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Decellularization of placentas: establishing a protocol.

L C P C Leonel1, C M F C Miranda1, T M Coelho2

  • 1Setor de Anatomia, Departamento de Cirurgia, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, São Paulo, SP, Brasil.

Brazilian Journal of Medical and Biological Research = Revista Brasileira De Pesquisas Medicas E Biologicas
|November 30, 2017
PubMed
Summary

Researchers optimized decellularization of canine placentas for tissue engineering. Protocol I better preserved extracellular matrix structure, while Protocol II more effectively removed cells and DNA using sodium dodecyl sulfate (SDS).

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Area of Science:

  • Biomaterials Science
  • Tissue Engineering
  • Veterinary Medicine

Background:

  • Biological biomaterials for tissue engineering are often derived from decellularized tissues and organs.
  • The extracellular matrix (ECM) must remain acellular while preserving its structural and protein integrity.
  • Canine placentas offer a readily available source of ECM, but established decellularization protocols are lacking.

Purpose of the Study:

  • To establish an effective decellularization method for both maternal and fetal portions of canine placentas.
  • To compare the efficacy of different detergents, freezing temperatures, and perfusion techniques.
  • To evaluate two distinct decellularization protocols for their impact on ECM preservation and cell removal.

Main Methods:

  • Ten preliminary tests evaluated various decellularization parameters.
  • Two optimized protocols were selected for detailed analysis.
  • Histology, scanning electron microscopy, immunofluorescence, and DNA quantification were employed for assessment.

Main Results:

  • Sodium dodecyl sulfate (SDS) proved to be the most effective detergent for cell removal.
  • Pre-freezing canine placentas increased the required incubation time for decellularization.
  • Both perfusion and immersion methods successfully removed cells.
  • Protocol I (1% SDS, 5 mM EDTA, 50 mM TRIS, 0.5% antibiotic) better preserved ECM structure.
  • Protocol II (1% SDS, 5 mM EDTA, 0.05% trypsin, 0.5% antibiotic) was more efficient in removing cells and DNA.

Conclusions:

  • Decellularization of canine placentas is feasible for generating biomaterials.
  • A balance exists between ECM structural preservation and complete cell/DNA removal.
  • Further refinement of decellularization protocols is necessary for optimal canine placental biomaterial development.