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Related Experiment Video

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Culturing Mammalian Cells in Three-dimensional Peptide Scaffolds
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Mesenchymal stem cells aligned and stretched in self-assembling peptide hydrogels.

Farzaneh Fouladgar1, Forough Ghasem Zadeh Moslabeh1, Yashesh Varun Kasani1

  • 1Department of Biomedical Engineering, University of North Texas, Texas, United States.

Heliyon
|January 18, 2024
PubMed
Summary

This study demonstrates that fluorenylmethoxycarbonyl-diphenylalanine (Fmoc-FF) peptide hydrogels effectively encapsulate and align mesenchymal stem cells (MSCs) under mechanical stretching, offering a promising biomaterial for tissue engineering.

Keywords:
HydrogelsMesenchymal stem cellsPeptideSelf-assembly

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Area of Science:

  • Biomaterials Science
  • Tissue Engineering
  • Stem Cell Biology

Background:

  • Peptide hydrogels offer tunable properties for mimicking the extracellular matrix.
  • Mesenchymal stem cells (MSCs) are crucial for regenerative medicine applications.
  • Controlling cell behavior within biomaterials is key for effective tissue regeneration.

Purpose of the Study:

  • To investigate the use of Fmoc-protected peptide hydrogels for encapsulating and mechanically stretching human MSCs.
  • To evaluate the influence of different solvents on peptide hydrogel properties and MSC behavior.
  • To establish conditions for successful MSC alignment and viability within peptide nanofibers for tissue engineering.

Main Methods:

  • Preparation of Fmoc-FF peptide hydrogels using DMSO, HFP, and DiH2O.
  • Characterization of hydrogel self-assembly, mechanical properties (compressive modulus), and morphology.
  • Encapsulation and mechanical stretching of human MSCs within the hydrogels using a custom device.
  • Assessment of MSC viability, morphology, and F-actin alignment via staining techniques.

Main Results:

  • Fmoc-FF hydrogels formed stiff, hydrated nanofiber networks (174-277 Pa modulus), mimicking extracellular matrix components.
  • MSCs encapsulated in Fmoc-FF/HFP and Fmoc-FF/DMSO hydrogels showed elongated morphology and aligned microfilament fibers after mechanical stretching.
  • A peptide concentration of 5 mM ensured 100% MSC viability.
  • Control cells maintained a round F-actin shape, indicating successful alignment induction by the hydrogel and stretching.

Conclusions:

  • Fmoc-FF peptide hydrogels provide a suitable microenvironment for MSC encapsulation and mechanical alignment.
  • Specific solvent choices (HFP, DMSO) and peptide concentrations are critical for achieving desired hydrogel properties and cell responses.
  • This approach presents a viable tissue engineering platform for regenerative medicine by leveraging peptide hydrogels and mechanical stimulation.