Nanostructured interfacial self-assembled peptide–polymer membranes for enhanced mineralization and cell adhesion

Abstract

Soft interfacial materials, such as self-assembled polymer membranes, are gaining increasing interest as biomaterials since they can provide selective barriers and/or controlled affinity interactions important to regulate cellular processes. Herein, we report the design and fabrication of multiscale structured membranes integrating selective molecular functionalities for potential applications in bone regeneration. The membranes were obtained by interfacial self-assembly of miscible aqueous solutions of hyaluronan and multi-domain peptides (MDPs) incorporating distinct biochemical motifs, including mineralizing (EE), integrin-binding (RGDS) and osteogenic (YGFGG) peptide sequences. Circular dichroism and Fourier transform infrared spectroscopy analyses of the MDPs revealed a predominant β-sheet conformation, while transmission electron microscopy (TEM) showed the formation of fibre-like nanostructures with different lengths. Scanning electron microscopy (SEM) of the membranes showed an anisotropic structure and surfaces with different nanotopographies, reflecting the morphological differences observed under TEM. All the membranes were able to promote the deposition of a calcium-phosphate mineral on their surface when incubated in a mineralizing solution. The ability of the MDPs, coated on coverslips or presented within the membranes, to support cell adhesion was investigated using primary adult periosteum-derived cells (PDCs) under serum-free conditions. Cells on the membranes lacking RGDS remained round, while in the presence of RGDS they appear to be more elongated and anchored to the membrane. These observations were confirmed by SEM analysis that showed cells attached to the membrane and exhibiting an extended morphology with close interactions with the membrane surface. We anticipate that these molecularly designed interfacial membranes can both provide relevant biochemical signals and structural biomimetic components for stem cell growth and differentiation and ultimately promote bone regeneration.

Supramolecular Peptide/Polymer Hybrid Hydrogels for Biomedical Applications

“…Several studies have reported the co‐assembly of peptide amphiphiles with polysaccharides of opposite charge (heparin, hyaluronic acid, alginate, carrageenan), which resulted in the formation of diverse type of biomaterials such as sacs, membranes, microcapsules, and microparticles . MDPs have been also self‐assembled with negatively charged biopolymers such as heparin to form hydrogels or with hyaluronic acid (HA) to create patterned and periosteum‐like membranes. From these studies, it is clear that PAs and MDPs form gels in the presence of heparin, while in the presence of other polysaccharides the dominant macroscopic structures are sacs, membranes, or capsules that are formed by interfacial self‐assembly.…”

Sequence length dependence in arginine/phenylalanine oligopeptides: Implications for self-assembly and cytotoxicity

“…[2,10] Interaction of cell membranes with -sheet nanofibers is a topic research field due to the high potential of peptide-drug amphiphiles for intracell delivery and appearance of these structures in a variety of biological processes such as cell adhesion, signaling and cytotoxicity. [11][12][13][14][15][16][17][18] Newcomb et al [11] investigated cytotoxicity onto surfaces coated with nanofibers obtained from two kinds of surfactant-like peptides; namely, C16H31O-A3G3K3 and C16H31O-V3A3K3 peptide amphiphiles (PAs). These PAs are characterized by cationic head groups (lysines) separated from alkyl tails by amino acid spacers able to drive the formation of -sheet in the resulting nanofibers.…”

Supramolecular Presentation of Hyaluronan onto Model Surfaces for Studying the Behavior of Cancer Stem Cells

“…[38] Pep-1 was capped at the N-terminal, either with an acetyl group ( Figure S2A, Supporting Information), by shaking twice for 20 min with 10% (v/v) acetic anhydride in DMF or with twofold excess of 3-(((4-methoxyphenyl) diphenylmethyl)thio)propanoic acid ( Figure S2B, Supporting Information) following the same coupling procedure as for the protected amino acids. [38] Pep-1 was capped at the N-terminal, either with an acetyl group ( Figure S2A, Supporting Information), by shaking twice for 20 min with 10% (v/v) acetic anhydride in DMF or with twofold excess of 3-(((4-methoxyphenyl) diphenylmethyl)thio)propanoic acid ( Figure S2B, Supporting Information) following the same coupling procedure as for the protected amino acids.…”