Poly(N-isopropylacrylamide-co-N-tert-butylacrylamide)- grafted hyaluronan as an injectable and self-assembling scaffold for cartilage tissue engineering

Novel poly(N-isopropylacrylamide-co-N-tert-butylacrylamide)-grafted hyaluronan [P(NIPAAm-co-NtBAAm)g-HA] has been developed as a modified derivative to improve phase-transition characteristics of PNIPAAmg-HA, which has a lower critical solution temperature (LCST) of approximately 32 ̊C. This promising selfassembling biomaterial has potential as an injectable scaffold for in situ cartilage tissue engineering. LCST of the P(NIPAAm-co-NtBAAm)-g-HA decreased to approximately 3.6 ̊C compared to that of the original PNIPAAm-g-HA. This modification enabled self-assembly at body temperatures lower than the temperature of the parental PNIPAAm-g-HA molecule. Cytotoxicity and acute systemic toxicity assays revealed that P(NIPAAm-co-NtBAAm)-g-HA was not hazardous. The DNA content of chondrogenic differentiated mesenchymal stem/stromal cells (MSCs) embedded in the gels was higher than that of biomaterial-free aggregates during the culture periods. Cartilage-related genes were also expressed in chondrogenic differentiated MSCs embedded in the P (NIPAAm-co-NtBAAm)-g-HA hydrogel. Specifically, an increased expression of SRY-related HMG boxcontaining gene 9 (Sox9) observed in the hydrogel group compared to controls. These data suggest that P(NIPAAm-co-NtBAAm)-g-HA is a promising injectable scaffold with thermoresponsive properties suitable for in situ cartilage tissue engineering.