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

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 (475%). 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.

The role of cell surface markers and enamel matrix derivatives on human periodontal ligament mesenchymal progenitor responses in vitro

“…Our data provide evidence that the cell division induced by EMD or rp(H)M180 is associated with an increase in the percentage of CD146(+) cells in the cultures. However, the increase in the number of CD146(+) hPDL cells was also reported for non-mitogenic molecules BMP-4 [59] and BMP2/7, suggesting that the regulation of CD146 expression may occur independent of cell division. Interestingly, treatment with rp(H)M180, rhBMP2/7 and EMD unbalanced the percentage of dividing cells in favor of CD146(+) subset rather than to favor their CD146(−) counterpart.…”

Vitamin C treatment promotes mesenchymal stem cell sheet formation and tissue regeneration by elevating telomerase activity

“…TERT was associated with highly osteogenic PDLSC clones (Sununliganon and Singhatanadgit, 2011). Furthermore, the combined ectopic expression of BMP4 and hTERT significantly enhanced the multipotent differentiation efficiency and capacity of human PDL fibroblasts, as shown by osteogenic, adipogenic and neurogenic differentiation in vitro, and cementum/PDL-like tissue regeneration in vivo (Mi et al, 2011). In consistent with previous reports, we found Vc increased hTERT activity, which might up-regulate COL I, integrin β1, fibronectin, Oct4, Sox2, Nanog, RUNX2, ALP and OCN, and enhance the proliferation and differentiation efficiency of PDLSCs.…”

Notch Signaling Is Involved in Neurogenic Commitment of Human Periodontal Ligament-Derived Mesenchymal Stem Cells

“…Regarding neuronal differentiation, our group and other investigators have illustrated that hPDLSCs were able to differentiate toward neurogenic lineage under specific conditions [4][5][6][7]. Neurite-like, long cellular processes and upregulation of neurogenic markers [b3-tubulin, nestin, and neurofilament (NF)] were noted after neurogenic induction [4,8].…”

“…These cells were able to differentiate into osteogenic/cementogenic, adipogenic, and neurogenic lineage in vitro [5]. In addition, human periodontal ligament-derived mesenchymal stem cells (hPDLSCs) could also differentiate into other specific cell types such as chondrocytes, cardiomyocytes, and vascular-like cells [6,7]. Upon subcutaneous implantation, hPDLSCs had the potential to generate cementum-periodontal ligament like tissues [1][2][3].…”