骨折固定後的修復需要一系列的炎症反應然後產生癒傷組織，骨頭癒合完成後便開始進行癒傷組織的重塑，骨折癒合是一個複雜的過程，有多個相互重疊的階段，包括發炎反應，軟骨形成與骨重塑，有許多內在或外在因素可能對骨折癒合造成影響，導致骨癒合延遲或無法癒合。雄性激素受體基因剔除的小鼠已經知道在軟骨內成骨的骨量減少，成骨細胞礦化能力降低，但對於骨折癒合時癒傷組織的形成所造成的影響仍不清楚。此篇研究主要是去探討小鼠骨折時，在骨折癒合的過程中，野生型小鼠和雄性激素受體基因剔除小鼠之雄性激素與雄性激素受體對於骨痂組織形成、礦化作用與骨重塑時所扮演的角色，因此，以動物實驗進行長期微型電腦斷層掃瞄的觀察，以確認雄性激素荷爾蒙調節作用在骨折癒合過程中其機制或流程之必要性與關聯性，並應用動物用體內微電腦斷層掃描儀，建立在不同的時間點，野生型小鼠和雄性激素受基因體剔除之小鼠在骨折癒合時的三維模型結構圖像，觀察癒合過程中骨痂組織微結構的發展情形，以達研究雄性激素與雄性激素受體在骨折癒合過程中，對於骨痂組織形成、礦化作用與骨重塑之微結構變化的影響，為進一步了解雄性激素和雄性激素受體在毎個過程中的影響，以組織切片染色觀察骨痂的組織型態變化，觀察骨痂中膠原組織，纖維組織與骨組織的分佈比例與位置。經實驗結果發現，雄性激素受體基因剔除小鼠較野生型小鼠的骨頭痊癒的速度慢，且年長的雄性激素受體基因剔除小鼠較年輕的雄性激素受體基因剔除小鼠之骨癒合的情況不好，去勢後使小鼠骨癒合的情況變更差，綜合以上結果，可能是性荷爾蒙在骨頭癒合的過程中所產生的影響。因此，雄性激素及雄性激素受體作用對於影響骨折癒合時，骨骼的修復和重建扮演著重要的調節者，往後可應用於治療骨質疏鬆症影響造成的骨折，對於骨折癒合相當具有效益。

Fracture healing requires a series of events including inflammatory response and callus formation, callus remodeling and bone healing. Fracture healing is a complex process, there are several overlapping phases , including inflammation , cartilage formation and bone remodeling, there are many internal or external factors could impact on fracture healing, leading to delayed bone healing or non healing. The global androgen receptor knockout (GARKO) mice has been know to reduce bone mass in endochondral bone and osteoblast mineralization, but the impact for callus formation in fracture healing is still unclear. The goals of study is to investigate the role of androgen and androgen receptor in wild-type (WT) mice and GARKO mice after fracture healing during callus formation and bone mineralization and bone remodeling. Therefore, long-term animal experiments observed by micro-computed tomography to study the roles of androgen and androgen receptor on the process and mechanisms of fracture healing is necessary. We applied in vivo micro-computer tomography (Micro-CT) to build up the three-dimensional model images at different time points for wild-type mice and GARKO mice after fracture healing and observe the bone healing process of micro-structure of the development of callus during fracture healing. The callus tissue morphology observed by histological staining to study the proportion and position of collagen, fibrous tissue and bone. The results show that the healing of WT mice is better than GARKO mice. GARKO mice develop smaller callus size and less bone volume and show delayed healing. In general, orchiectomy (ORX) decreases callus size in WT mice but not in GARKO mice. However, the healing rate of elderly GARKO mice is not obvious in comparison with young GARKO mice. Together, our study demonstrated that the androgen and androgen receptor regulate fracture healing and play an important role in bone repair and healing. Our mouse model may be used for the therapeutic drug screening of bone fractures caused by osteoporosis.

目錄
中文摘要..........................................................................................................................1
英文摘要..........................................................................................................................3
目錄.................................................................................................................................5
圖表目錄..........................................................................................................................7
符號及縮寫......................................................................................................................8
第一章 前言....................................................................................................................9
(一)骨頭的生成與代謝..........................................................................................9
(二)骨質疏鬆症....................................................................................................10
(三) 淺談骨折與骨癒合.......................................................................................11
(四) 雄性激素與雄性激素受體的作用途徑......................................................13
(五)研究目的........................................................................................................14
第二章 實驗材料與方法..............................................................................................15
第一節 實驗動物與切片製作..............................................................................15
第二節 一般組織染色..........................................................................................19
第三節 微電腦斷層掃描 (Micro computed tomography, Micro-CT)...........21
第四節 微電腦斷層掃描定量結果之統計分析..................................................23
第三章 結果..................................................................................................................24
第一節 比較野生型小鼠和雄性激素受體基因剔除之小鼠，在去勢前與去勢後，骨折後第7，14，21，28，42天骨癒合的二維模型結構圖像之結果..............................................................................................24
第二節 比較野生型小鼠和雄性激素受體基因剔除之小鼠，去勢前後，在骨折後第14，21，28，42天的骨癒合三維模型結構圖像與定量之結果......................................................................................................24
第三節 比較年輕與年老，去勢前後，雄性激素受體基因剔除之小鼠在骨折後第21，28的骨癒合三維模型結構圖像與定量之結果............26
第四節 比較野生型和雄性激素受體基因剔除之小鼠，去勢前後，在骨折後第21天的組織學染色結果............................................................28
第五節 比較年輕與年老雄性激素受體基因剔除之小鼠，去勢前後，在骨折後第28天的組織學染色結果........................................................29
第四章 討論..................................................................................................................31
雄性激素與雄性激素受體基因影響骨痂形成過程的探討................................31
骨折後骨痂形成過程的組織學形態變化............................................................32
骨痂形成過程相關生物標記的探討....................................................................33
動物實驗在骨折手術仍須克服的問題................................................................34
雄性激素與雄性激素受體基因在骨折後修復的運用........................................35
第五章 結論..................................................................................................................37
參考文獻........................................................................................................................38
附錄一 圖表..................................................................................................................50
附錄二 藥品..................................................................................................................59
附錄三 試劑配方..........................................................................................................60

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