Recently, cell therapy has been developed as a novel treatment for perinatal hypoxic-ischemic encephalopathy (HIE), which is an important cause of neurological disorder and death, and stem cells from human exfoliated deciduous teeth (SHED) express early markers for mesenchymal and neuroectodermal stem cells. We investigated the treatment effect of SHED for HIE in neonatal rats. Seven-day-old rats underwent ligation of the left carotid artery and were exposed to 8% hypoxic treatment. SHED (1 × 105 cells) were injected via the right external jugular vein 24 h after the insult. The effect of intravenous administration of SHED cells was evaluated neurologically and pathophysiologically. In the evaluation of engraftment using quantum dots 655, only a few SHED were detected in the injured cortex. In the immunohistological evaluation 24 h after injection, the numbers of positive cells of active caspase-3 and anti-4 hydroxynonenal antiserum were lower in the SHED group than in the vehicle group. The number of Iba-1+ cells in the cortex was higher in the SHED group. However, the proportion of M1 microglia (Iba-1+/ED-1+) was significantly decreased, whereas M2 microglia (Iba-1+/CD206+) tended to increase in the SHED group. In the behavioral tests performed 5 months after hypoxic treatment, compared to the vehicle group, the SHED group showed significant elongation of the endurance time in the rotarod treadmill test, significantly ameliorated proportion of using the impaired hand in the cylinder test, significantly lower ratio of right/left front paw area in gait analysis, and significantly higher avoidance rate in the active avoidance test. In the in vitro experiment with cultured neurons exposed to oxygen-glucose deprivation, we confirmed the neuroprotective effect of the condition medium of SHED. These results suggested that intravenous administration of SHED exerted a treatment effect both histologically and functionally, possibly via a paracrine effect.
Get full access to this article
View all access options for this article.
References
1.
KurinczukJJ, White-KoningM and BadawiN. (2010). Epidemiology of neonatal encephalopathy and hypoxic-ischaemic encephalopathy. Early Hum Dev, 86:329–338.
2.
SmithJ, WellsL and DoddK. (2000). The continuing fall in incidence of hypoxic-ischaemic encephalopathy in term infants. BJOG, 107:461–466.
3.
BadawiN, KurinczukJJ, KeoghJM, AlessandriLM, O'SullivanF, BurtonPR, PembertonPJ and StanleyFJ. (1998). Intrapartum risk factors for newborn encephalopathy: the Western Australian case-control study. BMJ, 317:1554–1558.
4.
ThornbergE, ThiringerK, OdebackA and MilsomI. (1995). Birth asphyxia: incidence, clinical course and outcome in a Swedish population. Acta Paediatr, 84:927–932.
5.
ShankaranS, WoldtE, KoepkeT, BedardMP and NandyalR. (1991). Acute neonatal morbidity and long-term central nervous system sequelae of perinatal asphyxia in term infants. Early Hum Dev, 25:135–148.
6.
TooleyJ, SatasS, EagleR, SilverIA and ThoresenM. (2002). Significant selective head cooling can be maintained long-term after global hypoxia ischemia in newborn piglets. Pediatrics, 109:643–649.
7.
TooleyJR, SatasS, PorterH, SilverIA and ThoresenM. (2003). Head cooling with mild systemic hypothermia in anesthetized piglets is neuroprotective. Ann Neurol, 53:65–72.
8.
WagnerBP, NedelcuJ and MartinE. (2002). Delayed postischemic hypothermia improves long-term Behavioral outcome after cerebral hypoxia-ischemia in neonatal rats. Pediatr Res, 51:354–360.
9.
AzzopardiDV, StrohmB, EdwardsAD, DyetL, HallidayHL, JuszczakE, KapellouO, LeveneM, MarlowN, et al. (2009). Moderate hypothermia to treat perinatal asphyxial encephalopathy. N Engl J Med, 361:1349–1358.
10.
