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Engineering Scaffold-free Dental Organoids Using Human Stem Cells from the Apical Papilla (SCAP)

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 (Unpublished)

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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:
CreatorsEmailPitt UsernameORCID
Meshry, Nadeen Mahmoudnadeen.meshry@gmail.comnmm1060000-0002-1489-440X
ETD Committee:
TitleMemberEmail AddressPitt UsernameORCID
Thesis AdvisorSyed-Picard, Fatima N.syedpicard@pitt.edusyedpicard0000-0002-8212-0254
Committee MemberBeniash, Eliaebeniash@pitt.eduebeniash0000-0001-9019-5160
Committee MemberTaboas, Juanjmt106@pitt.edujmt1060000-0003-0523-3359
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 2021 17:43
URI: http://d-scholarship.pitt.edu/id/eprint/41513

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