Decellularized cell sheet constructs for periodontal regeneration
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1
Menzies Health Institute Queensland. Griffith University, School of Dentistry and Oral Health, Australia
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2
Queensland University of Technology, Institute of Health and Biomedical Innovation, Australia
Decellularization aims to harness the regenerative properties of native extracellular matrix while removing immunogenic cellular material. The aim of this study was to evaluate decellularized cell sheet constructs for their ability to maintain extracellular matrix integrity, retain growth factors, support allogenic cell repopulation and facilitate periodontal regeneration in vivo. To this end, intact human periodontal ligament cell sheets were placed onto melt electrospun polycaprolactone (PCL) membrane that reinforced the cell sheets and facilitated the physical manipulation necessary during the decellularization protocol. These cell sheet constructs (CSCs) were decellularized under static and perfusion conditions using a variety of methods. The structural integrity of the decellularized CSCs was assessed using a collagen quantification assay, immunostaining of human collagen type I and scanning electron microscopy. Residual basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) were assessed with Bio-plex assays. The constructs were also assessed for their effect on cell differentiation markers that are relevant to periodontal ligament tissues using real time PCR. Differentiation was also assessed using STRO-1 immunostaining. Finally, the decellularized constructs were evaluated in an athymic rat periodontal defect model for their tissue integration and ability to promote periodontal regeneration. Treatment with NH4OH/Triton X-100 and DNase solution was the most efficient method for DNA removal and the preservation of extracellular matrix integrity and growth factors retention. The decellularized constructs were able to support re-population with allogenic cells, and promoted tissue specific cell differentiation compared to empty constructs. The decellularized constructs promoted similar levels of periodontal regeneration as cellularized constructs the in vivo periodontal defect model. The results of this study showed that the cell constructs could be efficiently decellularized while maintaing extracellular matrix integrity and growth factor retention. These constructs were able to facilitate allogenic cell repopulation and tissue specific differentiation in vitro, and support periodontal regeneration in vivo.
Keywords:
Cell Differentiation,
Extracellular Matrix,
Tissue Regeneration,
acellullar matrix
Conference:
10th World Biomaterials Congress, Montréal, Canada, 17 May - 22 May, 2016.
Presentation Type:
Poster
Topic:
Biomaterials in dental applications
Citation:
Amro
F,
Vaquette
C,
Hashimi
S,
Stephen
H,
Hutmacher
D and
Ivanovski
S
(2016). Decellularized cell sheet constructs for periodontal regeneration.
Front. Bioeng. Biotechnol.
Conference Abstract:
10th World Biomaterials Congress.
doi: 10.3389/conf.FBIOE.2016.01.02876
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Received:
27 Mar 2016;
Published Online:
30 Mar 2016.