ShankaranS, LaptookAR, EhrenkranzRA, TysonJE, McDonaldSA, DonovanEF, FanaroffAA, PooleWK, WrightLL, et al. (2005). Whole-body hypothermia for neonates with hypoxic-ischemic encephalopathy. N Engl J Med, 353:1574–1584.
11.
ShankaranS, PappasA, McDonaldSA, VohrBR, HintzSR, YoltonK, GustafsonKE, LeachTM, GreenC, et al. (2012). Childhood outcomes after hypothermia for neonatal encephalopathy. N Engl J Med, 366:2085–2092.
12.
SatoY, NakanishiK, HayakawaM, KakizawaH, SaitoA, KurodaY, IdaM, TokitaY, AonoS, et al. (2008). Reduction of brain injury in neonatal hypoxic-ischemic rats by intracerebroventricular injection of neural stem/progenitor cells together with chondroitinase ABC. Reprod Sci, 15:613–620.
13.
TsujiM, TaguchiA, OhshimaM, KasaharaY, SatoY, TsudaH, OtaniK, YamaharaK, IharaM, et al. (2014). Effects of intravenous administration of umbilical cord blood CD34 cells in a mouse model of neonatal stroke. Neuroscience, 263C:148–158.
14.
HattoriT, SatoY, KondoT, IchinohashiY, SugiyamaY, YamamotoM, KotaniT, HirataH, HirakawaA, et al. (2015). Administration of umbilical cord blood cells transiently decreased hypoxic-ischemic brain injury in neonatal rats. Dev Neurosci, 37:95–104.
15.
NakanishiK, SatoY, MizutaniY, ItoM, HirakawaA and HigashiY. (2017). Rat umbilical cord blood cells attenuate hypoxic-ischemic brain injury in neonatal rats. Sci Rep, 7:44111.
16.
ChenJ, LiY, WangL, ZhangZ, LuD, LuM and ChoppM. (2001). Therapeutic benefit of intravenous administration of bone marrow stromal cells after cerebral ischemia in rats. Stroke, 32:1005–1011.
17.
SchwarzEJ, AlexanderGM, ProckopDJ and AziziSA. (1999). Multipotential marrow stromal cells transduced to produce L-DOPA: engraftment in a rat model of Parkinson disease. Hum Gene Ther, 10:2539–2549.
18.
OrlicD, KajsturaJ, ChimentiS, JakoniukI, AndersonSM, LiB, PickelJ, McKayR, Nadal-GinardB, et al. (2001). Bone marrow cells regenerate infarcted myocardium. Nature, 410:701–705.
19.
UccelliA, MorettaL and PistoiaV. (2008). Mesenchymal stem cells in health and disease. Nat Rev Immunol, 8:726–736.
20.
Le Blanc K and PittengerM. (2005). Mesenchymal stem cells: progress toward promise. Cytotherapy, 7:36–45.
21.
SatoY. (2018). Other tissues-derived mesenchymal stem cells for perinatal brain injury. In: Cell Therapy for Perinatal Brain Injury. ShintakuH, OkaA and NabetaniM, eds. Springer Singapore, Singapore, pp 69–76.
22.
SatoY, UedaK, KondoT, HattoriT, MikrogeorgiouA, SugiyamaY, SuzukiT, YamamotoM, HirataH, et al. (2018). Administration of bone marrow-derived mononuclear cells contributed to the reduction of hypoxic-ischemic brain injury in neonatal rats. Front Neurol, 9:987.
23.
MikrogeorgiouA, SatoY, KondoT, HattoriT, SugiyamaY, ItoM, SaitoA, NakanishiK, TsujiM, et al. (2017). Dedifferentiated fat cells as a novel source for cell therapy to target neonatal hypoxic-ischemic encephalopathy. Dev Neurosci, 39:273–286.
24.
