Preparation and examination of BMP-2 loaded chitosan nanospheres in vitro

Abstract

Abstract: PURPOSE: To prepare chitosan nanospheres for loading of BMP-2 and to evaluate its size, zeta potential, appearance, degradation and release characteristic in vitro, and then to investigate its feasibility as a carrier for sustained release of BMP-2. METHODS: The BMP-2 loaded chitosan nanospheres were prepared using ionic crosslinking method with tripolyphosphate (TPP) and chitosan. Transmission electron microscope was used to evaluate the morphological properties, and laser particle size analyzer was used to analyze particle size, Zeta potential and distribution. Lysozyme degradation experiment was performed to assess the biodegradation behavior. ELISA assay was used to determine the loading efficiency, encapsulation efficiency and in vitro drug release kinetics. The data was analyzed by SPSS 19.0 software package. RESULTS: The BMP-2 loaded chitosan nanospheres were spherical in shape, smooth on surface and uniform dispersion without aggregation. The mean diameter was 150.85 nm. The dispersion index was 0.37, and zeta potential was +35.42 mV. The average loading efficiency and encapsulation efficiency were (56.83±2.26)% and (68.24±3.83)%, respectively. Release experiment in vitro showed that the releasing property of BMP-2 loaded chitosan nanospheres was consistent with two-phase kinetic regulation and BMP-2 was controlled to release from the chitosan nanospheres over 30 days. CONCLUSIONS: The BMP-2 loaded chitosan nanospheres prepared by ionic crosslinking method are successfully prepared which show a good controlled release property. It provides the basis for further application in bone tissue engineering.

Key words: Bone tissue engineering, Nanospheres, Chitosan, BMP-2, Delayed-release systems

CLC Number:

R782

LIU Feng,SUN Jian,LI Ya-li,CHEN Li-qiang,XIE Jing-sen,ZANG Xiao-long. Preparation and examination of BMP-2 loaded chitosan nanospheres in vitro[J]. Shanghai Journal of Stomatology, 2015, 24(2): 147-150.

References

[1] Schnürer SM, Gopp U, Kühn KD, et al. Bone substitutes [J]. Orthopade, 2003, 32(1): 2-10.
[2] Giannoudis PV, Dinopoulos H, Tsiridis E. Bone substitutes: an update [J]. Injury, 2005, 36(Suppl 3): S20-27.
[3] 周庆梅, 孙健, 李亚莉, 等. 生长因子缓释系统复合纳米支架材料修复犬下颌骨缺损 [J]. 中国组织工程研究, 2013, 17(21): 3816-3822.
[4] Kumar S, Wan C, Ramaswamy G, et al. Mesenehymal stem cells expressing osteogenic and angiogenic factors synergistically enhance bone formation in a mouse model of segmental bone defect [J]. Mol Ther, 2010, 18(5): 1026-1034.
[5] Calvo P, Remuñan-López C, Vila-Jato JL, et al. Chitosan and chitosan/ethylene oxide-propy1ene oxide block copolymer nanoparticles as novel carriers for proteins and vaccines [J].Pharm Res, 1997, 14(10): l431- 1436.
[6] Miyasono K, Kamiya Y, Mofikawa M. Bone morphogenetic protein receptors and signal transduction [J]. J Biochem, 2010, 147(1): 35-51.
[7] Li R, Stewart DJ, von Schroeder HP, et al. Effect of cell-based VEGF gene therapy on healing of a segmental bone defect [J]. J Orthop Res, 2009, 27(1): 8-14.
[8] 黄波, 廖琦. 负载骨生长因子的壳聚糖微球研究进展 [J].中外医学研究, 2012, 10(5): 157-158.
[9] 唐旭, 雷云坤, 孟增东. 壳聚糖/纳米羟基磷灰石﹙CS/nHA﹚复合材料修复骨缺损研究进展[J]. 生物骨科材料与临床研究, 2012, 9(5): 20-22.
[10] Aksungur P, Sungur A, Unal S, et al. Chitosan delivery systems for the treatment of oral mucositis: in vitro and in vivo studies [J]. J Control Release, 2004, 98(2): 2l1-215.
[11] Muzzarelli R. Biochemical significance of exogenous chitins and chitosans in animals and patients[J]. Carbohyd Polym,1993, 20(1): 7-l6.
[12] Tang ZX, Qian JQ, Shi LE. Preparation of chitosan nanoparticles as carrier for immobilized enzyme [J].Appl Biochem Biotechnol, 2007, 136(1): 77-96.
[13] Ozba-Turan S, Akbua J, Aral C. Controlled release of interleukin-2 from chitosan microsphere[J]. J Pharm Sci, 2002, 91(5): 1245-1251.
[14] Zheng AP, Wang JC, Lu WL, et al. Thymopentin-loaded pH-sensitive chitosan nanoparticles for oral administration: preparation, characterization and pharmacodynamics [J]. J Nanosci Nanotechnol, 2006, 6(9-10): 2936-2944.
[15] 梁恒伦, 童键, 李晶, 等. 多西紫杉醇壳聚糖纳米粒制备及其特性研究 [J]. 重庆医学, 2011, 40(31): 3154-3159.