The Influence of Type I Collagen on the Development and Maintenance of the Osteoblast Phenotype in Primary and Passaged Rat Calvarial Osteoblasts: Modification of Expression of Genes Supporting Cell Growth, Adhesion, and Extracellular Matrix Mineralization

Lynch, Michael J.; Stein, Janet L.; Lian, Jane B.

doi:10.1006/excr.1995.1005

Cited by 340 publications

(249 citation statements)

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“…3B). This result demonstrates that the collagen matrix used in this experiment stimulated osteocalcin expression in osteoblastic cells as previously described (10).…”

Section: Collagenase-3 Expression Is Increased By Culture On Collagensupporting

confidence: 87%

Regulation of Collagenase-3 and Osteocalcin Gene Expression by Collagen and Osteopontin in Differentiating MC3T3-E1 Cells

DʼAlonzo

Kowalski

Denhardt

et al. 2002

Journal of Biological Chemistry

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Both collagenase-3 and osteocalcin mRNAs are expressed maximally during the later stages of osteoblast differentiation. Here, we demonstrate that collagenase-3 mRNA expression in differentiating MC3T3-E1 cells is dependent upon the presence of ascorbic acid, is inhibited in the presence of the collagen synthesis inhibitor, 3,4-dehydroproline, and is stimulated by growth on collagen in the absence of ascorbic acid. Transient transfection studies show that collagenase-3 promoter activity increases during cell differentiation and requires the presence of ascorbic acid. Additionally, we show that, in differentiating MC3T3-E1 cells, collagenase-3 gene expression increases in the presence of an anti-osteopontin monoclonal antibody that binds near the RGD motif of this protein, whereas osteocalcin expression is inhibited. Furthermore, an RGD peptidomimetic compound, designed to block interaction of ligands to the ␣ v integrin subunit, increases osteocalcin expression and inhibits collagenase-3 expression, suggesting that the RGD peptidomimetic initiates certain ␣ v integrin signaling in osteoblastic cells. Overall, these studies demonstrate that stimulation of collagenase-3 expression during osteoblast differentiation requires synthesis of a collagenous matrix and that osteopontin and ␣ v integrins exert divergent regulation of collagenase-3 and osteocalcin expression during osteoblast differentiation.When primary or clonal osteoblastic cells, such as MC3T3-E1 cells, are cultured with ␤-glycerophosphate (␤-gly) 1 and ascorbic acid (AA), the cells undergo a differentiation process and eventually produce bone nodules resembling woven bone in vitro (1-4). Mineral deposition occurs in the final stages of osteoblast differentiation and is associated with maximal osteoblastic expression of osteocalcin and collagenase-3 (5-7). The mechanisms controlling osteoblast differentiation are unclear, but research suggests that a reciprocal interaction exists between the osteoblast and the surrounding extracellular matrix. For example, when osteoblasts or osteoblast-like cells are grown on plates pre-coated with collagen, they exhibit early signs of osteoblast maturation and osteoblastic gene expression (8 -12). Further accumulating data suggest that the extracellular matrix mediates many effects through activation of integrin signaling pathways in osteoblasts (12-14). Osteoblasts and osteoblast-like cells have been reported to express a wide variety of integrin receptor subunits, including ␣ 1 , ␣ 2 , ␣ 3 , ␣ 4 , ␣ 5 , ␣ v , ␤ 1 , and ␤ 3 (15-17).Osteocalcin (OC), or bone gla protein, is a calcium-binding protein found in bone extracellular matrix and expressed by osteoblasts, odontoblasts, and hypertrophic chondrocytes (reviewed in Ref. 18). It has an affinity for hydroxyapatite, suggesting a role in the regulation of mineralization. In fact, OCnull mutant mice exhibit increased bone density (19), although defects in mineralization have been detected (20). Recent evidence suggests that the up-regulation of OC late in osteoblast diff...

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“…3B). This result demonstrates that the collagen matrix used in this experiment stimulated osteocalcin expression in osteoblastic cells as previously described (10).…”

Section: Collagenase-3 Expression Is Increased By Culture On Collagensupporting

confidence: 87%

Regulation of Collagenase-3 and Osteocalcin Gene Expression by Collagen and Osteopontin in Differentiating MC3T3-E1 Cells

DʼAlonzo

Kowalski

Denhardt

et al. 2002

Journal of Biological Chemistry

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“…Osteoblasts maintain their phenotype when cultured in collagen type I containing ECM, while MSCs undergo osteogenesis (Lynch et al, 1995;Mizuno et al, 1997). The a2 integrin subunit is a component of a major collagen type I receptor a2b1 integrin.…”

