Table S1 from A sterilizable, biocompatible, tropoelastin surface coating immobilized by energetic ion activation

The fitting parameters for the curves shown in Fig. 1B

Molecular Orientation of Tropoelastin is Determined by Surface Hydrophobicity

Tropoelastin is the precursor of the extracellular protein elastin and is utilized in tissue engineering and implant technology by adapting the interface presented by surface-bound tropoelastin. The preferred orientation of the surface bound protein is relevant to biointerface interactions, as the C-terminus of tropoelastin is known to be a binding target for cells. Using recombinant human tropoelastin we monitored the binding of tropoelastin on hydrophilic silica and on silica made hydrophobic by depositing a self-assembled monolayer of octadecyl trichlorosilane. The layered organization of deposited tropoelastin was probed using neutron and X-ray reflectometry under aqueous and dried conditions. In a wet environment, tropoelastin retained a solution-like structure when adsorbed on silica but adopted a brush-like structure when on hydrophobized silica. The orientation of the surface-bound tropoelastin was investigated using cell binding assays and it was found that the C-terminus of tropoelastin faced the bulk solvent when bound to the hydrophobic surface, but a mixture of orientations was adopted when tropoelastin was bound to the hydrophilic surface. Drying the tropoelastin-coated surfaces irreversibly altered these protein structures for both hydrophilic and hydrophobic surfaces

Mechanism of EPC attachment to rhTE.

<p>(A) EPCs attached to 40 μg/ml rhTE in the presence of α-lactose, β-lactose, heparan sulfate, or EDTA. (B) Attachment of EPCs to 40 μg/ml rhTE in the presence of Ca²⁺, Mg²⁺, or Mn²⁺. (C) and (D) Inhibition of EPC attachment and spreading on 40 μg/ml rhTE using antibodies to integrins α₂β₁, α₅β₁, and α_vβ₃. Error bars represent S.E.M. of triplicate measurements.</p

Schematic diagram showing full-length rhTE and constructs N18, N25 and N10.

<p>Key domain features are indicated.</p

Mechanism of EPC attachment to truncated tropoelastin constructs.

<p>(A) and (B) EPCs attached to 40 μg/ml N25 and N18 respectively, in the presence of α-lactose, β-lactose, heparan sulfate or EDTA. (C) and (D) Attachment of EPCs to 40 μg/ml N25 and N18, respectively, in the presence of Ca²⁺, Mg²⁺, or Mn²⁺. (E) and (F), Inhibition of EPC spreading on 40 μg/ml N25 and N18 respectively, using antibody that inhibits binding to integrin α_vβ₃. Error bars represent S.E.M. of triplicate measurements.</p

OEC characterization by flow cytometry.

<p>A) Stained cells are shown as blue histograms, while unstained controls are shown in black. The percentage of positive cells is shown in the top right of each graph. The OECs are CD34/31/54/VEGFR2 positive and CD45/14 negative. B) Representative images of the binding of isothiocyanate-Ulex europaeus agglutinin I lectin binding (ULEX), uptake of acetylated low density lipoprotein (AcLDL) and staining for CD31 by EPCs (bottom row of panel) but not by fibroblasts (top row of panel). Together, these results are indicative of a positive endothelial cell phenotype.</p