Changes in Osteoblast Phenotype During Differentiation of Enzymatically Isolated Rat Calvaria Cells

Wada, Yasuhiro; Kataoka, Hiroko; Yokose, Satoshi; Ishizuya, Toshinori; Miyazono, Kohei; Gao, Yuhao; Shibasaki, Yoshinobu; Yamaguchi, Akira

doi:10.1016/s8756-3282(98)00039-8

Cited by 67 publications

(64 citation statements)

References 33 publications

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“…OC typically is expressed in cultures isolated from rat calvaria after the mineralization process has begun, with mRNA expression detectable after Ϸ10 days in culture and protein secretion detectable after Ϸ15 days in culture (24). Regardless of the adhesive region geometry and cell projected area, OC mRNA was observed in cells cultured on all patterned surfaces that limited cell spreading.…”

Section: Discussionmentioning

confidence: 98%

Engineering gene expression and protein synthesis by modulation of nuclear shape

Thomas

Collier

Sfeir

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

444

326

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The current understanding of the relationships between cell shape, intracellular forces and signaling, nuclear shape and organization, and gene expression is in its infancy. Here we introduce a method for investigating gene-specific responses in individual cells with controlled nuclear shape and projected area. The shape of the nuclei of primary osteogenic cells were controlled on microfabricated substrata with regiospecific chemistry by confining attachment and spreading of isolated cells on adhesive islands. Gene expression and protein synthesis were altered by changing nuclear shape. Collagen I synthesis correlated directly with cell shape and nuclear shape index (NSI), where intermediate values of nuclear distension (6 < NSI < 8) promoted maximum synthesis. Osteocalcin mRNA, a bone-specific differentiation marker, was observed intracellularly by using reverse transcription in situ PCR at 4 days in cells constrained by the pattern and not detected in unconstrained cells of similar projected area, but different NSI. Our data supports the concept of gene expression and protein synthesis based on optimal distortion of the nucleus, possibly altering transcription factor affinity for DNA, transport to the nucleus, or nuclear matrix organization. The combination of microfabricated surfaces, reverse transcription in situ PCR, and NSI measurement is an excellent system to study how transcription factors, the nuclear matrix, and the cytoskeleton interact to control gene expression and may be useful for studying a wide variety of other cell shape͞gene expression relationships.patterning ͉ cell size ͉ osteocalcin ͉ reverse transcription-PCR ͉ photolithography C ell morphology has a profound effect on a range of cellular events, such as proliferation (1-3), differentiation (4-7), cytoskeletal organization (8), and presumably gene expression. Changes to the cytoskeleton lead to altered stress levels imparted on the nucleus (9-11) and could affect organelle and DNA organization and distribution, ultimately altering cell function. For example, the rate of albumin secretion from hepatocytes can be altered by constraining cell size on patterned culture surfaces (3). In human epidermal kareatinocytes cell shape can modulate between terminal differentiation and proliferation (4). Furthermore, cells can be forced to enter the apoptotic cascade when the area on which the cell is allowed to spread is constrained (1). One proposed mechanism for the transduction of cell shape information into gene expression is through mechanical forces transmitted by means of the direct link of the cytoskeleton to the nucleus (9, 12), and in particular to nuclear matrix proteins (NMPs) such as NMP-1 and NMP-2 (13). These architectural transcription factors, which are components of the nuclear scaffold, induce changes in DNA supercoiling and can interact directly with gene promoter sequences (14). This interaction between the nuclear architecture and regulation of transcription or DNA topography raises the question of whether gross deformation of th...

