Dental follicle cell differentiation towards periodontal ligament-like tissue in a self-assembly three-dimensional organoid model

Periodontitis remains an unsolved oral disease, prevalent worldwide and resulting in tooth loss due to dysfunction of the periodontal ligament (PDL), a tissue connecting the tooth root with the alveolar bone. A scaffold-free three-dimensional (3D) organoid model for in vitro tenogenesis/ligamentogeneis has already been described. As PDL tissue naturally arises from the dental follicle, the aim of this study was to investigate the ligamentogenic differentiation potential of dental follicle cells (DFCs) in vitro by employing this 3D model. Human primary DFCs were compared, in both two- and three-dimensions, to a previously published PDL- hTERT cell line. The 3D organoids were evaluated by haematoxylin and eosin, 4′,6-diamidino-2-phenylindole and F-actin staining combined with detailed histomorphometric analyses of cell-row structure, angular deviation and cell density. Furthermore, the expression of 48 tendon/ligament- and multilineage-related genes was evaluated using quantitative polymerase chain reaction, followed by immunofluorescent analyses of collagen 1 and 3. The results showed that both cell types were successful in the formation of scaffold-free 3D organoids. DFC organoids were comparable to PDL-hTERT in terms of cell density; however, DFCs exhibited superior organoid morphology, cell-row organisation (p < 0.0001) and angular deviation (p < 0.0001). Interestingly, in 2 dimensions as well as in 3D, DFCs showed significantly higher levels of several ligament- related genes compared to the PDL-hTERT cell line. In conclusion, DFCs exhibited great potential to form PDL-like 3D organoids in vitro suggesting that this strategy can be further developed for functional PDL engineering.

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Agarose-based spheroid culture enhanced stemness and promoted odontogenic differentiation potential of human dental follicle cells in vitro

In Vitro Cellular & Developmental Biology - Animal ◽

10.1007/s11626-021-00591-5 ◽

2021 ◽

Author(s):

Min Li ◽

Tiwei Fu ◽

Sen Yang ◽

Lanlan Pan ◽

Jing Tang ◽

...

Keyword(s):

Follicle Cells ◽

Dental Follicle ◽

Differentiation Potential ◽

Spheroid Culture ◽

Dental Follicle Cells ◽

Odontogenic Differentiation

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Evaluation of Osteogenic and Cementogenic Potential of Periodontal Ligament Fibroblast Spheroids Using a Three-DimensionalIn VitroModel of Periodontium

International Journal of Dentistry ◽

10.1155/2015/605813 ◽

2015 ◽

Vol 2015 ◽

pp. 1-9 ◽

Cited By ~ 4

Author(s):

Zurairah Berahim ◽

Keyvan Moharamzadeh ◽

Adrian K. Jowett ◽

Andrew Rawlinson

Keyword(s):

Periodontal Ligament ◽

Three Dimensional ◽

Osteogenic Potential ◽

Collagen Membrane ◽

Positive Staining ◽

Differentiation Potential ◽

Membrane Complex ◽

3D Matrix ◽

Periodontal Ligament Fibroblast

The aim of this study was to develop a three-dimensionalin vitromodel of periodontium to investigate the osteogenic and cementogenic differentiation potential of the periodontal ligament fibroblast (PDLF) spheroids within a dentin-membrane complex. PDLFs were cultured in both spheroid forms and monolayers and were seeded onto two biological collagen-based and synthetic membranes. Cell-membrane composites were then transferred onto dentin slices with fibroblasts facing the dentin surface and further cultured for 20 days. The composites were then processed for histology and immunohistochemical analyses for osteocalcin, Runx2, periostin, and cementum attachment protein (CAP). Both membranes seeded with PDLF-derived cells adhered to dentin and fibroblasts were present at the dentin interface and spread within both membranes. All membrane-cell-dentine composites showed positive staining for osteocalcin, Runx2, and periostin. However, CAP was not expressed by any of the tissue composites. It can be concluded that PDLFs exhibited some osteogenic potential when cultured in a 3D matrix in the presence of dentin as shown by the expression of osteocalcin. However the interaction of cells and dentin in this study was unable to stimulate cementum formation. The type of membrane did not have a significant effect upon differentiation, but fibroblast seeded-PGA membrane demonstrated better attachment to dentin than the collagen membrane.

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Isolation, characterization and investigation of differentiation potential of human periodontal ligament cells and dental follicle progenitor cells and their response to BMP-7 in vitro

Odontology ◽

10.1007/s10266-015-0198-1 ◽

2015 ◽

Vol 104 (2) ◽

pp. 123-135 ◽

Cited By ~ 8

Author(s):

Yahya Açil ◽

Fan Yang ◽

Aydin Gulses ◽

Mustafa Ayna ◽

Jörg Wiltfang ◽

...

Keyword(s):

Progenitor Cells ◽

Periodontal Ligament ◽

Periodontal Ligament Cells ◽

Dental Follicle ◽

Differentiation Potential ◽

Human Periodontal Ligament Cells

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Similar Features, Different Behaviors: A Comparative In Vitro Study of the Adipogenic Potential of Stem Cells from Human Follicle, Dental Pulp, and Periodontal Ligament

Journal of Personalized Medicine ◽

10.3390/jpm11080738 ◽

2021 ◽

Vol 11 (8) ◽

pp. 738

Author(s):

Melissa D. Mercado-Rubio ◽

Erick Pérez-Argueta ◽

Alejandro Zepeda-Pedreguera ◽

Fernando J. Aguilar-Ayala ◽

Ricardo Peñaloza-Cuevas ◽

...

Keyword(s):

Stem Cells ◽

Dental Pulp ◽

Periodontal Ligament ◽

Adipogenic Differentiation ◽

In Vitro Study ◽

Mrna Levels ◽

Dental Tissue ◽

Periodontal Ligament Stem Cells ◽

Differentiation Potential

Dental tissue-derived mesenchymal stem cells (DT-MSCs) are a promising resource for tissue regeneration due to their multilineage potential. Despite accumulating data regarding the biology and differentiation potential of DT-MSCs, few studies have investigated their adipogenic capacity. In this study, we have investigated the mesenchymal features of dental pulp stem cells (DPSCs), as well as the in vitro effects of different adipogenic media on these cells, and compared them to those of periodontal ligament stem cells (PLSCs) and dental follicle stem cells (DFSCs). DFSC, PLSCs, and DPSCs exhibit similar morphology and proliferation capacity, but they differ in their self-renewal ability and expression of stemness markers (e.g OCT4 and c-MYC). Interestingly, DFSCs and PLSCs exhibited more lipid accumulation than DPSCs when induced to adipogenic differentiation. In addition, the mRNA levels of adipogenic markers (PPAR, LPL, and ADIPOQ) were significantly higher in DFSCs and PLSCs than in DPSCs, which could be related to the differences in the adipogenic commitment in those cells. These findings reveal that the adipogenic capacity differ among DT-MSCs, features that might be advantageous to increasing our understanding about the developmental origins and regulation of adipogenic commitment.

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