Correlation of nicotine dose and lumbar posterolateral fusion rate: imaging and biomechanical testing

doi:10.3969/j.issn.2095-4344.2016.48.016

Abstract

Abstract:

BACKGROUND: Smoking affects blood supply around the graft area, ultimately affecting spinal fusion. Few studies reported the effects of different doses of nicotine on spinal fusion.

OBJECTIVE: To evaluate nicotine effects on rabbit model of spinal fusion, and to clarify the correlation between the dose of nicotine and spinal fusion rate.

METHODS: Sixty healthy adult white rabbits aged 6-8 months were selected and randomly divided into four groups, with 15 rabbits in each group. Rabbits in the four groups received L_5/6 posterior lumbar intertransverse fusion under intramuscular anesthesia. On day 1 after surgery, rabbits in the control group were subcutaneously injected with physiological saline 1 mL/kg, three times a day. Rabbits in other groups received different doses of nicotine solution: 2, 4, 8 mg/kg through subcutaneous injection, three times a day. 12 weeks after surgery, effects of nicotine on the fusion rate of posterolateral lumbar interbody fusion surgery were determined by biomechanical testing, hand touch detection and imaging.

RESULTS AND CONCLUSION: (1) Hand touch detection and imaging: 2 mg/kg nicotine did not have significant effect on spinal fusion. 4 and 8 mg/kg nicotine blocked spinal fusion and decreased fusion rate. (2) Biomechanical testing: Significant differences were detected between nicotine groups and control group (P < 0.01). Significant differences were detectable among the three nicotine groups (P < 0.01). No significant difference was determined between 4 mg/kg nicotine and 8 mg/kg nicotine groups (P > 0.05). All doses of nicotine affected callus formation, reduced fusion rate; moreover, with the increased dose, the fusion rate obviously diminished. (3) These findings concluded that all doses of nicotine could decrease spinal fusion. Moreover, the effect of nicotine on spinal fusion was dose-dependent.

中国组织工程研究杂志出版内容重点：人工关节；骨植入物；脊柱；骨折；内固定；数字化骨科；组织工程

Key words: Spinal Fusion, Models, Animal, Nicotine, Biomechanics, Tissue Engineering

CLC Number:

R318

Yang Bao-jia, Yang Kai-shun, Yao Ru-bin. Correlation of nicotine dose and lumbar posterolateral fusion rate: imaging and biomechanical testing[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(48): 7251-7260.

