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Oral Biology pp 403-412 | Cite as

Fabrication and Characterization of Decellularized Periodontal Ligament Cell Sheet Constructs

  • Amro Farag
  • Cédryck Vaquette
  • Dietmar W. Hutmacher
  • P. Mark Bartold
  • Saso IvanovskiEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1537)

Abstract

Decellularized tissue-engineered constructs have the potential to promote regeneration by providing a biomimetic extracellular matrix that directs tissue-specific regeneration when implanted in situ. Recently, the use of cell sheets has shown promising results in promoting periodontal regeneration. Here, we describe the fabrication of decellularized periodontal cell sheets with intact extracellular matrix structural and biological properties. Melt electro spun polycaprolactone (PCL) scaffolds are used as a carrier for the inherently fragile cell sheets, to provide support during the processes of decellularization. An optimized decellularization method is outlined using perfusion with a combination of NH4OH and Triton X-100 together with a DNase treatment step for DNA removal. The maintenance of extracellular matrix structural and biological integrity is important, and here we describe the assessment of these properties using immunostaining for extracellular matrix proteins and ELISA for growth factor quantification.

Key words

Cell sheet Decellularization Tissue engineering Scaffolds Constructs Perfusion Polycaprolactone Periodontal regeneration 

Notes

Acknowledgments

This work was supported by Australia’s National Health and Medical Research Council project grant 1086181.

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Amro Farag
    • 1
  • Cédryck Vaquette
    • 2
  • Dietmar W. Hutmacher
    • 2
  • P. Mark Bartold
    • 3
  • Saso Ivanovski
    • 1
    • 4
    Email author
  1. 1.School of Dentistry and Oral Health, Regenerative Medicine CenterMenzies Health Institute QueenslandGold CoastAustralia
  2. 2.Queensland University of Technology (QUT)BrisbaneAustralia
  3. 3.Colgate Australian Clinical DentalResearch Centre, Dental SchoolUniversity of AdelaideAdelaideAustralia
  4. 4.Menzies Health Institute QueenslandGriffith UniversityGold CoastAustralia

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