Method for Detecting the Expression of Bone Matrix Protein by in situ Hybridization Using Decalcified Mineralized Tissue.

Nomura, Shosaku; Hirakawa, Kimiaki; Nagoshi, Junsuke; Hirota, Seiichi; Kim, Hyung-Min; Takemura, Teiji; Nakase, Takanobu; Takaoka, Kunio; Matsumoto, Sayuri; Nakajima, Yuko; Takebayashi, Kimiko; Takano‐Yamamoto, Teruko; Ikeda, Tohru; Kitamura, Yukihiko

doi:10.1267/ahc.26.303

Cited by 53 publications

(32 citation statements)

References 20 publications

(7 reference statements)

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“…The source of these proteins appeared to be through their synthesis and secretion by cells that had acquired an osteogenic phenotype. Osteopontin is a major non-collagenous protein in the bone matrix and is expressed by bone-related cells including osteoclasts, osteoblasts, and osteocytes (Nomura et al, 1993;Nanci et al, 1996). In the present study, osteopontin expression was recognized in the calcified matrix and in transforming cells after 4 weeks of transplantation.…”

Section: Discussionsupporting

confidence: 55%

Expression of osteogenic proteins during the intrasplenic transplantation of Meckel's chondrocytes: A histochemical and immunohistochemical study

Ishizeki

Kagiya

Fujiwara

et al. 2009

Archives of Histology and Cytology

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Section: Discussionsupporting

confidence: 55%

Expression of osteogenic proteins during the intrasplenic transplantation of Meckel's chondrocytes: A histochemical and immunohistochemical study

Ishizeki

Kagiya

Fujiwara

et al. 2009

Archives of Histology and Cytology

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“…13,14 The cRNA probes for in situ hybridisation of mouse ␣1 type-I collagen and ␣1 type-II collagen were given by Metsäranta et al 15 These were used after homology between mouse and rat cRNA had been confirmed by northern blotting. Type-X collagen cDNA was obtained by reverse transcription of mRNA from newborn rat bone tissue, followed by a polymerase chain reaction and subcloning into EcoRV site of pBluescript KS-(Stratagene, La Jolla, California).…”

Section: Methodsmentioning

confidence: 99%

Three Modes of Ossification During Distraction Osteogenesis in the Rat

Yasui¹,

Sato²,

Ochi³

et al. 1997

The Journal of Bone and Joint Surgery. British volume

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We developed a rat model of limb lengthening to study the basic mechanism of distraction osteogenesis, using a small monolateral external fixator. In 11-week-old male rats we performed a subperiosteal osteotomy in the midshaft of the femur with distraction at 0.25 mm every 12 hours from seven days after operation.Radiological and histological examinations showed a growth zone of constant thickness in the middle of the lengthened segment, with formation of new bone at its proximal and distal ends.Osteogenic cells were arranged longitudinally along the tension vector showing the origin and the fate of individual cells in a single section. Typical endochondral bone formation was prominent in the early stage of distraction, but intramembraneous bone formation became the predominant mechanism of ossification at later stages. We also showed a third mechanism of ossification, 'transchondroid bone formation'. Chondroid bone, a tissue intermediate between bone and cartilage, was formed directly by chondrocyte-like cells, with transition from fibrous tissue to bone occurring gradually and consecutively without capillary invasion.In situ hybridisation using digoxigenin-11-UTPlabelled complementary RNAs showed that the chondroid bone cells temporarily expressed type-II collagen mRNA. They did not show the classical morphological characteristics of chondrocytes, but were assumed to be young chondrocytes undergoing further differentiation into bone-forming cells.We found at least three different modes of ossification during bone lengthening by distraction osteogenesis. We believe that this is the first report of such a rat model, and have shown the validity of in situ hybridisation techniques for the study of the cellular and molecular mechanisms involved in distraction osteogenesis. J Bone Joint Surg [Br] 1997;79-B:824-30. Received 18 November 1996; Accepted after revision 21 March 1997The common principles of current techniques of limb lengthening are osteotomy and slow progressive distraction by an external fixation device.1,2 The type of osteotomy, the timing and rate of distraction, and the apparatus have varied considerably. 3-8 It has been established that slow distraction does not break the bony callus, but actually stimulates osteogenesis and the growth of surrounding soft tissues. The principle has been termed the 'law of tension-stress', 4,5 but the exact mechanism is not understood. We have previously reported the histological findings in lengthened segments in rabbits and concluded that new bone was formed predominantly by endochondral (indirect) ossification.9 Other reports of canine or ovine experiments have established that intramembranous (direct) bone formation is the main mechanism of ossification during distraction osteogenesis. 4,5,10,11 Factors such as the stability of fixation, the timing and rate of distraction, and speciesrelated differences may determine the relative share of endochondral and direct bone formation. Kossmann, Giebel and Glombitza 12 recently described a rat model of tibial lengthening...

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“…Sense and antisense digoxigenin-labeled riboprobes were used as previously described (Nomura et al, 1993;Sawitzke et al, 2005). Probes were synthesized from chick Pdgfra clones (ChEST55k2) and from chick Pdgf-a ligand clones (ChEST826l7).…”

Section: In Situ Hybridizationmentioning

confidence: 99%

Platelet‐derived growth factor is involved in the differentiation of second heart field‐derived cardiac structures in chicken embryos

Bax

Lie-Venema

Vicente‐Steijn

et al. 2009

Developmental Dynamics

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For the establishment of a fully functional septated heart, addition of myocardium from second heart field-derived structures is important. Platelet-derived growth factors (PDGFs) are known for their role in cardiovascular development. In this study, we aim to elucidate this role of PDGF-A, PDGF-C, and their receptor PDGFR-␣. We analyzed the expression patterns of PDGF-A, -C, and their receptor PDGFR-␣ during avian heart development. A spatiotemporal pattern of ligands was seen with colocalization of the PDGFR-␣. This was found in second heart field-derived myocardium as well as the proepicardial organ (PEO) and epicardium. Mechanical inhibition of epicardial outgrowth as well as chemical disturbance of PDGFR-␣ support a functional role of the ligands and the receptor in cardiac development. Developmental Dynamics

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Method for Detecting the Expression of Bone Matrix Protein by in situ Hybridization Using Decalcified Mineralized Tissue.

Cited by 53 publications

References 20 publications

Expression of osteogenic proteins during the intrasplenic transplantation of Meckel's chondrocytes: A histochemical and immunohistochemical study

Expression of osteogenic proteins during the intrasplenic transplantation of Meckel's chondrocytes: A histochemical and immunohistochemical study

Three Modes of Ossification During Distraction Osteogenesis in the Rat

Platelet‐derived growth factor is involved in the differentiation of second heart field‐derived cardiac structures in chicken embryos

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