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Patterning Bioactive Proteins or Peptides on Hydrogel Using Photochemistry for Biological Applications
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Chemically cross-linked hydrogels from repetitive protein arrays.

Rossana Boni1, Elizabeth A Blackburn2, Dirk-Jan Kleinjan3

  • 1Centre for Engineering Biology, Institute of Quantitative Biology, Biochemistry and Biotechnology, School of Biological Sciences, University of Edinburgh, Edinburgh, United Kingdom.

Journal of Structural Biology
|May 28, 2023
PubMed
Summary

Engineered protein hydrogels mimic native tissues for regeneration. This adaptable biomaterial allows precise control over physical properties and supports cell viability, advancing tissue engineering.

Keywords:
BiomaterialsHydrogelsProtein engineeringProtein rheologyRepetitive sequenceSpyTag SpyCatcherTissue engineering

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Area of Science:

  • Biomaterials Science
  • Protein Engineering
  • Tissue Engineering

Background:

  • Biomaterials for tissue regeneration require mimicking native physiological environments.
  • Protein engineering offers a method to create hydrogels with tailored biophysical properties.

Purpose of the Study:

  • To design and characterize novel protein hydrogels using repetitive engineered proteins.
  • To demonstrate the ability to control hydrogel properties by adjusting protein building block ratios.
  • To assess the biocompatibility and cell encapsulation capabilities of the developed hydrogels.

Main Methods:

  • Incorporation of SpyTag (ST) peptide and repetitive SpyCatcher (SC) protein units to form covalent networks.
  • Systematic variation of ST:SC ratios to modulate viscoelastic properties and gelation kinetics.
  • Assessment of hydrogel physical properties and biocompatibility using HepG2 cells expressing GFP.

Main Results:

  • Successfully engineered repetitive proteins formed covalent molecular networks with tunable physical characteristics.
  • Alteration of ST:SC ratios effectively controlled viscoelastic properties and gelation speeds.
  • Encapsulated HepG2 cells remained viable and expressed GFP, indicating successful biocompatibility and cell phenotype maintenance.

Conclusions:

  • Genetically encoded repetitive proteins provide a versatile platform for creating customizable biomaterials.
  • This approach bridges engineering biology and nanotechnology, enabling unprecedented biomaterial customization for tissue regeneration.
  • The developed hydrogels show significant potential for applications in liver tissue engineering and beyond.