Journal
ANNALS OF BIOMEDICAL ENGINEERING
Volume 44, Issue 6, Pages 1921-1930Publisher
SPRINGER
DOI: 10.1007/s10439-016-1622-6
Keywords
Hyaluronic acid; N-cadherin; Biomaterials; Tissue engineering
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Funding
- National Science Foundation
- National Institutes of Health [R01 EB008722, T32 AR007132]
- Department of Veterans Affairs [I01 RX000700]
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N-cadherin (Ncad) mediates cell-cell interactions, regulates beta-catenin (beta cat) signaling, and promotes the chondrogenic differentiation of mesenchymal lineage cells. Here, we utilized confocal imaging to investigate the influence of Ncad interactions on single mesenchymal stem cell (MSC) behavior within 3-dimensional hydrogel environments under conditions that promote chondrogenic differentiation. Human MSCs were photoencapsulated in hyaluronic acid hydrogels functionalized with Ncad mimetic peptides and compared to cells in environments with control non-active peptides (Ctrl). Using single-cell imaging, we observed a significant increase in membrane beta cat, nuclear beta cat, and cell roundness after 3 days in Ncad hydrogels compared to Ctrl hydrogels. The extent of membrane and nuclear beta cat localization and MSC roundness decreased to Ctrl hydrogel levels via pre-treatment with Ncad-specific antibodies prior to encapsulation in the Ncad hydrogels, confirming the activity of the peptide. Interestingly, there was a pronounced (> 80%) increase in beta cat nuclear localization in two-cell clusters within the Ctrl hydrogels, which was much greater than the increase (similar to 30%) in beta cat nuclear localization in two-cell clusters within the Ncad hydrogels. In summary, we utilized fluorescent imaging to demonstrate Ncad-mediated single cell responses to developmental cues within hydrogels towards chondrogenesis.
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