Decellularized Periodontal Ligament Cell Sheets with Recellularization Potential

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 perio...

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Bibliographic Details
Published in:Journal of dental research 2014-12, Vol.93 (12), p.1313-1319
Main Authors: Farag, A., Vaquette, C., Theodoropoulos, C., Hamlet, S.M., Hutmacher, D.W., Ivanovski, S.
Format: Article
Language:English
Subjects:
Acids
Ammonium Hydroxide - chemistry
Cell Culture Techniques
Collagen
Collagen (type I)
Collagen Type I - analysis
Deoxyribonuclease
Deoxyribonucleases - chemistry
DNA - analysis
Enzyme-linked immunosorbent assay
Extracellular matrix
Extracellular Matrix - chemistry
Fibroblast growth factor 2
Fibroblast Growth Factor 2 - analysis
Fibronectin
Fibronectins - analysis
Growth factors
Guided Tissue Regeneration, Periodontal - instrumentation
Hepatocyte growth factor
Hepatocyte Growth Factor - analysis
Humans
Ligaments
Membranes
Membranes, Artificial
Microscopy, Electron, Scanning
Octoxynol - chemistry
Perfusion
Periodontal ligament
Periodontal Ligament - cytology
Polycaprolactone
Polyesters - chemistry
Research Reports
Scanning electron microscopy
Tissue engineering
Tissue Engineering - methods
Tissue Scaffolds - chemistry
Vascular endothelial growth factor
Vascular Endothelial Growth Factor A - analysis
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Summary: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.
ISSN:0022-0345
1544-0591
DOI:10.1177/0022034514547762