本文關(guān)鍵詞： 內(nèi)撐器 股骨頭壞死 液氮冷凍 髓芯減壓 生物力學(xué) 壓縮強(qiáng)度 彈性模量 BMP-2 bFGF　出處：《中南大學(xué)》2014年博士論文　論文類(lèi)型：學(xué)位論文

【摘要】：目的：本研究將一種鈦合金股骨頭內(nèi)撐器植入通過(guò)改良的液氮冷凍法建立的比格犬股骨頭壞死模型,通過(guò)影像學(xué)和組織學(xué)檢查觀察其修復(fù)過(guò)程,并測(cè)量股骨頭軟骨下骨的最大壓縮強(qiáng)度和平均彈性模量,為使用股骨頭內(nèi)撐器治療股骨頭壞死提供理論基礎(chǔ)。 方法：1.比格犬股骨頭壞死模型的建立。26只健康成年比格犬(26髖)均采用改良的液氮冷凍法隨機(jī)側(cè)建立股骨頭壞死模型。采用髖關(guān)節(jié)后外側(cè)入路暴露髖關(guān)節(jié)后關(guān)節(jié)囊,“T”形切開(kāi)關(guān)節(jié)囊,切斷股骨頭圓韌帶,將股骨頭半脫位,先用無(wú)菌漏斗套住股骨頭,再用液氮澆灌冷凍股骨頭。術(shù)后2個(gè)月隨機(jī)選擇2只比格犬,行影像學(xué)檢查后處死動(dòng)物,再行組織學(xué)檢查檢驗(yàn)造模情況。 2.鈦合金股骨頭內(nèi)撐器治療比格犬股骨頭壞死的實(shí)驗(yàn)研究。驗(yàn)證造模成功后,將24只比格犬隨機(jī)分為A、B兩組(每組12只)。A組為壞死模型空白對(duì)照組；B組為髓芯減壓鈦合金股骨頭內(nèi)撐器植入組。兩組的對(duì)側(cè)均作為自身正常對(duì)照。分別于術(shù)后3、6個(gè)月時(shí),每組隨機(jī)選擇6只比格犬行影像學(xué)、組織學(xué)檢查及生物力學(xué)測(cè)定,并進(jìn)行組內(nèi)及組間比較分析。實(shí)驗(yàn)數(shù)據(jù)統(tǒng)計(jì)學(xué)分析采用兩樣本t檢驗(yàn)。 結(jié)果：1.造模術(shù)后2月行X線檢查示：模型側(cè)股骨頭骨質(zhì)密度不均勻,股骨頭外形欠規(guī)整,股骨頭內(nèi)可見(jiàn)囊性變,周?chē)琴|(zhì)可見(jiàn)硬化；MRI掃描結(jié)果顯示：股骨頭內(nèi)信號(hào)不均勻,T1WI上表現(xiàn)為點(diǎn)狀、細(xì)線狀、片狀中等信號(hào)灶,T2WI上表現(xiàn)為點(diǎn)狀、細(xì)線狀、片狀高信號(hào)灶,關(guān)節(jié)腔內(nèi)可見(jiàn)較多關(guān)節(jié)積液；組織學(xué)檢查中HE染色示模型側(cè)軟骨表面毛糙、變薄、變性、部分脫落,軟骨下骨壞死區(qū)可見(jiàn)增生結(jié)締組織,骨小梁變細(xì)、稀疏、斷裂、排列紊亂,骨細(xì)胞變性死亡,部分骨小梁斷裂不連續(xù),彌漫性的存在的空骨陷窩,髓腔內(nèi)造血細(xì)胞明顯減小,可見(jiàn)微小血管內(nèi)血栓形成。 2.內(nèi)撐器植入術(shù)后 1)內(nèi)撐器植入術(shù)后3個(gè)月： (1)影像學(xué)觀察。X線檢查及CT掃描示：A組股骨頭輪廓不規(guī)則,關(guān)節(jié)面欠光滑,承重區(qū)扁平塌陷,骨質(zhì)密度不均勻,關(guān)節(jié)間隙稍變窄。B組股骨頭外形基本正常,骨質(zhì)密度欠均勻,股骨頭無(wú)明顯塌陷,關(guān)節(jié)間隙無(wú)明顯變窄。 (2)生物力學(xué)檢測(cè)。A組、B組手術(shù)側(cè)和正常對(duì)側(cè)股骨頭軟骨下骨最大壓縮強(qiáng)度分別為：(49.38±2.37和91.30±2.21)、(81.78±2.36和91.16±2.32)Mpa；平均彈性模量分別為(78.86±2.24和153.42±3.36)、(139.90±3.49和151.96±3.75)MPa。A組、B組手術(shù)側(cè)股骨頭軟骨下骨的最大壓縮強(qiáng)度和平均彈性模量與正常對(duì)側(cè)比較,差異均有統(tǒng)計(jì)學(xué)意義(P0.05)；且A組與B組手術(shù)側(cè)進(jìn)行比較,差異也有統(tǒng)計(jì)學(xué)意義(P0.05)。 (3)HE染色組織學(xué)檢查。A組和B組手術(shù)側(cè)和正常對(duì)側(cè)股骨頭空骨陷窩計(jì)數(shù)百分比分別為：(22.44±1.34和11.52±1.29)、(15.44±1.47和12.54±1.61)%。血管計(jì)數(shù)分別為：(21.20±1.79和43.40±3.05)、(33.00±2.55和46.20±3.35)。A組、B組手術(shù)側(cè)股骨頭空骨陷窩計(jì)數(shù)百分比、血管計(jì)數(shù)與正常對(duì)側(cè)比較,差異均有統(tǒng)計(jì)學(xué)意義(P0.05)；且A組與B組手術(shù)側(cè)進(jìn)行比較,差異也有統(tǒng)計(jì)學(xué)意義(P0.05)。 (4)免疫組化染色。