<i>In vitro</i>
            generated extracellular matrix and fluid shear stress synergistically enhance 3D osteoblastic differentiation

Datta, Néha; Pham, Quynh P.; Sharma, Upma; Sikavitsas, Vassilios I.; Jansen, John A.; Mikos, Antonios G.

doi:10.1073/pnas.0505661103

Cited by 388 publications

(332 citation statements)

References 49 publications

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“…Collagen I comprises 90-95% of the organic matrix of bone, (Datta et al, 2006), is absolutely essential for bone formation (Plotkin, 2004), and associates tightly with collagen VI. Collagen VI is necessary for collagen I production in osteoblast like cells (Harumiya et al, 2002) and interlukin-4-dependent mineralization of human periosteal cells (Ishibashi et al, 1999).…”

Section: Discussionmentioning

confidence: 99%

Laminin-5 activates extracellular matrix production and osteogenic gene focusing in human mesenchymal stem cells

et al. 2007

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We recently reported that laminin-5, expressed by human mesenchymal stem cells (hMSC), stimulates osteogenic gene expression in these cells in the absence of any other osteogenic stimulus. Here we employ two dimensional liquid chromatography and tandem mass spectrometry, along with the Database for Annotation, Visualization and Integrated Discovery (DAVID), to obtain a more comprehensive profile of the protein (and hence gene) expression changes occurring during laminin-5-induced osteogenesis of hMSC. Specifically, we compare the protein expression profiles of undifferentiated hMSC, hMSC cultured on laminin-5 (Ln-5 hMSC), and fully differentiated human osteoblasts (hOST) with profiles from hMSC treated with well-established osteogenic stimuli (collagen I, vitronectin, or dexamethazone). We find a marked reduction in the number of proteins (e.g., those involved with calcium signaling and cellular metabolism) expressed in Ln-5 hMSC compared to hMSC, consistent with our previous finding that hOST express far fewer proteins than do their hMSC progenitors, a pattern we call "osteogenic gene focusing." This focused set, which resembles an intermediate stage between hMSC and mature hOST, mirrors the expression profiles of hMSC exposed to established osteogenic stimuli and includes osteogenic extracellular matrix proteins (collagen, vitronectin) and their integrin receptors, calcium signaling proteins, and enzymes involved in lipid metabolism. These results provide direct evidence that laminin-5 alone stimulates global changes in gene/protein expression in hMSC that lead to commitment of these cells to the osteogenic phenotype, and that this commitment correlates with extracellular matrix production.

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Section: Discussionmentioning

confidence: 99%

Laminin-5 activates extracellular matrix production and osteogenic gene focusing in human mesenchymal stem cells

et al. 2007

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“…About 100 mL of the rehydrated sample was then combined with 50 mL chloramine-T (Sigma) and 50 mL p-dimethylaminobenzaldehyde (Sigma) and incubated at 60°C for 30 min, as described previously. 43,44 Absorbance was measured at 570 nm in a plate reader with samples run in duplicate alongside hydroxyproline standards of known concentration. Absorbance correlated to the hydroxyproline content was then calculated as the amount of collagen per scaffold by correcting for dilution and scaffold pooling and then converting the amount of hydroxyproline per scaffold to collagen content using a previously reported 1:10 hydroxyproline to collagen ratio.…”

Section: Biochemical Assaysmentioning

confidence: 99%

Cell-Derived Polymer/Extracellular Matrix Composite Scaffolds for Cartilage Regeneration, Part 1: Investigation of Cocultures and Seeding Densities for Improved Extracellular Matrix Deposition

Levorson

Mountziaris

et al. 2014

Tissue Engineering Part C: Methods

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This study investigated the coculture of chondrocytes and mesenchymal stem cells (MSCs) on electrospun fibrous polymer scaffolds to produce polymer/extracellular matrix (ECM) hybrid constructs with the objective of reducing the number of chondrocytes necessary to produce ample cartilage-like ECM within the scaffolds. To generate these hybrid constructs, electrospun poly(e-caprolactone) fibrous scaffolds were seeded at both high and low initial densities with five different ratios of chondrocytes to MSCs: 1:0, 1:1, 1:3, 1:5, and 0:1, and cultured for 7, 14, and 21 days. Glycosaminoglycan production and distribution within the three coculture groups was similar to quantities generated by chondrocyte-only controls. Conversely, as the concentration of chondrocytes was increased, the collagen content of the constructs also increased at each time point, with a 1:1 chondrocyte to MSC ratio approximating the collagen production of chondrocytes alone. Histological staining suggested that cocultured constructs mimicked the well-distributed ECM patterns of chondrocyte generated constructs, while improving greatly over the restricted distribution of matrix within MSC-only constructs. These results support the capacity of cocultures of chondrocytes and MSCs to generate cartilaginous matrix within a polymeric scaffold. Further, the inclusion of MSCs in these cocultures enables the reduction of chondrocytes needed to produce cell-generated ECM.

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“…Mechanical forces have been imparted to multipotent cells using spinner flask [Meinel et al 2004], perfused cartridge [Meinel et al 2004], parallel flow chambers [Knippenberg et al 2005, Datta et al 2006, flexible membrane [Akimoto et al 2005, Jakkaraju et al 2005, Baksh et al 2006, David et al 2007], compressive gel [Takahashi et al 1998, Campbell et al 2006] as well as hydrostatic pressure based systems [Miyanishi et al 2006] (Table 1). Furthermore, a recent study maps the range of mechanical signals, time scale of mechanical stress application and resultant cell fates (Figure 7) [Anderson and Knothe Tate 2007a] from studies published to date.…”

Section: In Vitro Experimental Platforms To Elucidate Role Of Extrinsmentioning

confidence: 99%

Mechanical modulation of osteochondroprogenitor cell fate

Tate

Falls

McBride

et al. 2008

The International Journal of Biochemistry & Cell Biology

101

102

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Mesenchymal cells are natural tissue builders. They exhibit an extraordinary capacity to metamorphize into differentiated cells, using extrinsic spatial and temporal inputs and intrinsic algorithms, as well as to build and adapt their own habitat. In addition to providing a habitat for osteoprogenitor cells, tissues of the skeletal system provide mechanical support and protection for the multiple organs of vertebrate organisms. This review examines the role of mechanics on determination of cell fate during pre-, peri-and postnatal development of the skeleton as well as during tissue genesis and repair in postnatal life. The role of cell mechanics is examined and brought into context of intrinsic cues during mesenchymal condensation. Remarkable new insights regarding structure function relationships in mesenchymal stem cells, and their influence on determination of cell fate are integrated in the context of de novo tissue generation and postnatal repair. Key differences in the formation of osteogenic and chondrogenic condensations are discussed in relation to direct intramembranous and indirect endochondral ossification. New approaches are discussed to elucidate and exploit extrinsic cues to generate tissues in the laboratory and in the clinic.

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In vitro generated extracellular matrix and fluid shear stress synergistically enhance 3D osteoblastic differentiation

Cited by 388 publications

References 49 publications

Laminin-5 activates extracellular matrix production and osteogenic gene focusing in human mesenchymal stem cells

Laminin-5 activates extracellular matrix production and osteogenic gene focusing in human mesenchymal stem cells

Cell-Derived Polymer/Extracellular Matrix Composite Scaffolds for Cartilage Regeneration, Part 1: Investigation of Cocultures and Seeding Densities for Improved Extracellular Matrix Deposition

Mechanical modulation of osteochondroprogenitor cell fate

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