The effect of low temperature atmospheric nitrogen plasma on MC3T3-E1 preosteoblast proliferation and differentiation in vitro

Abstract

The aim of this work was to evaluate the impact of atmospheric pressure nitrogen plasma on viability, proliferation, and osteogenic differentiation of normal mouse calvarial preosteoblasts (MC3T3-E1 Subclone 4), which were maintained in Hanks’ balanced salt solution (HBSS) during plasma exposure. Obtained results clearly demonstrated that short-time (4, 8, and 16 s) nitrogen plasma treatment is non-toxic to the MC3T3-E1 cells, does not affect cell morphology, promotes preosteoblasts’ proliferation, enhances osteogenic differentiation by increasing bone alkaline phosphatase and osteocalcin concentration, but inhibits mineralization of extracellular matrix. The best results were achieved for 16 s exposure time and when the preosteoblasts were left in HBSS for 3 h after plasma treatment. Presented studies indicate great clinical potential of cold atmospheric nitrogen plasma for regenerative medicine applications to improve bone healing process.

Positive Effect of Cold Atmospheric Nitrogen Plasma on the Behavior of Mesenchymal Stem Cells Cultured on a Bone Scaffold Containing Iron Oxide-Loaded Silica Nanoparticles Catalyst

“…Furthermore, cells revealed higher viability after 16 s exposure to nitrogen plasma compared to the air plasma. In our another study [ 15 ], it was demonstrated that 16 s nitrogen plasma treatment promoted preosteoblasts proliferation and enhanced their osteogenic differentiation. Importantly, the best results were obtained when MC3T3-E1 preosteoblasts were left in Hanks’ Balanced Salt solution (HBSS) for 3 h after plasma exposure.…”

“…Stem cells-seeded biomaterials were exposed to cold nitrogen plasma for 16 s and left for 3 h in HBSS after plasma treatment. These conditions were demonstrated in our previous studies to be very effective in stimulation of proliferation and ECM synthesis by MC3T3-E1 preosteoblasts [ 15 ].…”

“…In our previous studies performed on MC3T3-E1 preosteoblasts, it was demonstrated that short-time (16 s) nitrogen plasma treatment of the cells significantly promoted cell proliferation and enhanced bALP and OC synthesis [ 15 ]. In this study, nitrogen plasma activation of control biomaterial (without NPs) seeded with ADSCs did not influence neither cell proliferation nor osteogenic differentiation.…”

“…Atmospheric pressure plasma was generated in GAD reactor presented in Figure 9 developed at Lublin University of Technology (LUT), consisting of two copper electrodes (1.5 mm thick, and 10 cm long with 12° angle between them), which has been described in detail in other related publications [ 15 , 48 ]. The smallest inter-electrode distance was 3 mm.…”

“…Since reactive species formed upon plasma treatment may have a positive effect on cell proliferation and differentiation, cold plasma has also attracted great attention in the regenerative medicine field. Possible applications of atmospheric pressure plasma have been reported for acceleration of wound healing [ 1 , 2 , 3 , 4 ] and bone regeneration [ 14 , 15 ]. The effectiveness of plasma technology application in the regenerative medicine field highly depends on the reactor design, the gas nature, the plasma power, and other characteristics of the treatment [ 11 ].…”