Human Osteoblasts Express a Repertoire of Cadherins, Which Are Critical for BMP-2–Induced Osteogenic Differentiation

Cheng, Su‐Li; Lecanda, Fernando; Davidson, Mari K.; Warlow, Pamela M.; Zhang, Shufang; Zhang, Liming; Suzuki, Shintaro; John, Tom St.; Civitelli, Roberto

doi:10.1359/jbmr.1998.13.4.633

Cited by 149 publications

(181 citation statements)

References 47 publications

Supporting

Mentioning

163

Contrasting

Unclassified

Order By: Relevance

“…The advantage of using cad-11, as opposed to a marker such as STRO-1, is the ability to identify osteoprogenitors, rather than mesenchymal progenitors, and the level of characterization the cell surface marker has previously received [39,46,73,74].…”

Section: Discussionmentioning

confidence: 99%

Comparison of Osteogenic Differentiation of Embryonic Stem Cells and Primary Osteoblasts Revealed by Responses to IL-1β, TNF-α, and IFN-γ

Sidney

Kirkham

Buttery

2014

Stem Cells and Development

View full text Add to dashboard Cite

There are well-established approaches for osteogenic differentiation of embryonic stem cells (ESCs), but few show direct comparison with primary osteoblasts or demonstrate differences in response to external factors. Here, we show comparative analysis of in vitro osteogenic differentiation of mouse ESC (osteo-mESC) and mouse primary osteoblasts. Both cell types formed mineralized bone nodules and produced osteogenic extracellular matrix, based on immunostaining for osteopontin and osteocalcin. However, there were marked differences in the morphology of osteo-mESCs and levels of mRNA expression for osteogenic genes. In response to the addition of proinflammatory cytokines interleukin-1b, tumor necrosis factor-a, and interferon-g to the culture medium, primary osteoblasts showed increased production of nitric oxide (NO) and prostaglandin E 2 (PGE 2 ) at early time points and decreases in cell viability. In contrast, osteo-mESCs maintained viability and did not produce NO and PGE 2 until day 21. The formation of bone nodules by primary osteoblasts was reduced markedly after cytokine stimulation but was unaffected in osteo-mESCs. Cell sorting of osteo-mESCs by cadherin-11 (cad-11) showed clear osteogenesis of cad-11+ cells compared to unsorted osteo-mESCs and cad-11 -cells. Moreover, the cad-11 + cells showed a significant response to cytokines, similar to primary osteoblasts. Overall, these results show that while osteo-mESC cultures, without specific cell sorting, show characteristics of osteoblasts, there are also marked differences, notably in their responses to cytokine stimuli. These findings are relevant to understanding the differentiation of stem cells and especially developing in vitro models of disease, testing new drugs, and developing cell therapies.

show abstract

Section: Discussionmentioning

confidence: 99%

Comparison of Osteogenic Differentiation of Embryonic Stem Cells and Primary Osteoblasts Revealed by Responses to IL-1β, TNF-α, and IFN-γ

Sidney

Kirkham

Buttery

2014

Stem Cells and Development

View full text Add to dashboard Cite

show abstract

“…We and others have previously demonstrated that uncommitted mesenchymal embryonic cells express several cadherins, and that commitment and differentiation into osteogenic, myogenic or adipogenic lineages is associated with regulated expression of specific cadherin repertoires (Kawaguchi et al, 2001b;Shin et al, 2000;Stains and Civitelli, 2005a). More specifically, cells of the osteoblast lineage are defined by the presence of two cadherins, N-cadherin and cadherin 11; and earlier in vitro data had suggested that cadherin 11 was specifically involved in osteoblast differentiation (Cheng et al, 1998;Kawaguchi et al, 2001b;Okazaki et al, 1994). In fact, overexpression of cadherin 11 in uncommitted embryonic cells gives rise to teratomas containing bone and cartilage (Kii et al, 2004).…”

Section: Introductionmentioning

confidence: 93%

N-cadherin and cadherin 11 modulate postnatal bone growth and osteoblast differentiation by distinct mechanisms

