2011-08-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/656423摘要：背景及構想: 在現代金屬對金屬介面全人工髖關節術後發生的骨溶蝕現象是一個新的課題，因為在金屬對金屬介面全人工髖關節磨損中所產生的金屬奈米粒子(50-nm (0.05-μm)大小)所引發的生物反應和傳統的金屬對高分子塑膠介面全人工髖關節磨損產生的高分子塑膠微粒(0.5-μm 大小)所引發的生物反應可能是完全不同的。區分這二種不同大小磨損粒子的生物相容性和所引發的不同生物反應是一個重要的課題。雖然雙磷酸鹽己有人嘗試用來治療全人工髖關節術後發生的骨溶蝕現象，但是其治療原理和效果仍有待被証實。而最近被用在成功治療骨質疏鬆症的鍶從未被用在治療全人工髖關節術後發生的骨溶蝕現象。在之前的研究中，我們已經成功地使用了air pouch發炎的小鼠模式來作為以高分子塑膠微粒來誘發骨溶蝕現象的動物模式，可供使用來作骨溶蝕現象的更進一步研究。特定目的: 我們研究目的要在已經成功建立air pouch發炎的小鼠模式中以金屬奈米微粒來誘發骨溶蝕現象，並使用微陣列來研究這一個以金屬奈米微粒誘發骨溶蝕現象的小鼠模式的mRNA基因表現，在mRNA和protein的層次來驗証其結果，以及運用這個動物模式來探討是否雙磷酸鹽和鍶能否被有效用來治療金屬奈米微粒所誘發的骨溶蝕現象。方法及實施進度: 在本研究中，我們以金屬奈米微粒使用在air pouch發炎的小鼠模式來建立一個誘發骨溶蝕現象的動物模式。我們將使用微陣列來研究在對照組小鼠和金屬奈米微粒刺激組小鼠其air pouch 組織的差異性mRNA 基因表現。我們將使用定量real-timereverse-transcriptase PCR來確認微陣列實驗的結果。我們也將使用air pouch組織的免疫組織生化染色分析來作進一步在protein 層次的確認。接下來我們將小鼠模式分為三組(第一組在day 0接受PBS注射、第二組在day 0接受金屬奈米微粒注射及day -3接受zoledronic acid注射，第三組在day 0接受金屬奈米微粒注射及day 5接受zoledronic acid注射)作實驗，以探討是否雙磷酸鹽能否被有效用來治療金屬奈米微粒所誘發的骨溶蝕現象。我們再將小鼠模式分為三組(第一組在day 0接受PBS注射、第二組在day 0接受金屬奈米微粒注射及day -3接受口服鍶之治療8個星期，第三組在day 0接受金屬奈米微粒注射及day 5接受口服鍶之治療8個星期)作實驗，以探討是否鍶能否被有效用來治療金屬奈米微粒所誘發的骨溶蝕現象。預期效果及其重要性: 在第一年的計畫中，我們預期建立一個以金屬奈米微粒誘發骨溶蝕現象的動物模式，並以微陣列來分析其基因表現。在第二年的計畫中，我們預期探討出是否雙磷酸鹽能否被有效用來治療金屬奈米微粒所誘發的骨溶蝕現象。在第三年的計畫中，我們預期探討出是否鍶能否被有效用來治療金屬奈米微粒所誘發的骨溶蝕現象。我們認為建立這個動物模式將使我們可以更進一步了解金屬奈米微粒誘發骨溶蝕現象的病理，並且可以將其使用在研發並驗証新的治療方法。我們也將知道是否雙磷酸鹽和鍶有無臨床潛力能運用在治療金屬奈米微粒所誘發的骨溶蝕現象。<br> Abstract: Objective, background and rationale: Periprosthetic osteolysis after modern metal-on-metal totalhip arthroplasty remains a specific clinical problem, and the biological reactions elicited inperiprosthetic osteolysis generated by metal-on-metal bearing couples may be different from thosegenerated by conventional metal-on-polyethylene joint replacements because of the bioreactivitydifference between 0.5-μm polyethylene particles versus 50-nm (0.05-μm) cobalt-chromium alloynanoparticles. Despite broad off-label use of bisphosphonates in patients with periprostheticosteolysis, studies are still needed to establish the rationale of the use of bisphosphonates forperiprosthetic osteolysis. A new class of anti-osteoporosis drug, strontium, has never been used inthe treatment of periprosthetic osteolysis. In the preliminary study, we had successfully usedmodification of the air pouch model of inflammation as an animal model of osteolysis induced bypolyethylene and can be utilized further in research of osteolysis.Specific aims: It’s our aim to establish an animal model of osteolysis induced by metalnanoparticles, to use microarrays to investigate the mRNA gene expression changes in this animalmodel of osteolysis and validate the findings in mRNA and protein levels, and to find out ifbisphosphonate (zoledronic acid) and strontium could be used in the treatment of the periprostheticosteolysis induced by metal nanoparticles.Methods and experimental plan: We will use metal nanoparticles in our established air pouchmodel of inflammation to create an osteolysis model. We will use microarrays to research thedifferential mRNA gene expression in the air-pouch model between the wild type mouse and themetal-nanoparticles-stimulated mouse. Quantitative real-time reverse-transcriptase polymerasechain reaction will be performed for confirmation of array findings. Further validation ofmicroarray finding in protein level will be performed by immunohistochemistry staining ofundecalcified frozen section of samples in the animal models of osteolysis. Then we will use threegroups of animal models (animals in group I underwent PBS sham injection only, animals in groupII received metal-nanoparticle injection at day 0 and treatment at day -3, and animals in group IIIreceived metal-nanoparticle injection at day 0 and treatment at day 5) in our experiments to evaluatethe therapeutic effect of zoledronic acid and strontium on osteolysis induced by metal nanoparticles.Expected results and significance: In the first year of the study, we aim to develop a animal modelof osteolysis caused by metal nanoparticles and evaluated the gene expression by microarray. In thesecond year of the study, we aim to evaluate the therapeutic effect of zoledronic acid on theosteolysis caused by metal nanoparticles. In the third year of the study, we aim to evaluate thetherapeutic effect of strontium on the osteolysis caused by metal nanoparticles. We expected todevelop an animal model of osteolysis caused by metal nanoparticles which is a valuable tool tofurther understand the biology of periprosthetic osteolysis. We also want to explore the possibilityof treating periprosthetic osteolysis in metal-on-metal joint arthroplasty with zoledronic acid andstrontium.