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Int. J. Mol. Sci. 2017, 18(8), 1646; doi:10.3390/ijms18081646

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

1
Orthopaedic Department, Jena University Hospital‐Friedrich Schiller University, Campus Eisenberg, Klosterlausnitzer Str. 81, 07607 Eisenberg, Germany
2
Institute for Biochemistry II, Jena University Hospital‐Friedrich Schiller University, Nonnenplan 4, 07743 Jena, Germany
*
Author to whom correspondence should be addressed.
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)
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Abstract

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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