Improvement of the Digestibility of Sulfated Hyaluronans by Bovine Testicular Hyaluronidase: A UV Spectroscopic and Mass Spectrometric Study

Abstract

Glycosaminoglycans (GAGs) such as hyaluronan (HA) and chondroitin sulfate (CS) are important, natural polysaccharides which occur in biological (connective) tissues and have various biotechnological and medical applications. Additionally, there is increasing evidence that chemically (over)sulfated GAGs possess promising properties and are useful as implant coatings. Unfortunately, a detailed characterization of these GAGs is challenging: although mass spectrometry (MS) is one of the most powerful tools to elucidate the structures of (poly)saccharides, MS is not applicable to high mass polysaccharides, but characteristic oligosaccharides are needed. These oligosaccharides are normally generated by enzymatic digestion. However, chemically modified (particularly sulfated) GAGs are extremely refractive to enzymatic digestion. This study focuses on the investigation of the digestibility of GAGs with different degrees of sulfation by bovine testicular hyaluronidase (BTH). It will be shown by using an adapted spectrophotometric assay that all investigated GAGs can be basically digested if the reaction conditions are carefully adjusted. However, the oligosaccharide yield correlates reciprocally with the number of sulfate residues per polymer repeating unit. Finally, matrix-laser desorption and ionization (MALDI) MS will be used to study the released oligosaccharides and their sulfation patterns.

Collagen/hyaluronan based hydrogels releasing sulfated hyaluronan improve dermal wound healing in diabetic mice via reducing inflammatory macrophage activity

“…In this respect, the use of sHA in immunomodulating hydrogels is a smart way to circumvent such undesired effects. In addition, sHA is hardly digested by HYAL and thus remains in its active form in the inflamed tissue [ 94 ].…”

Hyaluronan/Collagen Hydrogels with Sulfated Hyaluronan for Improved Repair of Vascularized Tissue Tune the Binding of Proteins and Promote Endothelial Cell Growth

“…The biodegradability is an important aspect for a future medical application of biomaterials. Since the degradation of HA via HYAL decreases with increasing degree of sulfation and HEP and CS are suggested to contribute to the regulation of the endoglycosidase activity of HYAL, the influence of solute sGAGs on the degradation of immobilized native HA via HYAL as well as the degradation profiles of the gels was studied in detail ( Figure 5 ). HA was degraded in a time‐dependent manner by HYAL.…”

Bioinspired Collagen/Glycosaminoglycan-Based Cellular Microenvironments for Tuning Osteoclastogenesis

“…For chemically modified HA derivatives no antibodies are available and due to the fact that the degradation via hyaluronidases is insufficient no fragment based discrimination is possible 44 . Also the standard assays for sulfated GAGs are not able to discriminate between these GAGs 35,36 .…”