Journal: Advanced therapeutics
Article Title: Bio-orthogonal, Site-Selective Conjugation of Recombinant Proteins to Microporous Annealed Particle Hydrogels for Tissue Engineering
doi: 10.1002/adtp.201900148
Figure Lengend Snippet: A) Representative live/dead cell viability image of human mesenchymal stem cells (hMSCs) within MAP hydrogels at 5 days. Average cell viability within MAP hydrogels was 83% ± 6% (n = 3). Scale bar = 200 μm. B) Representative hMSC spreading images within MAP hydrogels at 5 days overlayed with bright field of microparticles. Nuclear DAPI stain is represented in blue and cytoskeletal F-actin stain is represented in red. Average hMSC cell volume within MAP hydrogels was 11,200 μm3 ± 2,685 μm3 (n = 3). Scale bar = 200 μm. C) Schematic showing Tz-BMP2 functionalized MAP hydrogels with encapsulated hMSCs for a mineralization study. D) Tz-BMP2 and NF-BMP2 (5 μg/mL) functionalized MAP hydrogels compared using a modified surface ELISA (n = 3). Absorbance values demonstrate differences in ELISA development between NF- and Tz-BMP2 MAP hydrogels indicating differenes in BMP2 conjugation. The symbol ** indicates statistical significance between groups (p < 0.01). E) Comparison of MAP hydrogel-bound calcium mineralization after 21 days (n = 6 for BMP2 samples, n = 3 for media controls). Absorbance values show that only Tz-BMP2 functionalized MAP hydrogels induce hMSC osteogenic mineralization. The symbol * indicates statistical significance between all other experimental groups (p < 0.05).
Article Snippet: Mouse monoclonal, anti-human BMP2 antibody (Abcam, 100 μL, 0.1 μg mL −1 ) was incubated within each well for 3 hrs at room temperature.
Techniques: Staining, Modification, Enzyme-linked Immunosorbent Assay, Conjugation Assay, Comparison
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