Contribution of Fibroblasts Cultured on 3D Silica Nonwoven Fabrics to Cocultured Hepatocytes Function

Otsuka, Hidenori; Okimura, Saya; Nagamura, Masako; Watanabe, Rie; Kawabe, Mayumi

doi:10.1246/cl.130955

Cited by 6 publications

(26 citation statements)

References 20 publications

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“…In the present study, chondrogenic differentiation of human bone marrow-derived MSCs in 3D silica fabrics was investigated. For the chondrogenic differentiation of MSCs, we utilized 3D silica fabrics as previously reported, as it is a reasonable scaffold for tissue engineering [34][35][36]38]. Briefly, 3D silica fabrics were developed via the sol-gel process to create highly interconnected porous microstructures of electrospun fabrics for cell growth.…”

Section: Discussionmentioning

confidence: 99%

“…The random orientation of the silica fibers produces many interconnected pores that may promote cellular migration and tissue ingrowth and prevent shrinkage because of the sufficient mechanical strength of the fabrics [31,32]. As a result of these characteristics, fibroblasts migrated into and penetrated the 3D silica fabrics and showed remarkable growth rates compared with those in traditional 2D culture [33][34][35]. Moreover, the functions of hepatocytes co-cultured with fibroblasts in 3D silica fabrics were significantly enhanced compared with those of 2D fibroblasts because of the abundance of fibroblast-secreted soluble factors, which are important for maintaining hepatocytes [35].…”

Section: Introductionmentioning

confidence: 99%

“…As a result of these characteristics, fibroblasts migrated into and penetrated the 3D silica fabrics and showed remarkable growth rates compared with those in traditional 2D culture [33][34][35]. Moreover, the functions of hepatocytes co-cultured with fibroblasts in 3D silica fabrics were significantly enhanced compared with those of 2D fibroblasts because of the abundance of fibroblast-secreted soluble factors, which are important for maintaining hepatocytes [35]. We have recently demonstrated that osteogenic differentiation was significantly promoted in 3D silica fabrics.…”

Section: Introductionmentioning

confidence: 99%

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Cartilage Differentiation of Bone Marrow-Derived Mesenchymal Stem Cells in Three-Dimensional Silica Nonwoven Fabrics

et al. 2018

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In cartilage tissue engineering, three-dimensional (3D) scaffolds provide native extracellular matrix (ECM) environments that induce tissue ingrowth and ECM deposition for in vitro and in vivo tissue regeneration. In this report, we investigated 3D silica nonwoven fabrics (Cellbed®) as a scaffold for mesenchymal stem cells (MSCs) in cartilage tissue engineering applications. The unique, highly porous microstructure of 3D silica fabrics allows for immediate cell infiltration for tissue repair and orientation of cell–cell interaction. It is expected that the morphological similarity of silica fibers to that of fibrillar ECM contributes to the functionalization of cells. Human bone marrow-derived MSCs were cultured in 3D silica fabrics, and chondrogenic differentiation was induced by culture in chondrogenic differentiation medium. The characteristics of chondrogenic differentiation including cellular growth, ECM deposition of glycosaminoglycan and collagen, and gene expression were evaluated. Because of the highly interconnected network structure, stiffness, and permeability of the 3D silica fabrics, the level of chondrogenesis observed in MSCs seeded within was comparable to that observed in MSCs maintained on atelocollagen gels, which are widely used to study the chondrogenesis of MSCs in vitro and in vivo. These results indicated that 3D silica nonwoven fabrics are a promising scaffold for the regeneration of articular cartilage defects using MSCs, showing the particular importance of high elasticity.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Cartilage Differentiation of Bone Marrow-Derived Mesenchymal Stem Cells in Three-Dimensional Silica Nonwoven Fabrics

et al. 2018

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“…26 Furthermore, the functions of hepatocytes cocultured with the fibroblast in SNFs were significantly higher than those cocultured with the 2D fibroblast because of the abundance of fibroblast-secreted soluble factors, which is important for maintaining the function of hepatocytes. 26…”

Section: Introductionmentioning

confidence: 99%

“…SNFs have been applied for the construction of 3D tumor models 25 and a coculture system. 26 Cancer cells cultured in SNFs showed poorer response to anticancer agents by the upregulation of genes such as BCL2 , and potentials of SNFs for the construction of 3D tumor models in vitro were suggested. 25 We have clearly demonstrated the utility for a tissue-engineered construct: a fibroblast migrated and penetrated in SNFs and showed remarkable growth rates compared to the traditional 2D culture.…”

