Hyaluronic acid isolation

Papakonstantinou E, Roth M, Eickelberg O, Mirtsou-Fidani V, Mora M, Karakiulakis G. Hyaluronic acid isolated and purified from human atherosclerotic aortas is involved in the progression of atheromatosis. Biochem... 

The viability and function tests described above are used to evaluate the hepatocytes within the slice. Up to now, tests to measure the viability of the non-parenchymal cells have not been reported. The presence of the latter cell types is one of the conceptual advantages of slices as compared to isolated hepatocytes. As some drug targeting devices are designed to target non-parenchymal cells in the liver, the development of tests for the sinusoidal cell types deserves more attention. For example, the uptake of substrates such as succinylated human serum albumin (Suc-HSA,which is specifically endocytosed by endothelial cells [79]), or hyaluronic acid [80], can be used to assess the functionality of endocytotic pathways in the endothelial cells in the liver [81]. Other modified proteins that are specifically taken up by Kupffer cells such as mannosylated HSA, may be used to assess the functionality of the endocytotic pathway in Kupffer cells [79]. Another parameter which can be used to assess the functionality of these non-parenchymal liver cells, is the excretion of cytokines in response to pro-inflammatory stimuli. Non-parench5mal cell function in liver slices will be described in more detail in the Section 12.7. 

The same authors130 used gel chromatography on agarose for isolating the undegraded hyaluronic acid-protein complex from human synovial fluid. 

Agarose gels M sodium isolation of hyaluronic acid- 130... 

Compounds known to behave in this way in vivo are listed in recent reviews in this Series.1 2 The structures of some of the /3-D-glucopyranosiduronic acids isolated from urine have been proved by chemical synthesis.3 A few similar derivatives of flavones and triterpenes have been isolated from plants. D-Glucuronic acid also occurs in mammalian tissues as a constituent of acid mucopolysaccharides (aminodeoxypolysaccharides, containing uronic acid), such as hyaluronic acid, chondroitinsulfate, and heparin,4 and it is a direct precursor of L-ascorbic acid in plants and mammals.6 It is present in many of the plant polysaccharides classified as hemicelluloses6 and gums,7 and it has also been found in certain bacterial polysaccharides.4... 

Chemical composition and sedimentation coefficients (s 0) of proteoglycan monomer fractions from bovine articular cartilage (36). Column 1 shows the amino acid composition of the hyaluronic acid-binding region of proteoglycan monomer core protein, isolated... 

ISOLATION OF THE HYALURONIC ACID BINDING REGION OF PGS CORE PROTEIN... 

Rapport, M.M. et al., Isolation of a crystalline disaccharide, hyalobiuronic acid, from hyaluronic acid, Nature, 168, 996, 1951. 

Since protein complex formation and Ca2+ are critical to cell fixation within a tissue, dissociation media usually contain some type of proteolytic enzyme and the Ca2+ chelator, EDTA. The proteolytic enzyme can be of general specificity, such as trypsin, or can be a more targeted enzyme, such as a collagenase selective for the collagen-type characteristic of the tissue of interest. Hyaluronidase has been also used with matrix rich in hyaluronic acid, such as for isolation of duodenal entero-cytes. In all cases, the appropriate incubation times and concentrations to achieve cell dispersal, but retain high viability, need to be determined empirically. One factor... 

Hyaluronic acid is a component of the extracellular ground substance which surrounds the collagen and elastin fibres and cells of connective tissue [64], It is a member of the group of polysaccharides isolated from vertebrate connective tissues which were formerly called mucopolysaccharides and are now more commonly referred to as glycosaminoglycans [65,66], Glycosaminoglycans commonly occur in vivo as proteoglycans. 

Chondroitin 4- and 6-sulfates were originally designated as chondroitin sulfate A and C, respectively, and the close similarity in structure between them is demonstrated by the fact that both mucopolysaccharides yield the same disaccharide (chondrosine) on controlled, acidic hydrolysis together with sulfuric and acetic acids. Chondroitin 4-sulfate or chondroitin 6-sulfate, or both, occur in cartilage and adult bone, and the former mucopolysaccharide is a minor constituent of ligamentum nuchae and cornea (M17). Chondroitin 6-sulfate is a minor constituent of umbilical cord and occurs with dermatan sulfate (chondroitin sulfate B) and hyaluronic acid in heart valves and adult connective tissue (M17). An acid mucopolysaccharide isolated from human plasma resembles chondroitin 4-sulfate in its properties (S4). 

This mucopolysaccharide, possessing a strueture similar to those of chondroitin 4- and 6-sulfates but with a small content of sulfate, was isolated from bovine cornea (M16). Chondroitin resembles hyaluronic acid in its rate of hydrolysis by testicular and bacterial hyaluronidases, but was differentiated from hyaluronic acid ([a]n —65° to — 78°) by its optical rotation ( [o]d — 21°). Its structural similarity to chondroitin 4- and 6-sulfates was indicated by the fact that chondrosine was released in high yield on controlled, acidic hydrolysis (D3). The isolation of this mucopolysaccharide is of particular interest since it may be a precursor in the biosynthesis of chondroitin 4- and 6-sulfates. 

One of the manifestations of generalized or localized pretibial myxedema is a pronounced edema of the corium, with accumulation of mucinous material in the edema fluid. Both hyaluronic acid and a sulfated mucopolysaccharide fraction were isolated in increased quantity from the affected area of the skin of a patient with localized pretibial myxedema (Wl). Following this observation, the relationship between the functional status of the thyroid gland and the mucopolysaccharides of skin has been discussed in numerous publications. Analysis of the acid mucopolysaccharides of human skin has revealed the presence of dermatan sulfate, hyaluronic acid, and a small amount of chondroitin 4- or 6-sulfate (L9). 

B7. Bertelsen, S., and Marcher, K., Isolation of hyaluronic acid and chondroitin sulphuric acid from human aortae. Acta Pharmacol. Toxicol. 18, 1-9 (1961). 

W4. Weissmann, B., Meyer, K., Sampson, P., and Linker, A., Isolation of oligosaccharides enzymatically produced from hyaluronic acid. J. Biol. Chem. 208, 417-429 (1954). 

---