小尾寒羊颈椎前路椎间盘切除融合模型的建立及评估

Establishment and evaluation of anterior cervical discectomy fusion model in small-tailed Han sheep model

目的颈椎间盘突出症(cervical disc herniation,CDH)是骨科常见疾病之一,随着对该疾病研究的深入及颈椎内植物的发展,建立颈椎融合动物模型成为不可或缺的部分,目前国内对颈椎融合动物模型建立及评估的研究报道较少,本研究以期为颈椎融合相关研究提供完备的动物模型和内植物性能的评估方案。方法选择小尾寒羊,改良术式后行颈椎前路椎间盘切除融合术(anterior cervical discectomy and fusion,ACDF),将聚醚醚酮(polyetheretherketone,PEEK)椎间融合器(interbody fusion cage,Cage)(对照组)、3D打印钛合金Cage(实验组1)及新方法钛合金Cage(实验组2)分别植入每只羊的不同颈椎节段(C2/3~C4/5),术后行血液学检测、组织病理学分析评估手术恢复情况及材料生物安全性,利用X光、CT、Micro-CT及定量分析、硬组织切片染色、生物力学试验评估内植物的骨长入及骨融合情况。结果绵羊改良术式ACDF模型建立成功,血液学检测重要指标无显著性差异(P>0.05),组织病理学分析显示均无炎症细胞浸润等病理改变,内植物生物安全性良好,X光及CT显示内固定位置及椎间融合情况良好,术后3个月及6个月Micro-CT及定量分析表明,与PEEK Cage组相比,新方法钛合金Cage组及3D打印钛合金Cage组内部的骨体积/总体积、骨小梁数目显著性升高(P<0.01),骨小梁间距显著性降低(P<0.01),且新方法钛合金Cage组骨质长入更多(P<0.01),硬组织切片染色表明新方法钛合金Cage组及3D打印钛合金Cage组孔隙内有明显骨质长入且较为密实,结合较PEEK Cage组略好,生物力学试验显示,与PEEK Cage组相比,新方法钛合金Cage及3D打印钛合金Cage在一定程度上降低了颈椎屈伸、侧弯、扭转运动范围(P<0.05),同时增强了颈椎的稳定性,且新方法钛合金Cage更有优势(P<0.05)。结论建立绵羊改良术式ACDF模型后,利用合理有效的评估方法,证明了该模型的合理性及有效性,同时说明3种材料的Cage均显示出良好的生物安全性,新方法钛合金Cage及3D打印钛合金Cage较PEEK Cage的骨长入及骨融合性能更强,可增强颈椎的稳定性,且新方法钛合金Cage更有优势。

Objective Cervical disc herniation(CDH)is one of the common orthopaedic diseases.With the in-depth study of it and the development of cervical implants,the establishment of cervical fusion animal models has become an indispensable part.Notably however,studies of the establishment and evaluation of cervical fusion animal models in China are currently lacking.This study aimed to provide a suitable animal model and evaluation scheme for implants for cervical spine-related research.Methods Small-tailed Han sheep were chosen for anterior cervical discectomy fusion(ACDF)after modified surgery,and a polyetheretherketone(PEEK)interbody fusion cage(Cage)(control group),3D-printed Ti6Al4V Cage(group 1),and new method Ti6Al4V Cage(group 2)were implanted in different cervical segments(C2/3~C4/5)in each sheep,respectively.Hematology and histopathological analyses were carried out after surgery to evaluate recovery of sheep and the biosafety of the materials.Bone in-growth and bone fusion were assessed by X-ray,computed tomography(CT),Micro-CT and quantitative analysis,hard tissue section staining,and biomechanical tests.Results The modified ACDF ovine model was established successfully.There were no significant differences in important hematology indexes(P>0.05)and histopathological analysis showed no pathological changes,such as inflammatory cell infiltration.The implants had good biosafety.Furthermore,X-ray and CT examinations showed that the position of internal fixation and the interbody fusion were good.Micro-CT and quantitative analysis at 3 and 6 months after operation showed that compared with PEEK Cage group,the bone volume/total volume and trabecular number were significantly increased(P<0.01)while the trabecular spacing was significantly decreased in the new method Ti6Al4V and 3D-printed Ti6Al4V groups compared with the PEEK Cage group(P<0.01).Moreover,the new method new method Ti6Al4V Cage group had more bone growth(P<0.01).Hard tissue section staining demonstrated that the pores of the new method Ti6Al4V Cage and 3D-printed Ti6Al4V Cage had obvious bone growth and relatively dense pores in the new method Ti6Al4V and 3D-printed Ti6Al4V groups,and the combination was slightly better than that of PEEK Cage.Biomechanical evaluation indicated that the new method Ti6Al4V Cage and 3D-printed Ti6Al4V Cage reduced the range of cervical flexion-extension,lateral bending,and axial rotation(P<0.05)compared with the PEEK cage,as well as enhancing the stability of the cervical vertebra,and the new method Ti6Al4V Cage was more advantageous(P<0.05).Conclusions After the establishment of the modified ACDF ovine model,reasonable and effective assessment method were used to demonstrate the suitability and effectiveness of the model and the good biosecurity of all three Cage materials.Compared with the PEEK Cage,the new method Ti6Al4V Cage and 3D-printed Ti6Al4V Cages showed better performances in terms of bone growth and bone fusion,which could enhance the stability of the cervical vertebrae.The new method Ti6Al4V Cage was particularly advantageous.