The role of RhoA kinase (ROCK) in cell alignment on nanofibers

Andalib, Mohammad Nahid; Lee, Jeong Soon; Ha, Ligyeom; Dzenis, Yuris A.; Lim, Jung Yul

doi:10.1016/j.actbio.2013.04.013

Cited by 25 publications

(22 citation statements)

References 32 publications

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“…Vector control was produced by repeating the procedure but with green fluorescent protein (GFP) control plasmid (Santa Cruz Biotechnology, sc-108083). Silencing of FAK and ROCK was confirmed by RT-PCR and immunoblotting [18,19]. Examples of Western blotting data for silenced FAK and ROCK are shown in the electronic supplementary material, figure S14.…”

Section: Small Hairpin Rnamentioning

confidence: 99%

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Fluid-flow-induced mesenchymal stem cell migration: role of focal adhesion kinase and RhoA kinase sensors

Riehl

Lee

et al. 2015

J. R. Soc. Interface.

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The study of mesenchymal stem cell (MSC) migration under flow conditions with investigation of the underlying molecular mechanism could lead to a better understanding and outcome in stem-cell-based cell therapy and regenerative medicine. We used peer-reviewed open source software to develop methods for efficiently and accurately tracking, measuring and processing cell migration as well as morphology. Using these tools, we investigated MSC migration under flow-induced shear and tested the molecular mechanism with stable knockdown of focal adhesion kinase (FAK) and RhoA kinase (ROCK). Under steady flow, MSCs migrated following the flow direction in a shear stress magnitude-dependent manner, as assessed by root mean square displacement and mean square displacement, motility coefficient and confinement ratio. Silencing FAK in MSCs suppressed morphology adaptation capability and reduced cellular motility for both static and flow conditions. Interestingly, ROCK silencing significantly increased migration tendency especially under flow. Blocking ROCK, which is known to reduce cytoskeletal tension, may lower the resistance to skeletal remodelling during the flowinduced migration. Our data thus propose a potentially differential role of focal adhesion and cytoskeletal tension signalling elements in MSC migration under flow shear.

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Section: Small Hairpin Rnamentioning

confidence: 99%

“…Establishing MSCs with stable FAK or ROCK knockdown has been previously reported [18,19]. Briefly, murine C3H10T1/2 MSCs (ATCC, CCL-226) were transfected for 24 h with shRNA-FAK (sc-35353-SH) or shRNA-ROCK-I (sc-36432-SH), both from Santa Cruz Biotechnology, using lipofectamine 2000 (Invitrogen) transfection reagent.…”

Section: Small Hairpin Rnamentioning

confidence: 99%

Fluid-flow-induced mesenchymal stem cell migration: role of focal adhesion kinase and RhoA kinase sensors

Riehl

Lee

et al. 2015

J. R. Soc. Interface.

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“…The organization of the cell cytoskeleton actively participate in the ability of cells to sense and convert mechanical cues into biological responses. It cells display highly elongated cell morphologies when growing on aligned nanofibers and spread cell morphologies when growing on randomly distributed nanofibers [37,65]. These phenomena might be associated with the spatial distribution of the focal adhesion formation of the attached cells which arises from the "communication" between cells and diversely arranged nanofiber scaffolds.…”

Section: The Role Of Focal Adhesion Formation and Cellular Cytoskeletmentioning

confidence: 99%

“…Cells on fiber networks, developed longer and more concentrated focal adhesion clusters compared with cells on flat control substrates [69]. In addition, the highly elongated cell morphology also means the greater cytoskeletal tension, and relevant signaling such as ROCK may be up-regulated in cells on aligned nanofibers [65].…”

Section: The Role Of Focal Adhesion Formation and Cellular Cytoskeletmentioning

confidence: 99%

“…For example, the fiber diameters of electrospun nanofibers are in the range about 100 nm. The mineralized type I collagen fibrils, constituting $90 % of the bone structure, are nano-sized (50-500 nm in diameter) [65], and may thus be well mimicked by the synthetic nanofibers. In previous studies, it was demonstrated that the nanometer effects of nanofibrous scaffolds could induce up-regulated focal adhesion kinase signaling and increased cellular elastic modulus for osteoblastic cells and enhanced fate direction into osteogenesis for hBMSCs.…”