SugiyamaY, SatoY, KitaseY, SuzukiT, KondoT, MikrogeorgiouA, HorinouchiA, MaruyamaS, ShimoyamaY, et al. (2018). Intravenous administration of bone marrow-derived mesenchymal stem cell, but not adipose tissue-derived stem cell, ameliorated the neonatal hypoxic-ischemic brain injury by changing cerebral inflammatory state in rat. Front Neurol, 9:757.
25.
MiuraM, GronthosS, ZhaoM, LuB, FisherLW, RobeyPG and ShiS. (2003). SHED: stem cells from human exfoliated deciduous teeth. Proc Natl Acad Sci U S A, 100:5807–5812.
26.
SakaiVT, ZhangZ, DongZ, NeivaKG, MMachadoAAM, ShiS, SantosCF and NorJE. (2010). SHED differentiate into functional odontoblasts and endothelium. J Dent Res, 89:791–796.
27.
ChadipirallaK, YochimJM, BahuleyanB, HuangCYC, Garcia-GodoyF, MurrayPE and StelnickiEJ. (2010). Osteogenic differentiation of stem cells derived from human periodontal ligaments and pulp of human exfoliated deciduous teeth. Cell Tissue Res, 340:323–333.
28.
SakaiK, YamamotoA, MatsubaraK, NakamuraS, NaruseM, YamagataM, SakamotoK, TauchiR, WakaoN, et al. (2012). Human dental pulp-derived stem cells promote locomotor recovery after complete transection of the rat spinal cord by multiple neuro-regenerative mechanisms. J Clin Invest, 122:80–90.
29.
YamagataM, YamamotoA, KakoE, KanekoN, MatsubaraK, SakaiK, SawamotoK and UedaM. (2013). Human dental pulp-derived stem cells protect against hypoxic-ischemic brain injury in neonatal mice. Stroke, 44:551–554.
30.
NakamuraS, YamadaY, KatagiriW, SugitoT, ItoK and UedaM. (2009). Stem cell proliferation pathways comparison between human exfoliated deciduous teeth and dental pulp stem cells by gene expression profile from promising dental pulp. J Endod, 35:1536–1542.
31.
MartinelloK, HartAR, YapS, MitraS and RobertsonNJ. (2017). Management and investigation of neonatal encephalopathy: 2017 update. Arch Dis Child Fetal Neonatal Ed, 102:F346–F358.
32.
BashirRA, VayalthrikkovilS, EspinozaL, IrvineL, ScottJ and MohammadK. (2018). Prevalence and characteristics of intracranial hemorrhages in neonates with hypoxic ischemic encephalopathy. Am J Perinatol, 35:676–681.
33.
SatoY, ShinjyoN, SatoM, OsatoK, ZhuC, PeknaM, KuhnHG and BlomgrenK. (2013). Grafting of neural stem and progenitor cells to the hippocampus of young, irradiated mice causes gliosis and disrupts the granule cell layer. Cell Death Dis, 4:e591.
34.
YukawaH, WatanabeM, KajiN, OkamotoY, TokeshiM, MiyamotoY, NoguchiH, BabaY and HayashiS. (2012). Monitoring transplanted adipose tissue-derived stem cells combined with heparin in the liver by fluorescence imaging using quantum dots. Biomaterials, 33:2177–2186.
35.
YukawaH and BabaY. (2017). In vivo fluorescence imaging and the diagnosis of stem cells using quantum dots for regenerative medicine. Anal Chem, 89:2671–2681.
YamamotoA, SakaiK, MatsubaraK, KanoF and UedaM. (2014). Multifaceted neuro-regenerative activities of human dental pulp stem cells for functional recovery after spinal cord injury. Neurosci Res, 78:16–20.
38.
ZhaoY, WangL, JinY and ShiS. (2012). Fas ligand regulates the immunomodulatory properties of dental pulp stem cells. J Dent Res, 91:948–954.
39.
