High molecular weight hyaluronic acid (HMW-HA) and low molecular weight hyaluronic acid (LMW-HA) were mixed at different ratios and cross-linked with 1,4-butanediol diglycidyl ether (BDDE) to prepare five hyaluronic acid hydrogels A–E. Enzymolysis stability, swelling rate, crosslinking degree, rheological characteristics, BDDE residual rate, surface microstructure, and cytotoxicity of different hydrogels were investigated. The results showed that hydrogel B obtained by 10% HA (w/v, HMW-HA and LMW-HA having a mass ratio of 4 : 1) crosslinking with 1% BDDE (v/v) had stronger in vitro anti-degradation ability, better mechanical properties and lower cytotoxicity than those prepared by mixing in different proportions. Hydrogel B has potential applications in regenerative medicine and tissue engineering.

Hyaluronic acid (HA) was depolymerized by a combination of ultrasound, hydrogen peroxide and copper ion. The structures of high-molecular-weight hyaluronic acid (HMW-HA) and low-molecular-weight hyaluronic acid (LMW-HA) were determined by Fourier transform infrared (FTIR) spectroscopy, circular dichroism (CD) spectroscopy, and UV-VIS absorption spectroscopy. The degradations of HMW-HA using a physical method, a chemical method, and a combination of physical and chemical method were compared. The results show that HA can be effectively degraded by a combinatorial method involving ultrasound, hydrogen peroxide, and copper ion. Under the degradation conditions of 50 mM H₂O₂, 5.0 μM CuCl₂, 160 W, pH 4.0, and reaction at 50 °C for 30 min, the content of glucuronic acid was 36.56%, and the yield of LMW-HA was 81.71%. The FTIR, CD, and UV-VIS absorption spectra of HA did not change with the decrease in molecular weight, indicating that the structure of HA remained intact during the degradation.