Obtaining of Hydrolysates from Cartilage Tissue Biopolymers and Vitamin Mineral Complex to Prevent and Treat Joint Diseases

 

Joint arthritides and arthroses, destructions of intervertebral cartilage are widely spread chronic diseases. As degradation of connective tissue occurs, the disorders of musculoskeletal system are hardly treated by pharmacologic drugs. One of the problems of efficient search of preparations may be solved by development of the nutraceuticals derived from animals’ and hydrobionts’ tissues components. Provided that the preparations have biopharmaceutical effect on joints. Connective tissue formation arises from the substances that predominantly are orally administered in the form of nutrition. Complete set of biopolymers can be obtained from analogous tissues of farm animals. The basic structural elements of the cartilage tissue are collagen proteins and proteoglycans. Bioavailability of these biopolymers for absorption by human body is high after hydrolysis to the extent of amino acids peptides and glycosaminoglycans’ fragments. These low molecular components enter into blood-vascular system then into cells, they participate in the biosynthesis of macromolecules and in the formation of tissue matrix.

During diseases changes occur in all joints first of all in the hyaline cartilage. Thinning, separation and loss of elasticity of collagen fibrils take place. This is why we chosen material for study hyaline cartilages on the 1st stage of research. The cartilages were derived from the tracheae of bovine animals and pigs. As a rule, sets of enzymes, but not separate proteases, are used for the hydrolysis of proteins that are contained on the connective tissues. These enzymes have specific effect destroying certain links in the proteins. We used the following sets of enzymes: protepsin, phytopain, himopsin, papain, pancreatin, collagenase. We demonstrated that the highest extent of hydrolysis is observed under the effect of phytopain with the following conditions: рН 6,0, Т=55 оС, 10% of phytopain, time of hydrolysis is 3 hours. The analysis of qualitative composition of the hydrolysates was studied by the methods of nuclear magnetic resonance (NMR) and mass spectrometry. Signals of peptides that contain 10 to 50 amino-acid residues are identified in NMR spectra. The results of mass spectrometry demonstrate the presence of peptides having molecular weight 2000 - 4000 D and also fragments of hyaluronic acid and chondroitin-4-sulphate of molecular weight 80 - 1000 D.

As a result of enzymatic hydrolysis we made the basic composition of the nutraceuticals containing low molecular elements of matrix of the hyaline cartilage. Oral administration of these nutrients stimulates disordered during diseases biosynthesis of matrix macromolecules in cells. The efficiency of nutraceuticals increases if they contain vitamins, micro- and macroelements. We assembled the composition of them for activation of the enzymes that catalyze the process of collagen fibrils and proteoglycans’ formation. Such composition of substances is contained in marine algae fucus manufactured by LLC “Native” (Town of Pushchino, Moscow region). On the base of fucus and other vegetable components we obtained additional set of vitamins and minerals applicable to the basic one.