Time-dependent effect of electrical stimulation on osteogenic differentiation of bone mesenchymal stromal cells cultured on conductive nanofibers.

Bone tissue engineering using bone mesenchymal stromal cells (BMSCs) is a multidisciplinary strategy that requires biodegradable scaffold, cell, various promoting cues to work simultaneously. Electrical stimulation (ES) is known able to promote osteogenic differentiation of BMSCs, but it is interesting to know how can it play the strongest promotion effect. To strengthen local ES on BMSCs, parallel-aligned conductive nanofibers were electrospun from the mixtures of poly(L-lactide) (PLLA) and multi-walled carbon nanotubes (MWCNTs), and used for cell culture. Osteogenic differentiation of BMSCs was conducted by applying ES (direct current, 1.5 V, 1.5 h/day) perpendicular to the fiber direction during the day 1-7, day 8-14, or day 15-21 period of the osteoinductive culture. In comparison with ES-free groups, bone-related markers and genes were found significantly up-regulated when ES was applied on BMSCs growing on nanofibers having higher conductivity. When the ES was applied at the earlier stage of osteoinductive culture, the promotion effect on osteogenic differentiation would be stronger. In the presence of a BMP blocker, the down-regulated expressions of bone-related genes were able to be slightly recovered by ES, especially when the ES was applied at the beginning of osteoinductive culture (i.e. day 1-7). The promotion effect generated by ES in the early stage was found sustainable to later stages of differentiation, while those ES applied at later stages of differentiation should have missed the optimal point. In other words, later ES was not so necessary in inducing the osteogenic differentiation of BMSCs. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 3369-3383, 2017.