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.
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。