Evaluation of a vital staining protocol with 2,3,5-triphenyltetrazolium chloride for cancellous bone in a sheep model. 2017

René Schiffner, and Sabine J Bischoff, and Georg Matziolis, and Martin Schmidt
Orthopaedic Department, Jena University Hospital, Jena, Germany. Electronic address: rene.schiffner@med.uni-jena.de.

Decision making on the optimal surgical treatment of fractures often is hampered by the lack of a method for direct assessment of bone vitality. In various contexts, for example to determine the extents of cerebral insults or of myocardial infarctions in experimental studies, tetrazolium based staining procedures of vital cells are widely used. Here, we set out to test the applicability of tetrazolium based staining on bone samples. 8 brains and 26 femoral heads from sheep were used to prepare tissue slices, which were stained with 2,3,5-triphenyltetrazolium chloride (TTC) at various times (1 to 12h) after explantation. Staining of tissue slices was quantified by densitometric image analysis. Spectrophotometry was used for quantification in cultured cells. TTC-staining of tissue slices indicated detectability of vital cells in slices from both tissues up to 4h after explantation. Staining intensity at later time-points was indistinguishable from the staining of untreated samples or sodium azide treated (necrotic cells) controls. We provide experimental evidence that the choice of the optimal surgical approach for the treatment of fractures involving cancellous bone could be aided by a simple staining procedure for vital bone. However, the described procedure depends on the availability of bone specimens (slices). Therefore, search for an improved stain directly applicable to the bone surface is needed.

UI MeSH Term Description Entries
D001921 Brain The part of CENTRAL NERVOUS SYSTEM that is contained within the skull (CRANIUM). Arising from the NEURAL TUBE, the embryonic brain is comprised of three major parts including PROSENCEPHALON (the forebrain); MESENCEPHALON (the midbrain); and RHOMBENCEPHALON (the hindbrain). The developed brain consists of CEREBRUM; CEREBELLUM; and other structures in the BRAIN STEM. Encephalon
D004396 Coloring Agents Chemicals and substances that impart color including soluble dyes and insoluble pigments. They are used in INKS; PAINTS; and as INDICATORS AND REAGENTS. Coloring Agent,Dye,Dyes,Organic Pigment,Stain,Stains,Tissue Stain,Tissue Stains,Organic Pigments,Pigments, Inorganic,Agent, Coloring,Inorganic Pigments,Pigment, Organic,Pigments, Organic,Stain, Tissue,Stains, Tissue
D000071556 Cancellous Bone A type of osseous tissue which makes up the inner part of bone. It has a spongy, honeycomb-like structure with struts or trabecula and contains the BONE MARROW. It has higher rate of BONE REMODELING turnover than CORTICAL BONE. Spongy Bone,Trabecular Bone,Bone, Cancellous,Bone, Spongy,Bone, Trabecular,Bones, Cancellous,Bones, Spongy,Bones, Trabecular,Cancellous Bones,Spongy Bones,Trabecular Bones
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D012756 Sheep Any of the ruminant mammals with curved horns in the genus Ovis, family Bovidae. They possess lachrymal grooves and interdigital glands, which are absent in GOATS. Ovis,Sheep, Dall,Dall Sheep,Ovis dalli
D013194 Staining and Labeling The marking of biological material with a dye or other reagent for the purpose of identifying and quantitating components of tissues, cells or their extracts. Histological Labeling,Staining,Histological Labelings,Labeling and Staining,Labeling, Histological,Labelings, Histological,Stainings
D013778 Tetrazolium Salts Quaternary salts derived from tetrazoles. They are used in tests to distinguish between reducing sugars and simple aldehydes, for detection of dehydrogenase in tissues, cells, and bacteria, for determination of corticosteroids, and in color photography. (From Mall's Dictionary of Chemistry, 5th ed, p455) Tetrazolium Salt,Salt, Tetrazolium,Salts, Tetrazolium

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