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Decellularization of human amniotic membrane using detergent-free methods: Possibilities in tissue engineering.

Shahrzad Ashouri1, Seyede Atefe Hosseini2, Seyed Javad Hoseini3

  • 1Department of Anatomy and Cell Biology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.

Tissue & Cell
|May 17, 2022
PubMed
Summary

Detergent-free methods, including mechanical and physical treatments, offer simple and cost-effective ways to decellularize human amniotic membrane (HAM) for tissue engineering. These approaches preserve tissue integrity and biological properties, making d-HAM a viable option for wound dressings.

Keywords:
DecellularizationDetergent-free methodHuman amniotic membraneSkin tissue engineering

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Area of Science:

  • Biomaterials Science
  • Tissue Engineering
  • Regenerative Medicine

Background:

  • Human amniotic membrane (HAM) is a valuable natural scaffold in tissue engineering due to its beneficial biological properties.
  • Effective decellularization is crucial for utilizing HAM in regenerative medicine applications, minimizing immunogenicity while preserving structural integrity.

Purpose of the Study:

  • To develop simple, cost-effective, detergent-free protocols for decellularizing human amniotic membrane (HAM).
  • To compare the efficacy of mechanical (brushing) and physical (heating) methods against a chemical (EDTA + NaOH + NH4Cl) approach for HAM decellularization.
  • To evaluate the physico-chemical, mechanical, and biological properties of the resulting decellularized HAM (d-HAM).

Main Methods:

  • Decellularization of HAM using mechanical force (brushing), physical treatment (heating 45-55°C), and a chemical method (EDTA + NaOH + NH4Cl).
  • Characterization of d-HAM through DAPI staining for cell removal, DNA content analysis, histological evaluation (collagen, GAGs), mechanical testing (ultimate tensile stress), and hydroxyproline content measurement.
  • Assessment of cell attachment, proliferation, and migration on the prepared d-HAM samples.

Main Results:

  • Chemical method completely removed epithelial cells; mechanical and physical methods reduced cell numbers.
  • All methods significantly reduced DNA content (< 50 ng/mg) while preserving tissue integrity, collagen, and GAGs.
  • Mechanical method retained highest hydroxyproline levels, while physical method (heating) promoted cell proliferation; all d-HAMs supported cell attachment and migration.

Conclusions:

  • Detergent-free protocols (mechanical and physical) are effective for human amniotic membrane decellularization.
  • The developed d-HAM tissues maintain structural and biological integrity, suitable for tissue engineering.
  • These cost-effective d-HAMs represent promising, inexpensive wound dressings for various regenerative applications.