Section: Discussionmentioning

confidence: 99%

Chondrogenic differentiation of bovine bone marrow mesenchymal stem cells (MSCs) in different hydrogels: Influence of collagen type II extracellular matrix on MSC chondrogenesis

Bosnakovski

Mizuno

Kim

et al. 2006

Biotech & Bioengineering

432

320

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Bone marrow mesenchymal stem cells (MSCs) are candidate cells for cartilage tissue engineering. This is due to their ability to undergo chondrogenic differentiation after extensive expansion in vitro and stimulation with various biomaterials in three-dimensional (3-D) systems. Collagen type II is one of the major components of the hyaline cartilage and plays a key role in maintaining chondrocyte function. This study aimed at analyzing the MSC chondrogenic response during culture in different types of extracellular matrix (ECM) with a focus on the influence of collagen type II on MSC chondrogenesis. Bovine MSCs were cultured in monolayer as well as in alginate and collagen type I and II hydrogels, in both serum free medium and medium supplemented with transforming growth factor (TGF) b1. Chondrogenic differentiation was detected after 3 days of culture in 3-D hydrogels, by examining the presence of glycosaminoglycan and newly synthesized collagen type II in the ECM. Differentiation was most prominent in cells cultured in collagen type II hydrogel, and it increased in a timedependent manner. The expression levels of the of chondrocyte specific genes: sox9, collagen type II, aggrecan, and COMP were measured by quantitative ''Real Time'' RT-PCR, and genes distribution in the hydrogel beads were localized by in situ hybridization. All genes were upregulated by the presence of collagen, particularly type II, in the ECM. Additionally, the chondrogenic influence of TGF b1 on MSCs cultured in collagenincorporated ECM was analyzed. TGF b1 and dexamethasone treatment in the presence of collagen type II provided more favorable conditions for expression of the chondrogenic phenotype. In this study, we demonstrated that collagen type II alone has the potential to induce and maintain MSC chondrogenesis, and prior interaction with TGF b1 to enhance the differentiation.

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“…The biological significance of the proliferation/ differentiation relationship is also necessary for an understanding of the manner in which perturbations in specific genes may be related to a broad spectrum of skeletal disorders (15)(16)(17)(18). Gene expression inducement is associated with ECM maturation and mineralization.…”

Section: Introductionmentioning

confidence: 99%

A comparison of osteogenesis-related gene expression of mesenchymal stem cells during the osteoblastic differentiation induced by Type-I collagen and/or fibronectin

2003

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Mesenchymal stem cells (MSCs) contain multipotent cells and differentiate into osteoblasts, chondrocytes, and adiphocytes. Recently, MSCs have been used clinically in a tissue-engineering concept, but cell activity varies. It was found that the characteristics of extracellular matrices, type Tcollagen and/or fibronectin matrices, induced the osteoblastic differentiation of MSCs. Six weeks after cells were cultured with type I collagen and fibronectin, they formed mineralized tissue six times higher than other matrices. In this study, using "real time RT-PCR," we assessed the expression of osteogenesis-related genes (alkaline phosphatase, BMP-2, osteocalcin, and cbfa-1) during osteoblastic differentiation by measuring their mRNA, quantitatively. The expression of the alkaline phosphatase gene increased time-dependently during osteoblastic differentiation until day 12, but expression induced by type I collagen and type I collagen and fibronectin decreased gradually. The expression of osteocalcin and the BMP-2 gene increased time-dependently; osteocalcin gene expression induced by type I collagen and fibronectin was about 6.2 times higher than noninduced cells. Expression of the cbfa-1 gene was different from that of type I collagen, fibronectin, and non-induced cells from day 12 and increased timedependently. These findings were supportedby alkaline phosphatasestaining and immunofluoresence with an osteopontin antibody.

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The Influence of Type I Collagen on the Development and Maintenance of the Osteoblast Phenotype in Primary and Passaged Rat Calvarial Osteoblasts: Modification of Expression of Genes Supporting Cell Growth, Adhesion, and Extracellular Matrix Mineralization

Cited by 340 publications

References 0 publications

Regulation of Collagenase-3 and Osteocalcin Gene Expression by Collagen and Osteopontin in Differentiating MC3T3-E1 Cells

Regulation of Collagenase-3 and Osteocalcin Gene Expression by Collagen and Osteopontin in Differentiating MC3T3-E1 Cells

Chondrogenic differentiation of bovine bone marrow mesenchymal stem cells (MSCs) in different hydrogels: Influence of collagen type II extracellular matrix on MSC chondrogenesis

A comparison of osteogenesis-related gene expression of mesenchymal stem cells during the osteoblastic differentiation induced by Type-I collagen and/or fibronectin

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