Hemostatic property and cytotoxicity of the guanidine modified chitosan

doi:10.3969/j.issn.2095-4344.2017.06.015

Abstract

Abstract:

BACKGROUND: Chitosan is a kind of biomaterial with good hemostatic, antibacterial and absorption properties. The guanidine modified chitosan holding better antibacterial and absorption abilities is a commonly used chemical separation material. However, its coagulation property is still unclear.
OBJECTIVE: To evaluate the in vitro hemostatic property and cytotoxicity of the guanidine modified chitosan, and to explore its availability as a hemostatic material.
METHODS: The guanidine modified chitosan was synthesized with the raw materials of chitosan and arginine, and the coupling agents of EDC and NHS. The coagulation ability was evaluated preliminarily through the whole blood coagulation time, and three in vitro coagulation indexes (activated partial thromboplastin time, prothrombin time and thrombin time). The guanidine modified chitosan extracts were co-cultured with the mouse fibroblasts, and then the cell growth was observed by MTT assay and fluorescence microscope.
RESULTS AND CONCLUSION: The guanidine modified chitosan shortened the whole blood coagulation time, suggesting its hemostatic property. In vitro coagulation results indicated that the guanidine modified chitosan exerted the effect not through the traditional endogenous and extrinsic coagulation pathways. Guanidino group has not been proved to be hemostatic, so the chitosan may be the effective component. MTT assay showed that the cell proliferation rate was 87.8% and the toxicity level was grade 1. Fluorescent staining observed abundant live cells, and few dead cells, indicating that the guanidine modified chitosan exhibits low cytotoxicity.

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

Key words: Guanidine, Chitosan, Hemostasis, Cell Proliferation, Tissue Engineering

CLC Number:

R318

Yao Xin-pei, Jing Miao-lei, Guan Jing. Hemostatic property and cytotoxicity of the guanidine modified chitosan[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(6): 906-910.

Figures/Tables 4

References

[1]Elvin CM, Vuocolo T, Brownlee AG, et al. A highly elastic tissue sealant based on photopolymerised gelatin. Biomaterials. 2010;31(32):8323-8331.

[2]Wu Y, He J, Cheng W, et al. Oxidized regenerated cellulose-based hemostat with microscopically gradient structure. Carbohydrate Polymers. 2012; 88(3):1023-1032.

[3]Eldibany RM. Platelet rich fibrin versus Hemcon dental dressing following dental extraction in patients under anticoagulant therapy. Tanta Dental Journal. 2014; 11(2):75-84.

[4]Schmid BC, Rezniczek GA, Rolf N, et al. Postpartum hemorrhage: use of hemostatic combat gauze. Am J Obstet Gynecol. 2012;206(1):e12-13.

[5]蒋挺大. 壳聚糖[M]. 2版.北京:化学工业出版社, 2007.

[6]蒋小姝, 莫海涛, 苏海佳,等. 甲壳素及壳聚糖在农业领域方面的应用[J]. 中国农学通报, 2013, 29(6):170-174.

[7]乔德亮. 低聚壳聚糖制备及其在功能食品中应用[J]. 食品工业科技, 2007, 28(4):228-231.

[8]程志, 杜娜. 壳聚糖在医药领域应用研究进展[J]. 人民军医, 2013, (2):223-224.

[9]刘长岚, 崔文新, 李成彬. 壳聚糖及其衍生物在化工方面应用的研究[J]. 山东师范大学学报:自然科学版, 2003, 18(3):73-74.

[10]黄勇. 壳聚糖及其胍基化衍生物的合成、表征及抗菌性能研究[D]. 武汉:华中科技大学, 2013.

[11]Martins AF, Facchi SP, Follmann HD, et al. Antimicrobial activity of chitosan derivatives containing N-quaternized moieties in its backbone: a review. Int J Mol Sci. 2014;15(11): 20800-20832.

[12]Baker S, Wiesmann WP, Ryan S. Chitosan-derivative compounds and methods of controlling microbial populations. US, US8658775. 2014.

[13]Chan LW, Kim CH, Wang X, et al. PolySTAT-modified chitosan gauzes for improved hemostasis in external hemorrhage. Acta Biomater. 2016;31:178-185.

[14]Dowling MB, Smith W, Balogh P, et al. Hydrophobically-modified chitosan foam: description and hemostatic efficacy. J Surg Res. 2015;193(1):316-323.

[15]尹刚. 温敏性壳聚糖止血膜止血作用的实验研究[D]. 上海:第二军医大学, 2014.

[16]Gu BK, Park SJ, Kim MS, et al. Fabrication of sonicated chitosan nanofiber mat with enlarged porosity for use as hemostatic materials. Carbohydr Polym. 2013;97(1):65-73.

[17]奚宏伟, 魏长征, 王文斌, 等. 壳聚糖-三聚磷酸钠纳米粒对于肝组织严重出血模型的止血效果[J]. 现代生物医学进展, 2014, 14(15):2831-2834.

[18]谭奕哲, 吴国忠, 曾虹燕, 等. 抗菌性壳聚糖双胍醋酸盐的辐照辅助制备及其生物毒性[J]. 功能高分子学报, 2015, 28(2):183-187.

[19]Su YR, Yu SH, Chao AC, et al. Preparation and properties of pH-responsive, self-assembled colloidal nanoparticles from guanidine-containing polypeptide and chitosan for antibiotic delivery. Colloids & Surfaces A Physicochemical & Engineering Aspects. 2016; 494:9-20.

[20]Sun S, An Q, Li X, et al. Synergistic effects of chitosan-guanidine complexes on enhancing antimicrobial activity and wet-strength of paper.Bioresour Technol. 2010; 101(14):5693-5700.

[21]Ryu K, Kim K, Kim TI, et al. Synthesis and characterization of guanidinylated polyethylenimine-conjugated chitosan for gene delivery systems. Macromolecular Research. 2014; 22(3):264-271.

[22]He B, Shao Y, Liang M, et al. Biodiesel production from soybean oil by guanidinylated chitosan. Fuel. 2015; 159: 33-39.

[23]Luo Y, Zhai X, Ma C, et al. An inhalable β?-adrenoceptor ligand-directed guanidinylated chitosan carrier for targeted delivery of siRNA to lung. J Control Release. 2012;162(1):28-36.

[24]Chen CY, Lin HC, Huang YY, et al. ‘One-flask’ transformation of isocyanates and isothiocyanates to guanidines hydrochloride by using sodium bis(trimethylsilyl)amide. Tetrahedron. 2010; 66(10):1892-1897.

[25]Arafa RK, Ismail MA, Munde M, et al. Novel linear triaryl guanidines, N-substituted guanidines and potential prodrugs as antiprotozoal agents. Eur J Med Chem. 2008;43(12):2901-2908.

[26]杨健, 田丰, 陈世谦. 壳聚糖的止血机理和应用[J]. 国际生物医学工程杂志, 2001, 24(2):77-80.

[27]Di Cera E, Dang QD, Ayala YM. Molecular mechanisms of thrombin function. Cell Mol Life Sci. 1997;53(9):701-730.

[28]Matthias R, Wolfram R. Proteinase‐activated receptor activation by coagulation proteinases. Drug Development Research. 2003; 59(4):400-407.

[29]Klokkevold PR, Lew DS, Ellis DG, et al. Effect of chitosan on lingual hemostasis in rabbits. J Oral Maxillofac Surg. 1991; 49(8):858-863.

[30]Benesch J, Tengvall P. Blood protein adsorption onto chitosan. Biomaterials. 2002;23(12):2561-2568.