A high-glucose environment induces imbalance of mitochondrial homeostasis, and further result in the decreased autophagic capacity and energy metabolism of mitochondria. This mitochondrial dysfunction inhibits the regeneration of bone tissue. Electrical stimulation (ES) is an efficient strategy to rebalance the mitochondrial homeostasis to further accelerate the bone regeneration process. However, traditional ES strategies are invasive and easily cause secondary trauma, limiting its medical application. Here, we designed a new non-invasive direct-current electric field (DCEF) strategy. This strategy can provide ES in situ without implantation. Results show that this strategy can target and regulate mitochondrial homeostasis through the PI3K-AKT signaling pathway, activating mitochondrial autophagy, alleviate high-glucose-induced mitochondrial damage, and further promote osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). Consequently, this exogenous non-invasive ES strategy can effectively accelerate the repair of bone defects in a high-glucose environment through alleviating mitochondrial damage.