Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2023 Sep;56(9):1092-1107.
doi: 10.1111/iej.13943. Epub 2023 Jun 20.

Direct contact with endothelial cells drives dental pulp stem cells toward smooth muscle cells differentiation via TGF-β1 secretion

Yuchen Zhang  1 , Jialin Zhong  1 , Shulan Lin  1 , Mingxin Hu  1 , Junqing Liu  1 , Jun Kang  1 , Yubingqing Qi  1 , Mohammed S Basabrain  1 , Ting Zou  1 , Chengfei Zhang  1
Affiliations

Direct contact with endothelial cells drives dental pulp stem cells toward smooth muscle cells differentiation via TGF-β1 secretion

Yuchen Zhang et al. Int Endod J. 2023 Sep.

Abstract

Aim: Prevascularization is vital to accelerate functional blood circulation establishment in transplanted engineered tissue constructs. Mesenchymal stem cells (MSCs) or mural cells could promote the survival of implanted endothelial cells (ECs) and enhance the stabilization of newly formed blood vessels. However, the dynamic cell-cell interactions between MSCs, mural cells and ECs in the angiogenic processes remain unclear. This study aimed to explore the interactions of human umbilical vein ECs (HUVECs) and dental pulp stem cells (DPSCs) in an in vitro cell coculture model.

Methodology: Human umbilical vascular ECs and DPSCs were directly cocultured or indirectly cocultured with transwell inserts in endothelial basal media-2 (EBM-2) supplemented with 5% FBS for 6 days. Expression of SMC-specific markers in DPSCs monoculture and HUVEC+DPSC cocultures was assessed by western blot and immunofluorescence. Activin A and transforming growth factor-beta 1 (TGF-β1) in conditioned media (CM) of HUVECs monoculture (E-CM), DPSCs monoculture (D-CM) and HUVEC+DPSC cocultures (E+D-CM) were analysed by enzyme-linked immunosorbent assay. TGF-β RI kinase inhibitor VI, SB431542, was used to block TGF-β1/ALK5 signalling in DPSCs.

Results: The expression of SMC-specific markers, α-SMA, SM22α and Calponin, were markedly increased in HUVEC+DPSC direct cocultures compared to that in DPSCs monoculture, while no differences were demonstrated between HUVEC+DPSC indirect cocultures and DPSCs monoculture. E+D-CM significantly upregulated the expression of SMC-specific markers in DPSCs compared to E-CM and D-CM. Activin A and TGF-β1 were considerably higher in E+D-CM than that in D-CM, with upregulated Smad2 phosphorylation in HUVEC+DPSC cocultures. Treatment with activin A did not change the expression of SMC-specific markers in DPSCs, while treatment with TGF-β1 significantly enhanced these markers' expression in DPSCs. In addition, blocking TGF-β1/ALK5 signalling inhibited the expression of α-SMA, SM22α and Calponin in DPSCs.

Conclusions: TGF-β1 was responsible for DPSC differentiation into SMCs in HUVEC+DPSC cocultures, and TGF-β1/ALK5 signalling pathway played a vital role in this process.

Keywords: TGF-β1; dental pulp stem cells; differentiation; smooth muscle cells.

PubMed Disclaimer

Similar articles

See all similar articles

Cited by

References

REFERENCES

    1. Annes, J.P., Munger, J.S. & Rifkin, D.B. (2003) Making sense of latent TGFβ activation. Journal of Cell Science, 116(2), 217-224.
    1. Asahara, T. & Isner, J.M. (2002) Endothelial progenitor cells for vascular regeneration. Journal of Hematotherapy & Stem Cell Research, 11(2), 171-178.
    1. Atkins, S.K., Sonawane, A.R., Brouwhuis, R., Barrientos, J., Ha, A., Rogers, M. et al. (2022) Induced pluripotent stem cell-derived smooth muscle cells to study cardiovascular calcification. Frontiers in Cardiovascular Medicine, 9, 925777.
    1. Barbash, I.M., Chouraqui, P., Baron, J., Feinberg, M.S., Etzion, S., Tessone, A. et al. (2003) Systemic delivery of bone marrow-derived mesenchymal stem cells to the infarcted myocardium: feasibility, cell migration, and body distribution. Circulation, 108(7), 863-868.
    1. Beamish, J.A., Geyer, L.C., Haq-Siddiqi, N.A., Kottke-Marchant, K. & Marchant, R.E. (2009) The effects of heparin releasing hydrogels on vascular smooth muscle cell phenotype. Biomaterials, 30(31), 6286-6294.

Substances

LinkOut - more resources