钛合金股骨头内撑器治疗犬股骨头坏死的实验研究

发布时间：2018-02-26 00:21

本文关键词： 内撑器股骨头坏死液氮冷冻髓芯减压生物力学压缩强度弹性模量 BMP-2 bFGF　出处：《中南大学》2014年博士论文　论文类型：学位论文

【摘要】：目的：本研究将一种钛合金股骨头内撑器植入通过改良的液氮冷冻法建立的比格犬股骨头坏死模型,通过影像学和组织学检查观察其修复过程,并测量股骨头软骨下骨的最大压缩强度和平均弹性模量,为使用股骨头内撑器治疗股骨头坏死提供理论基础。方法：1.比格犬股骨头坏死模型的建立。26只健康成年比格犬(26髋)均采用改良的液氮冷冻法随机侧建立股骨头坏死模型。采用髋关节后外侧入路暴露髋关节后关节囊,“T”形切开关节囊,切断股骨头圆韧带,将股骨头半脱位,先用无菌漏斗套住股骨头,再用液氮浇灌冷冻股骨头。术后2个月随机选择2只比格犬,行影像学检查后处死动物,再行组织学检查检验造模情况。 2.钛合金股骨头内撑器治疗比格犬股骨头坏死的实验研究。验证造模成功后,将24只比格犬随机分为A、B两组(每组12只)。A组为坏死模型空白对照组；B组为髓芯减压钛合金股骨头内撑器植入组。两组的对侧均作为自身正常对照。分别于术后3、6个月时,每组随机选择6只比格犬行影像学、组织学检查及生物力学测定,并进行组内及组间比较分析。实验数据统计学分析采用两样本t检验。结果：1.造模术后2月行X线检查示：模型侧股骨头骨质密度不均匀,股骨头外形欠规整,股骨头内可见囊性变,周围骨质可见硬化；MRI扫描结果显示：股骨头内信号不均匀,T1WI上表现为点状、细线状、片状中等信号灶,T2WI上表现为点状、细线状、片状高信号灶,关节腔内可见较多关节积液；组织学检查中HE染色示模型侧软骨表面毛糙、变薄、变性、部分脱落,软骨下骨坏死区可见增生结缔组织,骨小梁变细、稀疏、断裂、排列紊乱,骨细胞变性死亡,部分骨小梁断裂不连续,弥漫性的存在的空骨陷窝,髓腔内造血细胞明显减小,可见微小血管内血栓形成。 2.内撑器植入术后 1)内撑器植入术后3个月： (1)影像学观察。X线检查及CT扫描示：A组股骨头轮廓不规则,关节面欠光滑,承重区扁平塌陷,骨质密度不均匀,关节间隙稍变窄。B组股骨头外形基本正常,骨质密度欠均匀,股骨头无明显塌陷,关节间隙无明显变窄。 (2)生物力学检测。A组、B组手术侧和正常对侧股骨头软骨下骨最大压缩强度分别为：(49.38±2.37和91.30±2.21)、(81.78±2.36和91.16±2.32)Mpa；平均弹性模量分别为(78.86±2.24和153.42±3.36)、(139.90±3.49和151.96±3.75)MPa。A组、B组手术侧股骨头软骨下骨的最大压缩强度和平均弹性模量与正常对侧比较,差异均有统计学意义(P0.05)；且A组与B组手术侧进行比较,差异也有统计学意义(P0.05)。 (3)HE染色组织学检查。A组和B组手术侧和正常对侧股骨头空骨陷窝计数百分比分别为：(22.44±1.34和11.52±1.29)、(15.44±1.47和12.54±1.61)%。血管计数分别为：(21.20±1.79和43.40±3.05)、(33.00±2.55和46.20±3.35)。A组、B组手术侧股骨头空骨陷窝计数百分比、血管计数与正常对侧比较,差异均有统计学意义(P0.05)；且A组与B组手术侧进行比较,差异也有统计学意义(P0.05)。 (4)免疫组化染色。BMP-2、bFGF的免疫组化染色阳性物质均呈棕黄色；免疫组化染色的阳性物质经MIAS (Micro Imagery Analysis System)图像分析系统量化分析：A组、B组手术侧与正常对侧的BMP-2平均灰度值分别为：(189.90±12.90和100.34±6.56)、(134.80±11.00和105.16±6.43)；bFGF平均灰度值分别为：(192.90±11.50和112.86±7.59)、(139.70±9.43和115.18±7.22)。A组和B组手术侧BMP-2、bFGF的表达与正常对侧比较,差异均有统计学意义(P0.05),且A组与B组手术侧进行比较,差异也有统计学意义(P0.05)。 2)内撑器治疗术后6个月： (1)影像学观察。X线检查及CT扫描示：A组股骨头可见塌陷较前明显加重,股骨头囊性变及骨质硬化,关节间隙变窄,而B组股骨头外形正常,骨质密度较均匀,关节间隙正常。 (2)生物力学检测。A组、B组手术侧和正常对侧股骨头软骨下骨最大压缩强度分别为：(53.44±2.45和91.50±2.02)、(88.78±2.45和91.94±2.99)Mpa；平均弹性模量分别为：(84.66±3.03和154.30±3.46)、(153.74±3.20和155.98±3.55)MPa。