Preparation and ectopic bone formation activity of recombinant human bone morphogenetic protein-2 loaded chitosan nanospheres in vivo

Abstract

Objective To prepare recombinant human bone morphogenetic protein-2(rhBMP-2) loaded chitosan nanospheres and to evaluate their ectopic bone formation activity in vivo. Methods The rhBMP-2 loaded chitosan nanospheres were prepaid using the ionic crosslinking method with TPP as a crosslinking agent.The morphological properties and particle size distribution were evaluated by transmission electron microcopy (TEM) and particle size analyzer.An intramuscular ectopic bone formation test was carried out to compare the ectopic bone formation activity in 24 SD rabbits that were randomized into 4 even groups.rhBMP-2 loaded chitosan nanospheres (1 mg),rhBMP-2 (1 mg),chitosan nanospheres and nothing were implanted into the muscle pouch at the left thigh,respectively,in groups A,B,C and D. Results The rhBMP-2 loaded chitosan nanospheres were well distributed and spherical in shape.The average particle diameter was 230.0 nm,the entrapment efficiency was 66.87％ ± 4.58％,and the loading rate was 33.44 ± 2.29 μg/mg.The mean ALP activity was respectively 1.94 ± 0.35 kat/g,1.48 ± 0.56 kat/g,0.20 ±0.07 kat/g and 0.18 ±0.06 kat/g in groups A,B,C and D,with a significant difference between the 4 groups ( F =42.959,P =0.000).The mean Ca2 + content was respectively 5.20 ± 1.42 μg/mg,3.80 ± 1.40 μg/mg,0.19 ± 0.08 μg/mgand 0.20 ± 0.08 μg/mg in groups A,B,C and D,with a significant difference between the 4 groups ( F =39.242,P =0.000).Groups A and B were obviously higher than groups C and D and group A was obviously higher than group B in terms of ALP activity and Ca2+ content. Conclusion The rhBMP-2 loaded chitosan nanospheres prepared by ionic crosslinking method show a good property and obvious ectopic bone formation activity. Key words: Bone morphogenetic proteins; Chitosan; Nanocomposites; Osteogenesis