In vitro degradation behavior of chitosan based hybrid microparticles

Ambalangodage C. Jayasuriya, Kristalyn J. Mauch

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DOI: 10.4236/jbise.2011.45048                

Abstract

The degradation properties of the MPs is important to the long-term benefits of the use of the chitosan (CS) based hybrid MPs in bone tissue-engineering, because the degradation kinetics could affect a multitude of processes within the cell, such as cell growth, tissue regeneration, and host response. The aim of this study was to investigate the degradation of solid, hybrid CS microparticles (MPs), CS-10% calcium phosphate (CaHPO₄, w/w), and CS-10% calcium carbonate (CaCO₃, w/w) MPs in phosphate buffered solution (PBS) over a 30-week period. The hybrid MPs were synthesized by emulsification technique, cross-linked with 64% sodium tripolyphosphate (TPP), purified and air dried overnight. Each sample had 30 mg of MPs was placed in a glass vial with 9 ml of PBS added and then the vial was closed to prevent evaporation. Every week 4 ml of the incubated solution was removed for sample measurement and all samples were replaced with an equivalent amount of fresh medium. The samples were maintained at 37^oC under continuous shaking. The hybrid MPs were measured for pH and calcium release, every week in triplicate. At 0, 5, 10, 15, 20, 25, and 30 weeks, surface and bulk morphology were analyzed with a scanning electron microscope (SEM). The degradation data suggested that the hybrid MPs were stable at least up to 25 week and maintain the physiologically relevant pH. Therefore, we can use these hybrid MPs to apply in the bone tissue engineering applications since they do not degrade within a short period.