Figures/Tables 3

2.1 实验动物数量分析纳入家兔60只，随机分为4组，每组15只，全部进入结果分析，无脱失。 2.2 手法触诊检查结果术后12周空气栓塞法处死家兔，截取L4-L7节段。由2位未参与该实验过程的骨科医师手触法检查。对照组4例标本检查发现活动融合节段有轻微松动，评定为不融合；其余11例融合节段结合紧密，触诊检查无活动，评定为融合。融合成功率为73%。实验组中，2 mg/(kg•次)尼古丁组有5例出现松动现象，认定为不融合，其余10例标本判定为融合，融合成功率为67%；4 mg/(kg•次)尼古丁组3例骨痂明显不连直接认定为不融合，有7例出现松动现象，认定为不融合，其余5例标本判定为融合，融合成功率为33%；8 mg/(kg•次)尼古丁组5例骨痂明显不连直接认定为不融合，有7例出现松动现象，认定为不融合，其余3例标本判定为融合，融合成功率为20%。对照组与2 mg/(kg•次)尼古丁组比较，2组差异无显著性意义(P > 0.05)，说明2 mg/(kg•次)的尼古丁对脊柱融合无明显影响。对照组与4 mg/(kg•次)尼古丁组比较，差异有显著性意义(P < 0.05)。对照组与8 mg/(kg•次)尼古丁组比较，差异有显著性意义(P < 0.05)。注射尼古丁的3组间采用行×列表资料χ2检验，差异有显著性意义(P < 0.05)，说明随着尼古丁剂量的增加，对脊柱融合产生了影响(见表1)。对照组融合标本骨痂体积大，硬度高，骨痂表面光滑(图5A)；8 mg/(kg•次)尼古丁组融合标本痂体积小，硬度低，骨痂表面粗糙(图5B)。 2.3 X射线检查结果所有标本经X射线检查发现：双侧横突间充满骨痂，手触法检查也为融合。单侧充满骨痂，对侧被吸收，手触测定也为不融合。对于双侧骨桥宽度不一致的标本，手触法与X射线检查结果不一致。部分手触法无明显骨痂形成的标本，X射线显示为横突间低密度影。对照组1例标本X射线检查发现单侧充满骨痂，对侧无明显骨痂，评定为不融合；其余14例双侧均充满骨痂，评定为融合，融合成功率为93%。实验组中，2 mg/(kg•次)尼古丁组有2例双侧无明显骨痂形成，认定为不融合；其余10例双侧均充满骨痂，3例双侧为骨桥相连，标本判定为融合，融合成功率为87%。4 mg/(kg•次)尼古丁组11例无明显骨痂形成，认定为不融合；其余4例标本判定为融合，融合成功率为27%。8 mg/(kg•次)尼古丁组12例无明显骨痂形成，认定为不融合；其余3例标本判定为融合，融合成功率为20%。对照组与2 mg/(kg.次)尼古丁组比较，2组差异无显著性意义(P > 0.05)，说明2 mg/(kg•次)的尼古丁对脊柱融合无明显影响。对照组与4 mg/(kg•次)尼古丁组比较，2组差异有显著性意义(P < 0.05)。对照组与 8 mg/(kg•次)尼古丁组比较，2组差异有显著性意义(P < 0.05)。注射尼古丁的3组间采用行×列表资料χ2检验，差异有显著性意义(P < 0.05)，说明随着尼古丁剂量的增加，对脊柱融合产生了影响(见表2)。 X射线表现对照组新生骨痂密度较高，皮质较厚，表面光滑(图6A)；尼古丁组随着剂量的增加植骨区骨痂较小，骨痂表面粗糙，不光滑(图6B-D)。 2.4 CT检查结果对照组3例标本CT扫描及三维重建发现骨痂表面粗糙，评定为不融合；其余12例双侧均充满骨痂，评定为融合，融合成功率为80%。实验组中，2 mg/(kg•次)尼古丁组有5例双侧无明显骨痂形成，认定为不融合；其余7例双侧均充满骨痂，3例双侧为骨桥相连，标本判定为融合，融合成功率为67%。4 mg/(kg•次)尼古丁组10例无明显骨痂形成，认定为不融合；其余5例标本判定为融合，融合成功率为33%。8 mg/(kg•次)尼古丁组11例无明显骨痂形成，认定为不融合，其余4例标本判定为融合，融合成功率为27%。图7为各组家兔融合节段CT重建图像。对照组与2 mg/(kg•次)尼古丁组比较，2组差异无显著性意义(P > 0.05)，说明2 mg/(kg•次)的尼古丁对脊柱融合无明显影响。对照组与4 mg/(kg•次)尼古丁组比较,2组差异有显著性意义(P < 0.05)。对照组与8 mg/(kg•次)尼古丁组比较，2组差异有显著性意义(P < 0.05)。注射尼古丁的3组间采用行×列表资料χ2检验，差异有显著性意义(P < 0.05)，说明随着尼古丁剂量的增加，对脊柱融合产生了影响(见表3)。 2.5 生物力学检查结果本实验通过液压万能力学试验机记录在0.4 N•m力作用下脊柱模型的最大位移，见表4。通过记录数据，利用SPSS 19.0软件包进行方差分析(表5-7)，评价不同剂量尼古丁对脊柱融合的生物力学影响。表5为各组数据的统计描述表，描述了对照组与不同剂量尼古丁实验组融合节段模型的集中趋势和离散趋势。从左到右依次为各组检测模型的例数(n)、位移数据的均数、标准差、标准误、总体均数95%可信区间、极小值和极大值。方差齐性检验结果显示，Levene统计量为0.668， ν1=3，ν2=25，P=0.58，可认为样本所有各总体的方差齐。表6为单因素方差分析的结果，F=368.976，P < 0.001，按α=0.05水准，拒绝H0，接收H1，差异有显著性意义。可认为不同剂量尼古丁对脊柱融合产生了影响。表7为采用LSD-t 检验进行两两比较的结果，均数差值用“*”标注者，表示所对应的2组均数差异有显著性意义(P < 0.01)。由表中可知：①所有实验组，无论低剂量或高剂量尼古丁组与对照组相比，差异均有显著性意义(P < 0.01)，说明尼古丁对脊柱融合产生了影响；②2 mg/(kg•次)尼古丁组与4 mg/(kg•次)尼古丁组及 8 mg/(kg•次)尼古丁组的差异均有显著性意义(P < 0.01)，说明低剂量和高剂量对脊柱融合产生的影响不一致；③4 mg/(kg•次)尼古丁组和8 mg/(kg•次)尼古丁组的差异无显著性意义(P > 0.05)。 "

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