BMP-2、bFGF的免疫組化染色陽(yáng)性物質(zhì)均呈棕黃色；免疫組化染色的陽(yáng)性物質(zhì)經(jīng)MIAS (Micro Imagery Analysis System)圖像分析系統(tǒng)量化分析：A組、B組手術(shù)側(cè)與正常對(duì)側(cè)的BMP-2平均灰度值分別為：(189.90±12.90和100.34±6.56)、(134.80±11.00和105.16±6.43)；bFGF平均灰度值分別為：(192.90±11.50和112.86±7.59)、(139.70±9.43和115.18±7.22)。A組和B組手術(shù)側(cè)BMP-2、bFGF的表達(dá)與正常對(duì)側(cè)比較,差異均有統(tǒng)計(jì)學(xué)意義(P0.05),且A組與B組手術(shù)側(cè)進(jìn)行比較,差異也有統(tǒng)計(jì)學(xué)意義(P0.05)。 2)內(nèi)撐器治療術(shù)后6個(gè)月： (1)影像學(xué)觀察。X線檢查及CT掃描示：A組股骨頭可見(jiàn)塌陷較前明顯加重,股骨頭囊性變及骨質(zhì)硬化,關(guān)節(jié)間隙變窄,而B(niǎo)組股骨頭外形正常,骨質(zhì)密度較均勻,關(guān)節(jié)間隙正常。 (2)生物力學(xué)檢測(cè)。A組、B組手術(shù)側(cè)和正常對(duì)側(cè)股骨頭軟骨下骨最大壓縮強(qiáng)度分別為：(53.44±2.45和91.50±2.02)、(88.78±2.45和91.94±2.99)Mpa；平均彈性模量分別為：(84.66±3.03和154.30±3.46)、(153.74±3.20和155.98±3.55)MPa。A組手術(shù)側(cè)股骨頭軟骨下骨最大壓縮強(qiáng)度和平均彈性模量與正常對(duì)側(cè)比較,差異有統(tǒng)計(jì)學(xué)意義(P0.05),B組手術(shù)側(cè)與正常對(duì)側(cè)比較,差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05),而A組與B組手術(shù)側(cè)比較,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。 (3)HE染色組織學(xué)檢查。A組、B組手術(shù)側(cè)和正常對(duì)側(cè)空骨陷窩計(jì)數(shù)百分比分別為：(19.58±1.41和12.42±1.40)、(12.46±1.51和11.48±1.54)%；血管計(jì)數(shù)分別為：(27.00±2.12和42.40±3.78)、(43.00±3.54和44.80±3.56)。A組手術(shù)側(cè)空骨陷窩計(jì)數(shù)百分比和血管計(jì)數(shù)與正常對(duì)側(cè)比較,差異有統(tǒng)計(jì)學(xué)意義(P0.05),B組手術(shù)側(cè)與正常對(duì)側(cè)比較,差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05),而A組與B組手術(shù)側(cè)比較,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。 (4)免疫組化染色。A組、B組手術(shù)側(cè)與正常對(duì)側(cè)的BMP-2平均灰度值為：(161.90±12.10和98.22±6.33)、(110.04±8.60和102.28±7.79)；bFGF平均灰度值分別為：(157.20±10.10和113.68±6.93)、(124.92±7.55和116.26±6.30)。A組手術(shù)側(cè)BMP-2、bFGF的表達(dá)與正常對(duì)側(cè)比較,差異有統(tǒng)計(jì)學(xué)意義(P0.05)；B組手術(shù)側(cè)與正常對(duì)側(cè)比較,差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05),而A組與B組手術(shù)側(cè)比較,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。 結(jié)論：1.本實(shí)驗(yàn)采用的改良液氮冷凍法可以成功制作出股骨頭壞死的動(dòng)物模型。該造模方法實(shí)驗(yàn)犬死亡率低,操作方便,可重復(fù)性高,創(chuàng)傷較小,不破壞髖關(guān)節(jié)的穩(wěn)定性。該動(dòng)物模型可用于臨床上探索治療股骨頭壞死的實(shí)驗(yàn)研究,同時(shí)也為我們后續(xù)的研究提供較為理想的動(dòng)物模型。 2.應(yīng)用鈦合金股骨頭內(nèi)撐器能有效恢復(fù)股骨頭的生物力學(xué)性能,促進(jìn)壞死區(qū)骨修復(fù),促進(jìn)新生血管形成,防止股骨頭塌陷。在植入術(shù)后三個(gè)月時(shí),內(nèi)撐器組各項(xiàng)指標(biāo)均優(yōu)于壞死空白對(duì)照組,但比自身正常對(duì)側(cè)差；在術(shù)后六個(gè)月時(shí),內(nèi)撐器組各項(xiàng)指標(biāo)均優(yōu)于壞死對(duì)照組,與自身正常對(duì)側(cè)比較,差異無(wú)統(tǒng)計(jì)學(xué)意義。有望為臨床上治療成人股骨頭壞死提供一種新的方法。
