Preparation of intervertebral disc degeneration models by percutaneous acupuncture rotary-cut under different anesthesia methods

doi:10.3969/j.issn.2095-4344.2017.24.012

Abstract

Abstract:

BACKGROUND: Different anesthesia methods significantly impact mean arterial pressure, systolic and diastolic blood pressure, arterial blood pH and blood viscosity, but what kind of anesthesia method is more suitable for preparation of animal model does not reach an agreement.

OBJECTIVE: To compare the effectiveness of local anesthesia and general anesthesia for preparation of rabbit intervertebral disc degeneration model in procession of percutaneous acupuncture rotary cut.

METHODS: A total of 48 New Zealand white rabbits were randomly divided into the local anesthesia group and the general anesthesia group. The local anesthesia group was anesthetized with 0.5% lidocaine. The general anesthesia group was intraperitoneally injected with 3% sodium pentobarbital (30 mg/kg). In the two groups, models of intervertebral disc degeneration were established by minimally invasive percutaneous acupuncture rotary cut at L_4/5, L_5/6 levels. Modeling time was observed in both groups. Disc degeneration was evaluated by the gross observation, MRI detection, and histopathological examination at 4, 8, 12, and 16 weeks after modeling.

RESULTS AND CONCLUSION: (1) General observation: the nucleus pulposus gradually with darker color and elasticity reduced in two groups after modeling. (2) MRI T2-weighted image results showed no significant changes in disc signal intensity at early stage, but the signal strength showed a decreasing trend with time. (3) Disc degeneration was evaluated according to Pfirrmann grading. Disc degeneration was gradually increased with time in two groups (P < 0.05), but there was no significant difference at postoperative time points in both groups (P > 0.05). (4) Masson staining showed that different degrees of annulus irregular arrangement appear in the two groups 8 and 12 weeks after modeling, but the structure was still intact. 16 weeks later, annulus disorganized, or even breakage in the two groups; no significant difference was detected between the two groups. (5) Safranin O staining showed that nucleus pulposus cells were not significantly reduced in the two groups at 4 weeks, but were significantly reduced at 16 weeks. (6) The average modeling time of local anesthesia group [(15.24±2.67) minutes] was significantly shorter than that in the general anesthesia group [(25.64±6.85) minutes] (P < 0.05). These results indicate that intervertebral disc degeneration model can be successfully established by minimally invasive percutaneous acupuncture rotary cut by using local anesthesia and general anesthesia; however, local anesthesia has a shorter operating time and simple anesthetic procedure. The efficacy was identical between the two methods.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Spine, Intervertebral Disk, Models, Animal, Anesthesia, Acupuncture

CLC Number:

R318

Lai Ju-yi, He Sheng-hua, Sun Zhi-tao, Wang Ye-guang, Wang Jian, Feng Hua-long, Huang Fei-qiang. Preparation of intervertebral disc degeneration models by percutaneous acupuncture rotary-cut under different anesthesia methods[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(24): 3839-3844.

Figures/Tables 1

2.1 实验动物数量分析实验选用新西兰大白兔48只，分为2组，局麻组造模后至实验结束未出现动物意外死亡，全麻组1只兔在穿刺操作过程死亡，1只兔于造模后1 d死亡(2只兔死亡后均立即按原实验设计进行补充)；进入结果分析48只。 2.2 动物一般情况 2组动物术后2 d四肢活动度好，精神状态恢复至造模前，2组均未见穿刺后并发症发生；局麻组的平均造模时间为(15.24±2.67) min，全麻组的平均造模时间为(25.64±6.85) min，造模时间2组相比较差异有显著性意义(t=6.66，P < 0.05)。 2.3 MRI观察 MRI T2加权像显示2组动物椎间盘信号强度造模后4周未见明显改变，8周后均略有减弱，造模后12，16周信号强度明显降低。2组动物造模椎间盘信号强度均随时间延长呈减弱趋势(图2)。根据Pfirrmann分级法对2组动物椎间盘退变程度进行统计对比发现，随着观察时间的延长，2组椎间盘退变程度均逐渐加重，但2组间比较差异无显著性意义(P > 0.05)，见表1。 2.4 椎间盘组织大体及组织学观察大体观察造模前2组兔髓核外观透亮、含水量高；造模后4周2组穿刺后的髓核颜色稍较造模前暗淡；造模后8，12周时髓核组织含水量减少较明显，并呈淡黄色；造模后16周时髓核含水量显著减少并呈干瘪样(图3)。组织学观察Masson染色显示2组纤维环在造模后4周呈序列整齐的同心圆形状分层结构；8周后2组染色结果均见部分纤维环排列紊乱，有少量裂隙存在；造模后12周时纤维环排列紊乱明显加重，裂隙增多(图4)；造模后16周时列紊乱程度进一步加重，甚有纤维环不连续现象。番红O染色结果发现2组椎间盘髓核细胞在造模4周后呈紫红色，分布情况均匀；至造模后8周时2组髓核细胞均有略减少；造模后12周髓核细胞染色变淡，分布不均；16周后2组髓核细胞均明显减少，染色更淡。"

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