Proteoglycans in articular cartilage revealed with a quick freezing and deep etching method.

Toriumi, Hiroshi; Nakagawa, Hiroaki; Ueda, Hideho; Leng, Chong-Guang; Fujii, Yasuhisa; Ohno, Shinichi

doi:10.1136/ard.55.7.466

Cited by 10 publications

(7 citation statements)

References 48 publications

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“…In this study, we observed granular electron-dense materials in the extracellular matrix by conventional electron microscopy, which were demonstrated by the QF-DE method as an artifact formed by accumulation of PG. The QF-DE method can remove artifacts due to usual chemical fixation for a conventional electron microscopy, and be used to visualize the three-dimensional ultrastructure in cartilage tissue in a more natural state as compared with conventional electron microscopy [12].…”

Section: Discussionmentioning

confidence: 99%

“…The former are collagen fibrils that are thought to be mostly type II collagen, and the latter is PG [13,14]. Toriumi et al [12] first examined the extracellular matrix in the normal hyaline cartilage of the rat by the QF-DE method. They demonstrated that the extracellular matrix had characteristic ultrastructure in three areas: the pericellular, territorial, and interterritorial matrices.…”

Section: Discussionmentioning

confidence: 99%

“…In the ground substance with myxoid changes and the cartilaginous matrix of the chondrosarcoma, the staining results were contrasted. An extracellular matrix of normal cartilage is usually divided into pericellular, territorial, and interterritorial matrices [12]. The pericellular and territorial matrices in the cartilaginous matrix of the chondrosarcoma were stained with Alcian blue and showed metachromasia by toluidine blue staining, but both staining reactions were weak in the interterritorial matrix.…”

Section: Histochemical Findingsmentioning

confidence: 99%

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Chondrosarcoma with Myxoid Change: A Study Using a Quick-freezing and Deep-etching Method

Hemmi

Osaka

Ohni

et al. 2007

Ultrastructural Pathology

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A middle-aged Japanese woman visited the Orthopedics Department of Nihon University Nerima Hikarigaoka Hospital complaining of pain in the left hip joint that had started approximately 8 months earlier. Following several examinations, including imaging diagnoses, an incisional biopsy demonstrated a malignant acetabular bone tumor, which was removed and examined by a quick-freezing and deep-etching (QF-DE) method, conventional electron microscopy, and light microscopy. Histologically, the tumor was a chondrosarcoma with marked myxoid changes. An interesting extracellular matrix was observed by the QF-DE method. The myxoid area consisted of a fine meshwork of proteoglycans (PG) without obvious aggrecans, which resembled that of PG usually present in the pericellular matrix of normal cartilage. Thin collagen fibrils with pleated surface structures of regular periodicity were also seen, which were sparsely distributed in wide areas except for the pericellular matrix. These collagen fibrils were of the type that are mainly located in the pericellular side of the territorial matrix in normal cartilage. A myxoid matrix consisting of thin collagen fibrils on the background of pericellular type PG suggested that the myxoid matrix in the chondrosarcoma resembled those of the pericellular and pericellular sides of the territorial matrices in normal cartilage.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Histochemical Findingsmentioning

confidence: 99%

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Chondrosarcoma with Myxoid Change: A Study Using a Quick-freezing and Deep-etching Method

Hemmi

Osaka

Ohni

et al. 2007

Ultrastructural Pathology

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“…The present experiment, as reported before (Toriumi et al 1996;Leng et al 1998;Watanabe et al 2000), was performed in accordance with the guidelines governing animal experiments within the University of Yamanashi.…”

Section: Methodsmentioning

confidence: 97%

“…For observation of the pure three-dimensional morphology, some of the QF-DE replica membranes with the specimens were floated on household bleach solution to dissolve the cartilaginous tissue components, as described before (Toriumi et al 1996;Watanabe et al 2000). The cleaned replica membranes were picked up onto Formvar-filmed grids and observed in a Hitachi H-7500 transmission electron microscope, as described above.…”

Section: Quick-freezing and Deep-etching Methodsmentioning

confidence: 99%

Replica immunoelectron microscopic study of the upper surface layer in rat mandibular condylar cartilage by a quick-freezing method

Zea-Aragón

Terada

Ohno

et al. 2004

Histochemistry and Cell Biology

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A quick-freezing and deep-etching method in combination with replica immunoelectron microscopy was applied for examining localization of hyaluronic acid and fibronectin on the upper surface layer of rat mandibular condylar cartilage. Rat temporomandibular joints were dissected with articular disks in order to leave the articular cartilage surface intact. The disks were slightly cut with razor blades for exposing the condylar articular cartilage surface. They were quickly frozen with the isopentane-propane cryogen (-193 degrees C) and prepared for freeze-fracturing and deep-etching replica membranes. They were additionally treated with 5% SDS and 0.5% collagenase to keep some antigens attached on the replica membranes. After such a treatment, a routine immunogold method was applied for clarifying the localization of hyaluronic acid and fibronectin in the upper surface layer. Small immunogold particles for hyaluronic acid were mainly localized around upper filamentous networks covered with amorphous materials, but large immunogold ones for fibronectin were localized on deep thicker fibrils. We have revealed the native architecture of the upper surface layer of mandibular condylar cartilage on the replica membranes and also three-dimensional localization of hyaluronic acid and fibronectin by the immunogold method.

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Ultrastructural Modifications of the Extracellular Matrix Upon Calcification of Growth Plate Cartilage as Revealed by Quick-Freeze Deep Etching Technique

Akisaka

Nakayama

Yoshida

et al. 1998

Calcified Tissue International

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The ultrastructural changes in cartilage matrix that occur during calcification have been examined in chick epiphyseal growth plate cartilage prepared by quick-frozen, deep-etched, and rotary shadowed replicas. The extracellular cartilage matrix contains a reticular network closely associated with an extensive network of collagen. The components of the reticular network, including thick and thin filaments, are attached directly to the cell membrane, matrix vesicle membrane, and collagen fibrils. This network, which interconnects the matrix vesicles and collagen, fills the extracellular matrix. The dimensions of the reticular network seem to remain almost constant in size from the reserve and proliferative zones to the calcifying zone. The collagen fibrils seem to consist of subfibrillar structures that branch and anastomose. In optimally quick-frozen, deep-etched, prepared collagen, a cross-banding pattern was exposed. Globular structures stud the collagen fibrils, which gradually diminish in number from the reserve zone down to the calcifying zone. The matrix vesicles, when fractured, showed a granular appearance. In most cases, the fracture plane passed through the bilayer of the matrix vesicle membrane. The true surface of the matrix vesicle membrane, therefore, was exposed after deep etching. At the calcifying zone, crystal deposition had occurred in needle-like and/or plate-like form within the membrane-bound matrix vesicles. The reticular network was still intact in the vicinity of the calcified matrix, but in the intercrystalline space, neither the reticular structure nor the globular structure was detectable. Within the calcified matrix, both reticular and granular structures had disappeared from the interfibrillar space of the collagen fibrils.

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Proteoglycans in articular cartilage revealed with a quick freezing and deep etching method.

Cited by 10 publications

References 48 publications

Chondrosarcoma with Myxoid Change: A Study Using a Quick-freezing and Deep-etching Method

Chondrosarcoma with Myxoid Change: A Study Using a Quick-freezing and Deep-etching Method

Replica immunoelectron microscopic study of the upper surface layer in rat mandibular condylar cartilage by a quick-freezing method

Ultrastructural Modifications of the Extracellular Matrix Upon Calcification of Growth Plate Cartilage as Revealed by Quick-Freeze Deep Etching Technique

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