The purpose of this study was to observe the effects of olfactory ensheathing cell conditioned medium (OECCM) on damaged astrocytes after exposure to H2O2 in vitro. OECCM was used to treat astrocytes after injury, which was induced by exposure to 500 μmol/L H2O2 for 20 min. The cell morphology was then observed under a light microscope, cell viability assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, cell ultrastructure observed with transmission electron microscopy (TEM), and apoptosis assessed by Annexin V staining followed by cytometry and Western blot. H2O2 induced severe damage to astrocytes as evidenced by decreased cell number, pathological changes in cell morphology, and significantly elevated cell apoptosis. Cells incubated with OECCM displayed significantly improved cell viability and decreased cell apoptotic rate. Under TEM, H2O2-treated cells showed partially broken plasma membranes, swollen rough endoplasmic reticula, visible vacuoles, and swollen or deformed mitochondria with ruptured cristae. Incubation with OECCM significantly ameliorated these pathological changes in astrocytes. These results suggest that OECCM may protect astrocytes from oxidative damage by promoting cell survival while reducing apoptosis of the damaged cells.
This is a preview of subscription content, log in to check access.
This study was supported by the grants from Jiangsu Province Health Department (KF200956).
Conflict of interest
The authors declared that they have no conflict of interest.
Chung RS et al (2004) Olfactory ensheathing cells promote neurite sprouting of injured axons in vitro by direct cellular contact and secretion of soluble factors. Cell Mol Life Sci 61:1238–1245PubMedCrossRefGoogle Scholar
De Keyser J et al (2008) Dysfunctional astrocytes as key players in the pathogenesis of central nervous system disorders. J Neurol Sci 267:3–16PubMedCrossRefGoogle Scholar
do Carmo Cunha J et al (2007) Responses of reactive astrocytes containing S100beta protein and fibroblast growth factor-2 in the border and in the adjacent preserved tissue after a contusion injury of the spinal cord in rats: implications for wound repair and neuroregeneration. Wound Repair Regen 15:134–146PubMedCrossRefGoogle Scholar
Faden AI (2002) Neuroprotection and traumatic brain injury: theoretical option or realistic proposition. Curr Opin Neurol 15:707–712PubMedCrossRefGoogle Scholar
Fauconneau B et al (2002) Induction of heat shock proteins (HSPs) by sodium arsenite in cultured astrocytes and reduction of hydrogen peroxide-induced cell death. J Neurochem 83:1338–1348PubMedCrossRefGoogle Scholar
Faulkner JR et al (2004) Reactive astrocytes protect tissue and preserve function after spinal cord injury. J Neurosci 24:2143–2155PubMedCrossRefGoogle Scholar
Feng L et al (2008) Olfactory ensheathing cells conditioned medium prevented apoptosis induced by 6-OHDA in PC12 cells through modulation of intrinsic apoptotic pathways. Int J Dev Neurosci 26:323–329PubMedCrossRefGoogle Scholar
Franssen EH et al (2007) Olfactory ensheathing glia: their contribution to primary olfactory nervous system regeneration and their regenerative potential following transplantation into the injured spinal cord. Brain Res Rev 56:236–258PubMedCrossRefGoogle Scholar
Friedman J (2011) Why is the nervous system vulnerable to oxidative stress? In: Gadoth N, Gobel HH (eds) Oxidative stress in applied basic research and clinical practice. The Humana Press, New York, pp 19–27Google Scholar
Garcia-Alias G et al (2004) Acute transplantation of olfactory ensheathing cells or Schwann cells promotes recovery after spinal cord injury in the rat. J Neurosci Res 75:632–641PubMedCrossRefGoogle Scholar
Granger N et al (2012) Autologous olfactory mucosal cell transplants in clinical spinal cord injury: a randomized double-blinded trial in a canine translational model. Brain 135:3227–3237PubMedCrossRefGoogle Scholar
Hertz L, Zielke HR (2004) Astrocytic control of glutamatergic activity: astrocytes as stars of the show. Trends Neurosci 27:735–743PubMedCrossRefGoogle Scholar
Huang H et al (2012) Long-term outcome of olfactory ensheathing cell therapy for patients with complete chronic spinal cord injury. Cell Transpl 21S:S23–S31CrossRefGoogle Scholar
Janeczko K (1989) Spatiotemporal patterns of the astroglial proliferation in rat brain injured at the postmitotic stage of postnatal development: a combined immunocytochemical and autoradiographic study. Brain Res 485:236–243PubMedCrossRefGoogle Scholar
Lakatos A et al (2000) Olfactory ensheathing cells and Schwann cells differ in their in vitro interactions with astrocytes. Glia 32:214–225PubMedCrossRefGoogle Scholar
Lakatos A et al (2003) Olfactory ensheathing cells induce less host astrocyte response and chondroitin sulphate proteoglycan expression than Schwann cells following transplantation into adult CNS white matter. Exp Neurol 184:237–246PubMedCrossRefGoogle Scholar
Li Y et al (2004) Interaction of transplanted olfactory-ensheathing cells and host astrocytic processes provides a bridge for axons to regenerate across the dorsal root entry zone. Exp Neurol 188:300–308PubMedCrossRefGoogle Scholar
McKeon RJ et al (1995) Injury-induced proteoglycans inhibit the potential for laminin-mediated axon growth on astrocytic scars. Exp Neurol 136:32–43PubMedCrossRefGoogle Scholar
O’Toole DA et al (2007) Effect of olfactory ensheathing cells on reactive astrocytes in vitro. Cell Mol Life Sci 64:1303–1309PubMedCrossRefGoogle Scholar
Raivich G et al (1999) Neuroglial activation repertoire in the injured brain: graded response, molecular mechanisms and cues to physiological function. Brain Res Brain Res Rev 30:77–105PubMedCrossRefGoogle Scholar
Verdu E et al (2001) Effects of ensheathing cells transplanted into photochemically damaged spinal cord. Neuroreport 12:2303–2309PubMedCrossRefGoogle Scholar
Wewetzer K et al (2001) In vitro expression and regulation of ciliary neurotrophic factor and its alpha receptor subunit in neonatal rat olfactory ensheathing cells. Neurosci Lett 306:165–168PubMedCrossRefGoogle Scholar
Woodhall E et al (2001) Cultured olfactory ensheathing cells express nerve growth factor, brain-derived neurotrophic factor, glia cell line-derived neurotrophic factor and their receptors. Brain Res Mol Brain Res 88:203–213PubMedCrossRefGoogle Scholar
Yu XD et al (2007) Effects of olfactory ensheathing cells on hydrogen peroxide-induced apoptosis in cultured dorsal root ganglion neurons. Chin Med J (Engl) 120:1438–1443Google Scholar