Section: Mechanisms Of Electrospun Nanofibrous Scaffolds-induced Cellmentioning

confidence: 99%

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Biological Events Occuring on the Biosis–Abiosis Interface: Cellular Responses Induced by Implantable Electrospun Nanofibrous Scaffolds

Deng

Wei

Zhang

et al. 2015

Interface Oral Health Science 2014

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Electrospun nanofibers have tremendous potential as novel scaffolds for tissue engineering of various soft and hard tissues because of thier high surface area, surface functional groups, interconnected pores, and nano-scaled size. In this chapter, we reviewed the types of the nanofibrous scaffolds that have been used as implantable biomedical devices and used electrospun nanofibrous guided tissue regeneration membrane as an example to illustrate how the physiochemical properties of nanofibrous scaffolds influenced the biological events occuring on the scaffolds-host interface. It could be concluded that physical and chemical stimuli caused by nanofibrous scaffold would support in vivo-like three-dimensional cell adhesion and activate cell-signaling pathways. In terms of physical stimulus, the process of mechanotransduction may play an important role in influencing cellular behaviors. A result, biological events such as cell-interface recognition, cell proliferation, and cell differentiation are altered. Nevertheless, the cellular and molecular mechanisms by which cells sense and respond to nanofiberous scaffolds remain poorly understood. More evidences are needed to reveal the underlying mechanisms whereby environmental cues alternated the cellular responses to the

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Structural and Biochemical Modification of a Collagen Scaffold to Selectively Enhance MSC Tenogenic, Chondrogenic, and Osteogenic Differentiation

Caliari

Harley

2014

Adv Healthcare Materials

102

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Biomaterial approaches for engineering orthopedic interfaces such as the tendon-bone junction (TBJ) are limited by a lack of understanding of how insoluble (microstructure, composition) and soluble regulators of stem cell fate work in concert to promote bioactivity and differentiation. One strategy for regenerating the interface is to design biomaterials containing spatially-graded structural properties sufficient to induce divergent mesenchymal stem cell (MSC) differentiation into multiple interface-specific phenotypes. This work explores the hypothesis that selective structural modification to a 3D collagen-glycosaminoglycan (CG) scaffold combined with biochemical supplementation can drive human bone marrow-derived MSC differentiation down tenogenic, osteogenic, and chondrogenic lineages. Tenogenic differentiation is enhanced in geometrically anisotropic scaffolds versus a standard isotropic control. Notably, blebbistatin treatment abrogates this microstructurally-driven effect. Further, enhanced osteogenic differentiation and new mineral synthesis is achieved by incorporation of a calcium phosphate mineral phase within the CG scaffold along with the use of osteogenic induction media. Finally, chondrogenic differentiation is optimally driven by combining chondrogenic induction media with a reduced density scaffold that promotes increased cellular condensation, significantly higher expression of chondrogenic genes, and increased GAG deposition. Together these data provide critical insight regarding design rules for elements of an integrated biomaterial platform for orthopedic interface regeneration.

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The role of RhoA kinase (ROCK) in cell alignment on nanofibers

Cited by 25 publications

References 32 publications

Fluid-flow-induced mesenchymal stem cell migration: role of focal adhesion kinase and RhoA kinase sensors

Fluid-flow-induced mesenchymal stem cell migration: role of focal adhesion kinase and RhoA kinase sensors

Biological Events Occuring on the Biosis–Abiosis Interface: Cellular Responses Induced by Implantable Electrospun Nanofibrous Scaffolds

Structural and Biochemical Modification of a Collagen Scaffold to Selectively Enhance MSC Tenogenic, Chondrogenic, and Osteogenic Differentiation

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