發(fā)布時(shí)間：2018-04-04 12:29

  本文選題：膝骨關(guān)節(jié)炎　切入點(diǎn)：微血管侵入　出處：《中國(guó)人民解放軍醫(yī)學(xué)院》2015年碩士論文

【摘要】：研究背景：膝骨關(guān)節(jié)炎(knee osteoarthritis, KOA)是老年人多種退行性病變中最常見(jiàn)的一種,膝關(guān)節(jié)活動(dòng)性疼痛、骨贅增生、內(nèi)翻畸形為常見(jiàn)癥狀。涉及KOA病變的因素有很多,體重超標(biāo)、外傷、劇烈活動(dòng)過(guò)多都與其有關(guān)。膝骨關(guān)節(jié)炎軟骨中可見(jiàn)微血管侵入,血管增生機(jī)制尚不清楚,各種調(diào)節(jié)因子作用機(jī)制、分子生物學(xué)水平有待進(jìn)一步研究。缺乏形象、直觀、特異性強(qiáng)的觀察方法觀測(cè)OA血管增生的部位與數(shù)量。本研究基于顯微CT的檢測(cè)人與大鼠膝骨性關(guān)節(jié)炎(OA)不同時(shí)期的軟骨、軟骨下骨的血管增生情況,探討骨關(guān)節(jié)炎不同階段、不同部位的血管增生變化,進(jìn)一步闡明軟骨血管侵入的機(jī)制,探索關(guān)節(jié)炎發(fā)病的機(jī)制及疼痛、炎癥與血管增生的關(guān)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(jī)數(shù),進(jìn)而將實(shí)驗(yàn)動(dòng)物分為實(shí)驗(yàn)組(16只)、假手術(shù)組(4只)。實(shí)驗(yàn)組行左膝前交叉韌帶切除、左膝半月板切除造模,假手術(shù)組只打開(kāi)關(guān)節(jié)囊,不切除前交叉韌帶、半月板,對(duì)照組不做任何處理,以亞納米級(jí)硫酸鋇與實(shí)驗(yàn)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(dòng)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(jìn)行造影劑的灌注。標(biāo)本處死后冷凍一夜固定,次日進(jìn)行標(biāo)本取材,實(shí)驗(yàn)組分別于術(shù)后3周、6周、9周、12周取材。取材部位位于大鼠膝關(guān)節(jié)標(biāo)本內(nèi)側(cè)脛骨平臺(tái)骨-軟骨單元,進(jìn)行大體觀察與組織學(xué)評(píng)分、Mirco-CT血管形態(tài)計(jì)量學(xué)分析、增生血管的三維重建、血管增生相關(guān)基因的PCR等檢測(cè)、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(guān)節(jié)置換術(shù)后的OA患者的脛骨平臺(tái)標(biāo)本50例,于-20°下保存,所取標(biāo)本均與患者簽署自愿捐贈(zèng)協(xié)議。根據(jù)軟骨評(píng)分將OA標(biāo)本共分為4組：A組(Ⅰ級(jí))、B組(Ⅱ級(jí))、C組(Ⅲ級(jí))、D組(Ⅳ級(jí))。進(jìn)行VEGF免疫組化染色、組織染色、PCR基因擴(kuò)增等檢測(cè),比較不同程度病變的血管增生情況。結(jié)果：(1)顯微CT顯示：造影劑充填脛骨平臺(tái)、股骨髁的微血管,顯影效果良好。手術(shù)組在切除前交叉韌帶與半月板三周后即出現(xiàn)脛骨平臺(tái)的軟骨退變,隨著時(shí)間的進(jìn)行,關(guān)節(jié)炎病變加重。根據(jù)大體觀察與組織學(xué)評(píng)分,術(shù)后3周：關(guān)節(jié)軟骨點(diǎn)狀損傷。術(shù)后6周：關(guān)節(jié)軟骨呈片狀退變,破壞范圍增大。術(shù)后9周：關(guān)節(jié)軟骨呈全層缺損。術(shù)后12周：有骨贅形成。利用顯微CT對(duì)脛骨平臺(tái)與股骨髁進(jìn)行掃描,通過(guò)設(shè)定閾值對(duì)增生的微血管進(jìn)行三維成像,手術(shù)組各期增生的微血管與假手術(shù)組相比,均有顯著差異(P0.05),手術(shù)各組增生的微血管與時(shí)間呈相關(guān)性變化,趨勢(shì)為先增加后減少,在術(shù)后9周達(dá)到高峰。(2)四組標(biāo)本中,C組標(biāo)本的VEGF免疫組化染色的陽(yáng)性率最高,各組進(jìn)行血管生成相關(guān)的VEGF基因擴(kuò)增檢測(cè),C組標(biāo)本高于其他各組。結(jié)論：(1)利用明膠配比硫酸鋇可以對(duì)大鼠進(jìn)行充分的血管充盈灌注；顯微CT可以對(duì)關(guān)節(jié)內(nèi)微血管進(jìn)行定性、定量的分析。在大鼠骨關(guān)節(jié)炎模型中,關(guān)節(jié)內(nèi)增生的微血管與時(shí)間呈相關(guān)性變化,先升高再降低,在關(guān)節(jié)炎中期,增生的微血管達(dá)到峰值(3)血管增生在人骨關(guān)節(jié)炎的發(fā)病的機(jī)制中至關(guān)重要,在病變?cè)缙?隨著關(guān)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(hào)：1709958