Time- and Dose-dependent Cytotoxicities of Ioxitalamate and Indigocarmine in Human Nucleus Pulposus Cells

Abstract

Introduction Ioxitalamate (Telebrix 300) is an ionic iodinated contrast medium (CM) commonly used for discography or PELD (percutaneous endoscopic lumbar discectomy), though it has side effects such as anaphylactic shock and renal toxicity. Indigocarmine is an organic compound dye with a distinctive blue color that is commonly used during PELD to stain the acidic, degenerated nucleus pulposus. Although ioxitalamate and indigocarmine are widely used in spinal surgery, there have been no reports on their effects on nucleus pulposus (NP) cells. We studied the toxicity of both ioxitalamate and indigocarmine to NP cells. Materials and Methods NP cells were obtained via discectomy from lumbar disc patients and isolated. NP cells were cultured in three-dimensional (3D) alginate beads with 0.001, 0.1, 10, and 100 mg/mL ioxitalamate, 0.00001, 0.001, 0.1 and 10 mg/mL indigocarmine, or a mixture of both for 1, 2, or 3 days. The living cells were analyzed with trypan blue staining. FACS analysis using Annexin V and propidium iodide (PI) and 3D alginate bead immunostaining was performed to identify live, apoptotic, and necrotic cells. Results Ioxitalamate, indigocarmine, and their combination induced statistically significant NP cell injury that was both time and dose dependent ( p < 0.05). Also, at the same concentration, ioxitalamate was more cytotoxic than was indigocarmine or the combination ( p < 0.05). All three treatments also showed dose-dependent cytotoxicity according to flow cytometry and immunostaining. Conclusion Ioxitalamate and indigocarmine are toxic to human NP cells in vitro in a time- and dose-dependent manner. We assume that ioxitalamate and indigocarmine may have similar effects in patients undergoing discography and PELD. Thus, we suggest that ioxitalamate and indigocarmine should be used carefully at low concentrations. Disclosure of Interest None declared References Stout A. Discography. Phys Med Rehabil Clin N Am 2010;21(4):859–867 Manchikanti L, Glaser SE, Wolfer L, Derby R, Cohen SP. Systematic review of lumbar discography as a diagnostic test for chronic low back pain. Pain Physician 2009;12(3):541–559 Carragee EJ, Tanner CM, Yang B, Brito JL, Truong T. False-positive findings on lumbar discography. Reliability of subjective concordance assessment during provocative disc injection. Spine 1999;24(23):2542–2547 Laslett M, Oberg B, Aprill CN, McDonald B. Centralization as a predictor of provocation discography results in chronic low back pain, and the influence of disability and distress on diagnostic power. Spine J 2005;5(4):370–380 Kim D, Wadley R. Variability in techniques and patient safety protocols in discography: a national multispecialty survey of International Spine Intervention Society members. J Spinal Disord Tech 2010;23(6):431–438 Carragee EJ, Tanner CM, Khurana S, et al. The rates of false-positive lumbar discography in select patients without low back symptoms. Spine 2000;25(11):1373-1380, discussion 1381 Paul MD, Tsou M. Posterolateral percutaneous endoscopic lumbar discectomy. Oper Tech Orthop 2003;13:207-213 Yeung AT. Selective endoscopic discectomy. In: Kambin P, ed. Arthroscopic and Endoscopic Spinal Surgery. Totowa, NJ: Humana Press; 2005:205–225 Mayer HM, Brock M. Percutaneous endoscopic discectomy: surgical technique and preliminary results compared to microsurgical discectomy. J Neurosurg 1993;78(2):216–225 Yeung AT, Tsou PM. Posterolateral endoscopic excision for lumbar disc herniation: Surgical technique, outcome, and complications in 307 consecutive cases. Spine 2002;27(7):722–731 Lee H, Sowa G, Vo N, et al. Effect of bupivacaine on intervertebral disc cell viability. Spine J 2010;10(2):159–166 Hsu SP, Tsai TJ, Chien CT. Ioxitalamate induces renal tubular apoptosis via activation of renal efferent nerve-mediated adrenergic signaling, renin activity, and reactive oxygen species production in rats. Toxicol Sci 2010;114(1):149–158 Lee HC, Chang JG, Yen HW, Liu IH, Lai WT, Sheu SH. Ionic contrast media induced more apoptosis in diabetic kidney than nonionic contrast media. J Nephrol 2011;24(3):376–380 Belhadjali H, Bouzgarrou L, Youssef M, Njim L, Zili J. DRESS syndrome induced by sodium meglumine ioxitalamate. Allergy 2008;63(6):786–787 Lacour B, Lacroix C, Jungers P, Bailly M. [Effects of intravenous urography using sodium and meglumine ioxitalamate (Telebrix 38) on blood creatinine and urinary excretion of N-acetyl-beta-D-glucosaminidase]. Ann Biol Clin (Paris) 1984;42(6):407–411