Antisenescence effect of REAC biomodulation to counteract the evolution of myelodysplastic syndrome

Abstract

About 30 percent of patients diagnosed with myelodysplastic syndromes (MDS) progress to acute myeloid leukemia (AML). The senescence of bone marrow?derived mesenchymal stem cells (BMSCs) seems to be one of the determining factors in inducing this drift. Research is continuously looking for new methodologies and technologies that can use bioelectric signals to act on senescence and cell differentiation towards the phenotype of interest. The Radio Electric Asymmetric Conveyer (REAC) technology, aimed at reorganizing the endogenous bioelectric activity, has already shown to be able to determine direct cell reprogramming effects and counteract the senescence mechanisms in stem cells. Aim of the present study was to prove if the anti-senescence results previously obtained in different kind of stem cells with the REAC Tissue optimization - regenerative (TO-RGN) treatment, could also be observed in BMSCs, evaluating cell viability, telomerase activity, p19ARF, P21, P53, and hTERT gene expression. The results show that the REAC TO-RGN treatment may be a useful tool to counteract the BMSCs senescence which can be the basis of AML drift. Nevertheless, further clinical studies on humans are needed to confirm this hypothesis.

Fig. 1

Increase of viability in REAC TO-RGN-treated cells. After plating (1 × 10⁶ cells/plate), the cells were cultured in the absence or presence of REAC TO-RGN treatment for 24, 48, and 72h. Then, BMSCs were collected and counted using an automatic cell counter. The percentage of vital cells was calculated as the number of positive cells divided by the total number of counted cells (mean ± SE; n = 6) (* p≤0.05).

Fig. 2

A) Effect of REAC TO-RGN in increasing the level of expression of TERT in the REAC TO-RGN-treated cells compared with the untreated control cells. For each timepoint, the mRNA level of the cells exposed to REAC TO-RGN treatment was significantly upregulated compared with the untreated controls (mean ± SE; n=6) (* p≤0.05) B) REAC TO-RGN treatment increases telomerase activity. The REAC TO-RGN-treated cells showed an increase in telomerase activity compared with the untreated control cells for each time of observation (mean ± SE; n=6; *p≤0.05; **p≤0.001).

Fig. 3

Gene expression of p19ARF (Panel A), p21 (Panel B), and p53 (Panel C) in BMSCs that were treated or not treated with REAC TO-RGN for 24, 48, or 72h. The expression of each gene increased for each time point in the untreated control cells, while it was significantly downregulated in the REAC TO-RGN-treated cells (mean ± SD; n=6) (* p≤0.05; ** p≤0.01).

Fig. 4

Effect of REAC TO-RGN in increasing the level of expression of Oct-4 (Panel A), Sox2 (Panel B), and NANOG (Panel C) in the REAC TO-RGN-treated cells compared with the untreated control cells. For each timepoint, the mRNA level of the cells exposed to REAC TO-RGN treatment was significantly upregulated compared with the untreated controls (mean ± SD; n=6) (* p≤0.05, **p≤0.001, ***p≤0.001).

Fig. 5

A) Senescence-associated β-galactosidase activity. B-galactosidase was evaluated in BMSCs treated or not with REAC for 72h. Blue arrows indicate the blue-positive cells. Scale bar=100 μm. B) The number of blue positive cells was calculated using ImageJ. Data are expressed as mean ± SD (n=6) (**p≤0,001).