Generally, the lysozyme degradation on chitosan (CTS) is slower than that of chitin (CT). The CTS can be fabricated in scaffold form but it is difficult to fabricate CT scaffold under mild conditions. The method for the preparation of scaffold from N-acetylated CTS (N-CTS) was investigated in this research. By using this method, the scaffolds could be fabricated chitosan to chitin with the degree of acetylation (DA) 18% - 70%. Among these scaffolds, the highest degradation of scaffold by lysozyme was observed on the N-CTS scaffold with DA 60%, which determined by examination of the reducing end contents in the degradation media and by measuring the weight loss of scaffolds. Moreover, the best condition for the degradation of N-CTS scaffold with DA70% by lysozyme was also investigated. The maximum degradation rate of the scaffold was observed on the treatment with lysozyme 500 mg/l of acetate buffer at pH 4.5, 37°C, 100 rpm and for 7 days.
Khor, E., & Lim, L.Y. (2003) Implantable applications of chitin and chitosan. Biomaterials, 24, 2339-2349. doi:10.1016/S0142-9612(03)00026-7
Kim, S., Jeon, Y., Byun, H. and Park, P. (2001) Subacute oral toxicity of chitosan oligosaccharide on Sprague Dawley rats. Proceedings of the 8th International Chitin and Chitosan Conference and 4th Asia Pacific Chitin and Chitosan Symposium, Kodansha Scientific Ltd, Tokyo, 276.
Shigemasa, Y., Morimoto, M., Saimoto, H., Okamoto, Y. and Minami, S. (1998) Applications of chitin and chitosan for biomaterials. Proceedings of the 3rd Asia Pacific Chitin and Chitosan Symposium, Taiwan, 47.
Sung, P.T., Chien, Y.H., Chung,Y.H., Da, M.W., Ling, H.L., Juin, Y.L. and Hsyue, J.H. (2007) Preparation and cell compatibility evaluation of chitosan/collagen composite scaffolds using amino acids as crosslinking bridges. Journal of Applied Polymer Science, 105, 1774-1785. doi:10.1002/app.26157
Tokura, S., Nishi, N., Itoyama, K., Shirai, A., Nishimura, S., Saiki, I. and Azuma, I. (1994) Chitin derivatives with biomedical functions. In: Karnicki, Z.S., Brzeski, M.M., Bykowski, P.J. and Wojtasz-Pajak, A., Eds., Wirtschaftsverlag NW: Verlag für neue Wissenschaft GmbH: Bremerhaven, Germany, 287.
Vande Vord, P.J., Matthew, H.W.T., De Silva, S.P., Mayton, L., Wu, B. and Wooley, P.H. (2002) Evaluation of the biocompatibility of a chitosan scaffold in mice. Journal of Biomedical Materials Research, 59, 585-590. doi:10.1002/jbm.1270
Hirano, S., Shigemasa, R. and Minami, S. (1995) Applications of chitin and chitosan for biomaterials. Biotechnology and Genetic Engineering Reviews, 13, 383-420.
Abia, S. (1992) Studies on chitosan: Lysozymic hydrolysis of partially N-acetylated chitosans. International Journal of Biological Macromolecules, 14, 225-228. doi:10.1016/S0141-8130(05)80032-7
Shashiwa, H., Saimoto, H., Shigemas, Y., Ogawa, R. and Tokura, S. (1990) Lysozyme susceptibility of partially deacetylated chitin. International Journal of Biological Macromolecules, 12, 295-296. doi:10.1016/0141-8130(90)90016-4
Nordtveit, R.J., Varum, K.M. and Smidsord, O. (1994) Degradation of fully water-soluble, partially N-acetylated chitosan with lysozyme. Carbohydrate Polymer, 23, 253-260. doi:10.1016/0144-8617(94)90187-2
Nordtveit, R.J., Varum, K.M. and Smidsord, O. (1996) Degradation of partially acetylated chitosans with hen egg white and human lysozyme. Carbohydrate Polymer, 29, 163-167. doi:10.1016/0144-8617(96)00003-3
Rothamel, D., Schwarz, F., Sculean, A., Herten, M., Scherbaum, W. and Becker, J. (2004) Biocompatibility of various collagen membranes in cultures of human PDL fibroblasts and human osteoblasts-like cells. Clinical Oral Implants Research, 15, 443-449. doi:10.1111/j.1600-0501.2004.01039.x
Varum, K.M., Myhr, M.M., Hjerde, R.J.N. and Smidsod, O. (1997) In-vitro degradation rates of partially N-acetylated chitosan in human serum. Carbohydrate Research, 299, 99-101. doi:10.1016/S0008-6215(96)00332-1
Woodard, J.R., Hilldore, A.J., Lan, S.K., Park, C.J., Morgan, A.W., Eurell, J.A.C., Clark, S.G., Wheeler, M.B., Jamison, R.D. and Johnson, A.J.W. (2007) The mechanical properties and osteoconductivity of hydroxyapatite bone scaffolds with multi-scale porosity. Biomaterials, 28, 45-54. doi:10.1016/j.biomaterials.2006.08.021
