Uticaj lasera niske snage i elektromagnetnog polja na morfologiju, proliferaciju, diferencijaciju i starenje humanih mezenhimalnih matičnih ćelija izolovanih iz masnog tkiva

Abstract

Poslednjih nekoliko godina otkriveno je da laser niske snage (engl. Low Level Laser, LLL) i elektromagnetno polje (engl. Electromagnetic Field, EMF) utiču na brojne biološke procese, rast i proliferaciju ćelija, a naročito na matične ćelije. Cilj ove studije bio je ispitivanje efekata LLL-a i EMF-a na karakteristike i potencijal humanih mezenhimalnih matičnih ćelija iz masnog tkiva (engl. human Adipose TissueDerived Mesenchymal Stem Cells, hAT-MSCs) i na njihovo prihvatanje nakon eventualne terapijske aplikacije. hAT-MSCs su izolovane iz masnog tkiva 12 osoba starosti 18- 65 godina. LLL tretman se primenjivao tokom 7 dana, jednom dnevno u različitom trajanju, u nekoliko različitih doza od 1 J/cm2, 3 J/cm2, 5 J/cm2 and 10 J/cm2, sa talasnom dužinom od 808 nm, snagom od 200 mW i gustinom energije od 0.2 W/cm2. EMF tretman se, takođe, primenjivao u trajanju od 7 dana, jednom dnevno u različitom trajanju, preko magnetnih jastučića, frekvencije od 50 Hz i različitim intenzitetom od 1 mT, 3 mT, 5 mT i 10 mT. Kontrolna grupa ćelija kultivisana je u istim uslovima, ali bez tretmana fizikalnim agensima. Sedam dana nakon tretmana utvrđene su pojedinačne optimalne doze LLL-a i EMF-a i ispitana je vijabilnost, morfologija, proliferacija, diferencijacija, starenje i oksidativni stres tretiranih ćelija. Rezultati su pokazali da je nakon 7 dana broj hAT-MSCs tretiranih EMF-om bio znatno veći od broja kontrolnih, netretiranih ćelija, hAT-MSCs tretirane LLL-om su bile brojnije od onih tretiranih EMF-om, a da su hAT-MSCs koje su stimulisane kombinovano LLL-om i EMF-om bile najbrojnije. LLL i/ili EMF tretman nije značajno uticao na vijabilnost tretiranih ćelija. Isti pozitivni efekti su registrovani i kada se ispitala diferencijacija. LLL i/ili EMF tretman nije uticao na starenje, niti je povećao oksidativni stres ćelija. Takođe su primećene promene u morfologiji ćelija u smislu povećanja površine ćelija i fraktalne dimenzije u grupama koje su tretirane EMF-om i kombinacijom EMF-a i LLL-a. U zaključku, LLL i/ili EMF tretman je ubrzao proliferaciju i poboljšao diferencijaciju hAT-MSCs, bez ugrožavanja njihove održivosti, bez uticaja na starenje i oksidativni stres, te predstavlja terapijski potencijal u ćelijskom inženjeringu.

In recent years, electromagnetic field (EMF) and low-level laser (LLL) have been found to affect various biological processes, the growth and proliferation of cells, and especially that of stem cells. The aim of this study was to investigate the effects of EMF and LLL on the characteristics and abilities of human adipose tissue-derived mesenchymal stem cells (hATMSCs) and thus to examine the impact of these therapeutic physical modalities on stem cell engraftment. hAT-MSCs were isolated from subcutaneous adipose tissue of 12 persons ranging in age from 18 to 65 years. LLL was applied also for 7 days, for varying periods of time, at different radiation energies of 1 J/cm2, 3 J/cm2, 5 J/cm2 and 10 J/cm2, with a wavelength of 808 nm, power output of 200 mW, and power density of 0.2 W/cm2. EMF was applied for a period of 7 days, once a day for varying periods of time, via a magnetic cushion surface at a frequency of 50 Hz and an different intensity of 1 mT, 3 mT, 5 mT and 10 mT. Nonexposed cells (control) were cultivated under the same culture conditions. Seven days after treatment, optimum doses of LLL and EMF are determined, and the cells were examined for cell viability, morphology, proliferation, differentiation, aging and oxidative stress. We found that after 7 days, the number of EMF-treated hATMSCs was significantly higher than the number of the untreated cells, LLL-treated hAT-MSCs were more numerous than EMFtreated cells, and hAT-MSCs that were treated with the combination of EMF and LLL were the most numerous. EMF and/or LLL treatment did not significantly affect hAT-MSC viability by itself. The same positive effects were registered also when differentiation was examined. LLL and/or EMF treatment did not affect aging and oxidative stress of hAT-MSCs. Changes in cell morphology were also observed, in terms of an increase in cell surface area and fractal dimension in hAT-MSCs treated with EMF and the combination of EMF and LLL. In conclusion, LLL and/or EMF treatment accelerated the proliferation and enhance differentiation of hAT-MSCs without compromising their viability, without affecting aging and oxidative stress, and therefore, they may be used in stem cell tissue engineering.