Biological Function And Biological Safety Research Of Sulfonated Chitin And Sulfonated Chitosan

Objective:The hydroxyl group on sugar residue of chitin, chitosan occuresterification reaction with an oxygen-containing inorganic acid; same time, theamino group of chitosan can also occur the esterification reaction. In a number of theesterification reaction of both chitin and chitosan, the most studies is sulfationreaction. The esterification reactions’ product, such as sulfonated chitin(S-CT) andsulfonated chitosan(S-CTS) have good water-solubility, film-forming property andbiological degradation, which have wide range of applications in the field of lightindustry, food industry, medicine industry, agriculture and other prospects. Studieshave shown that S-CT,S-CTS have good anticoagulant, antithrombotic, anti-oxidation,anti-viral, anti-atherogenic, anti-tumor and other biological activities. But there arevery little evaluation of their biological security. In this paper, it was synthesized bysulfonation reaction of the S-CT, S-CTS. Study the biological safety of the S-CT,S-CTS, explore its anticoagulant effect, study the effect of S-CT and S-CTS on the invitro proliferation of human umbilical vein endothelial cells (HUVEC), study theeffect of S-CT and S-CTS on the in vitro proliferation of human cervical cancer cells(HeLa), human laryngeal epidermoid carcinomas cells(HEp-2), human gastric cancercell (SGC-7901). For S-CT, S-CTS to provide experimental evidence in securityapplications.Methods:1Through the chitin/chitosan’s hydroxy (especially the C6-OH) with asulfonating agent (chlorosulfonic acid/concentrated sulfuric acid mixed acid) occurlow heterogeneous esterification reaction compose sulfonated chitin (S-CT)/sulfonated chitosan (S-CTS).Purification and Determination of physical and chemicalproperties of the S-CT, S-CTS. 2By stimulation within the rabbit, evaluate the body’s short-term reactionstrength of these two materials. Regard colloid and diaphragm as a carrier,respectively, to evaluate the compatibility and in vivo degradation of the twomaterials in subcutaneous and muscle tissues. By hemolysis rate measurement,evaluate the blood compatibility of these two materials.3Through the whole blood clotting test,it shows that S-CT and S-CTS have goodvitro anticoagulant biological function also. At the cellular level,combine themicroscopic observations with MTT method to explore the effect of S-CT S-CTS onthe in vitro growth of human vascular endothelial cells HUVEC, human cervicalcarcinoma cell line HeLa, human laryngeal epidermoid carcinoma HEp-2cells andhuman gastric cancer cell SGC-7901.Results:1IR spectra show two materials are sulfonated replaced, was measured both withhigher grafting yield.2The two materials intradermal stimulate reaction results are in line withnational requirements, exhibit good blood compatibility. In vivo degradation ofcolloid is fast, which with minimal inflammatory response. Diaphragm linear rate invivo degradation, and the inflammatory reaction is gradually reduced over time, thetwo materials as a whole exhibited good biological tissue compatibility and in vivodegradation.3S-CT, S-CTS have a significant role in anticoagulation within the concentrationof2.5-10mg/ml. S-CTS anticoagulant effect is superior to the S-CT. The twomaterials within the concentration of125-250μg/ml are good for human umbilicalvein endothelial cells’(HUVEC) in vitro growth, the effect of250mg/ml S-CT’spromoting is most significant treated for2d. The significant inhibitory effect of thesetwo materials within the concentration of62.5-1000μg/ml in vitro proliferation ofhuman cervical cancer cells (HeLa), human laryngeal epidermoid carcinoma (HEp-2),and human gastric cancer cells (SGC-7901). Maximal inhibition rate of S-CT is48.52%（6d）、35.73%（6d）、57.43%（6d）. Maximal inhibition rate of S-CTS is65.23% （6d）、43.19%（6d）、44.32%（4d）, and basically follow the inhibition rate increasedwith the increase of the duration of action, increasing concentrations of the basic trend,showing a dose-effect relationship.Conclusion:Both materials have good bio-security, good vivo degradation and the indicatorsare in line with the mandatory requirements of the state of the material safetyexperiment. In biological function, it shows good manifested significant anticoagulant.These two materials are good for human umbilical vein endothelial cells’(HUVEC) invitro growthy. The significant inhibitory effect of these two materials on the in vitroproliferation of there tumor cells, such as HeLa cell, HEp-2cell and SGC-7901cell.Provide some basis for further study of the biological functions of the two materialsand the possible future use of clinical safety.