A Novel Treatment with Stem Cells from Human Exfoliated Deciduous Teeth for Hypoxic-Ischemic Encephalopathy in Neonatal Rats
A Novel Treatment with Stem Cells from Human Exfoliated Deciduous Teeth for Hypoxic-Ischemic Encephalopathy in Neonatal Rats
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DOI：
10.1089/scd.2019.0221
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发表时间：
2020-01-15
影响因子：
4
通讯作者：
Hayakawa, Masahiro
中科院分区：
文献类型：
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作者：
Kitase, Yuma;Sato, Yoshiaki;Hayakawa, Masahiro
近年来，细胞疗法已发展成为治疗围产期缺氧缺血性脑病（hypoxic-ischemic encephalopathy，HIE）的一种新方法，HIE是神经系统疾病和死亡的重要原因，而来自人类脱落乳牙（exfoliated digny teeth，SHED）的干细胞表达间充质干细胞和神经外胚层干细胞的早期标志物。本实验观察了SHED对新生大鼠HIE的治疗作用。7日龄大鼠结扎左颈动脉，并暴露于8%的低氧治疗。损伤后24小时，通过右颈外静脉注射SHED（1 × 10（5）个细胞）。从神经学和病理生理学上评价静脉内施用SHED细胞的效果。在使用量子点655的植入评估中，在受损皮质中仅检测到少量SHED。在注射后24 h的免疫组织学评价中，活性半胱天冬酶-3和抗-4羟基壬烯醛抗血清的阳性细胞数在SHED组中低于溶剂组。SHED组皮质Iba-1（+）细胞数量较多。然而，在SH艾德组中，M1小胶质细胞的比例（Iba-1（+）/艾德-1（+））显著降低，而M2小胶质细胞的比例（Iba-1（+）/CD 206（+））倾向于增加。在低氧处理后5个月进行的行为测试中，与溶剂组相比，SHED组在旋转杆跑步机测试中显示出显著延长的耐受时间，在圆柱体测试中显示出显著改善的使用受损手的比例，在步态分析中显示出显著降低的右/左前爪面积比，并且在主动回避测试中显示出显著提高的回避率。在体外培养的神经元缺氧缺糖实验中，我们证实了SHED条件培养基的神经保护作用。这些结果表明，静脉注射SHED可能通过旁分泌效应在组织学和功能上产生治疗效果。
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 x 10(5) 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.