CN109682844B - 一种评估金属植入物内部或周围血管的检测方法 - Google Patents
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Abstract
本发明涉及一种评估金属植入物内部或周围血管的检测方法,其解决了现有金属植入体内部和周围的血管无法立体观测和定量的难题的技术问题,其利用金属氯化物凝胶复合物灌注血管方法进行血管填充,金属氯化物凝胶复合物由三氯化铬、明胶和磷酸盐缓冲溶液构成,金属氯化物凝胶复合物在40℃‑99℃呈液态,在4℃以下呈现凝胶态。本发明可广泛应于评估金属植入物内部或周围的血管。
Description
技术领域
本发明涉及金属植入物内部或周围血管检测评价技术领域,特别涉及一种利用金属氧化物氯化物凝胶复合物评估金属植入物内部或周围血管的无损检测方法。
背景技术
脊柱损伤、关节损伤等骨科疾病和口腔种植经常需要金属植入物进行填充或替换。为了保持植入物的稳固,植入后的骨修复是必不可少的。通常,骨修复的基本过程是修复区血管化、骨再生及骨整合。因为修复区血管化为骨再生和骨整合提供必要的原料、营养和氧气,维持整个骨修复微环境的动态平衡,因此修复区血管化既是骨修复的初始环节,也是骨修复的关键环节。
申请号201810541175名称为一种组织工程骨血管造影方法及血管插管的中国发明专利申请公开了一种组织工程骨血管造影方法,其采用氧化铅与明胶溶液注入动物血管内,在CT三维图像上同时显示出成骨和血管化情况。该方法中使用的氧化铅,按照国标GB5044-85,氧化铅为中度毒性物质,对生物体具有高度危害,经测定小鼠一次经口半数致死量(LD50)为180mg/kg;而且此物质水危害级别是3,也就是说氧化铅对水是极其危害的,即使是少量产品渗入地下也会对饮用水造成危害,而且对水中有机物有剧毒和危害,若无政府许可严禁将该类物质排入周围环境。因此从毒性和环保的角度来说氧化铅并不适合作为凝胶剂成份。
发明内容
本发明为了解决现有金属植入体内部和周围的血管无法立体观测和定量的难题,提供一种的评估金属植入物内部或周围血管的检测方法,优化了造影剂配方,降低造影剂毒性并提升环保性能。
本发明提供一种评估金属植入物内部或周围血管的检测方法,利用金属氯化物凝胶复合物灌注血管方法进行血管填充。
优选地,金属氯化物凝胶复合物由三氯化铬、明胶和磷酸盐缓冲溶液构成,所述金属氯化物凝胶复合物在40℃-99℃呈液态,在4℃以下呈现凝胶态。
优选地,金属氯化物凝胶复合物能够对高密度金属植入物周围血管进行显影。
优选地,血管填充后利用显微X射线计算机断层扫描技术进行血管无损检测。
优选地,显微X射线计算机断层扫描技术生成数据精确定量金属植入物内部或周围血管的总体积、血管管径以及不同血管管径区间内的血管体积,同时可提取出所述金属植入物内部或周围血管的三维图形。
优选地,血管填充后利用硬组织切磨系统制作硬组织切片,血管在硬组织切片中呈现绿色条带。
优选地,评估金属植入物内部或周围血管的检测方法,包括如下步骤:
(1)在40℃-60℃水浴锅中,向磷酸盐缓冲液中加入磷酸盐缓冲液质量2%-20%的明胶,缓缓搅拌,完全溶解,之后加入前述溶液质量5%-60%的三氯化铬粉末,搅拌混匀,保温待用;
(2)动物模型安乐死前,建立动脉流入、静脉流出的体外循环,通过体外循环灌注肝素生理盐水、组织固定液和步骤(1)中所得金属氯化物凝胶复合物;
(3)将步骤(2)中灌注完成的含有金属植入体的组织取材后置入4℃冰箱过夜,然后放入福尔马林固定液中固定;
(4)将步骤(3)中所得组织进行显微X射线计算机断层扫描技术扫描,记录样品断层扫描数据,扫描参数为,电压70kVp-140kVp,电流300μA-2000μA,曝光时间500ms-5000ms,有效像素值2μm-50μm,铝栅厚度0.1mm-5mm;
(5)将步骤(4)中所得显微X射线计算机断层扫描技术数据利用COBRA_Exxim软件进行数据重建和处理;
(6)将步骤(5)中所得重建数据利用图像分析和处理软件InveonResearchWorkplace进行分析,评估金属植入物内部或周围血管的体积和直径;
(7)将步骤(5)中所得重建数据利用图像分析和处理软件InveonResearchWorkplace进行三维模型重构,获得可视化的金属植入物内部或周围血管形态;
