Meshry, Nadeen Mahmoud
(2021)
Engineering Scaffold-free Dental Organoids Using Human Stem Cells from the Apical Papilla (SCAP).
Master's Thesis, University of Pittsburgh.
(Unpublished)
Abstract
The clinical dilemma of treating immature necrotic teeth has been the driving force behind the field of regenerative endodontics. Through evoking bleeding from the periapical tissues, clinicians have noticed root elongation and closure following this treatment. The periapical location of the apical papilla tissue renders it a very probable source of some of the progenitor cells that are homed to the canal space during this procedure. This study aims to generate an experimental model for studying the regenerative capacity of the apical papilla tissue, through engineering scaffold-free dental organoids (SFDO) using human stem cells from the apical papilla (SCAP).
The SFDOs were generated by culturing human SCAP to form a cell sheet, which later contracts resulting in the formation of a 3D construct which was cultured in osteogenic medium for 14 days before analyzing the results. Hematoxylin-and-Eosin-stained histological sections of the SFDOs revealed a densely cellular structure, with two distinct morphologies; spindle-shaped peripheral cells and distinctly more rounded cells in the center. Alizarin red staining for calcium deposition showed that the organoids had a triple mineralization pattern, with (1) an outermost thin rim of unmineralized tissue (corresponding to the flat peripheral cells) that encloses (2) a mineralized intermediate zone, which in turn exhibits a decreasing gradient of mineralization that ends with (3) an innermost core of unmineralized tissue. The resultant multi-tissue structure potentially resembles the organization seen in a tooth root (outer PDL, intermediate cementum/dentin, inner pulp). Immunofluorescent staining showed that expression of DSP (a dentinogenic marker) and asporin (a reported PDL marker) were localized to the inner bulk and the peripheral part of the SFDO respectively.
These results suggest that SCAP could have the potential to regenerate a full tooth root-like structure. As such, SCAP might be worth exploration as a potential cell-based endodontic treatment modality for promoting root elongation in cases of immature necrotic teeth.
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Details
| Item Type: |
University of Pittsburgh ETD
|
| Status: |
Unpublished |
| Creators/Authors: |
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| ETD Committee: |
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| Date: |
3 September 2021 |
| Date Type: |
Publication |
| Defense Date: |
14 July 2021 |
| Approval Date: |
3 September 2021 |
| Submission Date: |
28 July 2021 |
| Access Restriction: |
2 year -- Restrict access to University of Pittsburgh for a period of 2 years. |
| Number of Pages: |
72 |
| Institution: |
University of Pittsburgh |
| Schools and Programs: |
School of Dental Medicine > Dental Science |
| Degree: |
MS - Master of Science |
| Thesis Type: |
Master's Thesis |
| Refereed: |
Yes |
| Uncontrolled Keywords: |
Scaffold-free tissue engineering
Regenerative endodontics
Regeneration
Root elongation
Apical papilla
SCAP
Experimental model |
| Date Deposited: |
03 Sep 2021 17:43 |
| Last Modified: |
03 Sep 2023 05:15 |
| URI: |
http://d-scholarship.pitt.edu/id/eprint/41513 |
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