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Microsc Res Tech. 2015 Sep;78(9):792-800. doi: 10.1002/jemt.22542. Epub 2015 Jul 15.

Ultrastructural evaluation of mesenchymal stem cells from inflamed periodontium in different in vitro conditions.

Author information

1
Student, Faculty of Medicine, Department of Histology, Iuliu Haţieganu University of Medicine and Pharmacy, Cluj-Napoca, 400012, Romania.
2
Department of Molecular Biology and Biotechnologies, Faculty of Biology and Geology, Babeş-Bolyai University, Cluj-Napoca, 400006, Romania.
3
Department of Veterinary Reproduction, Obstetrics, and Gynecology, Faculty of Veterinary Medicine, University of Agricultural Sciences and Veterinary Medicine, Cluj-Napoca, 400372, Romania.
4
Department of Cell and Molecular Biology, Faculty of Medicine, Iuliu Haţieganu University of Medicine and Pharmacy, Cluj-Napoca, 400349, Romania.
5
Department of Periodontology, Faculty of Dental Medicine, Iuliu Haţieganu University of Medicine and Pharmacy, Cluj-Napoca, 400012, Romania.
6
Department of Histology, Faculty of Medicine, Iuliu Haţieganu University of Medicine and Pharmacy, Cluj-Napoca, 400349, Romania.

Abstract

This research aimed to observe the behavior of mesenchymal stem cells (MSCs) isolated from periodontal granulation tissue (gt) when manipulated ex vivo to induce three-dimensional (3D) spheroid (aggregates) formation as well as when seeded on two bone scaffolds of animal origin. Periodontal gt was chosen as a MSC source because of its availability, considering that it is eliminated as a waste material during conventional surgical therapies. 3D aggregates of cells were generated; they were grown for 3 and 7 days, respectively, and then prepared for transmission electron microscopic analysis. The two biomaterials were seeded for 72 h with gtMSCs and prepared for scanning electronic microscopic observation. The ultrastructural analysis of 3D spheroids remarked some differences between the inner and the outer cell layers, with a certain commitment observed at the inner cells. Both scaffolds showed a relatively smooth surface at low magnification. Macro- and micropores having a scarce distribution were observed on both bone substitutes. gtMSCs grew with relative difficulty on the biomaterials. After 72 h of proliferation, gtMSCs scarcely covered the surface of bovine bone scaffolds, demonstrating fibroblast-like or star-like shapes with elongated filiform extensions. Our results add other data on the possible usefulness of gtMSC and could question the current paradigm regarding the complete removal of chronically inflamed gts from the defects during periodontal surgeries. Until optimal protocols for ex vivo manipulation of MSCs are available for clinical settings, it is advisable to use biocompatible bone substitutes that allow the development of progenitor cells.

KEYWORDS:

adult; bone substitutes; brightfield microscopy; granulation tissue; scanning electron microscopy; transmission electron microscopy

PMID:
26179176
DOI:
10.1002/jemt.22542
[Indexed for MEDLINE]

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