(8)将步骤(3)中所得组织利用硬组织切磨系统制备硬组织切片,体视镜下观察即可看到清晰的血管形态。
优选地,步骤(6)和(7)中通过定义三维图形轮廓线的方式来选取计算机断层图像感兴趣区域。
优选地,步骤(7)中的三维模型重构包括图像的灰度量化,图像增强,图像锐化、平滑,图像轮廓跟踪,图像兴趣区域提取。
本发明的有益效果是:
1.本发明优化了金属化合物凝胶复合物的配方,采用三氯化铬替代四氧化三铅,在效果不受影响的前提下,其毒性显著降低而环保性大大提升;
2.本发明利用金属化合物凝胶复合物进行血管灌注并利用放射设备显影,金属植入体内部和周围的血管就避免了被金属植入物的金属伪影所遮挡的问题。这样就能清晰地观测到金属植入体内部和周围的血管;
3.本发明利用带颜色的金属化合物制成凝胶复合物进行血管灌注后,血管也会展现出该复合物的颜色,能够在硬组织切片中明显观测;
4.本发明利用利用图像分析和处理软件重建数据,解决了金属植入体内部和周围的血管无法立体观测和定量的难题。
附图说明
图1是采用本发明所示造影剂造影,显微X射线计算机断层扫描技术扫描重建后多孔钛合金支架内部和周围造影的血管分布立体图;
图2是采用本发明所示造影剂造影,硬组织切片所示多孔钛合金支架内部造影血管的分布;
图3是采用本发明所示造影剂造影,显微X射线计算机断层扫描技术扫描重建后钛合金植入体周围造影的血管分布立体图。
具体实施方式
下面结合附图和实施例对可本发明做进一步说明,以使本发明所属技术领域的技术人员能够容易实施本发明。
一种评估金属植入物内部或周围血管的检测方法,包括如下步骤:
(1)在40℃-60℃水浴锅中,向磷酸盐缓冲液中加入质量百分比2%-20%的明胶,缓缓搅拌,完全溶解,之后加入质量百分比5%-60%的三氯化铬粉末,搅拌混匀,保温待用;
(2)动物模型安乐死前,建立动脉流入、静脉流出的体外循环,通过体外循环灌注肝素生理盐水、组织固定液和步骤(1)中所得金属氯化物凝胶复合物;
(3)将步骤(2)中灌注完成的含有金属植入体的组织取材后置入4℃冰箱过夜,然后放入福尔马林固定液中固定;
(4)将步骤(3)中所得组织进行显微X射线计算机断层扫描技术扫描,记录样品断层扫描数据,扫描参数为:电压70kVp-140kVp,电流300μA-2000μA,曝光时间500ms-5000ms,有效像素值2μm-50μm,铝栅厚度0.1mm-5mm;
(5)将步骤(4)中所得显微X射线计算机断层扫描技术数据利用COBRA_Exxim软件进行数据重建和处理;
(6)将步骤(5)中所得重建数据利用图像分析和处理软件InveonResearchWorkplace进行分析,评估金属植入物内部或周围血管的体积和直径;
(7)将步骤(5)中所得重建数据利用图像分析和处理软件InveonResearchWorkplace进行三维模型重构,获得可视化的金属植入物内部或周围血管形态;
(8)将步骤(3)中所得组织利用硬组织切磨系统制备硬组织切片,体视镜下观察即可看到清晰的血管形态。
本发明采用的金属氯化物凝胶复合物在40-99℃时为混悬液,在4℃以下为凝胶态,由三氯化铬、明胶和磷酸盐缓冲溶液构成。其制作方法为:在40℃-60℃水浴锅中,向磷酸盐缓冲液中加入磷酸盐缓冲液质量2%-20%的明胶,缓缓搅拌,完全溶解;之后加入前述溶液质量10%-60%的三氯化铬粉末,搅拌混匀,保温待用。
本发明涉及的金属氯化物凝胶复合物用于体内软组织的血管显影,填充后的血管利用显微X射线计算机断层扫描技术(简称显微CT,下同)进行血管无损检测,精确定量金属植入物内部或周围的血管总体积,血管管径,以及不同血管管径区间内的血管体积,同时可提取出金属植入物内部或周围的血管三维图形。由于金属氯化物凝胶复合物高密度的特性,决定了该类复合物在放射设备照射时会产生强显影,如果用该类复合物进行血管灌注并利用放射设备显影,金属植入体内部和周围的血管就避免了被金属植入物的金属伪影所遮挡的问题,可以清晰地观测到金属植入体内部和周围的血管。而且利用带颜色的金属化合物制成凝胶复合物进行血管灌注后,血管也会展现出该复合物的颜色,可以在硬组织切片中明显观测。
