双向调控骨吸收和骨生成防治骨溶解疾病的新手段

发布时间：2018-04-15 03:03

本文选题：镁 + 穿心莲内酯　；参考：《上海交通大学》2015年博士论文

【摘要】：背景和目的无菌性松动是人工关节置换远期失效的主要原因,而磨损颗粒诱导的假体周围骨溶解则是造成无菌性松动的罪魁祸首。这一过程的发生与以下三个因素密切相关:炎性因子浸润、骨吸收亢进及骨生成障碍。三者相互促进,我们对任何一方的轻视,都将在无菌性松动的防治中付出代价。与此同时,感染也是关节置换的主要并发症之一,其结果具有灾难性,假体常需取出,并行二次翻修术。研究表明,内植物表面细菌黏附和生物膜的形成是假体感染形成的关键环节。而矛盾的是,骨内植物需满足良好的生物相容性,促进成骨细胞黏附以利于骨整合,但这无疑又会促进细菌黏附进而增加感染的机会。因此,理想的骨内植物材料需兼顾促进骨整合和抗感染活性。本课题拟针对磨损颗粒诱导骨溶解的发生机制,从对抗炎症因子释放、抑制骨吸收及促进骨形成的角度出发,探讨镁降解产生的局部微环境在防治假体无菌性松动中的潜在应用价值。并观察镁作为骨内植物的抗感染性能。同时,从调控机体全身骨代谢为出发点,探讨植物天然提取物穿心莲内酯(Andrographolide)的全身骨保护作用。以期从局部及全身出发,对抗磨损颗粒诱导骨溶解的发生,为防治假体无菌性松动提供新手段。内容与方法1.按照国标ISO10993浸泡镁(99.9%)模拟镁降解产物(Magnesium Degradation,Mg D),观察镁降解产物对单核-巨噬细胞(BMMS)增殖、破骨细胞分化及骨吸收功能的影响,探讨其引起的破骨细胞应答反应;建立磨损颗粒诱导小鼠颅骨溶解模型,利用micro-CT、组织形态计量等评价镁降解产物对骨溶解的抑制效果。观察镁降解产物对骨髓间充质干细胞(BMSC)粘附、铺展、增殖及向成骨细胞分化的影响,探讨其引起的h BMSC应答反应。2.体外观察镁(99.9%)的抗耐甲氧西林金黄色葡萄球菌(MRSA)效果,建立SD大鼠骨髓炎模型,将镁棒作为髓内钉放入大鼠髓腔内,观察对骨髓炎的治疗效果和周围骨量的改变,并探讨作用机制。3.通过观察Andrographolide对成骨细胞和破骨细胞分化及功能的影响,探讨其引起的骨应答反应,并阐述其作用机制;建立磨损颗粒诱导局部骨溶解模型和LPS诱导全身炎症性骨丢失模型,腹腔注射Andrographolide,利用micro-CT、组织形态计量等评价Andrographolide对小鼠局部和全身的骨保护作用。结果1.镁降解产物通过抑制破骨细胞特异性转录因子NFATc1的转录活性,进而显著抑制破骨细胞形成和骨吸收功能;在磨损颗粒诱导骨溶解小鼠体内,镁降解产物降低组织中活化破骨细胞数量和炎症因子TNF-?、IL-1β及RANKL水平,显著抑制骨丢失。此外,镁降解产物提高BMSC表面integrinα3/α5/β1表达水平,促进BMP2下游SMADs复合物核转位,增强BMSC增殖、黏附及成骨分化功能,但并不利于胞外钙沉积。2.镁在体外显著抑制MRSA的粘附和生物膜的形成,这可能得益于镁降解产生的碱性环境。同时,镁植入骨髓腔后显著抑制骨髓炎的进展,并有大量新生骨。3.体外研究发现,Andrographolide通过抑制RANKL下游p65入核和ERK磷酸化,进而发挥抑制破骨细胞功能的作用。与此同时,Andrographolide提高成骨标志性基因的表达和OPG/RANKL水平,促进成骨分化,促进骨代谢向骨生成方向发展。在LPS诱导的全身性骨丢失和磨损颗粒诱导局部骨溶解中,Andrographolide显著改善炎症浸润和骨溶解,骨小梁表面破骨细胞数量显著减少,显示出良好的骨保护作用。结论1.镁降解产生的微环境可抑制破骨细胞分化和骨吸收功能,促进BMSC增殖、粘附和成骨分化,降低炎症因子水平,对抗骨溶解发生机制,抑制磨损颗粒诱导的局部骨丢失。镁可能成为未来防治骨溶解疾病的新手段。2.镁在体外抑制MRSA粘附和生物膜形成,在体内抑制骨髓炎进展并减少感染周围骨丢失。3.Andrographolide抑制破骨细胞分化和骨吸收,促进成骨分化,在全身水平降低炎症水平,正向调节骨代谢,具有显著的全身骨保护作用。
[Abstract]:Background and objective: aseptic loosening is the main reason for the failure of artificial joint replacement in the long term, and the wear particle induced periprosthesis osteolysis is caused by arch-criminal aseptic loosening. This process is closely related to the following three factors: the infiltration of inflammatory factors, bone resorption and bone formation obstacles. Hyperthyroidism three promote each other we despise, to any party, will pay the price in the prevention and treatment of aseptic loosening. At the same time, one of the major complications of infection and joint replacement, the results are disastrous, often need to remove the prosthesis two times, parallel revision. The study shows that the formation of plant surface bacterial adhesion and biofilm is the key link of periprosthetic infection formation. Paradoxically, bone implants required good biocompatibility, promote osteoblast adhesion to bone integration, but this will undoubtedly promote the adhesion of bacteria and Increase the chance of infection. Therefore, the ideal bone implant materials should be taken into consideration to promote osseointegration and anti infection activity. This paper according to the mechanism of wear debris induced osteolysis, released from against inflammatory factors, inhibit bone resorption and promote bone formation in the perspective of local magnesium produced by the degradation of micro environment in aseptic loosening the prevention of prosthesis in potential applications. And to observe the bone magnesium as plant anti infective properties. At the same time, from the regulation of bone metabolism as a starting point to explore the natural plant extract andrographolide (Andrographolide) of the bone protective effect. In order from the perspective of local and systemic confrontation, wear particles induced osteolysis, provide a new method for prevention and treatment of aseptic loosening of the prosthesis. The contents and methods in accordance with the national standard ISO10993 1. for magnesium (99.9%) magnesium degradation products (Magnesium Degradation simulation, Mg, D) on magnesium reduction The hydrolysate