In vitro chondrogenic differentiation of human articular cartilage derived chondroprogenitors using pulsed electromagnetic field

BackgroundThe ability to grow new cartilage remains the standard goal of any treatment strategy directed at cartilage repair. Chondroprogenitors have garnered interest due to their applicability in cell therapy. Pulsed electromagnetic field (PEMF) favors chondrogenesis by possible upregulation of genes belonging to TGFβ superfamily. Since TGFβ is implicated in chondrogenic signalling, the aim of the study was to evaluate the ability of PEMF to induce chondrogenesis via endogenous TGFβ production in chondroprogenitors vs differentiation using chondrogenic medium inclusive of TGFβ.MethodsChondroprogenitors were harvested from three non-diseased human knee joints via fibronectin assay. Passage 3 pellets were subjected to four different culture conditions: a) negative control contained chondrogenic medium without TGFβ2, b) positive control contained medium with TGFβ2, c) PEMF 1 contained medium of negative control plus single exposure to PEMF and d) PEMF 2 contained medium of negative control plus multiple exposures to PEMF. Following differentiation (day 21), pellets were assessed for gene expression of ACAN, SOX9, COL2A1, TGFβ1, TGFβ2, and TGFβ3. Alcian blue staining to detect glycosaminoglycan deposition was also performed. Medium supernatant was used to detect endogenous latent TGF-β1 levels using ELISA.ResultsAll study arms exhibited comparable gene expression without any significant difference. Although positive control and PEMF study arms demonstrated notably better staining than negative control, the level of latent TGF-β1 was seen to be significantly high in supernatant from positive control (P < 0.05) when compared to other groups.ConclusionOur results indicate that PEMF induced chondrogenesis might involve other signalling molecules, which require further evaluation.