Abstract
The development of versatile bioactive surfaces able to emulate in vivo conditions is of enormous importance to the future of cell and tissue therapy. Tuning cell behaviour on two-dimensional surfaces so that the cells perform as if they were in a natural three-dimensional tissue represents a significant challenge, but one that must be met if the early promise of cell and tissue therapy is to be fully realised. Due to the inherent complexities involved in the manufacture of biomimetic three-dimensional substrates, the scaling up of engineered tissue-based therapies may be simpler if based upon proven two-dimensional culture systems. In this work, we developed new coating materials composed of the self-assembling peptide amphiphiles (PAs) C16G3RGD (RGD) and C16G3RGDS (RGDS) shown to control cell adhesion and tissue architecture while avoiding the use of serum. When mixed with the C16ETTES diluent PA at 13 : 87 (mol mol-1) ratio at 1.25 × 10-3 M, the bioactive PAs were shown to support optimal adhesion, maximal proliferation, and prolonged viability of human corneal stromal fibroblasts (hCSFs), while improving the cell phenotype. These PAs also provided stable adhesive coatings on highly-hydrophobic surfaces composed of striated polytetrafluoroethylene (PTFE), significantly enhancing proliferation of aligned cells and increasing the complexity of the produced tissue. The thickness and structure of this highly-organised tissue were similar to those observed in vivo, comprising aligned newly-deposited extracellular matrix. As such, the developed coatings can constitute a versatile biomaterial for applications in cell biology, tissue engineering, and regenerative medicine requiring serum-free conditions.
Highlights
It is well understood that the three-dimensional structure of tissues is essential for the maintenance of cellular function and the development of physiologically relevant structures, and that the extracellular matrix (ECM) provides the structural and organizational guides for this tissue development.[1]
We developed new coating materials composed of the self-assembling peptide amphiphiles (PAs) C16G3RGD (RGD) and C16G3RGDS (RGDS) shown to control cell adhesion and tissue architecture while avoiding the use of serum
(corresponding to the expression of ACTA2) or protein levels. These results indicated that the RGDS:ETTES coating increased cell adhesion and proliferation, and enhanced the molecular and morphological phenotypes characteristic of human corneal stromal fibroblasts (hCSFs) grown in serum-free conditions for long periods in culture
Summary
It is well understood that the three-dimensional structure of tissues is essential for the maintenance of cellular function and the development of physiologically relevant structures, and that the extracellular matrix (ECM) provides the structural and organizational guides for this tissue development.[1]. Unlike cultures in three-dimensional (3D) matrices, cells on at, non-functionalized surfaces do not exhibit native phenotypes, which affects their ability to form higher order structures.[2,3] a biomaterial through which one can translate sophisticated two-dimensional (2D) experimental design into complex cell-derived 3D structures is of great value to both basic scientists and industry researchers Such an approach may advance the understanding of tissue biology and disease progression as well as offer a solution to the existing paradox that cells can be grown quickly and on 2D substrates but lack the necessary hierarchical structure and phenotype to be used therapeutically as tissue engineered constructs. Synthetic 2D cell support surfaces have been developed that contain components from, or inspired by, the biological ECM.[1,4] The tripeptide Arg-Gly-Asp is the
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