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Tunable stiffness of graphene oxide/polyacrylamide composite scaffolds regulates cytoskeleton assembly.

ABSTRACT: The stiffness of the extracellular matrix (ECM) not only provides mechanical resistance to support the cellular shape, but also plays significant roles in many cell functions. However, it's difficult to utilize traditional substrate materials to investigate cell behaviors under physical microenvironments due to their unphysiological stiffness or intrinsic secondary effects. Herein, a stiffness-tunable graphene oxide/polyacrylamide composite scaffold was fabricated to investigate the effect of substrate stiffness on cytoskeleton assembly and specific gene expression during cell growth. In the composite structure, the polyacrylamide (PAAm) hydrogel plays an exceptional role in controlling the substrate stiffness; in contrast, graphene oxide (GO) sheets not only provide permissive surfaces for cell adhesion and growth, but also effectively eliminate the secondary effects of the PAAm hydrogel. It's found that substrate stiffness could affect cell morphology and cytoskeleton assembly via specific genetic pathways. Therefore, the composite structure can be considered an attractive candidate as a scaffold and provides potential to elucidate the disease association of ECMs.

SUBMITTER: Sun Y

PROVIDER: S-EPMC6115675 | biostudies-other | 2018 Aug

REPOSITORIES: biostudies-other

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Tunable stiffness of graphene oxide/polyacrylamide composite scaffolds regulates cytoskeleton assembly.

Sun Yupeng Y Zhang Kaixiang K Deng Ruijie R Ren Xiaojun X Wu Can C Li Jinghong J

Chemical science 20180702 31

The stiffness of the extracellular matrix (ECM) not only provides mechanical resistance to support the cellular shape, but also plays significant roles in many cell functions. However, it's difficult to utilize traditional substrate materials to investigate cell behaviors under physical microenvironments due to their unphysiological stiffness or intrinsic secondary effects. Herein, a stiffness-tunable graphene oxide/polyacrylamide composite scaffold was fabricated to investigate the effect of su ...[more]

PMID: 30310582

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Project description:The precise mechanisms that lead to orthopedic implant failure are not well understood; it is believed that the micromechanical environment at the bone-implant interface regulates structural stability of an implant. In this work, we seek to understand how the 3D mechanical environment of an implant affects bone formation during early osteointegration. We employed two-photon lithography (TPL) direct laser writing to fabricate 3-dimensional rigid polymer scaffolds with tetrakaidecahedral periodic geometry, herewith referred to as nanolattices, whose strut dimensions were on the same order as osteoblasts' focal adhesions (?2?m) and pore sizes on the order of cell size, ?10?m. Some of these nanolattices were subsequently coated with thin conformal layers of Ti or W, and a final outer layer of 18nm-thick TiO2 was deposited on all samples to ensure biocompatibility. Nanomechanical experiments on each type of nanolattice revealed the range of stiffnesses of 0.7-100MPa. Osteoblast-like cells (SAOS-2) were seeded on each nanolattice, and their mechanosensitve response was explored by tracking mineral secretions and intracellular f-actin and vinculin concentrations after 2, 8 and 12days of cell culture in mineralization media. Experiments revealed that the most compliant nanolattices had ?20% more intracellular f-actin and ?40% more Ca and P secreted onto them than the stiffer nanolattices, where such cellular response was virtually indistinguishable. We constructed a simple phenomenological model that appears to capture the observed relation between scaffold stiffness and f-actin concentration. This model predicts a range of optimal scaffold stiffnesses for maximum f-actin concentration, which appears to be directly correlated with osteoblast-driven mineral deposition. This work suggests that three-dimensional scaffolds with titania-coated surfaces may provide an optimal microenvironment for cell growth when their stiffness is similar to that of cartilage (?0.5-3MPa). These findings help provide a greater understanding of osteoblast mechanosensitivity and may have profound implications in developing more effective and safer bone prostheses. STATEMENT OF SIGNIFICANCE:Creating prostheses that lead to optimal bone remodeling has been a challenge for more than two decades because of a lack of thorough knowledge of cell behavior in three-dimensional (3D) environments. Literature has shown that 2D substrate stiffness plays a significant role in determining cell behavior, however, limitations in fabrication techniques and difficulties in characterizing cell-scaffold interactions have limited our understanding of how 3D scaffolds' stiffness affects cell response. The present study shows that scaffold structural stiffness affects osteoblasts cellular response. Specifically this work shows that the cells grown on the most compliant nanolattices with a stiffness of 0.7MPa expressed ?20% higher concentration of intracellular f-actin and secreted ?40% more Ca and P compared with all other nanolattices. This suggests that bone scaffolds with a stiffness close to that of cartilage may serve as optimal 3D scaffolds for new synthetic bone graft materials.

Dataset Information

Tunable stiffness of graphene oxide/polyacrylamide composite scaffolds regulates cytoskeleton assembly.

Publications

Tunable stiffness of graphene oxide/polyacrylamide composite scaffolds regulates cytoskeleton assembly.

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