本发明中显微CT数据利用COBRA_Exxim软件进行数据重建和处理,重建数据利用图像分析和处理软件InveonResearchWorkplace进行分析,评估金属植入物内部或周围血管的体积和直径,通过定义三维图形轮廓线的方式来选取计算机断层图像感兴趣区域。同时利用图像分析和处理软件InveonResearchWorkplace进行三维模型重构,获得可视化的金属植入物内部或周围血管形态,将三维模型重构包括图像的灰度量化,图像增强,图像锐化、平滑,图像轮廓跟踪,图像兴趣区域提取。
本发明优化了金属化合物凝胶复合物的配方,采用三氯化铬替代氧化铅,在效果不受影响的前提下,其毒性显著降低而环保性大大提升。按照国标GB5044-85规定,三氯化铬为低毒性物质,对生物体具有轻度危害且并未列入常见毒物名单,经测定小鼠一次经口LD50为2136mg/kg,远低于氧化铅LD50;而且三氯化铬水危害级别是1,也就是说三氯化铬通常对水体是稍微有害的,不将未稀释或大量产品接触地下水,水道或污水系统即可。
实施例1
(1)在40℃水浴锅中,向磷酸盐缓冲液中加入磷酸盐缓冲液质量5%的明胶粉末,边加入边缓缓搅拌,使之完全溶解,之后加入前述溶液质量12%的三氯化铬粉末,搅拌混匀,继续在水浴锅中保温待用。
(2)兔称体重,耳源静脉麻醉后,皮下给予肝素钙抗凝剂。剔除腹部毛发并消毒后,打开左右腿腹股沟,找到股动脉和静脉,便于一会观察灌注效果。打开腹腔,找到腹主动脉和静脉,小心去掉周围结缔组织,将两者从体壁分开,止血钳夹住两血管近心端;将留置针小心从结扎部位远心端刺入腹主动脉,拔出钢针,边拔边将留置针软针往动脉深处插入,直到完全没入;然后将蝶形针插入留置针注射口中,胶带固定留置针和蝶形针;小心在静脉结扎处远心端位置用眼科镊挑开一个小口,建立体外循环;接着用预热的肝素生理盐水持续灌洗血管网,直至回流液清亮接近无色;然后给予质量百分比浓度(下同)10%福尔马林,再接着注入步骤(1)中所得金属氯化物凝胶复合物。
(3)将步骤(2)中灌注的家兔模型安乐死,仔细取下股骨标本,放入4℃冰箱过夜;之后把标本置入10%福尔马林固定液中固定48小时。
(4)将步骤(3)中所得组织进行显微CT扫描,记录样品断层扫描数据。本实施例采用的显微CT型号为西门子公司生产的Inveon,该仪器探测器单元数为3072,像素数为3072×3072pixels,有效像素值为8.89μm,电压为80kVp,电流为500μA,曝光时间3.5s,机架旋转角度360°,旋转步数360步,1°一步,旋转停留时间0.25s;设置好参数后进行旋转中心校准,并将高精X光球管的铝栅设为1.5mm,以过滤低能射线,减少伪影。
(5)将步骤(4)中所得显微CT数据利用COBRA_Exxim软件进行数据重建和处理,重建体素值为:8.89μm×8.89μm×8.89μm,重建层数为3072×3072×3072,重建时进行射线硬度校准,校准参数为Coeff0(a0):0;Coeff0(a1):0.797;Coeff0(a2):0.3525;Coeff0(a3):-0.0396。
(6)将步骤(5)中所得重建数据利用图像分析和处理软件InveonResearchWorkplace进行查看和分析;将重建好的数据导入该软件,使用Generalanalysis模块打开数据,评估金属植入物内部或周围血管的体积和直径。
(7)将步骤(5)中所得重建数据利用图像分析和处理软件InveonResearchWorkplace进行三维重构,根据图像设定植入物区域的灰阶阈值,根据阈值对重建数据进行血管三维形貌的提取,获得获得可视化的金属植入物内部或周围血管形态。如图1所示为本发明实施例中显微CT扫描重建后多孔钛合金支架内部和周围造影的血管分布立体图,白色箭头所示为部分血管。
(8)将步骤(3)中所得组织利用硬组织切磨系统制备硬组织切片,甲苯胺蓝染色,体视镜下观察即可看到清晰的血管形态,呈绿色条带状。如图2所示为为本发明实施例中硬组织切片所示多孔钛合金支架内部造影血管的分布,白色箭头所示深色条带为支架内部分新生血管。
实施例2
(1)在50℃水浴锅中,向磷酸盐缓冲液中加入磷酸盐缓冲液质量10%的明胶粉末,边加入边缓缓搅拌,使之完全溶解;之后加入前述溶液质量30%的三氯化铬粉末,搅拌混匀,继续在水浴锅中保温待用。
