Evaluation of hyaluronan from different sources: Streptococcus zooepidemicus, rooster comb, bovine vitreous, and human umbilical cord

Department of Surgery, University of Washington Seattle, Seattle, Washington, United States
Biomacromolecules (Impact Factor: 5.79). 10/2004; 5(6):2122-7. DOI: 10.1021/bm0498427
Source: PubMed

ABSTRACT Sodium hyaluronate (HA) is widely distributed in extracellular matrixes and can play a role in orchestrating cell function. Consequently, many investigators have looked at the effect of exogenous HA on cell behavior in vitro. HA can be isolated from several sources (e.g., bacterial, rooster comb, umbilical cord) and therefore can possess diverse impurities. This current study compares the measured impurities and the differences in biological activity between HA preparations from these sources. It was demonstrated that nucleic acid and protein content was highest in human umbilical cord and bovine vitreous HA and was low in bacterial and rooster comb HA. Macrophages exposed to human umbilical cord HA produced significantly higher amounts of TNF-alpha relative to control or bacterial-derived HA. These results indicate that the source of HA should be considered due to differences in the amounts and types of contaminants that could lead to widely different behaviors in vitro and in vivo.

  • [Show abstract] [Hide abstract]
    ABSTRACT: In previous publications, we showed that extracellular glycosaminoglycans reduced the membrane potential, caused cell blebbing and swelling and decreased the intracellular pH independently of cell surface receptors. These phenomena were explained by Donnan effects. The effects were so large that they could not be attributed to glycosaminoglycans in solution. Therefore, we tested the hypothesis that glycosaminoglycans were concentrated on the cell membrane and analysed the mechanism of adsorption by fluorescent hyaluronan, chondroitin sulphate and heparin. The influence of the CD44 receptor was evaluated by comparing CD44 expressing human fibroblasts with CD44 deficient HEK cells. Higher amounts of glycosaminoglycans adsorbed to fibroblasts than to HEK cells. When the membrane potential was annihilated by substituting NaCl by KCl in the medium, adsorption was reduced and intracellular pH decrease was abolished. To eliminate other cellular interfering factors, potential-dependent adsorption was demonstrated for hyaluronan which adsorbed to inert gold foils in physiological salt concentrations at pH 7.2 and surface potentials up to 120 mV. From these results, we conclude that large cellular Donnan effects of glycosaminoglycans results from receptor mediated, hydrophobic and ionic adsorption to cell surfaces. J. Cell. Biochem. 9999: XX-XX, 2014. © 2013 Wiley Periodicals, Inc.
    Journal of Cellular Biochemistry 07/2014; 115(7). DOI:10.1002/jcb.24791 · 3.37 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: The cetyltrimethylammonium bromide turbidimetric method (CTM) has been developed to quantify the hyaluronic acid (HA) in complex media to overcome the lack of selectivity and specificity of the standard carbazole method. The objective of this work is to assess the potential application of CTM to determine HA concentration. Factors such as duration of incubation, linearity range, HA size and form (natural linear HA or cross linked HA), pH and ionic environment impact were investigated. The incubation time was set to 10 min and the calibration curve was linear up to 0.6 g L−1. The quantitative method was relevant whatever the HA size and form, and also for a wide range of conditions. The robustness of the CTM added to its high specificity and simplicity demonstrated that the CTM is a valuable method that would be an interesting substitute to the carbazole assay for HA quantification.
    Carbohydrate Polymers 11/2014; 112:102–108. DOI:10.1016/j.carbpol.2014.05.039 · 3.92 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: The encapsulation of living mammalian cells within a semi-permeable hydrogel matrix is an attractive procedure for many biomedical and biotechnological applications, such as xenotransplantation, maintenance of stem cell phenotype and bioprinting of three-dimensional scaffolds for tissue engineering and regenerative medicine. In this review, we focus on naturally derived polymers that can form hydrogels under mild conditions and that are thus capable of entrapping cells within controlled volumes. Our emphasis will be on polysaccharides and proteins, including agarose, alginate, carrageenan, chitosan, gellan gum, hyaluronic acid, collagen, elastin, gelatin, fibrin and silk fibroin. We also discuss the technologies commonly employed to encapsulate cells in these hydrogels, with particular attention on microencapsulation.
    Journal of The Royal Society Interface 11/2014; 11(100). DOI:10.1098/rsif.2014.0817 · 3.86 Impact Factor

Full-text (2 Sources)

Available from
Jun 10, 2014