A组手术侧股骨头软骨下骨最大压缩强度和平均弹性模量与正常对侧比较,差异有统计学意义(P0.05),B组手术侧与正常对侧比较,差异无统计学意义(P0.05),而A组与B组手术侧比较,差异有统计学意义(P0.05)。 (3)HE染色组织学检查。A组、B组手术侧和正常对侧空骨陷窝计数百分比分别为：(19.58±1.41和12.42±1.40)、(12.46±1.51和11.48±1.54)%；血管计数分别为：(27.00±2.12和42.40±3.78)、(43.00±3.54和44.80±3.56)。A组手术侧空骨陷窝计数百分比和血管计数与正常对侧比较,差异有统计学意义(P0.05),B组手术侧与正常对侧比较,差异无统计学意义(P0.05),而A组与B组手术侧比较,差异有统计学意义(P0.05)。 (4)免疫组化染色。A组、B组手术侧与正常对侧的BMP-2平均灰度值为：(161.90±12.10和98.22±6.33)、(110.04±8.60和102.28±7.79)；bFGF平均灰度值分别为：(157.20±10.10和113.68±6.93)、(124.92±7.55和116.26±6.30)。A组手术侧BMP-2、bFGF的表达与正常对侧比较,差异有统计学意义(P0.05)；B组手术侧与正常对侧比较,差异无统计学意义(P0.05),而A组与B组手术侧比较,差异有统计学意义(P0.05)。结论：1.本实验采用的改良液氮冷冻法可以成功制作出股骨头坏死的动物模型。该造模方法实验犬死亡率低,操作方便,可重复性高,创伤较小,不破坏髋关节的稳定性。该动物模型可用于临床上探索治疗股骨头坏死的实验研究,同时也为我们后续的研究提供较为理想的动物模型。 2.应用钛合金股骨头内撑器能有效恢复股骨头的生物力学性能,促进坏死区骨修复,促进新生血管形成,防止股骨头塌陷。在植入术后三个月时,内撑器组各项指标均优于坏死空白对照组,但比自身正常对侧差；在术后六个月时,内撑器组各项指标均优于坏死对照组,与自身正常对侧比较,差异无统计学意义。有望为临床上治疗成人股骨头坏死提供一种新的方法。
[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 卤 2.37 and 91.30 卤 2.21 ) , ( 81.78 卤 2.36 and 91.16 卤 3.36 ) , ( 139.90 卤 3.49 and 151.96 卤 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 卤 1.34 and 11.52 卤 1.29 ) , ( 15.44 卤 1.47 and 12.54 卤 3.35 ) , ( 33.00 卤 2.55 and 46.20 卤 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 卤 12.90 and 100.34 卤 6.56 ) , ( 139.70 卤 11.00 and 105.16 卤 7.22 ) , ( 139.70 卤 9.43 and 11.18 卤 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 卤 2.45 and 91.94 卤 2.99 ) Mpa , ( 88.78 卤 3.03 and 154.30 卤 3.46 ) , ( 153.74 卤 3.20 and 155.98 卤 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 卤 1.41 and 12.42 卤 1.40 ) , ( 12.46 卤 1.54 and 11.48 卤 3.56 ) , ( 43.00 卤 3.54 and 44.80 卤 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 卤 12.10 and 98.22 卤 6.33 ) , ( 110.04 卤 8.60 and 102.28 卤 7.79 ) , respectively . The average gray values of bFGF in group A were ( 157.20 卤 10.10 and 13.68 卤 6.93 ) , ( 124.92 卤 7.55 and 1160.26 卤 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 .

【学位授予单位】：中南大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R318.17

【参考文献】