(2)大鼠腹腔麻醉后,皮下给予肝素钙抗凝剂;剔除胸部毛发并消毒后,打开胸腔,找到心脏;左心室插入蝶形针作为注入端,右心房剪一小口,建立体外循环;将蝶形针与注射器连接,用预热的肝素生理盐水持续灌洗血管网,直至回流液清亮接近无色;然后给予10%福尔马林组织固定液,再接着注入步骤(1)中所得金属氯化物凝胶复合物。
(3)将步骤(2)中灌注的大鼠模型安乐死,仔细取下股骨标本,放入4℃冰箱过夜;之后10%福尔马林组织固定液固定48小时。
(4)将步骤(3)中所得组织进行显微CT扫描,记录样品断层扫描数据。本实施例采用的显微CT型号为西门子公司生产的Inveon,该仪器探测器单元数为3072,像素数为3072×3072pixels,有效像素值为8.89μm,电压为80kVp,电流为500μA,曝光时间2.5s,机架旋转角度360°,旋转步数360步,1°一步,旋转停留时间0.15s;设置好参数后进行旋转中心校准,并将高精X光球管的铝栅设为1mm,以过滤低能射线,减少伪影。
(5)将步骤(4)中所得显微CT数据利用COBRA_Exxim软件进行数据重建和处理,重建体素值为:8.89μm×8.89μm×8.89μm,重建层数为2048×2048×3072,重建时进行射线硬度校准,校准参数为Coeff0(a0):0;Coeff0(a1):0.797;Coeff0(a2):0.3525;Coeff0(a3):-0.0396。
(6)将步骤(5)中所得重建数据利用图像分析和处理软件InveonResearchWorkplace进行查看和分析;将重建好的数据导入该软件,使用Generalanalysis模块打开数据,评估金属植入物内部或周围血管的体积和直径。
(7)将步骤(5)中所得重建数据利用图像分析和处理软件InveonResearchWorkplace进行三维重构,根据图像设定植入物区域的灰阶阈值,根据阈值对重建数据进行血管三维形貌的提取,获得获得可视化的金属植入物内部或周围血管形态。图3为本发明实施例中显微CT扫描重建后钛合金植入体周围造影的血管分布立体图,白色箭头所示为部分血管。
以上所述仅对本发明的优选实施例而已,并不用于限制本发明,对于本领域的技术人员来说,本发明可以有各种更改和变化。凡是在本发明的权利要求限定范围内,所做的任何修改、等同替换、改进等,均应在本发明的保护范围之内。
Claims (3)
1.一种评估金属植入物内部或周围血管的检测方法,其特征是包括如下步骤:
(1)在40℃-60℃水浴锅中,向磷酸盐缓冲液中加入磷酸盐缓冲液质量2%-20%的明胶,缓缓搅拌,完全溶解,之后加入前述溶液质量5%-60%的三氯化铬粉末,搅拌混匀,保温待用;
(2)动物模型安乐死前,建立动脉流入、静脉流出的体外循环,通过体外循环灌注肝素生理盐水、组织固定液和步骤(1)中所得金属氯化物凝胶复合物;
(3)将步骤(2)中灌注完成的含有金属植入体的组织取材后置入4℃冰箱过夜,然后放入福尔马林固定液中固定;
(4)将步骤(3)中所得组织进行显微X射线计算机断层扫描技术扫描,记录样品断层扫描数据,扫描参数为,电压70kVp-140kVp,电流300μA-2000μA,曝光时间500ms-5000ms,有效像素值2μm-50μm,铝栅厚度0.1mm-5mm;
(5)将步骤(4)中所得显微X射线计算机断层扫描技术数据利用COBRA_Exxim软件进行数据重建和处理;
(6)将步骤(5)中所得重建数据利用图像分析和处理软件InveonResearchWorkplace进行分析,评估金属植入物内部或周围血管的体积和直径;
(7)将步骤(5)中所得重建数据利用图像分析和处理软件InveonResearchWorkplace进行三维模型重构,获得可视化的金属植入物内部或周围血管形态;
(8)将步骤(3)中所得组织利用硬组织切磨系统制备硬组织切片,体视镜下观察即可看到清晰的血管形态。
2.根据权利要求1所述的评估金属植入物内部或周围血管的检测方法,其特征在于,所述步骤(6)和(7)中通过定义三维图形轮廓线的方式来选取计算机断层图像感兴趣区域。
3.根据权利要求1所述的评估金属植入物内部或周围血管的检测方法,其特征在于,所述步骤(7)中的三维模型重构包括图像的灰度量化,图像增强,图像锐化、平滑,图像轮廓跟踪,图像兴趣区域提取。
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