Benedetto

Watkins

Grimston

et al. 2010

Journal of Cell Science

Self Cite

102

116

View full text Add to dashboard Cite

SummaryWe have previously shown that targeted expression of a dominant-negative truncated form of N-cadherin (Cdh2) delays acquisition of peak bone mass in mice and retards osteoblast differentiation; whereas deletion of cadherin 11 (Cdh11), another osteoblast cadherin, leads to only modest osteopenia. To determine the specific roles of these two cadherins in the adult skeleton, we generated mice with an osteoblast/osteocyte specific Cdh2 ablation (cKO) and double Cdh2 +/-;Cdh11 -/-germline mutant mice. Age-dependent osteopenia and smaller diaphyses with decreased bone strength characterize cKO bones. By contrast, Cdh2 +/-;Cdh11 -/-exhibit severely reduced trabecular bone mass, decreased in vivo bone formation rate, smaller diaphyses and impaired bone strength relative to single Cdh11 null mice. The number of bone marrow immature precursors and osteoprogenitor cells is reduced in both cKO and Cdh2 +/-;Cdh11 -/-mice, suggesting that N-cadherin is involved in maintenance of the stromal cell precursor pool via the osteoblast. Although Cdh11 is dispensable for postnatal skeletal growth, it favors osteogenesis over adipogenesis. Deletion of either cadherin reduces -catenin abundance and -catenin-dependent gene expression, whereas N-cadherin loss disrupts cell-cell adhesion more severely than loss of cadherin 11. Thus, Cdh2 and Cdh11 are crucial regulators of postnatal skeletal growth and bone mass maintenance, serving overlapping, yet distinct, functions in the osteogenic lineage.

show abstract

“…(28) Protein concentration in the extract was measured using a Bio-Rad DC kit. Samples containing equal amount of proteins were subjected to SDS-PAGE directly or were immunoprecipitated with P1F6 anti-␣v␤5 antibody (Chemicon International) or control antibody (Santa Cruz).…”

Section: Immunoprecipitation and Western Blot Analysismentioning

confidence: 99%

Four and Half Lim Protein 2 (FHL2) Stimulates Osteoblast Differentiation

Lai

Bai

Uthgenannt

et al. 2006

Journal of Bone and Mineral Research

Self Cite

View full text Add to dashboard Cite

FHL2, a molecule that interacts with many integrins and transcription factors, was found to play an important role in osteoblast differentiation. Overexpression of FHL2 increases the accumulation of osteoblast differentiation markers and matrix mineralization, whereas FHL2 deficiency results in inhibition of osteoblast differentiation and decreased bone formation.Introduction: Integrin-matrix interaction plays a critical role in osteoblast function. It has been shown that the cytoplasmic domains of integrin ␤ subunits mediate signal transduction induced by integrin-matrix interaction. We reasoned that the identification of proteins interacting with ␤-cytoplasmic tails followed by analysis of the function of these proteins would enhance our understanding on integrin signaling and the roles of these proteins in osteoblast activities. Materials and Methods: Yeast two hybrid assay was used to identify proteins interacting with the cytoplasmic domain of integrin ␤5 subunit. The association of these proteins with integrin ␣v␤5 was confirmed by confocal analysis and co-immunoprecipitation. A stable MC3T3-E1 cells line overexpressing Four and Half Lim Protein 2 (FHL2) and mouse osteoblasts deficient in FHL2 were used to study the roles of FHL2 in osteoblast differentiation and bone formation. Matrix protein expression was determined by mRNA analysis and Western blotting. Matrix mineralization was detected by Alizarin red staining. Alkaline phosphatase activity was also measured. CT was used to determine bone histomorphometry. Results and Conclusions: FHL2 and actin-binding proteins, palladin and filamin A, were identified as proteins interacting with ␤5 cytoplasmic domain. FHL2 co-localized with ␣v␤5 at the focal adhesion sites in association with palladin and filamin A. FHL2 was also present in nuclei. Osteoblasts overexpressing FHL2 exhibited increased adhesion to and migration on matrix proteins. Conversely, FHL2 stimulation of CREB activity was dependent on integrin function because it was inhibited by Gly-Arg-Gly-Asp-Ser (GRGDS) peptide. The expression of osteoblast differentiation markers and Msx2 was upregulated, and bone matrix mineralization was increased in FHL2 overexpressing cells. In contrast, FHL2-deficient bone marrow cells and osteoblasts displayed decreased osteoblast colony formation and differentiation, respectively, compared with wildtype cells. Moreover, FHL2-deficient female mice exhibited greater bone loss than the wildtype littermates after ovariectomy. Thus, FHL2 plays an important role in osteoblast differentiation and bone formation.

show abstract

Human Osteoblasts Express a Repertoire of Cadherins, Which Are Critical for BMP-2–Induced Osteogenic Differentiation

Abstract: ABSTRACT

Cited by 149 publications

References 47 publications

Comparison of Osteogenic Differentiation of Embryonic Stem Cells and Primary Osteoblasts Revealed by Responses to IL-1β, TNF-α, and IFN-γ

Comparison of Osteogenic Differentiation of Embryonic Stem Cells and Primary Osteoblasts Revealed by Responses to IL-1β, TNF-α, and IFN-γ

N-cadherin and cadherin 11 modulate postnatal bone growth and osteoblast differentiation by distinct mechanisms

Four and Half Lim Protein 2 (FHL2) Stimulates Osteoblast Differentiation

Contact Info

Product

Resources

About