Journal
ACS APPLIED MATERIALS & INTERFACES
Volume 10, Issue 31, Pages 26016-26027Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acsami.8b07632
Keywords
Michael addition; tetrazine ligation; hydrogel; synthetic extracellular matrices; mesenchymal stem cells
Funding
- National Institutes of Health (NIH) [R01DC011377, R01DC014461, R01DE022969]
- National Science Foundation (NSF) [DMR 1506613]
- Delaware Bioscience Center for Advanced Technology (DE CAT)
- W. L. Gore and Associates
- NSF IGERT Fellowship
- NIH [P20GM104316, P30GM110758, S10RR026962, S10OD016267]
- NSF [CHE-0840401, CHE-1229234, IIA-1301765]
- NIH-NIGMS [P20 GM103446]
- State of Delaware
- Division Of Materials Research
- Direct For Mathematical & Physical Scien [1506613] Funding Source: National Science Foundation
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Chemical modification of engineered microenvironments surrounding living cells represents a means for directing cellular behaviors through cell-matrix interactions. Presented here is a temporally controlled method for modulating the properties of biomimetic, synthetic extracellular matrices (ECM) during live cell culture employing the rapid, bioorthogonal tetrazine ligation with trans-cyclooctene (TCO) dienophiles. This approach is diffusion-controlled, cytocompatible, and does not rely on light, catalysts, or other external triggers. Human bone-marrow-derived mesenchymal stem cells (hMSCs) were initially entrapped in a hydrogel prepared using hyaluronic acid carrying sulfhydryl groups (HA-SH) and a hydrophilic polymer bearing both acrylate and tetrazine groups (POM-AT). Inclusion of a matrix metalloprotease (MMP)-degradable peptidic cross-linker enabled hMSC-mediated remodeling of the synthetic environment. The resultant network displayed dangling tetrazine groups for subsequent conjugation with TCO derivatives. Two days later, the stiffness of the matrix was increased by adding chemically modified HA carrying multiple copies of TCO (HA-TCO) to the hMSC growth media surrounding the cell-laden gel construct. In response, cells developed small processes radially around the cell body without a significant alteration of the overall shape. By contrast, modification of the 3D matrix with a TCO-tagged cell-adhesive motif caused the resident cells to undergo significant actin polymerization, changing from a rounded shape to spindle morphology with long cellular processes. After additional 7 days of culture in the growth media, quantitative analysis showed that, at the mRNA level, RGD tagging upregulated cellular expression of MMP1, but downregulated the expression of collagen I/III and tenascin C. RGD tagging, however, was not sufficient to induce the classic osteoblastic, chondrogenic, adipogenic, or fibroblastic/myofibroblastic differentiation. The modular approach allows facile manipulation of synthetic ECM to modulate cell behavior, thus potentially applicable to the engineering of functional tissues or tissue models.
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