Effect of electrical stimulation on chondrogenic differentiation of mesenchymal stem cells cultured in hyaluronic acid - Gelatin injectable hydrogels

Electrical stimulation (ES) has provided enhanced chondrogenesis of mesenchymal stem cells (MSCs) cultured in micro-mass without the addition of exogenous growth factors. In this study, we demonstrate for the first time that ES of MSCs encapsulated in an injectable hyaluronic acid (HA) - gelatin (GEL) mixture enhances the chondrogenic potential of the hydrogel. Samples were stimulated for 21 days with 10 mV/cm at 60 kHz, applied for 30 min every 6 h a day. Mechanical properties of hydrogels were higher if the precursors were dissolved in Calcium-Free Krebs Ringer Buffer (G' = 1141 +/- 23 Pa) compared to those diluted in culture media (G' = 213 +/- 19 Pa). Cells within stimulated hydrogels were rounder (55%) than non-stimulated cultures (32%) (p = 0.005). Chondrogenic markers such as SOX-9 and aggrecan were higher in stimulated hydrogels compared to controls. The ES demonstrated that normalized content of glycosaminoglycans and collagen to DNA was slightly higher in stimulated samples. Additionally, collagen type II normalized to total collagen was 2.43 times higher in stimulated hydrogels. These findings make ES a promising tool for enhancing articular cartilage tissue engineering outcomes by combining hydrogels and MSCs. (C) 2020 Elsevier B.V. All rights reserved.