Laminin-5 induces osteogenic gene expression in human mesenchymal stem cells through an ERK-dependent pathway. Mol Biol Cell

Department of Biology, Rensselaer Polytechnic Institute, Troy, NY 12180-3596, USA.
Molecular Biology of the Cell (Impact Factor: 4.47). 03/2005; 16(2):881-90. DOI: 10.1091/mbc.E04-08-0695
Source: PubMed


The laminin family of proteins is critical for managing a variety of cellular activities including migration, adhesion, and differentiation. In bone, the roles of laminins in controlling osteogenic differentiation of human mesenchymal stem cells (hMSC) are unknown. We report here that laminin-5 is found in bone and expressed by hMSC. hMSC isolated from bone synthesize laminin-5 and adhere to exogenous laminin-5 through alpha3beta1 integrin. Adhesion to laminin-5 activates extracellular signal-related kinase (ERK) within 30 min and leads to phosphorylation of the osteogenic transcription factor Runx2/CBFA-1 within 8 d. Cells plated on laminin-5 for 16 d express increased levels of osteogenic marker genes, and those plated for 21 d deposit a mineralized matrix, indicative of osteogenic differentiation. Addition of the ERK inhibitor PD98059 mitigates these effects. We conclude that contact with laminin-5 is sufficient to activate ERK and to stimulate osteogenic differentiation in hMSC.

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Available from: Adele L Boskey, Oct 03, 2015
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    • "To investigate the effect of ECM components on hMSCs behavior, a range of proteins, including fibronectin (FN, 40 μg/ml), laminin (LN, 40 μg/ml) and osteocalcin (OCN, 20 μg/ml & 40 μg/ml) were entrapped within the GE hydrogels (Figure b). The selected proteins are well known for their roles in osteogenic differentiation of hMSCs333435363738 and their affinity towards collagen I and IV394041, from which GE is derived. We also examined the effects of growth factors, using bone morphogenic proteins, (BMP2 and & BMP5, 50 ng/ml), which promote osteogenesis via the hedgehog pathway42. "
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    ABSTRACT: Development of three dimensional (3D) microenvironments that direct stem cell differentiation into functional cell types remains a major challenge in the field of regenerative medicine. Here, we describe a new platform to address this challenge by utilizing a robotic microarray spotter for testing stem cell fates inside various miniaturized cell-laden gels in a systematic manner. To demonstrate the feasibility of our platform, we evaluated the osteogenic differentiation of human mesenchymal stem cells (hMSCs) within combinatorial 3D niches. We were able to identify specific combinations, that enhanced the expression of osteogenic markers. Notably, these 'hit' combinations directed hMSCs to form mineralized tissue when conditions were translated to 3D macroscale hydrogels, indicating that the miniaturization of the experimental system did not alter stem cell fate. Overall, our findings confirmed that the 3D cell-laden gel microarray can be used for screening of different conditions in a rapid, cost-effective, and multiplexed manner for a broad range of tissue engineering applications.
    Scientific Reports 01/2014; 4:3896. DOI:10.1038/srep03896 · 5.58 Impact Factor
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    • "Elucidation of these mechanisms could improve the ability to promote the osteogenic differentiation of BMSCs for the purposes of cell therapy and tissue engineering. As one of the major components of the MAPK family, ERK1/2 has been associated with cellular survival, proliferation, and differentiation and especially the osteoblastic differentiation of BMSCs [51–53]. Thus, the objective of this study was to investigate whether activation of the ERK1/2 pathway was involved in the differentiation of BMSCs cultured on differentially modified surfaces into an osteogenic lineage. "
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    ABSTRACT: The current study examined the influence of culture substrates modified with the functional groups -OH, -COOH, -NH2, and -CH3 using SAMs technology, in conjunction with TAAB control, on the osteogenic differentiation of rabbit BMSCs. The CCK-8 assay revealed that BMSCs exhibited substrate-dependent cell viability. The cells plated on -NH2- and -OH-modified substrates were well spread and homogeneous, but those on the -COOH- and -CH3-modified substrates showed more rounded phenotype. The mRNA expression of BMSCs revealed that -NH2-modified substrate promoted the mRNA expression and osteogenic differentiation of the BMSCs. The contribution of ERK1/2 signaling pathway to the osteogenic differentiation of BMSCs cultured on the -NH2-modified substrate was investigated in vitro. The -NH2-modified substrate promoted the expression of integrins; the activation of FAK and ERK1/2. Inhibition of ERK1/2 activation by PD98059, a specific inhibitor of the ERK signaling pathway, blocked ERK1/2 activation in a dose-dependent manner, as revealed for expression of Cbf α -1 and ALP. Blockade of ERK1/2 phosphorylation in BMSCs by PD98059 suppressed osteogenic differentiation on chemical surfaces. These findings indicate a potential role for ERK in the osteogenic differentiation of BMSCs on surfaces modified by specific chemical functional groups, indicating that the microenvironment affects the differentiation of BMSCs. This observation has important implications for bone tissue engineering.
    08/2013; 2013(5411):361906. DOI:10.1155/2013/361906
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    • "In a follow-up study, Biggs et al. investigated the effect nanotopographies have on the ERK/MAPK signalling pathway for STRO-1+ human MSCs [37]. The signalling cascade, extracellular signal-regulated kinase (ERK), is a known member of the MAPK pathway, shown to play a significant role in MSC differentiation [38] [39] [40]. Intricate signalling pathways, such as the ERK/MAPK pathway, play a vital role in the cellular differentiation of MSCs through translating tensions applied at the tissue level to cellular functions. "
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    ABSTRACT: Stem cells continue to receive widespread attention due to their potential to revolutionise treatments in the fields of both tissue engineering and regenerative medicine. Adult stem cells, specifically mesenchymal stromal cells (MSCs), play a vital role in the natural events surrounding bone healing and osseointegration through being stimulated to differentiate along their osteogenic lineage and in doing so, they form new cortical and trabecular bone tissue. Understanding how to control, manipulate, and enhance the intrinsic healing events modulated through osteogenic differentiation of MSCs by the use of modified surfaces and biomaterials could potentially advance the fields of both orthopaedics and dentistry. This could be by either using surface modification to generate greater implant stability and more rapid healing following implantation or the stimulation of MSCs ex vivo for reimplantation. This review aims to gather publications targeted at promoting, enhancing, and controlling the osteogenic differentiation of MSCs through biomaterials, nanotopographies, and modified surfaces for use in implant procedures.
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