[Abstract]:Objective : To establish a model of femoral head necrosis of femoral head , established by modified liquid nitrogen freezing method , and to measure the maximum compressive strength and average modulus of elasticity of the lower bone of femoral head and to provide a theoretical basis for the treatment of femoral head necrosis with femoral head . Methods : 1 . The necrosis model of femoral head in Beagle dogs was established . 26 healthy adult dogs ( 26 hips ) were randomly divided into femoral head necrosis model by modified liquid nitrogen freezing method . 2 . The experimental study on the treatment of femoral head necrosis of dogs with titanium alloy femoral head was conducted . Twenty - four Beagle dogs were randomly divided into two groups : group A and group B ( 12 in each group ) . Results : 1 . The X - ray examination showed that the bone density of the femoral head in the model side was not uniform , the shape of the femoral head was irregular , the visible cystic changes in the femoral head and the surrounding bone were visible . 2 . Post - stent implantation 1 ) 3 months after stent implantation : ( 1 ) Imaging observation . X - ray examination and CT scanning showed that the profile of femoral head in group A was irregular , the articular surface was not smooth , the bearing area was flat and collapsed , the bone density was not uniform , the joint gap was slightly narrowed . The shape of femoral head in group B was basically normal , the bone density was not uniform , the femoral head did not collapse obviously , and the joint clearance was not significantly narrowed . ( 2 ) The maximum compressive strength of bone in group A , group B and normal contralateral femoral head were ( 49.38 鹵 2.37 and 91.30 鹵 2.21 ) , ( 81.78 鹵 2.36 and 91.16 鹵 3.36 ) , ( 139.90 鹵 3.49 and 151.96 鹵 3.75 ) MPa , respectively ( P0.05 ) . ( 3 ) The histological examination of HE staining showed that the percentages of the operative side and the normal contralateral femoral head in group A and group B were ( 22.44 鹵 1.34 and 11.52 鹵 1.29 ) , ( 15.44 鹵 1.47 and 12.54 鹵 3.35 ) , ( 33.00 鹵 2.55 and 46.20 鹵 3.35 ) respectively . The blood vessel counts were statistically significant ( P0.05 ) . The average gray values of BMP - 2 and bFGF in group A , group B and normal contralateral BMP - 2 were ( 189.90 鹵 12.90 and 100.34 鹵 6.56 ) , ( 139.70 鹵 11.00 and 105.16 鹵 7.22 ) , ( 139.70 鹵 9.43 and 11.18 鹵 7.22 ) , respectively . 