本文選題：膝骨關節(jié)炎　切入點：微血管侵入　出處：《中國人民解放軍醫(yī)學院》2015年碩士論文

【摘要】：研究背景：膝骨關節(jié)炎(knee osteoarthritis, KOA)是老年人多種退行性病變中最常見的一種,膝關節(jié)活動性疼痛、骨贅增生、內(nèi)翻畸形為常見癥狀。涉及KOA病變的因素有很多,體重超標、外傷、劇烈活動過多都與其有關。膝骨關節(jié)炎軟骨中可見微血管侵入,血管增生機制尚不清楚,各種調(diào)節(jié)因子作用機制、分子生物學水平有待進一步研究。缺乏形象、直觀、特異性強的觀察方法觀測OA血管增生的部位與數(shù)量。本研究基于顯微CT的檢測人與大鼠膝骨性關節(jié)炎(OA)不同時期的軟骨、軟骨下骨的血管增生情況,探討骨關節(jié)炎不同階段、不同部位的血管增生變化,進一步闡明軟骨血管侵入的機制,探索關節(jié)炎發(fā)病的機制及疼痛、炎癥與血管增生的關系。方法：(1)將20只SD大鼠(雌雄不限,體重300g左右)編碼,利用電腦產(chǎn)生隨機數(shù),進而將實驗動物分為實驗組(16只)、假手術組(4只)。實驗組行左膝前交叉韌帶切除、左膝半月板切除造模,假手術組只打開關節(jié)囊,不切除前交叉韌帶、半月板,對照組不做任何處理,以亞納米級硫酸鋇與實驗用明膠為原料,在65℃-70℃的水浴鍋中配置成所需造影劑,以大鼠的腹主動脈為注入段,下腔靜脈為流出段,在用肝素鈉注射液與生理鹽水混合置換出下肢血液,然后利用甲醛溶液固定血管形態(tài),進行造影劑的灌注。標本處死后冷凍一夜固定,次日進行標本取材,實驗組分別于術后3周、6周、9周、12周取材。取材部位位于大鼠膝關節(jié)標本內(nèi)側(cè)脛骨平臺骨-軟骨單元,進行大體觀察與組織學評分、Mirco-CT血管形態(tài)計量學分析、增生血管的三維重建、血管增生相關基因的PCR等檢測、VEGF因子的免疫組化染色。(2)取2013年5月-2014年5月行全膝人工關節(jié)置換術后的OA患者的脛骨平臺標本50例,于-20°下保存,所取標本均與患者簽署自愿捐贈協(xié)議。根據(jù)軟骨評分將OA標本共分為4組：A組(Ⅰ級)、B組(Ⅱ級)、C組(Ⅲ級)、D組(Ⅳ級)。進行VEGF免疫組化染色、組織染色、PCR基因擴增等檢測,比較不同程度病變的血管增生情況。結果：(1)顯微CT顯示：造影劑充填脛骨平臺、股骨髁的微血管,顯影效果良好。手術組在切除前交叉韌帶與半月板三周后即出現(xiàn)脛骨平臺的軟骨退變,隨著時間的進行,關節(jié)炎病變加重。根據(jù)大體觀察與組織學評分,術后3周：關節(jié)軟骨點狀損傷。術后6周：關節(jié)軟骨呈片狀退變,破壞范圍增大。術后9周：關節(jié)軟骨呈全層缺損。術后12周：有骨贅形成。利用顯微CT對脛骨平臺與股骨髁進行掃描,通過設定閾值對增生的微血管進行三維成像,手術組各期增生的微血管與假手術組相比,均有顯著差異(P0.05),手術各組增生的微血管與時間呈相關性變化,趨勢為先增加后減少,在術后9周達到高峰。(2)四組標本中,C組標本的VEGF免疫組化染色的陽性率最高,各組進行血管生成相關的VEGF基因擴增檢測,C組標本高于其他各組。結論：(1)利用明膠配比硫酸鋇可以對大鼠進行充分的血管充盈灌注；顯微CT可以對關節(jié)內(nèi)微血管進行定性、定量的分析。在大鼠骨關節(jié)炎模型中,關節(jié)內(nèi)增生的微血管與時間呈相關性變化,先升高再降低,在關節(jié)炎中期,增生的微血管達到峰值(3)血管增生在人骨關節(jié)炎的發(fā)病的機制中至關重要,在病變早期,隨著關節(jié)炎的癥狀加重,軟骨內(nèi)增生的血管增多,在病變晚期,軟骨全層缺損,骨贅形成,血管增生減少。
[Abstract]:Background: osteoarthritis of the knee (knee osteoarthritis KOA) is one of the most common variety of elderly degenerative diseases, pain of knee joint activity, osteophyma, varus deformity is a common symptom. There are many factors involved in KOA lesions of the weight exceed the standard, severe trauma, excessive activity is related. Cartilage of knee osteoarthritis in the visible microvascular invasion, angiogenesis mechanism is not clear, various factors regulating mechanism, further study the level of molecular biology. The lack of image, intuitive, specificity observation observation method of OA blood vessel increase position and quantity of students. This research is based on the micro CT detection of human and rat knee osteoarthritis (OA) in different stages of cartilage, subchondral bone vascular hyperplasia, to explore the different stages of osteoarthritis, vascular proliferation of different parts, to further elucidate the mechanism of cartilage vascular invasion, explore arthritis The incidence and mechanism of pain, inflammation and vascular proliferation. Methods: (1) 20 SD rats (male or female, weight about 300g) encoding, random numbers generated by computer, and then the experimental animal were divided into experimental group (16 rats), sham operation group (4 rats) in experimental group. Left knee anterior cruciate ligament resection, left knee meniscectomy model, sham operation group only open the joint capsule, without resection of anterior cruciate ligament, meniscus, the control group without any treatment, with sub nanometer barium sulfate and experiment with gelatin as raw materials, into the desired contrast agent configuration in a water bath at 65 -70 C. in the rat abdominal aorta for injection, inferior vena cava is the outflow segment in Heparin Sodium Injection, and injected with the replacement of blood, and then use Formaldehyde Solution for fixed vascular morphology, contrast agent perfusion. Specimens were frozen overnight and fixed, the specimens were drawn, experimental group 鍒簬鏈悗3鍛，

本文編號：1709958