Fullerene mediates proliferation and cardiomyogenic differentiation of adipose-derived stem cells via modulation of MAPK pathway and cardiac protein expression

Abstract

Zero-dimensional fullerenes can modulate the biological behavior of a variety of cell lines. However, the effects and molecular mechanisms of proliferation and cardiomyogenic differentiation in brown adipose-derived stem cells (BADSCs) are still unclear. In this study, we report the initial biological effects of fullerene-C₆₀ on BADSCs at different concentrations. Results suggest that fullerene-C₆₀ has no cytotoxic effects on BADSCs even at a concentration of 100 μg/mL. Fullerene-C₆₀ improves the MAPK expression level and stem cell survival, proliferation, and cardiomyogenesis. Further, we found that the fullerene-C₆₀ modulates cardiomyogenic differentiation. Fullerene-C₆₀ improves the expression of cardiomyocyte-specific proteins (cTnT and α-sarcomeric actinin). At elevated concentration, fullerene-C₆₀ reduces the incidence of diminished spontaneous cardiac differentiation of BADSCs with time. At the genetic level, fullerene-C₆₀ (5 μg/mL) also improves the expression of cTnT. In addition, fullerene-C₆₀ promotes the formation of gap junction among cells. These findings have important implications for clinical application of fullerenes in the treatment of myocardial infarction.

Keywords:

• C₆₀
• BADSCs
• molecular mechanisms
• myocardium

Supplementary materials

Figure S1 Flow cytometric analysis of BADSCs.

Note: The cells are positive for CD29 and CD90 for characteristic of mesenchymal stem cells, but negative for CD34, CD45, and CD133.

Abbreviations: BADSCs, brown adipose-derived stem cells; FITC, fluorescein isothiocyanate; M1, number of positive cells.

Figure S2 Alizarin Red staining images of BADSCs cultured without (A) or with (B) osteogenic-inducing microenvironment at day 14.

Abbreviation: BADSCs, brown adipose-derived stem cells.

Figure S3 Oil Red O staining images of BADSCs cultured without (A) or with (B) adipogenic-inducing microenvironment at day 7.

Abbreviation: BADSCs, brown adipose-derived stem cells.

Figure S4 The TEM images of fullerene-C₆₀ nanoparticles.

Notes: (A) Morphology image of fullerene-C₆₀. (B) The zoom of selected area in (A).

Abbreviation: TEM, transmission electron microscopy.

Acknowledgments

This work was supported by the National Natural Science Funds for Distinguished Young Scholar (No 31125013), National Key Basic Research and Development Program of China (No 2011CB606206), International Cooperation and Exchanges NSFC (No 31320103914), Key Program of National Natural Science Foundation of China (No 31030032), National Natural Science Foundation of China (No 31370975, No 31200733, No 31100674, No 31370987, No 31470940), and National High Technology Research and Development Program of China (No SS2015AA020304).

Disclosure

The authors report no conflicts of interest in this work.