Abstract：
Chromodiscography is an integral part of full-endoscopic discectomy (FED), comprising ordinary discography with radiopacity produced by contrast medium and intradiscal stain for visualizing annular defects in the endoscopic field. Nevertheless, concerns remain about the cytotoxicity of the stains used. The study of their staining efficacy is also lacking.
To evaluate the feasibility of methylene blue, patent blue, and indigo carmine for intradiscal injection, investigate the effectiveness of each dye, and define critical concentration with adequate staining efficacy and tolerable cytotoxicity for use in chromodiscography during FED.
An experimental in vitro study.
Dye stock solutions were prepared from powder. The stock was diluted with culture medium or balanced saline and used for cytotoxicity or intervertebral disc staining assays, respectively. Bovine tails were obtained from the local slaughterhouse and functional spine units of intervertebral discs were acquired by transverse incision at the disc level. Each disc was punctured over the posterolateral aspect using a surgical knife to simulate an annular defect. The intradiscal injection was performed with each dye at different concentrations using a 22G needle from the contralateral aspect of the punctured site. Staining efficacy was quantified using ImageJ software. Primary cells of bovine tails were cultivated in each dye at different concentrations. Cytotoxicity was assessed 24 hours after stain exposure using the CCK-8 toxicity assay.
Staining efficacy and cytotoxicity were proportional to the concentration of tested dyes. Lower limits of concentration producing significant staining efficacy of indigo carmine, methylene blue, and patent blue were 0.25 mg/mL, 0.25 mg/mL, and 0.05 mg/mL, respectively. Compared with controls, concentrations showing significant toxicity for indigo carmine, methylene blue, and patient blue were 1 mg/mL, 0.5 mg/mL, and 2.5 mg/mL, respectively.
Patent blue can serve as a more suitable tissue stain than either indigo carmine or methylene blue due to the widest range of tradeoff concentration within 0.05 to 2.5 mg/mL.
Patent blue with the characteristic of good staining efficacy and lower cytotoxicity may be a promising option for chromodiscography during FED.
To evaluate the feasibility of methylene blue, patent blue, and indigo carmine for intradiscal injection, investigate the effectiveness of each dye, and define critical concentration with adequate staining efficacy and tolerable cytotoxicity for use in chromodiscography during FED.
An experimental in vitro study.
Dye stock solutions were prepared from powder. The stock was diluted with culture medium or balanced saline and used for cytotoxicity or intervertebral disc staining assays, respectively. Bovine tails were obtained from the local slaughterhouse and functional spine units of intervertebral discs were acquired by transverse incision at the disc level. Each disc was punctured over the posterolateral aspect using a surgical knife to simulate an annular defect. The intradiscal injection was performed with each dye at different concentrations using a 22G needle from the contralateral aspect of the punctured site. Staining efficacy was quantified using ImageJ software. Primary cells of bovine tails were cultivated in each dye at different concentrations. Cytotoxicity was assessed 24 hours after stain exposure using the CCK-8 toxicity assay.
Staining efficacy and cytotoxicity were proportional to the concentration of tested dyes. Lower limits of concentration producing significant staining efficacy of indigo carmine, methylene blue, and patent blue were 0.25 mg/mL, 0.25 mg/mL, and 0.05 mg/mL, respectively. Compared with controls, concentrations showing significant toxicity for indigo carmine, methylene blue, and patient blue were 1 mg/mL, 0.5 mg/mL, and 2.5 mg/mL, respectively.
Patent blue can serve as a more suitable tissue stain than either indigo carmine or methylene blue due to the widest range of tradeoff concentration within 0.05 to 2.5 mg/mL.
Patent blue with the characteristic of good staining efficacy and lower cytotoxicity may be a promising option for chromodiscography during FED.
摘要：
背景：椎间盘造影是全内窥镜椎间盘切除术（FED）的组成部分，包括由造影剂和椎间盘内染色产生的具有射线不透性的普通椎间盘造影，用于可视化内窥镜视野中的环形缺损。然而，仍然担心使用的染色剂的细胞毒性。还缺乏对它们的染色功效的研究。
目的：为了评价亚甲蓝的可行性，专利蓝,和靛蓝Carmine用于椎间盘内注射，调查每种染料的有效性，并定义临界浓度，具有足够的染色功效和可耐受的细胞毒性，可用于FED期间的色盘照相术。
方法：体外实验研究。
方法:由粉末制备染料储备溶液。原液用培养基或平衡盐水稀释，用于细胞毒性或椎间盘染色测定。分别。从当地的屠宰场获得牛尾巴，并通过椎间盘水平的横向切口获得椎间盘的功能性脊柱单元。使用手术刀在后外侧上穿刺每个椎间盘以模拟环形缺损。使用22G针从穿刺部位的对侧进行不同浓度的每种染料的盘内注射。使用ImageJ软件定量染色功效。在不同浓度的每种染料中培养牛尾的原代细胞。使用CCK-8毒性测定在染色暴露后24小时评估细胞毒性。
结果:染色效力和细胞毒性与测试染料的浓度成比例。产生显著染色效果的浓度下限靛蓝,亚甲蓝,和专利蓝为0.25毫克/毫升，0.25mg/mL,和0.05mg/mL，分别。与对照组相比，对靛蓝有显著毒性的浓度，亚甲蓝,患者蓝色为1毫克/毫升，0.5mg/mL,和2.5mg/mL，分别。
结论：由于在0.05-2.5mg/mL范围内的最大权衡浓度范围，与靛蓝或亚甲蓝相比，专利蓝可以用作更合适的组织染色剂。
结论：专利蓝具有良好的染色效果和较低的细胞毒性的特征，可能是FED中显色的一种有希望的选择。
目的：为了评价亚甲蓝的可行性，
方法：体外实验研究。
方法:由粉末制备染料储备溶液。
结果:染色效力和细胞毒性与测试染料的浓度成比例。
结论：由于在0.05-2.5mg/mL范围内的最大权衡浓度范围，与靛蓝或亚甲蓝相比，专利蓝可以用作更合适的组织染色剂。
结论：专利蓝具有良好的染色效果和较低的细胞毒性的特征，可能是FED中显色的一种有希望的选择。
