Journal: ACS Applied Materials & Interfaces
Article Title: Hybrid Micro-/Nanoprotein Platform Provides Endocrine-like and Extracellular Matrix-like Cell Delivery of Growth Factors
doi: 10.1021/acsami.4c01210
Figure Lengend Snippet: MSCs differentiation in cultures over artificial hFGF2-H6 SGs PEA–FN surfaces. (A) Imaging of MSCs by fluorescence microscopy in the presence of artificial hFGF2-H6 SGs at 50 ng/mL for 14 days. Actin, vinculin, vimentin, and YAP were selected as cell markers. Merge refers to actin and vinculin combined fluorescence signals. White squares highlight cell focal adhesions. Close-up pictures of focal adhesions are displayed on the right panels. (B) Fold change on mRNA content (meaning RUNX2, OSN; osteonectin, and OPN; osteopontin gene expression) in MSCs upon incubation with soluble hFGF2-H6 (dark blue), artificial hFGF2-H6 SGs (pale blue), and free Zn 2+ (gray) at 50 ng/mL for 14 days. (C) In-cell Western (ICW) immunodetection of RUNX2, OSN, and OSP proteins in MSCs extracts upon incubation over soluble hFGF2-H6, artificial hFGF2-H6 SGs, and free Zn 2+ (gray) at 50 ng/mL for 14 days. The protein signal is displayed in green, and the cell signal is in red. (D) Statistical analysis of protein signal (green from panel C) expressed as fluorescence per cell and cm 2 in absorbance units). (E) Statistical analysis of cell signal (red from panel C) expressed as fluorescence per cm 2 in absorbance units (au). Peak numbers correspond to the increased percentage of cell growth comparing artificial hFGF2-H6 SGs (pale blue) with soluble hFGF2-H6 (dark blue), free Zn 2+ (gray), and control MSCs (black). Data are expressed as mean ± SEM, and statistical significance is achieved when p < 0.05 is represented as (*). Control refers to MSCs seeded on top of FN-PEA surfaces.
Article Snippet: Cells were incubated with monoclonal primary Abs (1:200) in blocking buffer (PBS/1% milk protein) at room temperature for 2.5 h, respectively; Runx2 (Santa Cruz Biotechnology, C1319), osteonectin (Santa Cruz Biotechnology, SC398419), and osteopontin (Santa Cruz Biotechnology, B1218).
Techniques: Imaging, Fluorescence, Microscopy, Gene Expression, Incubation, In-Cell ELISA, Immunodetection, Control
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