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Section: Discussionmentioning

confidence: 98%

Engineering gene expression and protein synthesis by modulation of nuclear shape

Thomas

Collier

Sfeir

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

444

326

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“…(64) reported that glucocorticoidinduced formation of bone nodules in fetal rat calvarial osteoblasts was mediated by BMP-6: glucocorticoids preferentially increased expression of BMP-6 mRNA, and the antisense oligonucleotide corresponding to BMP-6 strongly inhibited formation of bone nodules. We demonstrated that BMP receptors such as BMPR-IA, BMPR-IB and BMPR-II were preferentially expressed at the sites of nodule formation in calvarial culture ( 36 ) . Taken together, these observations indicate that BMPs play important roles in the process of bone nodule formation through their receptors in a paracrine and/or autocrine fashion.…”

Section: B Role Of Bmps In Differentiation Of Osteoblast Lineagementioning

confidence: 88%

“…Osteoblastic cells isolated from the calvariae of newborn rats (33) or the bone marrow of adult rats (34) (primary osteoblasts) provide a suitable model to explore the bone formation process in vitro , because these cells generate numerous mineralized bone nodules when cultured in the presence of β-glycerophosphate and ascorbic acid. Since only a limited number of clonal cell lines retain the capacity to form mineralized bones in vitro , primary osteoblasts are important tools for analyzing the differentiation process of osteoblasts from osteoprogeni- 2 Yamaguchi Regulatory mechanism of osteoblast differentiation and bone formation tors to bone-forming mature osteoblasts (35,36). The osteogenic potential of bone marrow stromal cells has been demonstrated.…”

Section: Useful Cell Lines For Analyzing the Differentiation Pathmentioning

confidence: 99%

“…To investigate the roles of BMPs in formation of bone nodules, we investigated the distributions of BMPs and their receptors in osteoblastic cells isolated from newborn rat calvariae (36). In situ hybridization studies detected strong signals for BMP-2 and BMP-4 mRNAs in bone nodule-forming cells, but not in the cells located in inter-nodular regions.…”

Section: B Role Of Bmps In Differentiation Of Osteoblast Lineagementioning

confidence: 99%

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Regulatory Mechanism of Osteoblast Differentiation and Bone Formation.

Yamaguchi

2000

Oral Med Pathol

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IntroductionOsteoblasts , chondrocytes , myoblasts , and bone marrow stromal cells including adipocytes are believed to derive from common mesenchymal progenitors ( 1-7 ) called pluripotent mesenchymal stem cells (5-7). These progenitors exhibit specific phenotypes depending on the maturational stages during their differentiation pro− cesses. In the case of osteoblasts, they express phenotypic characteristics such as high alkaline phosphatase (ALP) activity, and synthesize collagenous and non-collagenous bone matrix proteins including osteocalcin (4). Formation of mineralized bones is the most important function of osteoblasts . This phenotypic expression is regulated by various hormones including parathyroid hormone (PTH) (8,9), 1α, 25 (OH)2D3] (10) and estrogen (11,12). Various local factors produced by osteoblasts also regulate their own differentiation in a paracrine and/or an autocrine fashion (4). To investigate the roles of these hormones and local factors in osteoblast differentiation, various osteoblastic cell lines have been successfully established (13)(14)(15)(16)(17). Experiments using these in vitro assay systems have contributed to understanding the regulation of osteoblast differentiation by various hormones and local factors.In the process of cell differentiation , many transcription factors are involved in the regulatory mechanism of each cell type, and cell lineage-specific transcription factors play crucial roles in determining each cell fate. These transcription factors have been identified in several cell lineages such as myoblasts and adipocytes: MyoD family in myoblasts ( 18 ) and PPARγ-2 in adipocytes (19). Recently, three research groups independently reported that Cbfa 1, which belongs to the runt-domain gene family , is an essential transcription factor for osteoblast differentiation and bone formation (20)(21)(22).This review describes recent advances regarding the mechanism of osteoblast differentiation and its regulation by the systemic factors and local factors including Correspondence: Akira Yamaguchi, Department of Oral Pathology, Nagasaki University School of Dentistry, 1-7-1 Sakamoto, Nagasaki, 852-8588, Japan Phone/Fax +81-95-849-7644 E-mail akirayam@net.nagasaki-u.ac.jp estrogen, parathyroid hormone ( PTH) , bone morphogenetic proteins (BMPs), hedgehogs, and the transcription factor Cbfa1. I. Differentiation of osteoblastsOsteoprogenitors are present in both skeletal and extraskeletal tissues in the post-natal state. In the skeletal tissues, osteoprogenitors locate in bone marrow and the periosteum. Friedenstein and colleagues first demonstrated that bone marrow cells harvested from confluent in vitro cultures of marrow cells retained the ability to form bones when cultured in vivo within diffusion chambers (23). They also showed that single cell-derived fibroblastic colonies, which have been called colony forming units-fibroblastic (CFU-F), retained osteogenic potential. Intermittent injection of PTH increased the number of CFU-F, which explain a potential mechanism involve...

