邻近节段融合退变动物模型的建立及相关实验研究。
Establishment of an animal model of adjacent segment degeneration after interbody fusion and related experimental studies.
发表日期:2023 Sep 07
作者:
Song Chen, Shiqi Suo, Zhitao Xie, Jinglan He, Jiaqi Li, Dengke Duan, Guoyong Qiao, Wei Zhang
来源:
Bone & Joint Journal
摘要:
退行性脊柱病变是常见而频繁的临床疾病,脊柱融合术(SF)后的邻近节段疾病(ASD)是脊柱融合术(SF)后常见的并发症。本研究建立了一种ASD 的动物模型,观察邻近节段(AS)椎间盘的形态学变化,并确定肿瘤坏死因子-α(TNF-α)和白细胞介素-1β(IL-1β)在ASD组织中的表达和意义,为临床预防和治疗椎间盘融合术后ASD提供良好的实验基础和参考。将36只2.0-2.5千克的雄性和雌性新西兰白兔随机分为对照组(A组)和实验组(B组,C组,和D组),每组9只白兔,其中B组,C组和D组分别为4,8和12周组。自体髂骨移植物被用作骨移植材料。在实验组中,在C2-C3椎间隙进行脊柱融合术。检查C3-4邻近段。实验组的动物在手触摸检查,X射线检查,手触检查,并且术后4,8和12周接受显微计算机断层扫描检查(显微CT)。使用光学显微镜观察对照组和实验组椎间盘区段的微观形态学变化。免疫组织化学和Western blotting用于检测在与对照组和实验组进行植入体融合术后的AS组织中TNF-α和IL-1β的表达。兔子颈椎骨结构的测量数据表明,从第二至第六颈椎的椎体长度和椎体下端矢状直径逐渐减小,差异有统计学意义(P<0.05)。椎体下端横径的差异无统计学意义(P>0.05),椎体下端斜径的变化波动,差异有统计学意义(P<0.05)。手触检查显示,B组、C组、D组脊柱融合率分别为22.2%(2/9)、55.6%(5/9)和88.9%(8/9)。B、C、D组的骨体积/总体积(BV/TV)和X射线评分的差异具有统计学意义(P<0.05)。与A组相比,B组、C组、D组出现了明显的退变。与A组相比,实验组的椎间盘组织中TNF-α和IL-1β的表达明显升高(P<0.05),并随时间增加。本研究成功建立了兔子椎间盘融合术后ASD的动物模型。术后影像学和手触检查显示新椎间骨数量与融合程度呈正相关,随时间逐渐增加。免疫组织化学和Western blot结果显示,在椎间盘融合术后,实验组AS组织中TNF-α和IL-1β的表达显著增高,并且椎间盘退变程度与椎间盘融合术后时间呈正相关。(© 2023. BioMed Central Ltd., part of Springer Nature.)
Degenerative spine conditions are common and frequent clinical diseases, and adjacent segment disease (ASD) after spinal fusion (SF) is a common complication after spinal fusion (SF). In this study, we established an animal model of ASD after interbody fusion to observe the morphologic changes of adjacent segment (AS) disks and to determine the expression and significance of tumor necrosis factor-alpha (TNF-α) and interleukin-1beta (IL-1β) in ASD tissues to provide a good experimental basis and reference for clinical prevention and treatment of ASD after interbody fusion.Thirty-six male and female New Zealand rabbits weighing 2.0-2.5 kg were randomly divided into control group (group A) and experimental groups (groups B, C, and D), with 9 rabbits in each group, of which groups B, C, and D were the 4-, 8-, and 12-week groups, respectively. Autologous iliac bone grafts were used as the bone graft material. In the experimental groups, a SF was performed on the C2-C3 intervertebral space. The C3-4 adjacent segments were examined. In the experimental group, the animals were subjected to gross observation, X-ray examination, hand touch inspection, and micro-computed tomography (micro-CT) 4, 8, and 12 weeks after surgery. The micromorphologic changes of the cervical disks in the segments of the control group and experimental groups were observed under light microscopy. Immunohistochemistry and Western blotting were used to detect the expression of TNF-α and IL-1β in the AS tissues after interbody fusion in the control and experimental groups.The measurement data of the rabbit cervical spine bony structures indicated that the length of the vertebral body and the sagittal diameter of the lower end of the vertebral body decreased gradually from the 2nd-6th cervical vertebrae, and the difference was statistically significant (P < 0.05). The difference in the transverse diameter of the lower end of the vertebral body was not statistically significant (P > 0.05), the change in the oblique diameter of the lower end of the vertebral body fluctuated, and the difference was statistically significant (P < 0.05). The fusion rate of the cervical spine by hand touch inspection was 22.2% (2/9), 55.6% (5/9), and 88.9% (8/9) in groups B, C, and D, respectively. The differences in bone volume-to-total volume (BV/TV) and X-ray scores were statistically significant in groups B, C, and D (P < 0.05). Significant degeneration occurred in groups B, C, and D compared with group A. The expression of TNF-α and IL-1β in the intervertebral disk tissue was significantly higher in groups B, C, and D compared with group A (P < 0.05), and increased with time.In this study, an animal model of ASD after interbody fusion fixation in rabbits was successfully established. Postoperative imaging and hand touch inspection showed a positive correlation between the amount of new intervertebral bone and the degree of fusion with time. The results of immunohistochemistry and Western blot showed that TNF-α and IL-1β were highly expressed in the AS tissues of the experimental group after interbody fusion, and the degree of disk degeneration was positively correlated with the time after interbody fusion.© 2023. BioMed Central Ltd., part of Springer Nature.