on mononuclear macrophage (BMMS) proliferation, osteoclast differentiation and bone resorption of osteoclasts, which caused the response; establish mice induced by skull dissolution model, wear particles by micro-CT, the inhibitory effect of histomorphometric evaluation of magnesium degradation products on osteolysis. Observe the degradation products of magnesium bone marrow mesenchymal stem cells (BMSC) adhesion, spreading, proliferation and osteogenic differentiation in vitro, observe the magnesium in vitro h BMSC response induced by the.2. (99.9%) of methicillin resistant Staphylococcus aureus (MRSA) effect, establish the SD rat model of osteomyelitis, the magnesium stick as intramedullary nail into the medullary cavity of rats, to observe the therapeutic effect and osteomyelitis of the bone mass around the change, and to investigate the mechanism of.3. by observing the effect of Andrographolide on osteoblast and osteoclast differentiation and function, to explore its causes The bone response, and discuss the mechanism; establish local model and LPS induced osteolysis induced systemic inflammatory bone loss model of wear particles, intraperitoneal injection of Andrographolide, using micro-CT, bone histomorphometry and evaluate the protective effect of Andrographolide on local and systemic mice. Results 1. mg degradation products by inhibiting transcriptional activity of osteoclast specific the transcription factor NFATc1, and inhibit osteoclast formation and bone resorption in vivo; wear particles induced osteolysis, osteoclast number and inflammatory factor TNF- activation of magnesium degradation products reduce the organization?, IL-1 beta and RANKL level, inhibit bone loss. In addition, the degradation products of magnesium increased BMSC surface integrin alpha 3/ alpha 5/ beta 1 expression, promote BMP2 downstream SMADs complex nuclear translocation, enhanced BMSC proliferation, adhesion and osteogenic differentiation and function, but unfavorable to extracellular calcium deposition. The formation of 2. mg in vitro significantly inhibited MRSA adhesion and biofilm alkaline environment, this may be due to the degradation of magnesium production. At the same time, the progress of magnesium implanted bone marrow cavity significantly inhibited osteomyelitis, and a large amount of new bone.3. in vitro, Andrographolide through inhibition of RANKL downstream nuclear translocation of p65 and ERK phosphorylation, and play the inhibition of osteoclast function. At the same time, improve the Andrographolide of osteoblast marker gene expression and OPG/RANKL levels, and promote osteogenic differentiation, promote bone metabolism to bone formation and lose direction. The wear particle induced osteolysis in local systemic bone induced by LPS, Andrographolide significantly improved the inflammatory infiltration and bone osteolysis on the surface of the number of osteoclasts were significantly decreased, bone showed good protective effect. The micro environment of the conclusion of the 1. mg degradation can inhibit osteoclast differentiation and bone resorption, To promote BMSC proliferation, adhesion and osteogenic differentiation, decrease the level of inflammatory factors and mechanism against osteolysis, wear particle induced inhibition of local bone loss. Magnesium may become the new direction of prevention and treatment of diseases of.2. magnesium osteolysis inhibited MRSA adhesion and biofilm formation in vitro, inhibition of osteomyelitis progress and reduce infection inhibit the differentiation and bone loss around.3.Andrographolide bone resorption in osteoclasts in vivo, promote osteogenic differentiation, reduce the level of inflammation at the systemic level, positive regulation of bone metabolism, bone has a significant protective effect.

【学位授予单位】：上海交通大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R687.4

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