title: Decellularized periodontal ligament cell sheets with recellularization potential.
abstract: The periodontal ligament is the key tissue facilitating periodontal regeneration. This study aimed to fabricate decellularized human periodontal ligament cell sheets for subsequent periodontal tissue engineering applications. The decellularization protocol involved the transfer of intact human periodontal ligament cell sheets onto melt electrospun polycaprolactone membranes and subsequent bi-directional perfusion with NH4OH/Triton X-100 and DNase solutions. The protocol was shown to remove 92% of DNA content. The structural integrity of the decellularized cell sheets was confirmed by a collagen quantification assay, immunostaining of human collagen type I and fibronectin, and scanning electron microscopy. ELISA was used to demonstrate the presence of residual basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), and hepatocyte growth factor (HGF) in the decellularized cell sheet constructs. The decellularized cell sheets were shown to have the ability to support recellularization by allogenic human periodontal ligament cells. This study describes the fabrication of decellularized periodontal ligament cell sheets that retain an intact extracellular matrix and resident growth factors and can support repopulation by allogenic cells. The decellularized hPDL cell sheet concept has the potential to be utilized in future "off-the-shelf" periodontal tissue engineering strategies.
authors.affiliation: Griffith Health Institute, Regenerative Medicine Center, School of Dentistry and Oral Health, Griffith University, Gold Coast, Australia Institute of Health and Biomedical Innovation, Kelvin Grove, Brisbane, Australia.
authors.forename: A
authors.initials: A
authors.lastname: Farag
authors.affiliation: Institute of Health and Biomedical Innovation, Kelvin Grove, Brisbane, Australia.
authors.forename: C
authors.initials: C
authors.lastname: Vaquette
authors.affiliation: Institute of Health and Biomedical Innovation, Kelvin Grove, Brisbane, Australia.
authors.forename: C
authors.initials: C
authors.lastname: Theodoropoulos
authors.affiliation: Griffith Health Institute, Regenerative Medicine Center, School of Dentistry and Oral Health, Griffith University, Gold Coast, Australia.
authors.forename: S M
authors.initials: SM
authors.lastname: Hamlet
authors.affiliation: Institute of Health and Biomedical Innovation, Kelvin Grove, Brisbane, Australia.
authors.forename: D W
authors.initials: DW
authors.lastname: Hutmacher
authors.affiliation: Griffith Health Institute, Regenerative Medicine Center, School of Dentistry and Oral Health, Griffith University, Gold Coast, Australia s.ivanovski@griffith.edu.au.
authors.forename: S
authors.initials: S
authors.lastname: Ivanovski
mesh_terms: D064753:Ammonium Hydroxide / Q000737:chemistry; D018929:Cell Culture Techniques; D024042:Collagen Type I / Q000032:analysis; D004247:DNA / Q000032:analysis; D003851:Deoxyribonucleases / Q000737:chemistry; D005109:Extracellular Matrix / Q000737:chemistry; D016222:Fibroblast Growth Factor 2 / Q000032:analysis; D005353:Fibronectins / Q000032:analysis; D016557:Guided Tissue Regeneration, Periodontal / Q000295:instrumentation; D017228:Hepatocyte Growth Factor / Q000032:analysis; D006801:Humans; D008567:Membranes, Artificial; D008855:Microscopy, Electron, Scanning; D017830:Octoxynol / Q000737:chemistry; D010513:Periodontal Ligament / Q000166:cytology*; D011091:Polyesters / Q000737:chemistry; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds / Q000737:chemistry; D042461:Vascular Endothelial Growth Factor A / Q000032:analysis
chemical_list: D024042:Collagen Type I; D005353:Fibronectins; C066855:HGF protein, human; D008567:Membranes, Artificial; D011091:Polyesters; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D016222:Fibroblast Growth Factor 2; C016240:polycaprolactone; D064753:Ammonium Hydroxide; D017228:Hepatocyte Growth Factor; D017830:Octoxynol; D004247:DNA; D003851:Deoxyribonucleases
keywords: collagen; extracellular matrix; fibroblast growth factor; polycaprolactone; tissue engineering; vascular endothelial growth factor