Influence of bio-electrospraying voltage (a), flow rate (b) and distance (c) on viability of MSC. Number of recovered MSC after BES (d) (n = 4). (*P < 0.05 and ***P < 0.001 using ANOVA followed by Tukey post-test)
Influence of bio-electrospraying time on viability of cells. (n = 4) No significant differences were observed using Anova followed by Tukey post test.
Morphology of bio-electrosprayed mesenchymal stem cells after 1 day of culture: control group (a) and 15 min (b), 30 min (c), and 60 min (d) of BES. Scale bars represent 1 μm. The images were taken using 200× magnification.
MTT assay showing no differences in the metabolism/proliferation between the control group and the bio-electrosprayed MSC after 1, 7, and 15 days in culture. The bio-electrosprayed MSC showed a gradual and significant increase of viable cells over 15 days. (n = 5) (***P < 0.001 using two-way ANOVA)
Chondrogenic, adipogenic and osteogenic differentiation of MSC after staining with specific dyes for each differentiation protocol. The results showed no differences in the plasticity of bio-electrosprayed MSC at any time during analysis, compared to the positive control group (n = 3). Black arrows indicate fat deposits. Scale bars represent 1 μm. The pictures were taken using 400× of magnification.
DNA damage evaluation in MSC after 15, 30, and 60 min of BES (n = 3). Cultured cells for 1 h before performing the comet assay (a), cultured cells for 3 h before performing the comet assay (b) and cultured cells for 5 h before performing the comet assay (c). Comparison between time of culture before performing the comet assay (d). The DI reduction suggests DNA repair. (P < 0,05, **P < 0,01 and ***P < 0,001 using ANOVA followed by Tukey post-test)
Immunophenotypic profile of bio-electrosprayed MSC (n = 3). Results less than 0.1% were considered 0%.
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