Effects of Indigo Carmine on Human Chondrocytes In Vitro

Timo Zippelius, Arnd Hoburg, Bernd Preininger, Pauline Vörös, Carsten Perka, Georg Matziolis, Eric Röhner*
Department of Traumatology and Orthopedics, Charité - Universitätsmedizin, Berlin, Germany

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© Zippelius et al.; Licensee Bentham Open.

open-access license: This is an open access article distributed under the terms of the Creative Commons Attribution License ( which permits unrestrictive use, distribution, and reproduction in any medium, provided the original work is properly cited.

* Address correspondence to this author at the Center for Musculoskeletal Surgery, Orthopedic Department, Charité University Hospital, Berlin, Charitéplatz 1, D-10117 Berlin, Germany; Tel: ++49 (0)30 450 652 463; Fax: ++49 (0)30 450 515 911; E-mail:


Joint infections following or accompanying superficious soft tissue infections are severe complication in orthopedic surgery. The use of intra-articular blue staining is a helpful method to visualize a fistula and to differentiate between superficial and intra-articular infections. Regarding this clinical implication data about the effects of indigo carmine, a frequently used blue staining substance, on cartilage is missing. The hypothesis of this study was that indigo carmine damages human chondrocytes in a time and concentration dependent manner. Human chondrocytes were isolated from donors with osteoarthritis who were treated with TKA. Cells were cultivated and treated with different concentrations of indigo carmine for 5 and 10 minutes. Morphologic damage was examined by light microscopy. Toxicity was quantified by counting vital cell number and lactate dehydrogenase (LDH) expression. Analysis by light microscopy showed defected cell structure and loss of cell number after treatment with 100% indigo carmine for 10 minutes. Treatment with 10% and 1% indigo carmine showed no significant cell defects and loss of cells. Counting vital cell number showed loss of vital cells after treatment with 100% and 10% indigo carmine for 10 minutes. LDH expression was significantly increased after treatment with 100% indigo carmine.

Toxic effects were shown after treatment with indigo carmine. Therefore, it should be used in 1:100 dilution. This is both, sufficient for visualizing a fistula in a possible clinical application and could be protective for chondrocytes.

Keywords: Indigo carmine, human chondrocytes, toxicity.