The Demonstration of Bone and Cartilage Remodelling Using Alcian Blue and Hematoxylin

Sayers, D.C.J.; Volpin, Gershon; Bentley, G.

doi:10.3109/10520298809107161

Cited by 14 publications

(9 citation statements)

References 4 publications

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“…To visualize any bone bridges and newly formed cartilage or the remaining adjacent cartilage at the injury site, histological staining was performed tising the Barbara's stain method which can effectively distinguish bone from cartilage [19]. Briefly, de-waxed sections were immersed in acid alcohol (10 mM HCI in 70% ethanol) for 30 s and then stained for 30 min with an Alcian Blue haematoxylin solution containing I'X, Alcian Blue, 0.1 5% haematoxylin, 0.05'%1 iodine and 7% aluminium amonium sulphate.…”

Section: Methodsmentioning

confidence: 99%

Intramembranous ossification mechanism for bone bridge formation at the growth plate cartilage injury site

Chen

Zhou

McCarty

et al. 2004

Journal Orthopaedic Research

133

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Salter's type I11 and type IV growth plate injuries often induce bone bridge formation at the injury site. To understand the cellular mechanisms, this study characterized proximal tibia1 transphyseal injury in rats. Histologically, bony bridge trabeculae appeared on day 7, increased on day 10, and became well-constructed on day 14 with marrow. Prior to and during bone bridging, there was no cartilage proteoglycan metachromatic staining and no collagen-)< immunostaining at the injury site, nor was there any up-regulation of BrdU-labelled chondrocyte proliferation at the adjacent physeal cartilage, suggesting no new cartilage formation at the injury site. However, infiltration of vimentin-immunopositive mesenchymal cells from metaphysis and epiphysis was apparent on day 3, with the mesenchymal population being prominent on days 7 and 10 and subsided on day 14. Among these infiltrates were osteoprogenitor precursors expressing osteoblast differentiation factor (cbf-al ) on day 3, along with some cbf-a1 osteoblast-like cells lining bone trabeculae on days 7 and 10. Some mesenchymal cells and trabecula-lining cells were also alkaline phosphataseimmunopositive, further suggesting their osteoblast differentiation. From day 7 onwards, some trabecula-lining cells became osteocalcin-producing mature osteoblasts. These results suggest that bone bridge formation after growth plate injury occurs directly via intramembranous ossification through recruitment of marrow-derived osteoprogenitor cells.Crown

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Section: Methodsmentioning

confidence: 99%

Intramembranous ossification mechanism for bone bridge formation at the growth plate cartilage injury site

Chen

Zhou

McCarty

et al. 2004

Journal Orthopaedic Research

133

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“…The characteristics of thionin in GMA and in paraffin embedded cartilage sections were comparable to safranin 0 for staining negatively charged GAG, although safranin 0 stains chondroitin sulfate more specifically [Cole 1984, Cole and Kamal 1984, Dingerkus and Uhler 1977, Goldstein and Horobin 1974, Kincaid and Evander 1985, Kiviranta et al 1985, Poole 1970, Sayers et al 1987. The absence of metachromasia and its staining properties with cartilage matrix make thionin a favorable stain for the assessment of the quality of cartilage embedded in GMA.…”

Section: Discussionmentioning

confidence: 96%

“…The network plays a n important role in differentiation and maturation of cartilage (Hjertquist Thionin Staining ot Paraffin and Plastic Embedded Cartilage 27 1972. Mallinger and Stockinger 1987, Sayers et al 1987, Shuckett and Malemud 1990. Stockwell and Scott 1967.…”

Section: Discussionmentioning

confidence: 99%

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Thionin Staining of Paraffin and Plastic Embedded Sections of Cartilage

Bulstra

Drukker

Kuijer

et al. 1993

Biotechnic & Histochemistry

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The usefulness of thionin for staining cartilage sections embedded in glycol methacrylate (GMA) and the effect of decalcification on cartilage sections embedded in paraffin and GMA were assessed. Short decalcification periods using 5% formic acid or 10% EDTA did not influence the staining properties or the morphology of cartilage matrix and chondrocytes. The standard stain safranin O-fast green for differential staining of cartilage was used as control in these experiments. Prolonged exposure of safranin O stained sections to fast green resulted in disappearance of the safranin O stained matrix, thereby hampering the quantitative measurement of negatively charged glycosaminoglycans (GAG). Thionin stained evenly throughout all cartilage layers, independent of the staining times. In contrast to safranin O, thionin did not show metachromasia in nondehydrated cartilage sections, which made it more suitable for assessing cartilage quality in GMA embedded cartilage. To evaluate the selectivity of thionin staining in cartilage, chondroitinase ABC and trypsin digestions were carried out. Thionin staining was prevented by these enzymes in the territorial matrix, representing the interlacunar network and the chondrocyte capsule. Staining with thionin of the interterritorial matrix was only slightly reduced, possibly representing keratan sulfate and hyaluronic acid in cartilage of elderly patients. Comparison of thionin stained GMA embedded cartilage with safranin O stained paraffin embedded sections showed significant similarity in optical densitometry, indicative of the specificity of thionin bound to negatively charged GAG in cartilage. In GMA embedded cartilage morphology was relatively intact compared to paraffin embedded sections due to less shrinkage of chondrocytes and the interlacunar network.

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“…Sections were stained as described previously. 30 Briefly, the tissue slides were dewaxed with 100% xylene and rehydrated in a series of baths with descending concentrations of alcohol. After hydration, the sections were dipped in 1% acid alcohol (1% hydrochloric acid mixed with 70% ethyl alcohol) for 30 seconds, drained well, and then stained with an alcian blue/hematoxylin solution (0.15% hematoxylin, 7% aluminum ammonium sulfate, 0.05% sodium iodate, and 1% alcian blue) for 30 minutes at 25°C.…”

Section: Alcian Blue Stainingmentioning

confidence: 99%

Low-Intensity Pulsed Ultrasound-Promoted Bone Healing Is Not Entirely Cyclooxgenase 2 Dependent

Huang

Tang

Chen

et al. 2008

Journal of Ultrasound in Medicine

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The Demonstration of Bone and Cartilage Remodelling Using Alcian Blue and Hematoxylin

Cited by 14 publications

References 4 publications

Intramembranous ossification mechanism for bone bridge formation at the growth plate cartilage injury site

Intramembranous ossification mechanism for bone bridge formation at the growth plate cartilage injury site

Thionin Staining of Paraffin and Plastic Embedded Sections of Cartilage

Low-Intensity Pulsed Ultrasound-Promoted Bone Healing Is Not Entirely Cyclooxgenase 2 Dependent

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