2 ) 6 months after stent treatment : ( 1 ) Imaging observation . X - ray examination and CT scan showed that the collapse of femoral head in group A was significantly heavier than that before , the cystic degeneration of femoral head and osteoporosis , the narrowing of joint space , and the normal shape of femoral head in group B , the bone density was more uniform , and the joint clearance was normal . ( 2 ) The maximum compressive strength of subcortical bone in group A , group B and normal contralateral femoral head were ( 53.44 鹵 2.45 and 91.94 鹵 2.99 ) Mpa , ( 88.78 鹵 3.03 and 154.30 鹵 3.46 ) , ( 153.74 鹵 3.20 and 155.98 鹵 3.55 ) MPa , respectively . ( 3 ) histological examination of HE staining . The percentages of the operative side and the normal contralateral empty fossa in group A and group B were ( 19.58 鹵 1.41 and 12.42 鹵 1.40 ) , ( 12.46 鹵 1.54 and 11.48 鹵 3.56 ) , ( 43.00 鹵 3.54 and 44.80 鹵 3.56 ) respectively . The difference was statistically significant ( P0.05 ) . ( 4 ) The mean gray values of BMP - 2 and bFGF in group A , group B and normal contralateral side were ( 161.90 鹵 12.10 and 98.22 鹵 6.33 ) , ( 110.04 鹵 8.60 and 102.28 鹵 7.79 ) , respectively . The average gray values of bFGF in group A were ( 157.20 鹵 10.10 and 13.68 鹵 6.93 ) , ( 124.92 鹵 7.55 and 1160.26 鹵 6.30 ) respectively . There was no significant difference between the expression of BMP - 2 and bFGF in group A and normal contralateral side ( P0.05 ) . Conclusion : 1 . The modified liquid nitrogen freezing method used in this experiment can successfully produce the animal model of femoral head necrosis . The animal model has low mortality , convenient operation , high repeatability and less trauma . The animal model can be used to study the experimental study of femoral head necrosis in clinic , and provide an ideal animal model for our subsequent study . 2 . The biological mechanical properties of the femoral head can be effectively recovered by using the titanium alloy femoral head inner brace , promoting the bone repair in the necrotic area , promoting the formation of the neovascularization and preventing the femoral head from collapsing .

【學(xué)位授予單位】：中南大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2014
【分類(lèi)號(hào)】：R318.17

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