Introduction and Aims. Undifferentiated mesenchymal stem cells (MSCs) from bone marrow express multiple functional K^+ channel currents and in a less extent Na^+ and L-type Ca^2+ current. These two latter currents increase in expression with MSCs proliferation and differentiation. Low laser intensity (LLL) therapy is described in the literature as causing increases in the amount of DNA synthesis, osteoclasts proliferation in bone defects tibia and osteoporosis. The aim of this study was to characterize the effects of LLL stimulation on functional K^+ and Ca^2+ channel currents in cultured MSCs. Material and methods. Mouse bone marrow mesenchymal stromal cells (MSCs) were isolated from femura and tibiae of male C2F1 mice . MSCs were cultured in growth medium (DMEM containing 1% Glutammine, 1% P/S and 20 % FBS), in the presence or absence of 0.5 mM Ba^2+, for 24 and 48 h. MSCs were stimulated for 10-26 s with a diode laser operating at a wavelength a 632 nm in continuous irradiation mode (power 89 mW; energy density dose of 0.26 J/cm^2; laser output was coupled to a 0.6-mm-diameter optical fiber). Cell proliferation was assayed by cell counting, EdU incorporation assay, and Notch-1 activation. Ionic currents were recorded with the whole-cell patch clamp technique. Experiments were made before (time 0) and 24 or 48 hours after laser treatment. Results. Laser treatment increased significantly MSC proliferation and up-regulated Notch-1 expression. These effects were paralleled by a reduced value of the cell membrane capacitance, index of a reduced surface membrane area, and an increase of the specific resting membrane conductance. Laser stimulated cells showed significant increase of K(Ca), Kir and T- and L-type Ca^2+ currents. Notably, the above changes were reduced in cells pretreated with Ba^2+. Conclusions. Taken together, these results, suggest a role for diode laser in the stimulation of MSC and underscore novel intriguing mechanisms responsible for laser biostimulation
Effects of diode laser 635 nm stimulation on K+ and Ca2+ channel currents in mesenchymal stem/stromal cells isolated from mouse bone marrow / F. Francini; C. Sassoli;D. Nosi;F. Chellini;M. Giannelli;R. Squecco. - ELETTRONICO. - (2012), pp. 1-1. (Intervento presentato al convegno 3rd Disputationes on Native and Induced Pluripotent Stem Cell Standardization tenutosi a Florence, Italy nel March 19–21, 2012).
Effects of diode laser 635 nm stimulation on K+ and Ca2+ channel currents in mesenchymal stem/stromal cells isolated from mouse bone marrow
FRANCINI, FABIO;SASSOLI, CHIARA;NOSI, DANIELE;CHELLINI, FLAMINIA;SQUECCO, ROBERTA
2012
Abstract
Introduction and Aims. Undifferentiated mesenchymal stem cells (MSCs) from bone marrow express multiple functional K^+ channel currents and in a less extent Na^+ and L-type Ca^2+ current. These two latter currents increase in expression with MSCs proliferation and differentiation. Low laser intensity (LLL) therapy is described in the literature as causing increases in the amount of DNA synthesis, osteoclasts proliferation in bone defects tibia and osteoporosis. The aim of this study was to characterize the effects of LLL stimulation on functional K^+ and Ca^2+ channel currents in cultured MSCs. Material and methods. Mouse bone marrow mesenchymal stromal cells (MSCs) were isolated from femura and tibiae of male C2F1 mice . MSCs were cultured in growth medium (DMEM containing 1% Glutammine, 1% P/S and 20 % FBS), in the presence or absence of 0.5 mM Ba^2+, for 24 and 48 h. MSCs were stimulated for 10-26 s with a diode laser operating at a wavelength a 632 nm in continuous irradiation mode (power 89 mW; energy density dose of 0.26 J/cm^2; laser output was coupled to a 0.6-mm-diameter optical fiber). Cell proliferation was assayed by cell counting, EdU incorporation assay, and Notch-1 activation. Ionic currents were recorded with the whole-cell patch clamp technique. Experiments were made before (time 0) and 24 or 48 hours after laser treatment. Results. Laser treatment increased significantly MSC proliferation and up-regulated Notch-1 expression. These effects were paralleled by a reduced value of the cell membrane capacitance, index of a reduced surface membrane area, and an increase of the specific resting membrane conductance. Laser stimulated cells showed significant increase of K(Ca), Kir and T- and L-type Ca^2+ currents. Notably, the above changes were reduced in cells pretreated with Ba^2+. Conclusions. Taken together, these results, suggest a role for diode laser in the stimulation of MSC and underscore novel intriguing mechanisms responsible for laser biostimulation
I documenti in FLORE sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
