title: Fabrication and Characterization of Decellularized Periodontal Ligament Cell Sheet Constructs.
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.
authors.affiliation: School of Dentistry and Oral Health, Regenerative Medicine Center, Menzies Health Institute Queensland, Gold Coast, QLD, Australia.
authors.forename: Amro
authors.initials: A
authors.lastname: Farag
authors.affiliation: Queensland University of Technology (QUT), Brisbane, QLD, Australia.
authors.forename: Cédryck
authors.initials: C
authors.lastname: Vaquette
authors.affiliation: Queensland University of Technology (QUT), Brisbane, QLD, Australia.
authors.forename: Dietmar W
authors.initials: DW
authors.lastname: Hutmacher
authors.affiliation: Colgate Australian Clinical DentalResearch Centre, Dental School, University of Adelaide, Frome Road, Adelaide, SA, 5005, Australia.
authors.forename: P Mark
authors.initials: PM
authors.lastname: Bartold
authors.affiliation: School of Dentistry and Oral Health, Regenerative Medicine Center, Menzies Health Institute Queensland, Gold Coast, QLD, Australia. s.ivanovski@griffith.edu.au.|Menzies Health Institute Queensland, Griffith University, Gold Coast Campus, Parklands Drive, Gold Coast, QLD, 4222, Australia. s.ivanovski@griffith.edu.au.
authors.forename: Saso
authors.initials: S
authors.lastname: Ivanovski
mesh_terms: D018929:Cell Culture Techniques; D002469:Cell Separation; D005109:Extracellular Matrix; D006801:Humans; D007150:Immunohistochemistry; D010513:Periodontal Ligament / Q000166:cytology*; D061251:Primary Cell Culture; D012038:Regeneration; D023822:Tissue Engineering; D054457:Tissue Scaffolds
keywords: Cell sheet; Constructs; Decellularization; Perfusion; Periodontal regeneration; Polycaprolactone; Scaffolds; Tissue engineering