The effect of increasing the sulfation level of chondroitin sulfate on anticoagulant specific activity and activation of the kinin system

Abstract

Oversulfated chondroitin sulfate (OSCS) was identified as a contaminant in certain heparin preparations as the cause of adverse reactions in patients. OSCS was found to possess both plasma anticoagulant activity and the ability to activate prekallikrein to kallikrein. Differentially sulfated chondroitin sulfates were prepared by synthetic modification of chondroitin sulfate and were compared to the activity of OSCS purified from contaminated heparin. Whilst chondroitin sulfate was found to have minimal anticoagulant activity, increasing sulfation levels produced an anticoagulant response which we directly show for the first time is mediated through heparin cofactor II. However, the tetra-sulfated preparations did not possess any higher anticoagulant activity than several tri-sulfated variants, and also had lower heparin cofactor II mediated activity. Activation of prekallikrein was concentration dependent for all samples, and broadly increased with the degree of sulfation, though the di-sulfated preparation was able to form more kallikrein than some of the tri-sulfated preparations. The ability of the samples to activate the kinin system, as measured by bradykinin, was observed to be through kallikrein generation. These results show that whilst an increase in sulfation of chondroitin sulfate did cause an increase in anticoagulant activity and activation of the kinin system, there may be subtler structural interactions other than sulfation at play given the different responses observed.

By-Products of Heparin Production Provide a Diverse Source of Heparin-like and Heparan Sulfate Glycosaminoglycans

“…Specific activities for each method were estimated using parallel line bioassay model in CombiStats. Plasma APTT, antithrombin dependent anti-IIa and anti-Xa and heparin cofactor II dependent anti-IIa assays were carried out as previously described 31 .…”

Glycosaminoglycan-Inspired Biomaterials for the Development of Bioactive Hydrogel Networks

“…(iv) Membranes formed by self-assembly of HBPA with HA and HEP showed FGF-2 and VEGF retention and enhanced angiogenesis in a CAM model. Membranes showed a hierarchical structure comprising a HBPA hydrogel region (1), a permeable membrane (2) and perpendicular nanofibers formed by GAG/peptide electrostatic interactions (3). (Reproduced with permission from [146] (i and ii), [153] (iii), [156] and [157] [129,130] (i) and [128] (ii), with permission.…”

“…GAG can occur with different sulfation degrees and patterns that affect their biological function [2], with exception of HA, the only non-sulfated GAG. Examples of biofunctional modulation include the increased anticoagulant activity of oversulfated CS [3] or distinct regulation of signaling pathways and cellular response according to GAG sulfation degree [4,5]. Differences in GAG chain elongation or molecular weight can also affect Herein, a major focus will be given to ECM GAG and their applications in the development of new biomaterials for Tissue Engineering (TE) and Regenerative Medicine (RM).…”

Galactosaminoglycans: Medical Applications and Drawbacks

“…As in the “ Heparin crisis ” of 2008 [64,225,226,227,228,229], highly sulfated CS from either from natural sources like CS-D, CS-D, CS-K, CS-L, and CS-M (Table 1) or from chemical reactions [230] are likely to be capable of triggering the same side effects as oversulfated chondroitin sulfate (OSCS) in the activation of factor XII and kinin-kallikrein system, which will result in bradykinin formation and severe hypotension. The ability of highly sulfated CS to activate the kinin-kallikrein system is likely to depend on both the extent and the pattern of sulfation [230].…”