1.Effects of electro-acupuncture combined with bone marrow stromal cell transplantation on recovery of neurological function in rats with spinal cord injury
Li ZHANG ; Donglin XIONG ; Kuihao CHEN ; Rongrong WANG ; Yingxiang MIAO
Chinese Journal of Tissue Engineering Research 2009;13(27):5217-5220
BACKGROUND: The study indicated that directed neural differentiation of bone marrow stromal cells (BMSCs) was associated with changes in local micro-environment in injured spinal cord following cerebral ischemia, in particular neurotrophic factor induction. Group pre-test has confirmed that acupuncture can increase the expression of vadous cytokines and neurotrophic factors to promote nerve regeneration and repair.OBJECTIVE: To observe the effects of electro-acupuncture combined with BMSC kansplantaUon on recovery of neurological function in rats with spinal cord injury.DESIGN, TIME AND SETTING: Randomized controlled animal experiments were performed at the Laboratory of Cell Biology,Harbin Medical University from March 2005 to July 2006.MATERIALS: A total of 80 pure healthy Sprague Dawley rats were selected. Eight were used for the isolation and culture of BMSCs, and the remaining 72 rats were equally and randomly divided into 4 groups: blank control group, cell transplantation group, electro-acupuncture group, and combination group. KWD-80811 pulse-type apparatus was produced by the third radio factory in Jiangsu Wujin production.METHODS: In vitro isolation and culture of the third passage of BMSCs were obtained and labeled by BrdU at hour 72 before transplantation, and BMSCs at 1 ×10 11/L were harvested for usa. Rat models of spinal cord injury were established in each group.Following model establishment, BMSC suspension was injected into the injured spinal cord near the junction of the gray matter and white matter in the cell transplantation group, totally 6 × 10<'5> cells. The same volume of phosphate buffer saline was infused into rats of the blank control group by the same method. In the electro-acupuncture group, at hour 24 after the successful model establishment, Jiaji Electroacupuncture Electroacupuncture instrument treatment was performed by pulse electro-acupuncture apparatus at 3.0-4.0 mm from median line in the upper and lower end of the spinous process vertebral body adjacent to open space for 20 minutes, once a day. In the combination group, rats underwent BMSC transplantation + Jiaji electro-acupuncture treatment. MAIN OUTCOME MEASURES: After transplantation for 3, 7, 14 days, neurological conditions were assessed by combined behavioral scores in rats with spinal cord injury. Glial fibrillary acidic protein and neuron-specific enolase expression was measured in BrdU-labeled BMSCs by immuno-double-labeling method.RESULTS: 3, 7, 14 days after injury, compared with the blank control group, significant differences in combined behavioral scores were determined in the call transplantation group, electro-acupuncture group, and combination group (P <0.05, 0.05, 0.01).The recovery of neurological function was significantly greater in the combination group than in the cell transplantation and electro-acupuncture groups (P < 0.05). No significant difference was detected between the cell transplantation group and the electro-acupunctura group (P > 0.05). Compared with blank control group, spinal cord structure was relatively complete in the cell transplantation group and electro-acupuncture group. The structure was more integrity in the combination group. Bone marrow stromal cell transplantation presented the organization BrdU-labeled cells in the lesion and the surrounding area, with obvious aggregation and survival. At day 14 following cell transplantation, positive rates of neuron-specific enolase and glial fibrillary acidic protein were respectively 7.2% and 1.5% in the call transplantation group, 7.9% and 2.1% in the combination group.CONCLUSION: The BMSCs after transplantation can survive in the host body. Electro-acupuncture could promote the differentiation of BMSCs into neural cells. Combination of electro-acupuncture and BMSC transplantation can significantly improve the motor and sensory function in rats with spinal cord injury.