植入物界面结核分枝杆菌生物膜形成的实验研究

发布时间：2018-08-19 09:16

【摘要】：【目的】研究不同植入物界面结核分枝杆菌的粘附能力及生物膜形成情况,同时观察抗结核药物利福平对结核分枝杆菌粘附能力及生物膜的影响,以及骨结核病灶内结核分枝杆菌在植入物界面形成生物被膜的情况,探讨一期内固定或关节成行技术治疗活动性骨与关节结核的可行性和安全性,为内固定术或人工关节成行术治疗活动性骨结核安全可靠的原因提供坚实的基础理论依据。【方法】将钛合金、钴铬钼合金、聚乙烯三种材料的植入物在无菌条件下分别置于用Middlebrook 7H9液体培养基制备的结核分枝杆菌悬浮液中,共同培养4周后,随机取出1块,再加入利福平溶液继续培养2周。样品经固定、干燥、喷金后,运用电子显微镜观察,并计数植入物界面单位面积粘附菌落数,比较结核分枝杆菌对不同植入物(钛合金、钴铬钼合金、聚乙烯)、界面(光滑面、粗糙面)的粘附能力,并观察利福平干预前后植入物界面生物膜结构。收集骨结核内固定术后复发患者再次手术时取出的内置物,经固定、干燥、喷金后,采用扫描电子显微镜观察取出内固定物界面结核生物被膜的形成情况。【结果】利福平干预前,聚乙烯界面黏附的菌落数显著多于钴铬钼界面,差异有统计学意义(P0.05);钴铬钼材料界面黏附的菌落数与钛合金材料界面相比较,差异无统计学意义(P0.05);利福平干预后,三种植入物界面黏附菌落数均显著减少,差异均有统计学意义。植入物界面粗糙度不同,结核分枝杆菌的粘附能力不同,更容易黏附于植入物的粗糙界面。植入物界面与材料属性之间具有一阶交互作用(P0.05)。结核分枝杆菌在钛合金、钴铬钼材料界面未见生物膜形成;在聚乙烯界面能见典型的生物膜形成,但利福平干预能抑制,甚至破坏结核生物膜。从骨结核病灶内取出的植入物界面仅见散在结核分枝杆菌粘附,或是干瘪或是裂解,未见结核生物膜形成。【结论】结核分支杆菌对植入物的粘附能力与植入物的材料属性、界面状态有关,可在植入物界面形成生物膜,但具有选择性和特异性。利福平干预能降低结核分支杆菌粘附植入物界面的能力,而且能抑制,甚至破坏结核生物膜。骨结核病灶内,未见结核分枝杆菌在植入物钛合金、钴铬钼材料的界面形成生物膜。结核分支杆菌对植入物界面的低粘附力、极少形成生物膜是活动性骨关节结核一期病灶清除内固定或关节成行技术安全可行的紧要因素。在全身和局部使用抗结核药物的前提下,选择性应用假体植入结核病灶,不会增加骨结核术后复发的风险。
[Abstract]:[objective] to study the adhesion ability and biofilm formation of Mycobacterium tuberculosis at the interface of different implants, and to observe the effect of rifampicin on the adhesion ability and biofilm of Mycobacterium tuberculosis. And the formation of biofilm of Mycobacterium tuberculosis in bone tuberculosis lesions at the implant interface, and to explore the feasibility and safety of one-stage internal fixation or joint formation technique in the treatment of active bone and joint tuberculosis. To provide a solid theoretical basis for the safe and reliable treatment of active bone tuberculosis by internal fixation or artificial articulation. [methods] Titanium alloy, cobalt-chromium-molybdenum alloy, The implants of three kinds of polyethylene materials were placed in the suspension of Mycobacterium tuberculosis prepared by Middlebrook 7H9 liquid medium in aseptic condition. After 4 weeks of co-culture, one piece was taken out at random, and then added rifampicin solution for 2 weeks. After fixing, drying and spraying gold, the samples were observed by electron microscope, and the number of colony adhesion per unit area of implant interface was counted. The effect of Mycobacterium tuberculosis on different implants (titanium alloy, cobalt-chromium-molybdenum alloy, polyethylene) and interface (smooth surface) were compared. The adhesiveness of rifampicin was observed before and after the intervention of rifampicin, and the biofilm structure of implant interface was observed. To collect the internal objects taken out by the patients with recurrent bone tuberculosis after reoperation, after fixation, drying, and spraying gold, The formation of biofilm was observed by scanning electron microscope. [results] before the intervention of rifampicin, the number of colonies adhered to the interface of polyethylene was significantly higher than that of the interface of cobalt, chromium and molybdenum. There was significant difference (P0.05); there was no significant difference in colony number between cobalt, chromium-molybdenum interfacial adhesion and titanium alloy interface (P0.05); after rifampicin intervention, the number of colonies at the interface of the three implants decreased significantly. The difference was statistically significant. The interface roughness of implants is different, and the adhesion ability of Mycobacterium tuberculosis is different, so it is easier to adhere to the rough interface of implants. There was a first order interaction between implant interface and material properties (P0.05). Mycobacterium tuberculosis did not form biofilm at the interface of cobalt, chromium and molybdenum, but typical biofilm formation could be seen at the interface of polyethylene, but rifampicin could inhibit or even destroy the biofilm. The interface of the implants extracted from the bone tuberculosis lesions was only scattered in the adhesion of Mycobacterium tuberculosis, either dried or cracked, and no biofilm formation was observed. [conclusion] the adhesion ability of Mycobacterium tuberculosis to the implants and the material properties of the implants were observed. The biofilm can be formed at the implant interface, but it is selective and specific. Rifampicin can reduce the ability of mycobacterium tuberculosis to adhere to the implant interface, and can inhibit or even destroy TB biofilm. In the focus of bone tuberculosis, no mycobacterium tuberculosis formed biofilm at the interface of the implanted titanium alloy and cobalt chromium-molybdenum material. The low adhesion of Mycobacterium tuberculosis to the implant interface and the rare formation of biofilm are the key factors for the primary debridement and internal fixation of active bone joint tuberculosis or for the safety and feasibility of joint formation. Under the premise of systemic and local use of antituberculous drugs, selective implant implantation of tuberculosis does not increase the risk of postoperative recurrence of bone tuberculosis.
【学位授予单位】：天津医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R687;R529.2

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