YamazaT, KentaroA, ChenC, LiuY, ShiY, GronthosS, WangS and ShiS. (2010). Immunomodulatory properties of stem cells from human exfoliated deciduous teeth. Stem Cell Res Ther, 1:5.
40.
ArteagaO, AlvarezA, RevueltaM, SantaolallaF, UrtasunA and HilarioE. (2017). Role of antioxidants in neonatal hypoxic-ischemic brain injury: new therapeutic approaches. Int J Mol Sci 18:pii:. E265.
41.
Rocha-FerreiraE and HristovaM. (2016). Plasticity in the neonatal brain following hypoxic-ischaemic injury. Neural Plast, 2016:4901014.
42.
LimoliCL, GiedzinskiE, RolaR, OtsukaS, PalmerTD and FikeJR. (2004). Radiation response of neural precursor cells: linking cellular sensitivity to cell cycle checkpoints, apoptosis and oxidative stress. Radiat Res, 161:17–27.
43.
IadecolaC and AnratherJ. (2011). The immunology of stroke: from mechanisms to translation. Nat Med, 17:796–808.
44.
KonsmanJP, DrukarchB and Van DamAM. (2007). (Peri)vascular production and action of pro-inflammatory cytokines in brain pathology. Clin Sci (Lond), 112:1–25.
45.
VarnumMM and IkezuT. (2012). The classification of microglial activation phenotypes on neurodegeneration and regeneration in Alzheimer's disease brain. Arch Immunol Ther Exp (Warsz), 60:251–266.
46.
OhtakiH, YlostaloJH, ForakerJE, RobinsonAP, RegerRL, ShiodaS and ProckopDJ. (2008). Stem/progenitor cells from bone marrow decrease neuronal death in global ischemia by modulation of inflammatory/immune responses. Proc Natl Acad Sci U S A, 105:14638–14643.
47.
NakajimaH, UchidaK, GuerreroAR, WatanabeS, SugitaD, TakeuraN, YoshidaA, LongG, WrightKT, JohnsonWE and BabaH. (2012). Transplantation of mesenchymal stem cells promotes an alternative pathway of macrophage activation and functional recovery after spinal cord injury. J Neurotrauma, 29:1614–1625.
48.
MatsubaraK, MatsushitaY, SakaiK, KanoF, KondoM, NodaM, HashimotoN, ImagamaS, IshiguroN, et al. (2015). Secreted ectodomain of sialic acid-binding Ig-like lectin-9 and monocyte chemoattractant protein-1 promote recovery after rat spinal cord injury by altering macrophage polarity. J Neurosci, 35:2452–2464.
49.
ItoT, IshigamiM, MatsushitaY, HirataM, MatsubaraK, IshikawaT, HibiH, UedaM, HirookaY, GotoH and YamamotoA. (2017). Secreted ectodomain of SIGLEC-9 and MCP-1 synergistically improve acute liver failure in rats by altering macrophage polarity. Sci Rep, 7:44043.
50.
KanoF, MatsubaraK, UedaM, HibiH and YamamotoA. (2017). Secreted ectodomain of sialic acid-binding ig-like lectin-9 and monocyte chemoattractant protein-1 synergistically regenerate transected rat peripheral nerves by altering macrophage polarity. Stem Cells, 35:641–653.
51.
TanakaE, OgawaY, MukaiT, SatoY, HamazakiT, Nagamura-InoueT, Harada-ShibaM, ShintakuH and TsujiM. (2018). Dose-dependent effect of intravenous administration of human umbilical cord-derived mesenchymal stem cells in neonatal stroke mice. Front Neurol, 9:133.
52.
ParkWS, SungSI, AhnSY, YooHS, SungDK, ImGH, ChoiSJ and ChangYS. (2015). Hypothermia augments neuroprotective activity of mesenchymal stem cells for neonatal hypoxic-ischemic encephalopathy. PLoS One, 10:e0120893.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
