{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T04:14:33Z","timestamp":1760242473197,"version":"build-2065373602"},"reference-count":36,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2017,7,28]],"date-time":"2017-07-28T00:00:00Z","timestamp":1501200000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>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.<\/jats:p>","DOI":"10.3390\/ijms18081646","type":"journal-article","created":{"date-parts":[[2017,7,28]],"date-time":"2017-07-28T10:14:34Z","timestamp":1501236874000},"page":"1646","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["A Simple Procedure for the Evaluation of Bone Vitality by Staining with a Tetrazolium Salt"],"prefix":"10.3390","volume":"18","author":[{"given":"Ren\u00e9","family":"Schiffner","sequence":"first","affiliation":[{"name":"Orthopaedic Department, Jena University Hospital\u2010Friedrich Schiller University, Campus Eisenberg, Klosterlausnitzer Str. 81, 07607 Eisenberg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Juliane","family":"Reiche","sequence":"additional","affiliation":[{"name":"Institute for Biochemistry II, Jena University Hospital\u2010Friedrich Schiller University, Nonnenplan 4, 07743 Jena, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Steffen","family":"Brodt","sequence":"additional","affiliation":[{"name":"Orthopaedic Department, Jena University Hospital\u2010Friedrich Schiller University, Campus Eisenberg, Klosterlausnitzer Str. 81, 07607 Eisenberg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Olaf","family":"Brinkmann","sequence":"additional","affiliation":[{"name":"Orthopaedic Department, Jena University Hospital\u2010Friedrich Schiller University, Campus Eisenberg, Klosterlausnitzer Str. 81, 07607 Eisenberg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Matthias","family":"Bungartz","sequence":"additional","affiliation":[{"name":"Orthopaedic Department, Jena University Hospital\u2010Friedrich Schiller University, Campus Eisenberg, Klosterlausnitzer Str. 81, 07607 Eisenberg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Georg","family":"Matziolis","sequence":"additional","affiliation":[{"name":"Orthopaedic Department, Jena University Hospital\u2010Friedrich Schiller University, Campus Eisenberg, Klosterlausnitzer Str. 81, 07607 Eisenberg, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3578-4338","authenticated-orcid":false,"given":"Martin","family":"Schmidt","sequence":"additional","affiliation":[{"name":"Institute for Biochemistry II, Jena University Hospital\u2010Friedrich Schiller University, Nonnenplan 4, 07743 Jena, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2017,7,28]]},"reference":[{"key":"ref_1","first-page":"933","article-title":"Quantitative analysis of acute myocardial infarct in rat hearts with ischemia-reperfusion using a high-resolution stationary SPECT system","volume":"43","author":"Liu","year":"2002","journal-title":"J. Nucl. Med."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1016\/j.brainresrev.2007.10.010","article-title":"Neuroprotection by pravastatin in acute ischemic stroke in rats","volume":"58","author":"Berger","year":"2008","journal-title":"Brain Res. Rev."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1304","DOI":"10.1161\/01.STR.17.6.1304","article-title":"Evaluation of 2,3,5-triphenyltetrazolium chloride as a stain for detection and quantification of experimental cerebral infarction in rats","volume":"17","author":"Bederson","year":"1986","journal-title":"Stroke"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1016\/j.rvsc.2017.03.013","article-title":"Evaluation of a vital staining protocol with 2,3,5-triphenyltetrazolium chloride for cancellous bone in a sheep model","volume":"114","author":"Schiffner","year":"2017","journal-title":"Re. Vet. Sci."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"152","DOI":"10.1302\/0301-620X.98B2.36614","article-title":"Cost effectiveness of surgical versus non-surgical treatment of adults with displaced fractures of the proximal humerus: Economic evaluation alongside the PROFHER trial","volume":"98","author":"Corbacho","year":"2016","journal-title":"Bone Jt. J."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"E130","DOI":"10.1016\/j.jse.2015.11.016","article-title":"Intramedullary nailing of the proximal humerus: Evolution, technique, and results","volume":"25","author":"Dilisio","year":"2016","journal-title":"J. Shoulder Elbow Surg."