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Open AccessArticle

A Simple Procedure for the Evaluation of Bone Vitality by Staining with a Tetrazolium Salt

Orthopaedic Department, Jena University Hospital‐Friedrich Schiller University, Campus Eisenberg, Klosterlausnitzer Str. 81, 07607 Eisenberg, Germany
Institute for Biochemistry II, Jena University Hospital‐Friedrich Schiller University, Nonnenplan 4, 07743 Jena, Germany
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2017, 18(8), 1646;
Received: 23 June 2017 / Revised: 25 July 2017 / Accepted: 27 July 2017 / Published: 28 July 2017
(This article belongs to the Special Issue Advances in Bone and Cartilage Research)
Presently, no intra-operative method for a direct assessment of bone vitality exists. Therefore, we set out to test the applicability of tetrazolium-based staining on bone samples. The explanted femoral heads of 37 patients were used to obtain either cancellous bone fragments or bone slices. Samples were stained with 2,3,5-triphenyl-2H-tetrazolium chloride (TTC) or 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (thiazolyl blue, MTT) at different times (one to twelve hours) after explantation. Staining was quantified either spectrophotometrically after extraction of the dyes or by densitometric image analysis. TTC-staining of cancellous bone fragments and bone slices, respectively, indicated the detectability of vital cells in both types of samples in a window of up to six hours after explantation. Staining intensity at later time-points was indistinguishable from the staining of untreated samples or sodium azide treated samples, which represent dead cells. In contrast, MTT-staining of bone slices revealed intense unspecific staining, which obscured the evaluation of the vitality of the samples. The lack of a detectable increase of colour intensity in TTC-stained bone samples, which were treated more than six hours after explantation, corresponds to reduced fracture healing. The described simple procedure could provide a basis for an intraoperative decision by the orthopaedic surgeon. View Full-Text
Keywords: TTC; TTC staining; osteonecrosis; vital staining; bone vitality TTC; TTC staining; osteonecrosis; vital staining; bone vitality
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Schiffner, R.; Reiche, J.; Brodt, S.; Brinkmann, O.; Bungartz, M.; Matziolis, G.; Schmidt, M. A Simple Procedure for the Evaluation of Bone Vitality by Staining with a Tetrazolium Salt. Int. J. Mol. Sci. 2017, 18, 1646.

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