Tissues engineering has gained a lot of interest, since this approach has potential to restore lost tooth-supporting structures, which is one of the biggest challenges for periodontal treatment. In this study, we aimed to develop an in situ hydrogel that could conceivably support and promote the regeneration of lost periodontal tissues. The hydrogel was fabricated from methacrylated hyaluronic acid (MeHA). Fragment/short-chain hyaluronic acid (sHA) was incorporated in this hydrogel to encourage the bio-synergistic effects of two different molecular weights of hyaluronic acid. The physical properties of the hydrogel system, including gelation time, mechanical profile, swelling and degrading behavior, etc., were tested to assess the effect of incorporated sHA. Additionally, the biological properties of the hydrogels were performed in both in vitro and in vivo models. The results revealed that sHA slightly interfered with some behaviors of networking systems; however, the overall properties were not significantly changed compared to the base MeHA hydrogel. In addition, all hydrogel formulations were found to be compatible with oral tissues in both in vitro and in vivo models. Therefore, this HA-based hydrogel could be a promising delivery system for low molecular weight macromolecules. Further, this approach could be translated into the clinical applications for dental tissue regeneration.

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