Section: Introductionmentioning

confidence: 99%

Osteogenic Differentiation of Bone Marrow-Derived Mesenchymal Stem Cells in Electrospun Silica Nonwoven Fabrics

et al. 2018

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Silica nonwoven fabrics (SNFs) with enough mechanical strength are candidates as implantable scaffolds. Culture of cells therein is expected to affect the proliferation and differentiation of the cells through cell–cell and cell–SNF interactions. In this study, we examined three-dimensional (3D) SNFs as a scaffold of mesenchymal stem cells (MSCs) for bone tissue engineering applications. The interconnected highly porous microstructure of 3D SNFs is expected to allow omnidirectional cell–cell interactions, and the morphological similarity of a silica nanofiber to that of a fibrous extracellular matrix can contribute to the promotion of cell functions. 3D SNFs were prepared by the sol–gel process, and their mechanical properties were characterized by the compression test and rheological analysis. In the compression test, SNFs showed a compressive elastic modulus of over 1 MPa and a compressive strength of about 200 kPa. These values are higher than those of porous polystyrene disks used for in vitro 3D cell culture. In rheological analysis, the elastic modulus and fracture stress were 3.27 ± 0.54 kPa and 25.9 ± 8.3 Pa, respectively. Then, human bone marrow-derived MSCs were cultured on SNFs, and the effects on proliferation and osteogenic differentiation were evaluated. The MSCs seeded on SNF proliferated, and the thickness of the cell layer became over 80 μm after 14 days of culture. The osteogenic differentiation of MSCs on SNFs was induced by the culture in the commercial osteogenic differentiation medium. The alkaline phosphatase activity of MSCs on SNFs increased rapidly and remained high up to 14 days and was much higher than that on two-dimensional tissue culture-treated polystyrene. The high expression of RUNX2 and intense staining by alizarin red s after differentiation supported that SNFs enhanced the osteogenic differentiation of MSCs. Furthermore, permeation analysis of SNFs using fluorescein isothiocyanate-dextran suggested a sufficient permeability of SNFs for oxygen, minerals, nutrients, and secretions, which is important for maintaining the cell viability and vitality. These results suggested that SNFs are promising scaffolds for the regeneration of bone defects using MSCs, originated from highly porous and elastic SNF characters.

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Three‐dimensional co‐culture model employing silica nonwoven fabrics to enhance cell‐to‐cell communication of paracrine signaling between hepatocytes and fibroblasts

Ishikawa

Iijima

Kawabe³

et al. 2023

Biotech & Bioengineering

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The realization that soluble factors secreted by heterotypic cells play an importanta role in paracrine signaling, which facilitates intercellular communication, enabled the development of physiologically relevant co‐culture models for drug screening and the engineering of tissues, such as hepatic tissues. The most crucial issues confronting the use of conventional membrane inserts in segregated co‐culture models that are used to study paracrine signaling between heterotypic cells have been identified as long‐term viability and retention of cell‐specific functions, especially when isolated primary cells are used. Herein, we present an in vitro segregated co‐culture model consisting of a well plate incubated with rat primary hepatocytes and normal human dermal fibroblasts which were segregated using a membrane insert with silica nonwoven fabric (SNF) on it. SNF, which mimics a physiological environment much more effectively than a two‐dimensional (2D) one, promotes cell differentiation and resultant paracrine signaling in a manner that is not possible in a conventional 2D culture, owing to high mechanical strength generated by its inorganic materials and interconnected network structure. In segregated co‐cultures, SNF clearly enhanced the functions of hepatocytes and fibroblasts, thereby showing its potential as a measure of paracrine signaling. These results may advance the understanding of the role played by paracrine signaling in cell‐to‐cell communication and provide novel insights into the applications of drug metabolism, tissue repair, and regeneration.

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Contribution of Fibroblasts Cultured on 3D Silica Nonwoven Fabrics to Cocultured Hepatocytes Function

Cited by 6 publications

References 20 publications

Cartilage Differentiation of Bone Marrow-Derived Mesenchymal Stem Cells in Three-Dimensional Silica Nonwoven Fabrics

Cartilage Differentiation of Bone Marrow-Derived Mesenchymal Stem Cells in Three-Dimensional Silica Nonwoven Fabrics

Osteogenic Differentiation of Bone Marrow-Derived Mesenchymal Stem Cells in Electrospun Silica Nonwoven Fabrics

Three‐dimensional co‐culture model employing silica nonwoven fabrics to enhance cell‐to‐cell communication of paracrine signaling between hepatocytes and fibroblasts

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