Osteodystrophy, osteoporosis, bone injury or defect are common clinically, mainly manifested as stunting, osteomalacia, bone deformation, long-term pain and limited mobility, which seriously affect the lives of patients. Chondroitin sulfate is generally extracted from cartilage tissues such as throat bone, nasal bone and trachea of pig or cow. It is mainly composed of the copolymer of N- acetylgalactosamine (2- acetamide -2 deoxy -β-D- galactopyranose) and D- glucuronic acid, and the hexose in the copolymer is alternately connected by β-1,3 and β-1,4 glycosidic bonds. At present, chondroitin sulfate has been applied as a medicine or nutritional supplement for treating joint diseases, or used in the preparation of biomaterials. However, the preparation technology of chondroitin sulfate with different molecular weights and its activity on osteoblasts have not been deeply studied. In this work, macromolecule chondroitin sulfate was separated and purified by enzymolysis-membrane separation-column separation coupling technique from shark cartilage, with purity of 98.89±0.140% and weight average molecular weight of 81812±99 Da. Chondroitin sulfate with medium molecular weight and small molecular weight were obtained by H2O2 oxidation degradation method, and the weight average molecular weights were 8424 ±67 Da and 2091±85 Da, respectively. The spectra of UV-vis and IR demonstrated that the structure of chondroitin sulfate with different molecular weights remained intact, and the characteristic groups such as acetamido, carboxyl, sulfate and sugar ring did not change during H2O2 oxidative degradation. MTT assay showed that chondroitin sulfate with different molecular weights could significantly promote the proliferation and migration of mouse preimplantation osteoblasts MC3T3-E1 (10-500 μg/mL). The results of real-time qPCR exhibited that chondroitin sulfate with different molecular weights promoted the expression of differentiation gene such as ALP, OPN, OCN and RUNX2 in MC3T3-E1 cells. The activity of low molecular weight chondroitin sulfate is significantly higher than that of medium molecular weight and high molecular weight chondroitin sulfate, indicating that low molecular weight chondroitin sulfate is more easily absorbed and utilized by cells and plays a key role. The experimental results suggest that small molecule shark chondroitin sulfate has more potential for bone nutrition and bone repair, and subsequent in vivo experiments are under way.

Application of shark chondroitin sulfate with different molecular weights in bone nutrition and bone tissue repair

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