50 mm × 50 g) to injure the posterior portion of the spinal cord at T-10. In the methylprednisolone therapy group, monkeys were intravenously infused with methylprednisolone (30 mg/kg) immediately after SCI. All animals were intravenously infused with 5-bromo-2-deoxyuridine (BrdU) (50 mg/kg/day) for 3 days prior to study end point. The small intestine was dissected for immunohistochemical examination. After 3, 7, and 14 days, the spinal cord segments of the control and SCI groups were dissected to prepare frozen and paraffin sections. The proliferation of ENSCs was evaluated using BrdU and nestin immunofluorescence staining.

RESULTS

Histological examination showed that a larger number of mucosa epithelial cells in the small intestine of all groups were BrdU positive. Nestin-positive ependymal cells are increased around the central canal after SCI. After 3, 7, and 14 days of SCI, BrdU-positive ependymal cells in the SCI group were significantly increased compared with the control group, and the percentage of BrdU-positive cells in the left/right ventral horns and dorsal horn was significantly higher than that of the control group. Seven days after SCI, the percentages of both BrdU-positive ependymal cells around the central canal and BrdU– and nestin–double positive cells in the left/right ventral horns and dorsal horn were significantly lower in the methylprednisolone therapy group than in the SCI group.

CONCLUSIONS

While ENSCs proliferate significantly after SCI in nonhuman primates, methylprednisolone can inhibit the proliferation of ependymal cells after SCI.

ABBREVIATIONS BrdU = 5-bromo-2-deoxyuridine; ENSC = endogenous neural stem cell; ESC = embryonic stem cell; MP = methylprednisolone; NPC = neural precursor cell; NSC = neural stem cell; PBS = phosphate-buffered saline; SCI = spinal cord injury.

Article Information

Correspondence Huiyong Shen: Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, China. shyshenhuiyong@163.com.

INCLUDE WHEN CITING Published online May 18, 2018; DOI: 10.3171/2017.12.SPINE17669.

J.Y. and Y.Q. contributed equally to this work.

Disclosures The authors report no conflict of interest concerning the materials or methods used in this study or the findings specified in this paper.

© AANS, except where prohibited by US copyright law.

Headings

Figures

  • View in gallery

    The histopathology of spinal cords in control and SCI animals. Dense and normal spinal cord structures were observed in the control animals. The damage to the spinal cord of the SCI group was located on the dorsal side on the center of the spinal cord. H & E, original magnification ×100. Figure is available in color online only.

  • View in gallery

    BrdU immunohistochemical staining of small intestinal mucosal epithelia of SCI animals. Most (> 80%) of the mucosal epithelial cells in the small intestine were BrdU positive with brown nuclei staining. Original magnification ×100. Figure is available in color online only.

  • View in gallery

    Hoechst staining and nestin immunofluorescent staining of ependymal cells in control (A–C) and SCI (D–F) animals. Nestin-positive ependymal cells around the central canal were observed under a light microscope. Ependymal cells in both the control and SCI animals are ENSCs or NPCs. However, ependymal cells in the SCI animals exhibited stronger nestin staining than those of the control animals. Original magnification ×400. Figure is available in color online only.

  • View in gallery

    BrdU immunohistochemical staining of ependymal cells in control animals (A) and SCI animals on days 3 (B), 7 (C), and 14 (D). This staining is confined to the nucleus of ependymal cells. Scattered BrdU-positive ependymal cells were observed around the central canal of the spinal cord of the control animals. More BrdU-positive ependymal cells were observed around the central canal of the spinal cord of the SCI animals on days 3, 7, and 14 after SCI. Original magnification ×400. Figure is available in color online only.

  • View in gallery

    Nestin-positive cells in the ventral and dorsal horns of spinal cord gray matter. The number of nestin-positive cells in the right ventral and dorsal horns of the SCI group (C and D) on day 7 after SCI was significantly higher than that in the ventral and dorsal horns of control animals (A and B). Original magnification ×100. Figure is available in color online only.

  • View in gallery

    BrdU-positive cells in the ventral and dorsal horns of spinal cord gray matter. The number of BrdU-positive cells in the right ventral and dorsal horns of the SCI group (C and D) on day 7 after SCI was significantly higher than that in the ventral and dorsal horns of control animals (A and B). Original magnification ×100. Figure is available in color online only.

  • View in gallery

    Nestin/BrdU–double positive ependymal cells in ventral and dorsal horns of spinal cord in SCI and MP animals. The number of nestin/BrdU–double positive ependymal cells was significantly decreased in the MP group compared with the SCI group. *p < 0.05 versus SCI group.

