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Novel cationic tannin/glycosaminoglycan-based polyelectrolyte multilayers promote stem cells adhesion and proliferation.

ABSTRACT: Modifying materials with biocompatible surface coatings is an important method for controlling cell responses to biomaterials. In this work, tanfloc (TN), a cationic tannin-derivative polymer was assembled with heparin (HEP) and chondroitin sulfate (CS), using the layer-by-layer (LbL) approach, to build polyelectrolyte multilayers (PEMs) and to design cytocompatible coatings. LbL deposition was monitored through Fourier-transform surface plasmon resonance, and characterized by X-ray photoelectron spectroscopy, atomic force microscopy, and contact angle measurements. The response of human adipose-derived stem cells (ADSCs) was evaluated in vitro. All of the TN-containing PEMs exhibit cytocompatibility and support adhesion, proliferation, and the spreading of ADSCs after 7 days of culture. HEP-TN PEM assembly with 11 layers (HEP-TN₁₁) supports the greatest rate of cell proliferation. When TN is the terminal layer of the PEM, the surfaces promote the spreading of ADSCs, indicating that the surface charge and PEM terminal layer are key determinants of the microenvironmental niche that control cellular response. The promotion of stem cell attachment and proliferation makes these surface coatings potentially useful for biomedical implants and regenerative medicine.

SUBMITTER: da Camara PCF

PROVIDER: S-EPMC9070077 | biostudies-literature | 2019 Aug

REPOSITORIES: biostudies-literature

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Novel cationic tannin/glycosaminoglycan-based polyelectrolyte multilayers promote stem cells adhesion and proliferation.

da Câmara Paulo C F PCF Balaban Rosangela C RC Hedayati Mohammadhasan M Popat Ketul C KC Martins Alessandro F AF Kipper Matt J MJ

RSC advances 20190819 44

Modifying materials with biocompatible surface coatings is an important method for controlling cell responses to biomaterials. In this work, tanfloc (TN), a cationic tannin-derivative polymer was assembled with heparin (HEP) and chondroitin sulfate (CS), using the layer-by-layer (LbL) approach, to build polyelectrolyte multilayers (PEMs) and to design cytocompatible coatings. LbL deposition was monitored through Fourier-transform surface plasmon resonance, and characterized by X-ray photoelectro ...[more]

PMID: 35530064

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Project description:Polyelectrolyte multilayers (PEMUs) are tunable thin films that could serve as coatings for biomedical implants. PEMUs built layer by layer with the polyanion poly(acrylic acid) (PAA) modified with a photosensitive 4-(2-hydroxyethoxy) benzophenone (PAABp) group and the polycation poly(allylamine hydrochloride) (PAH) are mechanically tunable by UV irradiation, which forms covalent bonds between the layers and increases PEMU stiffness. PAH-terminated PEMUs (PAH-PEMUs) that were uncrosslinked, UV-crosslinked to a uniform stiffness, or UV-crosslinked with an edge mask or through a neutral density optical gradient filter to form continuous compliance gradients were used to investigate how differences in PEMU stiffness affect the adhesion and migration of epithelial cell sheets from scales of the fish Poecilia sphenops (Black Molly) and Carassius auratus (Comet Goldfish). During the progressive collective cell migration, the edge cells (also known as 'leader' cells) in the sheets on softer uncrosslinked PEMUs and less crosslinked regions of the gradient formed more actin filaments and vinculin-containing adherens junctions and focal adhesions than formed in the sheet cells on stiffer PEMUs or glass. During sheet migration, the ratio of edge cell to internal cell (also known as 'follower' cells) motilities were greater on the softer PEMUs than on the stiffer PEMUs or glass, causing tension to develop across the sheet and periods of retraction, during which the edge cells lost adhesion to the substrate and regions of the sheet retracted toward the more adherent internal cell region. These retraction events were inhibited by the myosin II inhibitor Blebbistatin, which reduced the motility velocity ratios to those for sheets on the stiffer PEMUs. Blebbistatin also caused disassembly of actin filaments, reorganization of focal adhesions, increased cell spreading at the leading edge, as well as loss of edge cell-cell connections in epithelial cell sheets on all surfaces. Interestingly, cells throughout the interior region of the sheets on uncrosslinked PEMUs retained their actin and vinculin organization at adherens junctions after treatment with Blebbistatin. Like Blebbistatin, a Rho-kinase (ROCK) inhibitor, Y27632, promoted loss of cell-cell connections between edge cells, whereas a Rac1 inhibitor, NSC23766, primarily altered the lamellipodial protrusion in edge cells. Compliance gradient PAH-PEMUs promoted durotaxis of the cell sheets but not of individual keratocytes, demonstrating durotaxis, like plithotaxis, is an emergent property of cell sheet organization.

Project description:We report an approach to the design of multilayered polyelectrolyte thin films (or 'polyelectrolyte multilayers', PEMs) that can be used to provide tunable control over the release of plasmid DNA (or multiple different DNA constructs) from film-coated surfaces. Our approach is based upon methods for the layer-by-layer assembly of DNA-containing thin films, and exploits the properties of a new class of cationic 'charge-shifting' polymers (amine functionalized polymers that undergo gradual changes in net charge upon side chain ester hydrolysis) to provide control over the rates at which these films erode and release DNA. We synthesized two 'charge-shifting' polymers (polymers 1 and 2) containing different side chain structures by ring-opening reactions of poly(2-alkenyl azlactone)s with two different tertiary amine functionalized alcohols (3-dimethylamino-1-propanol and 2-dimethylaminoethanol, respectively). Subsequent characterization revealed large changes in the rates of side chain ester hydrolysis for these two polymers; whereas the half-life for the hydrolysis of the esters in polymer 1 was ~200 days, the half-life for polymer 2 was ~6 days. We demonstrate that these large differences in side chain hydrolysis make possible the design of PEMs that erode and promote the surface-mediated release of DNA either rapidly (e.g., over ~3 days for films fabricated using polymer 2) or slowly (e.g., over ~1 month for films fabricated using polymer 1). We demonstrate further that it is possible to design films with release profiles that are intermediate to these two extremes by fabricating films using solutions containing different mixtures of these two polymers. This approach can thus expand the usefulness of these two polymers and achieve a broader range of DNA release profiles without the need to synthesize polymers with new structures or properties. Finally, we demonstrate that polymers 1 and 2 can be used to fabricate multilayered films with hierarchical structures that promote the sequential release of two different DNA constructs with separate and distinct release profiles (e.g., the release of a first construct over a period of ~3 days, followed by the sustained release of a second for a period of ~70 days). With further development, this approach could contribute to the design of functional thin films and surface coatings that provide sophisticated control over the timing and the order of the release of two or more DNA constructs (or other agents) of interest in a range of biomedical contexts.

Dataset Information

Novel cationic tannin/glycosaminoglycan-based polyelectrolyte multilayers promote stem cells adhesion and proliferation.

Publications

Novel cationic tannin/glycosaminoglycan-based polyelectrolyte multilayers promote stem cells adhesion and proliferation.

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