抗结核药缓释涂层在兔脊柱结核模型体内的抗结核性能及组织相容性观察

Antituberculosis performance and histocompatibility of anti-tuberculosis drug slow-release coating in rabbit spinal tuberculosis model

目的:观察抗结核药异烟肼(H)、利福平(R)、吡嗪酰胺(Z)缓释涂层在兔脊柱结核模型体内的抗结核性能及组织相容性。方法:选取健康新西兰兔构建L4/5脊柱结核模型,将建模成功的60只模型兔随机分为A、B、C三组,每组20只,其中A组(实验组)病灶清除术后植入包裹HRZ缓释涂层的自体髂骨;B组(对照组)病灶清除术后植入包裹空白涂层的自体髂骨;C组(空白对照组)病灶清除术后单纯植入自体髂骨。术后观察实验兔一般状况及伤口情况;造模前1d、病灶清除术前及术后1d、3d、7d、14d、28d、56d、84d、112d进行血沉(ESR)、C反应蛋白(CRP)检测;术前以及术后14d、28d、56d进行肝肾功能检测;术后28d时A组随机处死5只兔,取心脏、肝脏、脊髓、脾脏及肾脏组织制作病理切片观察;术后56d行脊柱X线片检查后,每组随机处死5只兔在植骨部位局部取材进行骨-材料界面组织病理学观察。结果:术后B组1只兔切口感染,C组1只兔死亡、1只兔截瘫,其余兔术后一般状况良好。三组造模前和手术前的ESR和CRP均无统计学差异(P>0.05);三组手术前的ESR和CRP均较造模前显著性升高(P<0.01)。三组术后1d时ESR和CRP均较术前显著性升高,3d时CRP达到峰值,7d时ESR达到高峰,此后逐渐下降;A组至术后56d时CRP降至造模前水平,84d时ESR降至造模前水平;B、C组至112d时CRP和ESR均仍显著性高于造模前水平(P<0.05)。A组术后7d、56d、84d及112d的ESR和CRP均低于同时间点B、C组(P<0.05),B组与C组同时间点无显著性差异(P>0.05)。三组间术前和术后14d、28d、56d的肝、肾功能指标比较均无统计学差异(P>0.05);三组术后14d、28d、56d的肝、肾功能指标与术前比较均无统计学差异(P>0.05)。术后28d时A组心脏、肝脏、脾脏、脊髓及肾脏组织未见器质性改变及组织损伤征象。术后56d时A组植骨融合情况优于B组及C组(P<0.05),骨-材料界面组织病理切片骨小梁面积显著性高于B组及C组(P<0.01)。结论:HRZ缓释涂层在兔体内具有优良的抗结核性能和组织相容性。

Objectives:The anti-tuberculosis drugs isoniazid(H),rifampicin(R)and pyrazinamide(Z)sustainedrelease coatings were observed in rabbit spinal tuberculosis model for anti-tuberculosis performance and tissue compatibility.Methods:Healthy New Zealand rabbits were selected to construct L4/5 spinal tuberculosis models,and 60 successfully constructed rabbits were randomly divided into 3 groups of 20 rabbits.Group A(ex-perimental group),implanted autologous iliac bone with adhesive HRZ coating after removing the lesion;group B(control group),implanted autologous iliac bone with blank coating after lesion removal;group C(blank con-trol group),implanted autologous iliac bone after lesion removal.The general conditions and wounds of experi-mental rabbits after operation were observed.ESR and CRP were performed 1 d before modeling,preoperationand 1 d,3 d,7 d,14 d,28 d,56 d,84 d,and 112 d after operation.Liver and kidney function tests were performed before and 14 d,28 d,and 56 d after operation.5 rabbits in group A were randomly executed at 28 d after surgery,and their hearts,livers,spinal cord,spleens,and kidney tissues were excised to make pathological sections for observation.Spinal X-ray examination was performed at 56 d after operation,and 5 rabbits were randomly executed in each group to collect material samples from the bone graft site for histopathological observation of the bone-material interface.Results:One rabbit in group B was infected with incision,one rabbit in group C died,one rabbit was paraplegic,and the remaining rabbits were generally in good conditions after operation.The ESR and CRP of the three groups before modeling and before surgery were not statistically different(P>0.05);the ESR and CRP of the three groups before surgery were significantly higher than before modeling(P<0.01).The ESR and CRP of the three groups were significantly higher at 1 d after operation,and the CRP peaked at 3 d,the ESR peaked at 7 d,and then gradually decreased.In group A,the CRP decreased to the pre-model level at 56 d after surgery,and the ESR decreased to the pre-model level at 84 d;in groups B and C,the CRP and ESR were still significantly higher than the pre-model level at 112 d(P<0.05).The ESR and CRP in group A at 7 d,56 d,84 d,and 112 d after operation were lower than those at the same time in groups B and C(P>0.05),and there was no significant difference between groups B and C at the same time(P>0.05).There was no statistically significant difference in liver and kidney function indexes between the three groups before operation and 14 d,28 d,and 56 d(P>0.05);of all three groups,the liver and kidney function indexes at 14 d,28 d,and 56 d after operation were with no statistical difference comparing with those before operation.At 28 d after surgery,there were no organic changes and signs of tissue damage in the heart,liver,spleen,spinal cord and kidney tissues of group A.At 56 d after surgery,the bone graft fusion in group A was better than that in groups B and C(P<0.05),and the area of bone-material interface histopathological sections of group A was significantly bigger than that of groups B and C(P<0.01).Conclusions:HRZ slow-release coating had excellent anti-tuberculosis performance and tissue compatibility.