

Cell-instructive hydrogels are attractive for skin repair and regeneration, serving as interactive matrices to promote cell adhesion, cell-driven remodeling and de novo deposition of extracellular matrix components. This paper describes the synthesis and photocrosslinking of cell-instructive pectin hydrogels using cell-degradable peptide crosslinkers and integrin-specific adhesive ligands. Protease-degradable hydrogels obtained by photoinitiated thiol-norbornene click chemistry are rapidly formed in the presence of dermal fibroblasts, exhibit tunable properties and are capable of modulating the behavior of embedded cells, including the cell spreading, hydrogel contraction and secretion of matrix metalloproteases. Keratinocytes seeded on top of fibroblast-loaded hydrogels are able to adhere and form a compact and dense layer of epidermis, mimicking the architecture of the native skin. Thiol-ene photocrosslinkable pectin hydrogels support the in vitro formation of full-thickness skin and are thus a highly promising platform for skin tissue engineering applications, including wound healing and in vitro testing models. Statement of Significance Photopolymerizable hydrogels are attractive for skin applications due to their unique spatiotemporal control over the hydrogel formation. This study reports the design of a promising photo-clickable pectin hydrogel which biophysical and biochemical properties can be independently tailored to control cell behavior. A fast method for the norbornene-functionalization of pectin was developed and hydrogels fabricated through UV photoinitiated thiol-norbornene chemistry. This one-pot click reaction was performed in the presence of cells using cell-adhesive and matrix metalloproteinase-sensitive peptides, yielding hydrogels that support extensive cell spreading. Keratinocytes seeded on top of the fibroblast-loaded hydrogel formed a compact epidermis with morphological resemblance to human skin. This work presents a new protease-degradable hydrogel that supports in vitro skin formation with potential for skin tissue engineering. © 2017 Acta Materialia Inc.
| EMTREE drug terms: | integrinmatrix metalloproteinasenorbornene derivativepectinpeptideproteinasethiol2-norbornenearginyl-glycyl-aspartic acidcross linking reagentmatrix metalloproteinasenorbornane derivativeoligopeptidepectinthiol derivative |
|---|---|
| EMTREE medical terms: | Articlecell adhesioncell spreadingcell viabilityclick chemistrycross linkingdegradationenzyme releaseepidermisextracellular matrixfibroblasthumanhuman cellhuman tissuehydrogelin vitro studykeratinocytephotoclick chemistrypriority journalprotein expressionskinskin structuresynthesistissue engineeringwound healingartificial skincell motionchemistryclick chemistrycytologydrug effecthydrogellightmalemetabolismpolymerizationproceduresskin |
| MeSH: | Cell AdhesionCell MovementClick ChemistryCross-Linking ReagentsFibroblastsHumansHydrogelsLightMaleMatrix MetalloproteinasesNorbornanesOligopeptidesPectinsPolymerizationSkinSkin, ArtificialSulfhydryl CompoundsTissue Engineering |
pectin, 9000-69-5; proteinase, 9001-92-7; thiol derivative, 13940-21-1;
2-norbornene; arginyl-glycyl-aspartic acid; Cross-Linking Reagents; Hydrogels; Matrix Metalloproteinases; Norbornanes; Oligopeptides; pectin; Pectins; Sulfhydryl Compounds
| Funding sponsor | Funding number | Acronym |
|---|---|---|
| Fundação Portugal Telecom | ||
| Fuel Cell Technologies Program | FCT | |
| Irene W. and C.B. Pennington Foundation | SFRH/BD/91151/2012 | |
| POCI,2020 | ||
| IF/00296/2015 | ||
| European Regional Development Fund | ||
| NORTE 2020 |
This work was supported by the European Regional Development Fund (ERDF) through the COMPETE 2020 - Operational Programme for Competitiveness and Internationalization (POCI), Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, and by Portuguese funds through Portuguese Foundation for Science and Technology (FCT) in the framework of the project Ref. PTDC/BBB-ECT/2145/2014. R.P. and C.B. thank FCT for the doctoral grant SFRH/BD/91151/2012 and the FCT Investigator research position IF/00296/2015 (FCT and POPH/ESF), respectively. The authors acknowledge Mariana Andrade from CEMUP (Centro de Materiais da Universidade do Porto) for the 1 H NMR analyses, Dr Frederico Silva from B2Tech (Biochemical and Biophysical Technologies, i3S) for the support with HPLC analyses, and Dr Silvia Bidarra from Biomaterials for Multistage Drug & Cell Delivery Group (i3S) for the support with gelatin zymography. Appendix A
Bártolo, P.J.; School of Mechanical, Aerospace and Civil Engineering, University of Manchester, United Kingdom;
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