Keywords

Degradation; In Vitro; Microparticles; pH; Chitosan; Phosphate Buffered Saline

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Kakar, S. and Einhorn, T.A. (2005) Chapter 11: Tissue engineering of bone. In: T. Einhorn & J. O. Hollinger, Eds., Bone Tissue Engineering, Boca Raton, FL: CRC Press, 277-302.
[2]	Kruyt, M.C., Persson, C., Johansson, G., Dhert, W.J. and De Bruijn, J.D. (2006) Towards injectable cell-based tissue-engineered bone: The effect of different calcium phosphate mps and pre-culturing. Tissue Engineering, 12, 309-317. doi:10.1089/ten.2006.12.309
[3]	Zhang, Y. and Zhang, M. (2001) Synthesis and characterization of macroporous CS/calcium phosphate composite scaffolds for tissue engineering. Journal of Biomedical Materials Research, 55, 304-312. doi:10.1002/1097-4636(20010605)55:3<304::AID-JBM1018>3.0.CO;2-J
[4]	Kim, I.Y., Seo, S.J., Moon, H.S., Yoo, M.K., Park, I.Y. and Kim, B.C. et al. (2008) Chitosan and its derivatives for tissue engineering applications. Biotechnology Advances, 26, 1-21. doi:10.1002/1097-4636(20010605)55:3<304::AID-JBM1018>3.0.CO;2-J
[5]	Jiang, T., Kumbar, S.G., Nair, L.S. and Laurencin, C.T. (2008) Biologically active CS systems for tissue engineering and regenerative medicine. Current Topics in Medicinal Chemistry, 8, 354-364. doi:10.2174/156802608783790974
[6]	Khor, E. (2001). Chitin: Fulfilling a biomaterials promise. Elsevier, New York.
[7]	Shi, C., Zhu, Y., Ran, X., Wang, M., Su, Y. and Cheng, T. (2006) Therapeutic potential of CS and its derivatives in regenerative medicine. Journal of Surgical Research, 133, 185-192. doi:10.1016/j.jss.2005.12.013
[8]	Kato, Y., Onishi, H. and Machida, Y. (2003) Application of chitin and CS derivatives in the pharmaceutical field. Current Pharmaceutical Biotechnology, 4, 303-309. doi:10.2174/1389201033489748
[9]	Abdel-Fattah, W.I., Jiang, T., El-Bassyouni, G.E.T. and Laurencin, C.T. (2007) Synthesis, characterization of CSs and fabrication of sintered CS microsphere matrices for bone tissue engineering. Acta Biomaterialia, 3, 503-514. doi:10.1016/j.actbio.2006.12.004
[10]	Freier, T., Koh, H.S., Kazazian, K. and Shoichet, M.S. (2005) Controlling cell adhesion and degradation of CS films by N-acetylation. Biomaterials, 26, 5872-5878.
[11]	doi:10.1016/j.biomaterials.2005.02.033 Couto, D.S., Hong, Z. and Mano, J.F. (2009) Development of bioactive and biodegradable CS-based injectable systems containing bioactive glass nanoparticles. Acta Biomaterialia, 5, 115-123. doi:10.1016/j.biomaterials.2005.02.033
[12]	Di Martino, A., Sittinger, M. and Risbud, M.V. (2005) Chitosan: A versatile biopolymer for orthopaedic tissue-engineering. Biomaterials, 26, 5983-5990. doi:10.1016/j.biomaterials.2005.03.016
[13]	Francis Suh, J.K. and Matthhew, H.W. (2000) Application of CS-based polysaccharide biomaterials in cartilage tissue engineering: A review. Biomaterials, 21, 2589- 598. doi:10.1016/S0142-9612(00)00126-5
[14]	Anal, A.K., Stevens, W.F. and Remunan-Lopez, C. (2006) Ionotropic cross-linked CS microspheres for controlled release of ampicillin. International Journal of Pharmaceutics, 312, 166-173. doi:10.1016/j.ijpharm.2006.01.043
[15]	Costantino, P.D., Hiltzik, D., Govindaraj, S. and Moche, J. (2002) Bone healing and bone substitutes. Facial Plastic Surgery, 18, 13-26. doi:10.1055/s-2002-19823
[16]	Jayasuriya, A.C. and Bhat, A. (2009) Optimization of scaled-up Chitosan Microparticles for Bone Regeneration, Biomedical Materials; 4, 55006. doi:10.1088/1748-6041/4/5/055006
[17]	Jayasuriya, A.C. and Bhat, A. (2010) Fabrication and characterization of novel hybrid organic/inorganic microparticles to apply in bone regeneration. Journal of Biomedical Materials Research A, 93A, 1280-1288.
[18]	Widmaier, E.P., Raff, H. and Strang, K.T. (2006). Chapter 2: Chemical composition of the body. In: Vander, Ed., Human Physiology: the Mechanisms of Body Function, McGraw-Hill, Boston, pp. 21-46.
[19]	Seltzer, M.H., Rosato, F.E. and Fletcher, M.J. (1970) Serum and tissue in human breast carcinoma. Cancer Research, 30, 615-616.
[20]	Chesnutt, B.M., Viano, A.M., Yuan, Y., Yang, Y., Guda, T., Appleford, M.R. et al. (2009) Design and characterization of a novel CS/nanocrystalline calcium phosphate composite scaffold for bone regeneration. Journal of Biomedical Materials Research Part A, 88A, 491-502. doi:10.1002/jbm.a.31878
[21]	Varum, K.M., Myhr, M.M., Hjerde, R.J., Smidsrod, O. (1997) In vitro degradation rates of partially N-acetylated CSs in human serum. Carbohydrate Research, 299, 99-101. doi:10.1016/S0008-6215(96)00332-1
[22]	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/S0008-6215(96)00332-1
[23]	Tigli, R.S., Karakecili, A. and Gümüsderelioglu, M. (2007) In vitro characterization of CS scaffolds: influence of composition and deacetylation degree. Journal of Materials Science: Materials in Medicine, 18, 1665-1674. doi:10.1007/s10856-007-3066-x
[24]	Young, M.A., Ravishanker, G., Beveridge, D.L. and Berman, H.M. (1995) Analysis of local helix bending in crystal structures of DNA oligonucleotides and DNA- rotein complexes. Biophysical Journal, 68, 2454-2468. doi:10.1016/S0006-3495(95)80427-3