Sufentanil combined with propofol anesthesia as an aid to umbilical cord blood mesenchymal stem cell transplantation for the treatment of spinal cord injury

doi:10.3969/j.issn.2095-4344.2016.45.008

Abstract

Abstract:

BACKGROUND: Sufentanil and propofol are both found to have good neuroprotective effects on neurological damage in clinical practice.
OBJECTIVE: To investigate the effect of propofol combined with sufentanil in umbilical cord blood mesenchymal stem cells transplantation for the treatment of spinal cord injury.
METHODS: Sixty-five Wistar rats were selected to make animal models of acute spinal cord injury using Allen’s method. Six hours after modeling, these rats were randomly assigned into combined group (injection of 2×10⁷/L human umbilical cord blood mesenchymal stem cell suspension (0.5 mL) plus injection of 1.0-1.5 mg/kg propofol and 0.5 μg/kg sufentanil via the tail vein), stem cell group (injection of 2×10⁷/L human umbilical cord blood mesenchymal stem cell suspension (0.5 mL) via the tail vein), or control group (injection of 30 μL of LDMEM containing 5% fetal bovine serum). S100β protein level in serum was detected in each group at 15 and 60 minutes after injection. Motor function of rat in each group was assessed by Basso Beattie Bresnahan (BBB) scoring and incline plane test at 1, 2, 4, 6, 8 weeks after modeling. Pathological changes of the spinal cord were observed at 4 weeks after modeling. Expression of vascular endothelial growth factor was detected using western blot assay at 1 and 2 weeks after modeling.
RESULTS AND CONCLUSION: After 15 and 60 minutes of intervention, S100β protein level was lowest in the combined group followed by the stem cell and control groups (P < 0.05). At 2, 4, 6, 8 weeks after modeling, scores on the incline plane test and BBB were ranked as follows: combined group > stem cell group > model group (P < 0.05). At 4 weeks after modeling, severe damage to the spinal cord and few nerve fibers were found in the control group; spinal cord hyperplasia and a few of regenerated axons and PKH-26-positive stem cells appeared in the stem cell group; while in the combined group, there were a large amount of PKH-26-positive stem cells and nerve axon-like structures. At 1 and 2 weeks after modeling, the highest protein level of vascular endothelial growth factor was found in the combined group followed by the stem cell group and control group (P < 0.05). To conclude, these findings indicate that propofol and sufentanil in umbilical cord blood mesenchymal stem cell transplantation therapy can promote the recovery of hindlimb function after spinal cord injury, thereby promoting the functional recovery of rats from spinal cord injury.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Stem Cells, Cord Blood Stem Cell Transplantation, Propofol, Sufentanil, Tissue Engineering

CLC Number:

R394.2

Li Wen-liang, Tong Dong-yi. Sufentanil combined with propofol anesthesia as an aid to umbilical cord blood mesenchymal stem cell transplantation for the treatment of spinal cord injury[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(45): 6739-6745.

Figures/Tables 2

2.1 脐血间充质干细胞的形态及标记效果培养四五天后，细胞呈长梭形或多角形并出现有丝分裂，部分细胞克隆呈集落状生长，形态呈单一的长梭形，胞体中央膨大；培养八九天，细胞铺满培养瓶底面，成漩涡状生长，漩涡中心的细胞为复层生长，周围大部分细胞胞体呈单层的漩涡状或辐射状；PKH-26标记的脐血间充质干细胞在荧光显微镜下呈红色荧光，FCM检测细胞标记率达99%，见图1。 2.2 造模过程中动物一般情况造摸后死亡动物3只，重新给予补足。 2.3 各组血清S100β蛋白水平比较干预后15，60 min时，联合组血清S100β蛋白水平低于干细胞组、对照组 (P < 0.05)，干细胞组低于对照组(P < 0.05)，见表1。 2.4 各组斜板实验、BBB评分结果斜板实验运动功能评分：脊髓损伤后2，4，6，8周，联合组评分高于干细胞组、对照组(P < 0.05)，干细胞组高于对照组(P < 0.05)，见表2。 BBB运动功能评分：脊髓损伤后1周，联合组评分高于干细胞组、对照组(P < 0.05)，干细胞组与对照组比较差异无显著性意义；脊髓损伤后2，4，6，8周，联合组评分高于干细胞组、对照组(P < 0.05)，干细胞组高于对照组(P < 0.05)，见表3。 2.5 各组脊髓组织组织学变化苏木精-伊红染色后可观察到，对照组脊髓损伤区部分组织坏死，脊髓结构破坏较为严重，灰白质结构紊乱，细胞及血管周围间隙扩大，神经纤维排列稀疏甚至断裂，有少量瘢痕形成；干细胞组脊髓损伤区有许多增生的脊髓组织充填，脊髓组织中空泡小而少，损伤区近端有较多再生轴突，并开始向损伤区伸展；联合组可见较多的神经轴索样结构，而干细胞组和对照组则少见，见图2；干细胞组及联合组荧光显微镜下均可见散在PKH-26染色的阳性细胞数，且联合组大于干细胞组[(12.87±2.39)%，(41.04± 5.17)%，P < 0.05]，对照组未见PKH-26染色的阳性细胞数，见图3。 2.6 各组血管内皮生长因子蛋白表达情况脊髓损伤后1，2周，对照组脊髓损伤处血管内皮生长因子蛋白相对表达量逐渐减少，干细胞组、联合组逐渐增多；联合组不同时间点的血管内皮生长因子蛋白相对表达量高于干细胞组、对照组(P < 0.05)，干细胞组不同时间点的血管内皮生长因子蛋白相对表达量高于对照组(P < 0.05)，见表4。"

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