组织工程化脂肪基质细胞诱导脊柱融合的实验研究

目的 探讨组织工程化脂肪基质细胞诱导脊柱融合的能力.方法 从成年大鼠腹股沟处无菌获取脂肪组织,消化分离培养出脂肪基质细胞.在大鼠脊柱融合模型中将32只大鼠随机分为4组,每组8只,分别植入骨形态发生蛋白-2(BMP-2)基因重组的腺病毒载体转染的脂肪细胞(A组)、LacZ基因重组的腺病毒载体转染的脂肪细胞(B组)、未转染脂肪细胞(C组)、胶原海绵对照(D组).测定A组和B组的BMP-2的定量表达.比较4组术后4、6、8周X线片评分.8周后处死大鼠,进行MicroCT扫描分析和组织学观察.结果 ELISA法结果显示A组BMP-2表达水平为3.46 ng/mL,而B组无BMP-2表达.A组4、6、8周X线片评分分别为(4.56±1.01)分、(4.72±0.24)分、(4.92±0.13)分,均明显高于其他组,差异有统计学意义(P＜0.05).Micro CT和组织学切片均显示A组全部获骨性融合,而其他组均未融合.结论 组织工程化的脂肪基质细胞能够很好地诱导脊柱融合,有望成为一种新颖、有效的治疗方法.

Abstract：

Objective To assess the capability of inducing spinal fusion by adipose derived stromal cells in a rat model. Methods Adipose derived stromal cells were isolated from inguinal fat pads in rats.In the rat model of spinal fusion,32 rats were randomized into 4 groups.In group A,the cells infected with AdBMP-2 were implanted.In group B,cells infected with AdLacZ were implanted.In group C,uninfected cells were implanted.In group D,only collagen sponges were implanted.Expressions of BMP-2 were quantified in groups A and B.The radiographic scores were compared at weeks 4,6 and 8 among the 4 groups.After sacrifice at week 8,the spine samples were assessed by radiographs,MicroCT,and histological analysis. Results ELISA revealed a 3.46 ng/mL expression of BMP-2 in group A but no expression in group B.The mean radiographic scores for group A at weeks 4,6 and 8 were 4.56 ± 1.01,4.72 ± 0.24,4.92 ±0.13,significantly higher than those for the other groups ( P ＜ 0.05) .MicroCT and histological analysis revealed spinal fusion in all the rats in group A.None of the rats in the control groups developed fusion.Conclusion Because tissue engineered adipose derived stromal cells can induce spinal fusion,they may be a promising strategy for spinal fusion in the future.

Tissue engineered adipose derived stromal cells induce spinal fusion

Objective To assess the capability of inducing spinal fusion by adipose derived stromal cells in a rat model. Methods Adipose derived stromal cells were isolated from inguinal fat pads in rats.In the rat model of spinal fusion,32 rats were randomized into 4 groups.In group A,the cells infected with AdBMP-2 were implanted.In group B,cells infected with AdLacZ were implanted.In group C,uninfected cells were implanted.In group D,only collagen sponges were implanted.Expressions of BMP-2 were quantified in groups A and B.The radiographic scores were compared at weeks 4,6 and 8 among the 4 groups.After sacrifice at week 8,the spine samples were assessed by radiographs,MicroCT,and histological analysis. Results ELISA revealed a 3.46 ng/mL expression of BMP-2 in group A but no expression in group B.The mean radiographic scores for group A at weeks 4,6 and 8 were 4.56 ± 1.01,4.72 ± 0.24,4.92 ±0.13,significantly higher than those for the other groups ( P ＜ 0.05) .MicroCT and histological analysis revealed spinal fusion in all the rats in group A.None of the rats in the control groups developed fusion.Conclusion Because tissue engineered adipose derived stromal cells can induce spinal fusion,they may be a promising strategy for spinal fusion in the future.