References

  • 1

    Breslin KAgrawal D: The use of methylprednisolone in acute spinal cord injury: a review of the evidence, controversies, and recommendations. Pediatr Emerg Care 28:123812482012

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 2

    Caroni P: Neuro-regeneration: plasticity for repair and adaptation. Essays Biochem 33:53641998

  • 3

    Del Bigio MR: The ependyma: a protective barrier between brain and cerebrospinal fluid. Glia 14:1131995

  • 4

    Di Giovanni SKnoblach SMBrandoli CAden SAHoffman EPFaden AI: Gene profiling in spinal cord injury shows role of cell cycle in neuronal death. Ann Neurol 53:4544682003

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 5

    Djebaili MGuo QPettus EHHoffman SWStein DG: The neurosteroids progesterone and allopregnanolone reduce cell death, gliosis, and functional deficits after traumatic brain injury in rats. J Neurotrauma 22:1061182005

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 6

    Enzmann GUBenton RLTalbott JFCao QWhittemore SR: Functional considerations of stem cell transplantation therapy for spinal cord repair. J Neurotrauma 23:4794952006

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 7

    Fehlings MGVawda R: Cellular treatments for spinal cord injury: the time is right for clinical trials. Neurotherapeutics 8:7047202011

  • 8

    Fujimoto YAbematsu MFalk ATsujimura KSanosaka TJuliandi B: Treatment of a mouse model of spinal cord injury by transplantation of human induced pluripotent stem cell-derived long-term self-renewing neuroepithelial-like stem cells. Stem Cells 30:116311732012

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 9

    Garbossa DBoido MFontanella MFronda CDucati AVercelli A: Recent therapeutic strategies for spinal cord injury treatment: possible role of stem cells. Neurosurg Rev 35:2933112012

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 10

    Gu HYu SPGutekunst CAGross REWei L: Inhibition of the Rho signaling pathway improves neurite outgrowth and neuronal differentiation of mouse neural stem cells. Int J Physiol Pathophysiol Pharmacol 5:11202013

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 11

    Hatami MMehrjardi NZKiani SHemmesi KAzizi HShahverdi A: Human embryonic stem cell-derived neural precursor transplants in collagen scaffolds promote recovery in injured rat spinal cord. Cytotherapy 11:6186302009

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 12

    Hofstetter CPHolmström NALilja JASchweinhardt PHao JSpenger C: Allodynia limits the usefulness of intraspinal neural stem cell grafts; directed differentiation improves outcome. Nat Neurosci 8:3463532005

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 13

    Horner PJPower AEKempermann GKuhn HGPalmer TDWinkler J: Proliferation and differentiation of progenitor cells throughout the intact adult rat spinal cord. J Neurosci 20:221822282000

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 14

    Ishikawa MYoshitomi TZorumski CFIzumi Y: Neurosteroids are endogenous neuroprotectants in an ex vivo glaucoma model. Invest Ophthalmol Vis Sci 55:853185412014

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 15

    Johansson CBMomma SClarke DLRisling MLendahl UFrisén J: Identification of a neural stem cell in the adult mammalian central nervous system. Cell 96:25341999

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 16

    Kalyani AHobson KRao MS: Neuroepithelial stem cells from the embryonic spinal cord: isolation, characterization, and clonal analysis. Dev Biol 186:2022231997

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 17

    Kehn MKroll T: Reporting trends of spinal cord injury research representation: a media content analysis. Disabil Health J 4:1211282011

  • 18

    Kennea NLMehmet H: Neural stem cells. J Pathol 197:5365502002

  • 19

    Kim YJPark HJLee GBang OYAhn YHJoe E: Neuroprotective effects of human mesenchymal stem cells on dopaminergic neurons through anti-inflammatory action. Glia 57:13232009

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 20

    Leal-Filho MB: Spinal cord injury: from inflammation to glial scar. Surg Neurol Int 2:1122011

  • 21

    Liu YTan BWang LLong ZLi YLiao W: Endogenous neural stem cells in central canal of adult rats acquired limited ability to differentiate into neurons following mild spinal cord injury. Int J Clin Exp Pathol 8:383538422015

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 22

    Logan AAhmed ZBaird AGonzalez AMBerry M: Neurotrophic factor synergy is required for neuronal survival and disinhibited axon regeneration after CNS injury. Brain 129:4905022006

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 23

    Lu PJones LLSnyder EYTuszynski MH: Neural stem cells constitutively secrete neurotrophic factors and promote extensive host axonal growth after spinal cord injury. Exp Neurol 181:1151292003

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 24

    Magnus TRao MS: Neural stem cells in inflammatory CNS diseases: mechanisms and therapy. J Cell Mol Med 9:3033192005

  • 25

    McClellan AD: Functional axonal regeneration following spinal cord injury. Brain Res Bull 50:4034041999

