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Versatile click alginate hydrogels with protease-sensitive domains as cell responsive/instructive 3D

Mariana I Neves1, Mariana V Magalhães1, Sílvia J Bidarra2

  • 1i3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, R. Alfredo Allen 208, 4200-135 Porto, Portugal; INEB - Instituto de Engenharia Biomédica, Universidade do Porto, Portugal; FEUP - Faculdade de Engenharia, Universidade do Porto, Portugal.

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Summary

This study developed new alginate hydrogels that mimic the body's extracellular matrix. These smart biomaterials degrade in response to enzymes, enabling better cell growth and tissue regeneration for soft tissue engineering.

Keywords:
Bio-orthogonalBioactiveMatrix remodelingProtease sensitive

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Area of Science:

  • Biomaterials Science
  • Tissue Engineering
  • Biotechnology

Background:

  • Alginate (ALG) is a common biomaterial for tissue engineering but lacks degradation properties.
  • Imparting proteolytic sensitivity to alginate hydrogels enhances their function as artificial extracellular matrices (ECM).

Purpose of the Study:

  • To develop enzyme-degradable alginate hydrogels using click chemistry for improved tissue engineering applications.
  • To create cell-permissive, multifunctional 3D matrices for soft tissue regeneration.

Main Methods:

  • Utilized strain-promoted azide-alkyne cycloaddition (SPAAC) to conjugate cyclooctyne-modified alginate (ALG-K) with matrix metalloproteinase (MMP)-sensitive PVGLIG peptides.
  • Grafted ALG-K with both PVGLIG and cell-adhesive RGD peptides for creating multifunctional matrices.
  • Formed cell-laden hydrogels via secondary ionic crosslinking immediately after SPAAC conjugation.

Main Results:

  • Successfully incorporated PVGLIG peptides into alginate, demonstrating crosslinking ability and enzymatic degradability.
  • Hydrogels with intermediate PVGLIG concentration (125 μM) showed increased stiffness, promoting cell spreading and cell-cell connections.
  • Embedded cells remodeled the hydrogel network, expressed MMPs, and secreted enzymes that degraded the PVGLIG peptides.

Conclusions:

  • Developed enzyme-degradable alginate-based hydrogels that mimic key ECM features.
  • These novel hydrogels support cell activity, pericellular matrix production, and soft tissue regeneration.
  • The rapid SPAAC-based method offers a streamlined approach for creating advanced biomaterials.