},{"key":"ref_7","first-page":"CD000434","article-title":"Interventions for treating proximal humeral fractures in adults","volume":"11","author":"Handoll","year":"2015","journal-title":"Cochrane Database Syst. Rev."},{"key":"ref_8","first-page":"2254","article-title":"Treatment of femoral neck fractures in young adults","volume":"60","author":"Ly","year":"2009","journal-title":"JBJS"},{"key":"ref_9","first-page":"251","article-title":"Femoral neck fractures in adults: Joint sparing operations. I. Results of an AO collective study with 328 patients","volume":"98","author":"Kuner","year":"1995","journal-title":"Der. Unfallchirurg"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1097\/00003086-200206000-00005","article-title":"Algorithms for the treatment of femoral neck fractures","volume":"399","author":"Shah","year":"2002","journal-title":"Clin. Orthop. Relat. Res."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1926","DOI":"10.1016\/j.injury.2013.05.002","article-title":"Clinical experience, primary results and pitfalls in the treatment of intracapsular hip fractures with the Targon(R) FN locking plate","volume":"44","author":"Korver","year":"2013","journal-title":"Injury"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"459","DOI":"10.1016\/j.injury.2014.12.025","article-title":"Timing of internal fixation of femoral neck fractures. A systematic review and meta-analysis of the final outcome","volume":"46","author":"Papakostidis","year":"2015","journal-title":"Injury"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"S58","DOI":"10.1016\/S0020-1383(15)30019-X","article-title":"Timing of surgery for open reduction and internal fixation of displaced proximal humeral fractures","volume":"46","author":"Siebenburger","year":"2015","journal-title":"Injury"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1016\/0020-1383(89)90152-6","article-title":"Significance of urgent (within 6 h) internal fixation in the management of fractures of the neck of the femur","volume":"20","author":"Manninger","year":"1989","journal-title":"Injury"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Ono, T., and Takayanagi, H. (2017). Osteoimmunology in Bone Fracture Healing. Curr. Osteoporos. Rep.","DOI":"10.1007\/s11914-017-0381-0"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1016\/j.bone.2014.07.033","article-title":"Bone fracture healing: Cell therapy in delayed unions and nonunions","volume":"70","author":"Rosset","year":"2015","journal-title":"Bone"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"311","DOI":"10.4248\/BR201304002","article-title":"Skeletal Blood Flow in Bone Repair and Maintenance","volume":"1","author":"Tomlinson","year":"2013","journal-title":"Bone Res."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"119","DOI":"10.1016\/j.bone.2016.02.020","article-title":"Inflammation, fracture and bone repair","volume":"86","author":"Loi","year":"2016","journal-title":"Bone"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1016\/j.bonr.2017.03.002","article-title":"Bone fracture healing in mechanobiological modeling: A review of principles and methods","volume":"6","author":"Ghiasi","year":"2017","journal-title":"Bone Rep."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Pigossi, S.C., Medeiros, M.C., Saska, S., Cirelli, J.A., and Scarel-Caminaga, R.M. (2016). Role of Osteogenic Growth Peptide (OGP) and OGP(101\u20134) in Bone Regeneration: A Review. Int. J. Mol. Sci., 17.","DOI":"10.3390\/ijms17111885"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Thurairajah, K., Broadhead, M.L., and Balogh, Z.J. (2017). Trauma and Stem Cells: Biology and Potential Therapeutic Implications. Int. J. Mol. Sci., 18.","DOI":"10.3390\/ijms18030577"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Valenti, M.T., Dalle Carbonare, L., and Mottes, M. (2016). Osteogenic Differentiation in Healthy and Pathological Conditions. Int. J. Mol. Sci., 18.","DOI":"10.3390\/ijms18010041"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Yu, W., Zhu, C., Xu, W., Jiang, L., and Jiang, S. (2016). Neuropeptide Y1 Receptor Regulates Glucocorticoid-Induced Inhibition of Osteoblast Differentiation in Murine MC3T3-E1 Cells via ERK Signaling. Int. J. Mol. Sci., 17.","DOI":"10.3390\/ijms17122150"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Hong, G., Zhou, L., Shi, X., He, W., Wang, H., Wei, Q., Chen, P., Qi, L., Tickner, J., Lin, L., and