  • 26

    McDonald JWLiu XZQu YLiu SMickey SKTuretsky D: Transplanted embryonic stem cells survive, differentiate and promote recovery in injured rat spinal cord. Nat Med 5:141014121999

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 27

    Muheremu APeng JAo Q: Stem cell based therapies for spinal cord injury. Tissue Cell 48:3283332016

  • 28

    Nash HHBorke RCAnders JJ: Ensheathing cells and methylprednisolone promote axonal regeneration and functional recovery in the lesioned adult rat spinal cord. J Neurosci 22:711171202002

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 29

    Nemati SNJabbari RHajinasrollah MZare Mehrjerdi NAzizi HHemmesi K: Transplantation of adult monkey neural stem cells into a contusion spinal cord injury model in rhesus macaque monkeys. Cell J 16:1171302014

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 30

    Nishimura SYasuda AIwai HTakano MKobayashi YNori S: Time-dependent changes in the microenvironment of injured spinal cord affects the therapeutic potential of neural stem cell transplantation for spinal cord injury. Mol Brain 6:32013

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 31

    Panayiotou EMalas S: Adult spinal cord ependymal layer: a promising pool of quiescent stem cells to treat spinal cord injury. Front Physiol 4:3402013

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 32

    Picard-Riera NNait-Oumesmar BBaron-Van Evercooren A: Endogenous adult neural stem cells: limits and potential to repair the injured central nervous system. J Neurosci Res 76:2232312004

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 33

    Piechota MKorostynski MGolda SFicek JJantas DBarbara Z: Transcriptional signatures of steroid hormones in the striatal neurons and astrocytes. BMC Neurosci 18:372017

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 34

    Rhodes KEMoon LDFawcett JW: Inhibiting cell proliferation during formation of the glial scar: effects on axon regeneration in the CNS. Neuroscience 120:41562003

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 35

    Ronaghi MErceg SMoreno-Manzano VStojkovic M: Challenges of stem cell therapy for spinal cord injury: human embryonic stem cells, endogenous neural stem cells, or induced pluripotent stem cells? Stem Cells 28:93992010

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 36

    Schröter ALustenberger RMObermair FJThallmair M: High-dose corticosteroids after spinal cord injury reduce neural progenitor cell proliferation. Neuroscience 161:7537632009

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 37

    Shibuya SMiyamoto OAuer RNItano TMori SNorimatsu H: Embryonic intermediate filament, nestin, expression following traumatic spinal cord injury in adult rats. Neuroscience 114:9059162002

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 38

    Shihabuddin LSHorner PJRay JGage FH: Adult spinal cord stem cells generate neurons after transplantation in the adult dentate gyrus. J Neurosci 20:872787352000

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 39

    Silver JR: A systematic review of the therapeutic interventions for heterotopic ossification after spinal cord injury. Spinal Cord 49:4824842011 (Letter)

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 40

    Sofroniew MVVinters HV: Astrocytes: biology and pathology. Acta Neuropathol 119:7352010

  • 41

    Sulzer DJoyce MPLin LGeldwert DHaber SNHattori T: Dopamine neurons make glutamatergic synapses in vitro. J Neurosci 18:458846021998

  • 42

    Takahashi MArai YKurosawa HSueyoshi NShirai S: Ependymal cell reactions in spinal cord segments after compression injury in adult rat. J Neuropathol Exp Neurol 62:1851942003

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 43

    Tetzlaff WOkon EBKarimi-Abdolrezaee SHill CESparling JSPlemel JR: A systematic review of cellular transplantation therapies for spinal cord injury. J Neurotrauma 28:161116822011

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 44

    Wallace MCTator CHLewis AJ: Chronic regenerative changes in the spinal cord after cord compression injury in rats. Surg Neurol 27:2092191987

  • 45

    Wang GAo QGong KZuo HGong YZhang X: Synergistic effect of neural stem cells and olfactory ensheathing cells on repair of adult rat spinal cord injury. Cell Transplant 19:132513372010

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 46

    Yan JWelsh AMBora SHSnyder EYKoliatsos VE: Differentiation and tropic/trophic effects of exogenous neural precursors in the adult spinal cord. J Comp Neurol 480:1011142004

    • Crossref
    • PubMed
    • Search Google Scholar
    • Export Citation
  • 47

    Yu SPatchev AVWu YLu JHolsboer FZhang JZ: Depletion of the neural precursor cell pool by glucocorticoids. Ann Neurol 67:21302010

Metrics

Metrics

All Time Past Year Past 30 Days
Abstract Views 223 223 33
Full Text Views 114 114 1
PDF Downloads 161 161 2
EPUB Downloads 0 0 0

PubMed

Google Scholar