Abstract
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.
Highlights
IntroductionHigh-molecular-weight hyaluronic acid (HMW-Hyaluronic acid (HA)) inhibits cell proliferation, migration of vascular endothelial cell, and angiogenesis, whereas low-molecular-weight hyaluronic acid (LMW-HA) with completely different physiological functions promotes the adhesion and proliferation of endothelial cells [3,4,5,6]
Hyaluronic acid (HA) is a linear polysaccharide consisting of two alternating units, β-1,4-D-glucuronic acid and β-1,3-N-acetyl-D-glucosamine; HA is the main component of extracellular matrix [1,2].HA is present in the skin, joints, and cornea at high concentrations.HA is a homogeneous polymer, and the molecular size is widely distributed (105 –107 Da).High-molecular-weight hyaluronic acid (HMW-HA) inhibits cell proliferation, migration of vascular endothelial cell, and angiogenesis, whereas low-molecular-weight hyaluronic acid (LMW-HA) with completely different physiological functions promotes the adhesion and proliferation of endothelial cells [3,4,5,6].The molecular weight of hyaluronic acid usually exceeds 106 Da
Using a system containing H2 O2 plus CuCl2, the dynamic viscosity of the HA solution indicated that the degradation of biopolymer decreased faster than the system containing only H2 O2 [13]
Summary
High-molecular-weight hyaluronic acid (HMW-HA) inhibits cell proliferation, migration of vascular endothelial cell, and angiogenesis, whereas low-molecular-weight hyaluronic acid (LMW-HA) with completely different physiological functions promotes the adhesion and proliferation of endothelial cells [3,4,5,6]. The molecular weight of hyaluronic acid usually exceeds 106 Da. Various methods have been reported to produce LMW-HA, including chemical degradation, physical degradation, and enzymatic degradation [7,8,9]. Methods of chemical degradation mainly include acid hydrolysis, alkaline hydrolysis, and oxidative degradation [10,11,12]. Alkali, and oxidant react with HMW-HA at different reaction sites. Ultrasonication is a common method of physical degradation to obtain LMW-HA
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