Xu, J. (2017). Bajijiasu Abrogates Osteoclast Differentiation via the Suppression of RANKL Signaling Pathways through NF-kappaB and NFAT. Int. J. Mol. Sci., 18.","DOI":"10.3390\/ijms18010203"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Baek, J.M., Kim, J.Y., Lee, C.H., Yoon, K.H., and Lee, M.S. (2017). Methyl Gallate Inhibits Osteoclast Formation and Function by Suppressing Akt and Btk-PLCgamma2-Ca2+ Signaling and Prevents Lipopolysaccharide-Induced Bone Loss. Int. J. Mol. Sci., 18.","DOI":"10.3390\/ijms18030581"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Shi, G.X., Zheng, X.F., Zhu, C., Li, B., Wang, Y.R., Jiang, S.D., and Jiang, L.S. (2017). Evidence of the Role of R-Spondin 1 and Its Receptor Lgr4 in the Transmission of Mechanical Stimuli to Biological Signals for Bone Formation. Int. J. Mol. Sci., 18.","DOI":"10.3390\/ijms18030564"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Hoff, P., Gaber, T., Strehl, C., Jakstadt, M., Hoff, H., Schmidt-Bleek, K., Lang, A., Rohner, E., Huscher, D., and Matziolis, G. (2017). Pronounced Inflammatory Activity Characterizes the Early Fracture Healing Phase in Immunologically Restricted Patients. Int. J. Mol. Sci., 18.","DOI":"10.3390\/ijms18030583"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Qiu, Z.C., Dong, X.L., Dai, Y., Xiao, G.K., Wang, X.L., Wong, K.C., Wong, M.S., and Yao, X.S. (2016). Discovery of a New Class of Cathepsin K Inhibitors in Rhizoma Drynariae as Potential Candidates for the Treatment of Osteoporosis. Int. J. Mol. Sci., 17.","DOI":"10.3390\/ijms17122116"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"413","DOI":"10.1002\/jor.1100070314","article-title":"Laser Doppler flowmetry for bone blood flow measurements: Helium-neon laser light attenuation and depth of perfusion assessment","volume":"7","author":"Notzli","year":"1989","journal-title":"J. Orthop. Res."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"427","DOI":"10.1016\/j.jse.2004.01.034","article-title":"Predictors of humeral head ischemia after intracapsular fracture of the proximal humerus","volume":"13","author":"Hertel","year":"2004","journal-title":"J. Shoulder Elbow Surg."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"319","DOI":"10.1016\/0014-5793(69)80346-7","article-title":"On the state of mitochondria in perfused liver: Action of sodium azide on respiratory carriers and respiration","volume":"5","author":"Sies","year":"1969","journal-title":"FEBS Lett."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"251","DOI":"10.1016\/S0091-679X(08)61583-6","article-title":"Measurement of cell death","volume":"57","author":"Loo","year":"1998","journal-title":"Methods Cell Biol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"297","DOI":"10.1007\/BF02556147","article-title":"Calcitonin has direct effects on 3[H]-thymidine incorporation and alkaline phosphatase activity in human osteoblast-line cells","volume":"48","author":"Farley","year":"1991","journal-title":"Calcif. Tissue Int."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1016\/S0079-6336(11)80060-4","article-title":"The tetrazolium-formazan system: Design and histochemistry","volume":"24","author":"Seidler","year":"1991","journal-title":"Prog. Histochem. Cytochem."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"676","DOI":"10.1038\/nmeth.2019","article-title":"Fiji: An open-source platform for biological-image analysis","volume":"9","author":"Schindelin","year":"2012","journal-title":"Nat. Methods"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1007\/s00402-015-2375-7","article-title":"S3-Guideline non-traumatic adult femoral head necrosis","volume":"136","author":"Roth","year":"2016","journal-title":"Arch. Orthop. Trauma Surg."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/18\/8\/1646\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T18:44:22Z","timestamp":1760208262000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/18\/8\/1646"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,7,28]]},"references-count":36,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2017,8]]}},"alternative-id":["ijms18081646"],"URL":"https:\/\/doi.org\/10.3390\/ijms18081646","relation":{},"ISSN":["1422-0067"],"issn-type":[{"type":"electronic","value":"1422-0067"}],"subject":[],"published":{"date-parts":[[2017,7,28]]}}}