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Highly efficient multipotent differentiation of human periodontal ligament fibroblasts induced by combined BMP4 and hTERT gene transfer

Gene Therapy volume 18, pages 452–461 (2011)Cite this article

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Abstract

Because periodontal ligament (PDL) cells are reported to contain progenitor or stem cell populations, they are considered a beneficial cell source for clinical periodontal regeneration. Both bone morphogenetic protein 4 (BMP4) and human telomerase reverse transcriptase (hTERT) have essential roles in the modulation of stem cell properties. In this study we report for the first time that the combined ectopic expression of BMP4 and hTERT significantly enhanced the multipotent differentiation efficiency and capacity of human PDL fibroblasts (PFs), as shown by osteogenic, adipogenic and neurogenic differentiation in vitro, and cementum/PDL-like tissue regeneration in vivo. These findings may be attributed, at least in part, to the original upregulation of important stem cell markers, such as scleraxis, Stro-1 and CD146, and the extremely lowered threshold for BMP concentration to activate BMP signaling by enhanced basal phosphorylation levels of Smad 1/5/8. In addition, the significantly reduced expression levels of CD146 and CD90 with the presence of Noggin confirms the direct effect of BMP4 on the stem cell-like phenotype of genetically modified PF cells (BT-PFs). Furthermore, BT-PFs exhibited a high neural differentiation capacity (>75%). After transplantation into NOD/SCID mice, genetically modified-PFs generated cementum/PDL-like structures on the surface of the carrier. The multipotency of these modified cells potentially provides an attractive source of stem cells for therapeutic purposes and regenerative medicine.

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Abbreviations

oDM:

osteogenic differentiation medium

aDM:

adipogenic differentiation medium

nDM:

neuronal differentiation medium

CM:

regular medium

Col I :

type I collagen

POSTN :

periostin

ALP :

alkaline phosphatase

BSP :

bone sialoprotein

LPL :

lipoprotein lipase

PPARγ2 :

peroxisome proliferator-activated receptor γ2

GFAP :

glial fibrillary acidic protein

MAP :

microtubule-associated protein 2

OPN :

osteopontin

MSC:

mesenchymal stem cell

RT-PCR:

reverse transcriptase PCR

HA/TCP:

hydroxyapatite/tricalcium phosphate

PDLSC:

PDL stem cell

DMEM:

Dulbecco’s modified Eagle’s medium

FBS:

fetal bovine serum

cDNA:

complementary DNA

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Acknowledgements

This study was supported in part by grants from the Taiwan National Health Research Institutes (NHRI) and grants from the National Taiwan University, School of Dentistry and Graduate Institute of Clinical Dentistry.

Author ContributionHsin-Wu Mi: conception and design, collection and/or assembly of data and manuscript writing; Ming-Cheng Lee: provision of study material, collection and/or assembly of data; Earl Fu: data analysis and interpretation and manuscript writing; Lu-Ping Chow: data analysis and interpretation and manuscript writing; and Chun-Pin Lin: financial support, data analysis and interpretation and manuscript writing.

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Authors and Affiliations

  1. Graduate Institute of Clinical Dentistry, School of Dentistry, National Taiwan University and National Taiwan University Hospital, Taipei, Taiwan

    H-W Mi & C-P Lin

  2. Department of Internal Medicine, National Taiwan University Hospital, Taipei, Taiwan

    M-C Lee

  3. Department of Periodontology, School of Dentistry, Tri-Service General Hospital, National Defense University, Taipei, Taiwan

    E Fu

  4. Graduate Institute of Biochemistry and Molecular Biology, College of Medicine, National Taiwan University, Taipei, Taiwan

    L-P Chow

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Correspondence to C-P Lin.

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Mi, HW., Lee, MC., Fu, E. et al. Highly efficient multipotent differentiation of human periodontal ligament fibroblasts induced by combined BMP4 and hTERT gene transfer. Gene Ther 18, 452–461 (2011). https://doi.org/10.1038/gt.2010.158

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