Electrical stimulation confers pre-chondrogenic differentiation by modulating TGF-beta 1 levels in canine adipose-derived mesenchymal stem cells

Background \u0026 Aim Background Osteoarthritis (OA) is closely related to the extent of defective articular cartilage, and the targeting of this loss of chondrocytes has emerged as a promising therapeutic strategy. Aim Here, we investigate that electrical stimulation (ES) can drive direct reprogramming of adipose mesenchymal stem cells (MSCs) into hyaline chondrogenic cells. Methods, Results \u0026 Conclusion Methods Canine primary adipose-derived mesenchymal stem cells were formed in micromass for 90 min and stimulated under the electrical field for 3 days. Enzymatic digested ES mass and their 3D control were analyzed through western blot and flow cytometry using chondrogenic markers. In addition, secreted TGF-β1 and TGF-β1 inhibition assay were performed in ES samples. Results We found that TGF-beta1, a key requirement for the in vitro chondrogenic differentiation of MSCs, is highly expressed in micromass of MSCs under ES. TGF-beta1 is also an important regulator of specific cartilage genes during chondrogenesis. Notably, Sox9, a key transcriptional regulator of chondrogenesis, distinctively upregulated in ES-micromass in TGF-beta1 dependent manner. Conclusions We demonstrated that ES-micromass showed the features closely resembling those of chondrocytes in articular cartilage. Further understanding of these insights could serve as the basis for broadening the electrotherapeutic scope for cartilage repair using MSCs.