Almela, T., Al-Sahaf, S., Bolt, R. et al. (2 more authors) (2018) Characterization of multi-layered tissue engineered human alveolar bone and gingival mucosa. Tissue Engineering Part C: Methods, 24 (2). pp. 99-107. ISSN 1937-3384
Abstract
Advances in tissue engineering have permitted assembly of multi-layered composite tissue constructs for potential applications in the treatment of combined hard and soft tissue defects and as an alternative in vitro test model to animal experimental systems. The aim of this study was to develop and characterize a novel three-dimensional combined human alveolar bone and gingival mucosal model based on primary cells isolated from the oral tissues. Bone component of the model was engineered by seeding primary human alveolar osteoblasts (HAOBs) into a hydroxyapatite/tricalcium phosphate (HA/TCP) scaffold and culturing in a spinner bioreactor. The engineered bone was then laminated, using an adhesive tissue sealant, with tissue engineered gingival mucosa consisting of air/liquid interface-cultured normal human gingival keratinocytes on oral fibroblast-populated collagen gel scaffold. Histological characterization revealed a structure consisting of established epithelial, connective tissue, and bone layers closely comparable to normal oral tissue architecture. The mucosal component demonstrated a mature epithelium undergoing terminal differentiation similar to that characteristic of native buccal mucosa, as confirmed using cytokeratin 13 (CK13) and cytokeratin 14 (CK14) immunohistochemistry. Ultrastructural analysis confirmed the presence of desmosomes and hemi-desmosomes in the epithelial layer, a continuous basement membrane and newly synthesized collagen in the connective tissue layer. Quantitative PCR (qPCR) assessment of osteogenesis-related gene expression showed a higher expression of genes encoded Collagen I (COL1) and Osteonectin (ON) compared with Osteocalcin (OC), Osteopontin (OPN), and Alkaline phosphatase (ALP). ELISA quantification of COL1, ON, and OC confirmed a pattern of secretion which paralleled the model’s gene expression profile. We demonstrate here that replicating the anatomical setting between oral mucosa and the underlying alveolar bone is feasible and the developed model showed characteristics similar to those of normal tissue counterparts. This tri-layered model therefore offers great scope as an advanced, and anatomically-representative tissue-engineered alternative to animal models.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2017 Mary Ann Liebert, Inc. This is an author produced version of a paper subsequently published in Tissue Engineering Part C: Methods. Uploaded in accordance with the publisher's self-archiving policy. |
Dates: |
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Institution: | The University of Sheffield |
Academic Units: | The University of Sheffield > Faculty of Medicine, Dentistry and Health (Sheffield) > School of Clinical Dentistry (Sheffield) |
Depositing User: | Symplectic Sheffield |
Date Deposited: | 10 Nov 2017 11:35 |
Last Modified: | 09 Apr 2024 16:09 |
Status: | Published |
Publisher: | Mary Ann Liebert |
Refereed: | Yes |
Identification Number: | 10.1089/ten.TEC.2017.0370 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:123618 |