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“…Among all available growth factors, PDGF, IGF, VEGF, TGFβ and BMPs appear to have the closest association (Urist, 1965(Urist, , 1997Urist et al, 1979;Cheifetz et al, 1996;Yeh et al, 1997;Wada et al, 1998;Wozney and Rosen, 1998;Chen et al, 2001;Reddi, 2001) Epidermal growth factor (EGF) Stimulates chondrocyte proliferation while decreasing the ability of cells to synthesize matrix components (Caplan and Boyan, 1994) Basic fibroblast growth factor (bFGF) Mitogenic effects on cells from the mesenchymal lineage. Promotes proliferation and inhibits differentiation.…”

Section: Growth Factorsmentioning

confidence: 99%

Materials in particulate form for tissue engineering. 2. Applications in bone

Silva

Coutinho

Ducheyne

et al. 2007

J Tissue Eng Regen Med

100

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Materials in particulate form have been the subjects of intensive research in view of their use as drug delivery systems. While within this application there are still issues to be addressed, these systems are now being regarded as having a great potential for tissue engineering applications. Bone repair is a very demanding task, due to the specific characteristics of skeletal tissues, and the design of scaffolds for bone tissue engineering presents several difficulties. Materials in particulate form are now seen as a means of achieving higher control over parameters such as porosity, pore size, surface area and the mechanical properties of the scaffold. These materials also have the potential to incorporate biologically active molecules for release and to serve as carriers for cells. It is believed that the combination of these features would create a more efficient approach towards regeneration. This review focuses on the application of materials in particulate form for bone tissue engineering. A brief overview of bone biology and the healing process is also provided in order to place the application in its broader context. An original compilation of molecules with a documented role in bone tissue biology is listed, as they have the potential to be used in bone tissue engineering strategies. To sum up this review, examples of works addressing the above aspects are presented. JOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE R E V I E W A R T I C L E AbstractMaterials in particulate form have been the subjects of intensive research in view of their use as drug delivery systems. While within this application there are still issues to be addressed, these systems are now being regarded as having a great potential for tissue engineering applications. Bone repair is a very demanding task, due to the specific characteristics of skeletal tissues, and the design of scaffolds for bone tissue engineering presents several difficulties. Materials in particulate form are now seen as a means of achieving higher control over parameters such as porosity, pore size, surface area and the mechanical properties of the scaffold. These materials also have the potential to incorporate biologically active molecules for release and to serve as carriers for cells. It is believed that the combination of these features would create a more efficient approach towards regeneration. This review focuses on the application of materials in particulate form for bone tissue engineering. A brief overview of bone biology and the healing process is also provided in order to place the application in its broader context. An original compilation of molecules with a documented role in bone tissue biology is listed, as they have the potential to be used in bone tissue engineering strategies. To sum up this review, examples of works addressing the above aspects are presented.

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Changes in Osteoblast Phenotype During Differentiation of Enzymatically Isolated Rat Calvaria Cells

Cited by 67 publications

References 33 publications

Engineering gene expression and protein synthesis by modulation of nuclear shape

Engineering gene expression and protein synthesis by modulation of nuclear shape

Regulatory Mechanism of Osteoblast Differentiation and Bone Formation.

Materials in particulate form for tissue engineering. 2. Applications in bone

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