保留鈣化層結(jié)構(gòu)的豬股骨滑車全厚軟骨缺損模型建立
寧志剛 1 楊柳 1 王富友 1 張一 1 王恩洪 2
【 摘 要】 目的 建立保留鈣化層結(jié)構(gòu)的豬股骨滑車全厚軟骨缺損模型，為觀察組織工程軟骨在保留鈣化層的膝關(guān)節(jié)軟骨缺損模型中的修復(fù)效果提供良好的實(shí)驗(yàn)研究平臺(tái)。 方法 選取 6 月齡清潔級(jí)貴州小香豬 9 只，體重40 ～ 50 kg，用標(biāo)準(zhǔn)的軟骨缺損制作套件在其右后肢股骨滑車切跡旁制備直徑 6 mm、深 0.2 ～ 0.5 mm、不傷及鈣化層結(jié)構(gòu)的圓柱形全厚軟骨缺損模型。造模 4 周后行 3.0T MRI 觀察，取材后進(jìn)行大體、體視顯微鏡觀察及固綠 - 番紅O、阿利新藍(lán)、天狼星紅組織學(xué)染色觀察缺損處軟骨修復(fù)情況。 結(jié)果 造模后實(shí)驗(yàn)動(dòng)物均存活，術(shù)后切口無(wú)感染，無(wú)髕骨脫位；術(shù)后即可下地行走并部分負(fù)重，1 周后均能自由活動(dòng)，無(wú)跛行。造模后 4 周，MRI 檢查可見(jiàn)滑車處有明顯連續(xù)信號(hào)中斷，異常信號(hào)深及軟骨下骨，缺損周邊深層未見(jiàn)明顯信號(hào)異常。標(biāo)本大體觀察示缺損底部有少量填充物、出血點(diǎn)，與周圍正常軟骨界 限清楚。體式顯微鏡觀察示鈣化層基本完整，缺損局部軟骨下骨板有塌陷。普通顯微鏡下，固綠 - 番紅 O 及阿利新藍(lán)染色示缺損處無(wú)軟骨細(xì)胞及染料著色；偏光顯微鏡下，天狼星紅染色示缺損底部被連續(xù)、強(qiáng)折光性的纖維組織少量填充。結(jié)論 通過(guò)該造模方法制作的不傷及鈣化層結(jié)構(gòu)的豬股骨滑車全厚軟骨缺損模型，可用于骨關(guān)節(jié)炎早期軟骨病變修復(fù)的研究及豬軟骨鈣化層結(jié)構(gòu)作用研究的動(dòng)物模型。
【關(guān)鍵詞】 骨關(guān)節(jié)炎 關(guān)節(jié)軟骨缺損模型 鈣化層 豬
AN ANIMAL MODEL FOUNDATION OF ARTICULAR FULL-THICKNESS CARTILAGE DEFECT BY SAVING CALCIFIED CARTILAGE ZONE ON FEMORAL TROCHLEA IN PORCINE/NING Zhigang1, YANG Liu1, WANG Fuyou1,
ZHANG Yi1, WANG Enhong2. 1Center for Joint Surgery, 2Department of Ophthalmology, Affiliated Southwest Hospital, the Third
Military Medical University, Chongqing, 400038, P.R.China. Corresponding author: YANG Liu, E-mail: jointsurgery@163.com
【 Abstract 】 Objective To establish a porcine model of articular full-thickness cartilage defect characterized by remaining cartilage calcified zone on femoral trochlea, so as to provide a considerable ａnd comparative control group for investigating repair effects of tissue engineered scaffolds in articular cartilage defects with cartilage calcified zone remaining. Methods The full-thickness cartilage column defects (6 mm in diameter, 0.2-0.5 mm in depth) without damage on calcified cartilage zone were made on the femoral trochlea in 9 clean-grade 6-month-old Guizhou minipigs by standard cartilage-defect- making suites. Microscopical observation was performed after modeling. Scanning were made by 3.0T MRI at 4 weeks. Then general observation, stereomicroscope, ａnd histological staining were used to observe cartilage repair. Results All animals were alive. No infection of incisions or patellar dislocations occurred; they were able to walk with partial weight-bearing immediately after surgery ａnd could move freely without limp at 1 week. Obvious signal discontinuity in trochlea ａnd subchondral bone could be observed in MRI, without deep signal change in defects surrounding. Microscopical observation showed a few repair tissue ａnd petechia at base of the defect with clear boundary. Nearly intact calcified zone of cartilage ａnd zonal collapse of subchondral bone in defects could be observed with stereomicroscope. Under common microscope, no chondrocytes was found in defects, as well as negative staining of fast green-safranin O ａnd alcian blue. Under polarized microscope, the bottom of defects were filled with a little of fibrous tissue presenting continuous ａnd strong light-refraction by sirius red staining. Conclusion The animal model of articular full-thickness cartilage defect on femoral trochlea by standard cartilage-defect-making suites can be applied for the research of cartilage disease in early human osteoarthritis ａnd function of calcified cartilage zone in pig.
【 Key words 】 Osteoarthritis Articular cartilage defect model Calcified cartilage zone Porcine Foundation items: National Natural Science Foundation of China (31070865); China Postdoctoral Science Foundation
(2009451520)
基金項(xiàng)目：國(guó)家自然科學(xué)基金面上項(xiàng)目（31070865）；中國(guó)博士后科學(xué)基金資助項(xiàng)目（2009451520）
作者單位：第三軍醫(yī)大學(xué)附屬西南醫(yī)院（重慶，400038） 1 關(guān)節(jié)外科中心，2 眼科
通訊作者：楊柳，教授，博士生導(dǎo)師，研究方向：關(guān)節(jié)外科與運(yùn)動(dòng)創(chuàng)傷，E-mail: jointsurgery@163.com
網(wǎng)絡(luò)出版時(shí)間：2012-4-17 13:09；網(wǎng)絡(luò)出版地址：http://www.cnki. net/kcms/detail/51.1372.R.20120417.1309.015.html
應(yīng)用組織工程技術(shù)進(jìn)行關(guān)節(jié)軟骨再生修復(fù)，已成 為軟骨再生研究熱點(diǎn) [1-5]。目前文獻(xiàn)報(bào)道制備軟骨缺損動(dòng)物模型的方法較多，如在兔、鼠等動(dòng)物股骨髁上 用骨鉆鉆孔直接破壞關(guān)節(jié)軟骨及軟骨下骨等方法 [6-8]。與實(shí)驗(yàn)用豬相比，兔、鼠等動(dòng)物模型的生理及關(guān)節(jié)結(jié) 構(gòu)與人差異較大，不能很好應(yīng)用于組織工程軟骨產(chǎn) 品臨床前研究，因此這方面研究所選用的動(dòng)物模型
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原創(chuàng)作者：上海恒遠(yuǎn)生物技術(shù)發(fā)展有限公司
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