Yang, Y., Hu, W., Wang, X. and Gu, X. (2007) The controlling biodegradation of chitosan fibers by N-acetylation in vitro and in vivo. Journal of Materials Science: Materials in Medicine, 18, 2117-2121. doi:10.1007/s10856-007-3013-x
Hirano, S. and Midorikawa, T. (1998) A novel method for the preparation of N-acylchitosan fibre and N-acylchitosan-cellulose fibre. Biomaterials, 19, 293-297. doi:10.1016/S0142-9612(97)00216-0
Hirano, S. and Yagi, Y. (1989) The effects of N-substitution of chitosan and the physical form of the products on the rate of hydrolysis by chitinase form Streptomyces griseus. Carbohydrate Research, 83, 103-108. doi:10.1016/S0008-6215(00)85369-0
Aiba, S. (1994) Preparation of N-acetylchitooligosachrides from lysozymic hydrolysates of partially N-acetylated chitosans. Carbohydrate Research, 261, 297-306. doi:10.1016/0008-6215(94)84025-3
Kurita, K., Koyama, Y., Nishimura, S. and Kamiya, M. (1989) Facile preparation of water soluble from chitosan. Chemistry letters, 1597-1598. doi:10.1246/cl.1989.1597
Hirano, S., Ohe, Y. and Ono, H. (1976) Selective N-acylation of chitosan. Carbohydrate Research, 47, 315-320. doi:10.1016/S0008-6215(00)84198-1
Patent (WO/2003/037935).
Molly, S.S., Thomas, F., Hui, S.K. and Karineh, K. (2005) Controlling cell adhesion and degradation of chitosan films by N-acetylation. Biomaterials, 26, 5872-5878. doi:10.1016/j.biomaterials.2005.02.033
Dygert, S., Li, L.G., Floride, D. and Thomas, J.A. (1965) Determination of reducing sugar with improved precision. Analytical Biochemistry, 13, 367. doi:10.1016/0003-2697(65)90327-1
Sabnis, S. and Block, L.H. (1997) Improved infrared spectroscopic method for the analysis of degree of N-acetylation of chitosan. Polymer Bulletin, 39, 67-71. doi:10.1007/s002890050121
Hsiao, H.Y., Tsai, C.C., Chen, S., Hsieh, B.C. and Chen, R.L.C. (2004) Spectrophotometric determination of deacetylation degree of chitinous materials dissolved in phosphoric acid. Macromolecular Bioscience, 4, 919-921. doi:10.1002/mabi.200400084
Hein, S., Chuen-How, N., Stevens, W.F. and Wang, K. (2008) Selection of a practical assay for the determination of the entire range of acetyl content in chitin and chitosan: UV spectrophotometry with phosphoric acid as solvent. Journal of Applied Biomaterials, 86B, 558-586.
Tamura, H., Nagahama, H. and Tokura, S. (2006) Preparation of chitin hydrogel under mild conditions. Cellulose, 13, 357-364. doi:10.1007/s10570-006-9058-z
Nwe, N., Furuike, T. and Tamura, H. (2009) The mechanical and biological properties of chitosan scaffolds for tissue regeneration templates are significantly enhanced by chitosan from Gongronella butleri. Materials, 2, 379-398. doi:10.3390/ma2020374
Cohen, J.S. (1969) Proton magnetic resonance studies of human lysozyme. Nature, 223, 43-46. doi:10.1038/223043a0
Banjard, S.H., Blake, C.C.F. and Swan, I.D.A. (1974) In lysozyme, E. F. Osserman, R. E. Canfield and S. Beychok, (Ed.), Academic Press Inc., New York, 71-79.
Ren, D., Yi, H., Wang, W. and Ma, X. (2005) The enzymatic degradation and swelling properties of chitosan matrices with different degrees of N-acetylation. Carbohydrate Research, 340, 2403-2410. doi:10.1016/j.carres.2005.07.022
Thein-Han, W.W. and Kitiyanant, Y. (2006) Chitosan scaffold for in vitro buffalo embryonic stem-like cell culture: An approach to tissue engineering. Journal of Biomedical Materials Research, 80B, 92-101. doi:10.1002/jbm.b.30573
Amaral, I.F., Sampaio, P. and Barbosa, M.A. (2006) Three-dimensional culture of human osteoblastic cells in chitosan sponges: the effect of the degree of acetylation. Journal of Biomedical Materials Research, 76A, 335-346. doi:10.1002/jbm.a.30522
Hirano, S., Yamaguchi, Y. and Kamiya, M. (2003) Water-solube N-(n-fatty acyl) chitosans. Macromolecular Bioscience, 3, 629-631. doi:10.1002/mabi.200350029
Sannan, T., Kurita, K. and Iwakura, Y. (1976) Studies on chitin. 2. Effect of deacetylation on solubility. Makromolekular Chemistry, 177, 3589-3600. doi:10.1002/macp.1976.021771210