浓缩生长因子在大鼠椎间盘纤维环修复中的作用研究

Effect of concentrated growth factor on repair of fibrous annulus of intervertebral disc in rats

中文关键词:  浓缩生长因子  大鼠椎间盘破裂模型  椎间盘破损  椎间盘修复

英文关键词:Concentrated growth factors(CGF)  Rat intervertebral disc rupture model  Intervertebral disc injury  Disc annulus repair

中文摘要:

  【摘要】 目的：探究浓缩生长因子（concentrated growth factor，CGF）在大鼠椎间盘纤维环破损修复中的作用。方法：首先选取成年SD雌性大鼠4只，周龄6~7周，体重200~250g。麻醉后从大鼠的腹静脉取血，提取血液置于变速离心机中离心，提取获得CGF。然后选取同批成年SD雌性大鼠36只，随机分为空白组、手术组、CGF组，每组12只。空白组只暴露L4/5椎间盘不做其余处理；手术组大鼠L4/5行椎间盘穿刺造大鼠椎间盘破裂模型；CGF组大鼠造模后用CGF制作成膜后覆盖纤维环破口。造模后常规给予抗感染、止痛等处理。术后4、6周分批取材，取L4/5椎间盘观察，并行HE、Ⅱ型胶原免疫组化染色。结果：术后4、6周，基本保持髓核组织完整性情况下，手术组L4/5椎间盘纤维环对比空白组出现明显破损，CGF治疗组L4/5椎间盘纤维环对比手术组破损明显减少；椎间盘切片HE染色结果提示，与手术组相比，CGF组纤维环修复明显；术后6周，各组大鼠椎间盘Ⅱ型胶原酶蛋白免疫组化染色结果提示，手术组纤维环破损较CGF组更严重，Ⅱ型胶原酶蛋白表达集中在纤维环破损部位。与手术组相比，CGF组纤维环破损较轻，Ⅱ型胶原酶蛋白表达同样集中在破损部位。结论：CGF可有效促进大鼠椎间盘纤维环破损修复。

英文摘要:

  【Abstract】 Objectives: To investigate the effect of concentrated growth factor(CGF) on the repairation of damaged intervertebral disc annulus in rats. Methods: First, 4 adult Sprague-Dawley(SD) female rats were selected, 6-7 weeks old, weight 200-250g, blood was collected from the rats′ abdominal veins after anesthesia and processed through centrifugation to isolate CGF. Then 36 adult SD female rats were selected from the same batch rats, and were randomly assigned into three groups of control group, surgery group, and CGF group, with 12 rats per group. In the blank group, only the L4/5 discs were exposed and no other treatment was done. In the control group, only the L4/5 discs were exposed and no other treatment was done. The surgery group had a disc puncture performed at the L4/5 level to induce disc rupture; In the CGF group, after modeling, membrane was made with CGF to cover over the annulus fibrosus defect. Postoperative care included standard antiseptic and analgesic treatments. At 4 and 6 weeks postoperatively, tissues from the L4/5 level were harvested for morphological assessment via hematoxylin and eosin(HE) and type Ⅱ collagen immunohistochemistry staining. Results: Postoperatively at both 4 and 6 weeks, with the preservation of the nucleus pulposus′s integrity, the surgery group demonstrated pronounced annulus fibrosus damage at the L4/5 disc compared to the control group. In contrast, the CGF group showed a significant decrease in annulus fibrosus damage relative to the surgery group. HE staining of disc sections revealed that the CGF group had markedly improved annulus fibrosus healing compared to the surgery group. The immunohistochemistry of type Ⅱ collagenase protein of intervertebral disc in each group at 6 weeks after operation indicated that the damage of fibrous ring in the surgery group was more serious than that in the other two groups, and the expression of type Ⅱ collagenase protein was concentrated in the damaged site. Compared with the surgery group, the annulus fibrosus damage was lighter in the CGF group, and the expression of type Ⅱ collagenase protein was also concentrated in the damaged site. Conclusions: CGF can effectively promote the repair of damaged intervertebral disc annulus fibrosus.