Role of dexamethasone in the long‐term functional maturation of MSC‐laden hyaluronic acid hydrogels for cartilage tissue engineering

ABSTRACT The purpose of study was to investigate the maturation of mesenchymal stem cells (MSC) laden in HA constructs with various combinations of chemically defined medium (CM) components and determine the impact of dexamethasone and serum on construct properties. Constructs were cultured in CM with the addition or withdrawal of media components or were transferred to serum containing media that partially represents an in vivo‐like condition where pro‐inflammatory signals are present. Constructs cultured in CM+ (CM with TGF‐β3) and DEX− (CM+ without dexamethasone) conditions produced robust matrix, while those in ITS/BSA/LA− (CM+ without ITS/BSA/LA) and Serum+ (10% FBS with TGF‐β3) produced little matrix. While construct properties in DEX− were greater than those in CM+ at 4 weeks, properties in CM+ and DEX− reversed by 8 weeks. While construct properties in DEX− were greater than those in CM+ at 4 weeks, the continued absence or removal of dexamethasone resulted in marked GAG loss by 8 weeks. Conversely, the continued presence or new addition of dexamethasone at 4 weeks further improved or maintained construct properties through 8 weeks. Finally, when constructs were converted to Serum (in the continued presence of TGF‐β3 with or without dexamethasone) after pre‐culture in CM+ for 4 weeks, GAG loss was attenuated with addition of dexamethasone. Interestingly, however, collagen content and type was not impacted. In conclusion, dexamethasone influences the functional maturation of MSC‐laden HA constructs, and may help to maintain properties during long‐term culture or with in vivo translation by repressing pro‐inflammatory signals. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:1717–1727, 2018. MSCs were viable only in CM+ and DEX− and maintained their initial shape with robust matrix production. Conversely, cells in ITS/BSA/LA− and Serum+ did not remain viable, and these constructs contracted significantly over the culture period. Constructs cultured in Serum+ contracted very quickly, MSCs appeared to aggregate centrally, and this culminated in the formation of a central pellet‐like structure that separated from the HA hydrogel.