Medical/biological study (experimental study)

Synergic effects of extremely low-frequency electromagnetic field and betaine on in vitro osteogenic differentiation of human adipose tissue-derived mesenchymal stem cells med./bio.

By: Tabatabai TS, Haji Ghasem Kashani M, Maskani R, Nasiri M, Nabavi Amri SA, Atashi A, Bitaraf FS

Published in: In Vitro Cell Dev Biol Anim 2021; 57 (4): 468-476

Aim of study (acc. to author)

The effects of (co-)exposure to a 50 Hz magnetic field and betaine on the osteogenic differentiation of human adipose tissue-derived mesenchymal stem cells should be investigated.

Background/further details

Cells were divided into the following groups: 1) exposure to the magnetic field, 2) exposure to betaine (10 mM), 3) co-exposure to the magnetic field and betaine, 4) control group (only treatment with osteogenic differentiation medium), 5) negative control (no treatment). Groups 1-4 were treated with osteogenic differentiation medium. Betaine could be a promising therapeutic agent for osteoporosis as it stimulates different signaling pathways leading to enhanced bone formation.

Endpoint

cell viability/cell division/proliferation: osteogenic differentiation

Exposure

- 50/60 Hz
- magnetic field
- also other exposures without EMF
- co-exposure

Exposure	Parameters
Exposure 1: 50 Hz Exposure duration: 8 hours/day for up to 21 days	magnetic flux density: 1 mT

Exposure 1

Main characteristics

Frequency	50 Hz
Type	magnetic field
Waveform	sinusoidal
Exposure duration	8 hours/day for up to 21 days

Exposure setup

Exposure source	coil(s)
Setup	a uniform magnetic field was produced by a coil with dimensions of 68 x 100 mm², corresponding to the dimensions of the flask used for exposure of cells; the coil was made of 2200 turns of 0.4 mm diameter enameled wire wound around a plastic spool; a magnetic silicon alloy core was placed into the spool; to prevent heat from the eddy current, the magnetic core was made of a large number of very thin EI-shaped sheets; the whole coil was sealed with an air-during varnish; a 24-well plate cover with dimensions of 85 × 128 mm² was placed on the coil as the cell flask holder

Exposure source

coil(s)

Setup

a uniform magnetic field was produced by a coil with dimensions of 68 x 100 mm², corresponding to the dimensions of the flask used for exposure of cells; the coil was made of 2200 turns of 0.4 mm diameter enameled wire wound around a plastic spool; a magnetic silicon alloy core was placed into the spool; to prevent heat from the eddy current, the magnetic core was made of a large number of very thin EI-shaped sheets; the whole coil was sealed with an air-during varnish; a 24-well plate cover with dimensions of 85 × 128 mm² was placed on the coil as the cell flask holder

Parameters

Measurand	Value	Type	Method	Mass	Remarks
magnetic flux density	1 mT	-	measured	-	-

Reference articles

Luo F et al. (2012): Effects of pulsed electromagnetic field frequencies on the osteogenic differentiation of human mesenchymal stem cells

Exposed system:

intact cell/cell culture
human adipose tissue-derived mesenchymal stem cells

Methods Endpoint/measurement parameters/methodology

cell viability/cell division/proliferation: cell morphology and identification of mineralized matrix (Alizarin Red S stain, inverse microscopy) and calcium quantification (spectrophotometry) after 21 days, enzyme activity of alkaline phosphatase after 7 and 14 days (spectrophotometry)
molecular biosynthesis: gene expression of osteogenic differentiation related genes RUNX2 and OCN after 21 days (quantitative real-time RT-PCR)

Investigated system:

intact cell/cell culture
isolated bio./chem. substance
DNA/RNA
cell lysates and homogenates

Time of investigation:

after exposure

Main outcome of study (acc. to author)

A significant increase in calcium deposition was observed in the co-exposure group (group 3) compared to the other groups. The enzyme activity of alkaline phosphatase was significantly decreased in the magnetic field (group 1) and betaine (group 2) groups compared to the control group and but it was significantly higher in group 3 compared to group 1. The gene expression of RUNX2 and OCN was significantly reduced in group 1 compared to the control group but group 3 showed a significant increase in RUNX2 gene expression compared to group 1.
The authors concluded that co-exposure to a 50 Hz magnetic field and betaine might have synergistic effects on the osteogenic differentiation of human adipose tissue-derived mesenchymal stem cells.

Study character:

medical/biological study
experimental study
full/main study

Study funded by

Damghan University, Iran

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Ferroni L et al. (2016): Pulsed magnetic therapy increases osteogenic differentiation of mesenchymal stem cells only if they are pre-committed
Ardeshirylajimi A et al. (2015): Enhanced growth and osteogenic differentiation of Induced Pluripotent Stem cells by Extremely Low-Frequency Electromagnetic Field
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