Dentin-like tissue formation and biomineralization by multicellular human pulp cell spheres in vitro


Introduction:
Maintaining or regenerating a vital pulp is a preferable goal in current endodontic research. In this study, human dental pulp cell aggregates (spheres) were applied onto bovine and human root canal models to evaluate their potential use as pre-differentiated tissue units for dental pulp tissue regeneration.

Methods:
Human dental pulp cells (DPC) were derived from wisdom teeth, cultivated into three-dimensional cell spheres and seeded onto bovine and into human root canals. Sphere formation, tissue-like and mineralization properties as well as growth behavior of cells on dentin structure were evaluated by light microscopy (LM), confocal laser scanning microscopy (CLSM), scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDX).

Results:
Spheres and outgrown cells showed tissue-like properties, the ability to merge with other cell spheres and extra cellular matrix formation; CLSM investigation revealed a dense network of actin and focal adhesion contacts (FAC) inside the spheres and a pronounced actin structure of cells outgrown from the spheres. A dentin-structure-orientated migration of the cells was shown by SEM investigation. Besides the direct extension of the cells into dentinal tubules, the coverage of the tubular walls with cell matrix was detected. Moreover, an emulation of dentin-like structures with tubuli-like and biomineral formation was detected by SEM- and EDX-investigation.

Conclusions:
The results of the present study show tissue-like behavior, the replication of tubular structures and the mineralization of human dental pulp spheres when colonized on root dentin. The application of cells in form of pulp spheres on root dentin reveals their beneficial potential for dental tissue regeneration.

Keywords:
Biomineralization, Dental pulp cells, Tissue formation, Pulp spheres, Pulp tissue regeneration


Autoři: Jörg Neunzehn 1*;  Marie-Theres Weber 2;  Gretel Wittenburg 3;  Günter Lauer 3;  Christian Hannig 2;  And Hans-Peter Wiesmann 1
Působiště autorů: Technische Universität Dresden, Institute of Material Science, Chair for Biomaterials, Budapester Strasse 7, D-01069 Dresden, Germany. 1;  Department of Restorative and Pediatric Dentistry, University Hospital Carl Gustav Carus, Fetscherstrasse 74, D-01 07 Dresden, Germany. 2;  Department of Oral and Maxillofacial Surgery, University Hospital Carl Gustav Carus, Fetscherstrasse 74, D-01307 Dresden, Germany. 3
Vyšlo v časopise: Head & Face Medicine 2014, 10:25
Kategorie: Research
prolekare.web.journal.doi_sk: 10.1186/1746-160X-10-25

© 2014 Neunzehn et al.; licensee BioMed Central Ltd.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
The electronic version of this article is the complete one and can be found online at: http://www.head-face-med.com/content/10/1/25.

Souhrn

Introduction:
Maintaining or regenerating a vital pulp is a preferable goal in current endodontic research. In this study, human dental pulp cell aggregates (spheres) were applied onto bovine and human root canal models to evaluate their potential use as pre-differentiated tissue units for dental pulp tissue regeneration.

Methods:
Human dental pulp cells (DPC) were derived from wisdom teeth, cultivated into three-dimensional cell spheres and seeded onto bovine and into human root canals. Sphere formation, tissue-like and mineralization properties as well as growth behavior of cells on dentin structure were evaluated by light microscopy (LM), confocal laser scanning microscopy (CLSM), scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDX).

Results:
Spheres and outgrown cells showed tissue-like properties, the ability to merge with other cell spheres and extra cellular matrix formation; CLSM investigation revealed a dense network of actin and focal adhesion contacts (FAC) inside the spheres and a pronounced actin structure of cells outgrown from the spheres. A dentin-structure-orientated migration of the cells was shown by SEM investigation. Besides the direct extension of the cells into dentinal tubules, the coverage of the tubular walls with cell matrix was detected. Moreover, an emulation of dentin-like structures with tubuli-like and biomineral formation was detected by SEM- and EDX-investigation.

Conclusions:
The results of the present study show tissue-like behavior, the replication of tubular structures and the mineralization of human dental pulp spheres when colonized on root dentin. The application of cells in form of pulp spheres on root dentin reveals their beneficial potential for dental tissue regeneration.

Keywords:
Biomineralization, Dental pulp cells, Tissue formation, Pulp spheres, Pulp tissue regeneration


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