Ectopic osteogenesis evaluation of two kind of chitosan rhBMP-2 loaded microspheres in SD rats
YU Xiang, XIA Yuan-jun，ZHENG Xiao-hui, ZHANG Ying, CHEN Ze-peng, XIA Hong,YIN Qing-shui
Chinese Journal of Clinical Anatomy ›› 2016, Vol. 34 ›› Issue (4) : 412-418.
Ectopic osteogenesis evaluation of two kind of chitosan rhBMP-2 loaded microspheres in SD rats
Objective To explore the ectopic osteogenesis introducing ability of recombinant human bone morphogenetic protein-2/chitosan/dextran sulfate composite microspheres and recombinant human bone morphogenetic protein-2/chitosan composite microspheres on SD rats. Method 36 SD rats were randomly divided into three groups, in which the quadriceps muscle bag model of the rats were established; the bags were implanted respectively with rhBMP-2(group A), CS/DS microspheres (group B), and rhBMP-2/CS/DS microspheres (group C). Hardness of the implanted regions tissuesat the 4th,8th,12th weekend were observed, and four rats were sacrificed in each group, from which the ectopic bone tissues were harvested. The bone formation situation of plantation areas were generally observed, and the heterotopic ossification organization tissues were scanned by micro-CT, and were three-dimensionally reconstructed by using the Mimics software；parameters such astissue bone volume fraction (BVF), trabecular thickness (Tb.Th), bone mineral density (BMD) were detected; Some heterotopic ossification organization tissues were taken for histological observation and analysis of ALP activity and calcium content. Results At the 4th weekend, tissues around implanted regions of each group had slightly harder texture, and there was no significant difference among the three groups; at the 8th and 12th weekend, hardness of tissues around implanted regions of each group increased significantly, and the group C had harder tissues formed than group A and B. At the 4th weekend, HE staining showed that the three groups had a small amount of bone tissue formation, but not obviously; the BVF, Tb.Th, BMD, ALP activity and calcium levels of B, C groups were significantly higher than group A; there were no statistically significance on the above indicators of group B and C. At the 8th and 12th weekend, gradually-increased and marture bone tissues could be seen in the three groups under HE staining. However,there were more mature ectopic bone tissues in group B and C than group A; And the bone tissues of group C were more mature than group B. Meanwhile, the BVF, Tb.Th, BMD, ALP activity and calcium levels of group B and C were significantly higher than group A, and the above-mentioned indicators of group C were higher than that of group B. Conclusion rhBMP-2/CS/DS microspheres have stronger ability of inducing ectopic bone than rhBMP-2/CS microspheres and single rhBMP-2, and it mayhave good prospects for applicationin bone tissue engineering.
Bone morphogenetic proteins-2 / Chitosan / Dextran sulfate / Ectopic osteogenesis / Micro-CT / Three-dimensional reconstruction
[1] Urist MR. Bone formation by autoinduction[J].Science,1965,150(698):893-899.
[2] Martínez A, Arana P, Fernandez A, et al. Synthesis and characterisation of alginate/chitosan nanoparticles as tamoxifen controlled delivery systems[J].J Microencapsul, 2013,30(4):398-408.
[3] 杨强, 彭江, 卢世璧, 等. 软骨细胞外基质与壳聚糖复合制备组织工程软骨支架及其性能研究[J].中华骨科杂志, 2011, 31(8):904-910.
[4] Wink JD, Gerety PA, Sherif RD, et al. Sustained delivery of rhBMP-2 by means of Poly(Lactic-co-Glycolic Acid) microspheres: cranial bone regeneration without heterotopic ossification or craniosynostosis[J]. Plast Reconstr Surg, 2014, 134(1): 51-59.
[5] 王玮, 尹庆水, 张余. 重组人骨形态发生蛋白-2壳聚糖纳米微球的制备及检测[J].中国骨与关节损伤杂志, 2011，26(7):598-600.
[6] 夏远军, 章莹, 李丽华, 等. 重组人骨形态发生蛋白-2/壳聚糖/硫酸葡聚糖复合微球对SD大鼠骨髓基质干细胞诱导及分化作用实验研究[J].中华创伤骨科志, 2014, 16(5):421-426.
[7] 夏远军,余翔,章莹,等.BMP-2/壳聚糖/硫酸葡聚糖复合微球诱导异位成骨的Micro-CT评价[J].中国修复重建外科杂志, 2016, 30(3):286-291.
[8] Jun SH, Lee EJ, Jang TS, et al. Bone morphogenic protein-2 (BMP-2) loaded hybrid coating on porous hydroxyapatite scaffolds for bone tissue engineering[J].J Mater Sci Mater Med，2013, 24(3):773-782.
[9] Kanczler JM, Ginty PJ, Barry JA, et al. The effect of mesenchymal populations and vascular endothelial growth factor delivered from biodegradable polymer scaffolds on bone formation[J]. Biomaterials, 2008, 29(12): 1892-1900.
[10]Huang D, Zuo Y, Zou Q, et al. Reinforced nanohydroxyapatite/polyamide66 scaffolds by chitosan coating for bone tissue engineering[J]. J Biomed Mater Res B Appl Biomater, 2012，100(1):51-57.
[11]Palazzo B, Gallo A, Casillo A, et al. Fabrication, characterization and cell cultures on a novel composite chitosan-nano-hydroxyapatite scaffold[J].Int J Immunopathol Pharmacol, 2011, 24(1 Suppl 2):73-78.
[12]Patel ZS, Ya mamoto M, Ueda H, et al. Biodegradable gelatin microparticles as deliveiy systems for the controlled release of bone morphogenetic Protein-2[J]. Acta Bioniater, 2008, 4(5):l 126-1138.
[13]Guliyeva U, Oner F, Ozsoy S, et al. Chitosan microparticles containing plasmid DNA as potential oral gene delivery system[J].Eur J Pharm Biopharm, 2006, 62(1):17-25.
[14] Shah RK, Moncayo VM, Smitson RD, et al. Recombinant human bone morphogenetic protein 2-induced heterotopic ossification of the retroperitoneum, psoas muscle, pelvis and abdominal wall following lumbar spinal fusion[J]. Skeletal Radiol, 2010, 39(5):501-504.
[15] Inoda H, Yanmmoto G, Hattori T. rhBMP2 induced ectopic bone for grafting critical size defects: a preliminary histological evaluation in rat calvariae[J]. Int J Oral Maxillofac Surg, 2007, 36(1): 39-44.
[16]Iwata H, Sakano S, Itoh T, et al. Demineralized bone matrix and native bone morphgenetic protein in orthpadic surgery[J] .Clin Orthop, 2002, 395:99-109.
[17]Nakagawa T, Tagawa T. UItrastructural study of direct bone formation induced by Uitrastructural study of direct bone formation induced by BMPs-collagen complex implanted into an ecropic site[J].Oral Dis, 2000, 6(3):172-179.
[18]Vehof JW, Takita H, Kuboki Y, et al. Histological characterization of the carly stages of bone morphpgenetic protein-induced osteogencsis[J].J Biomed Mater Res, 2002, 61(3):440-449.
[19] Sullad AG，Manjeshwar LS，Aminabhavi TM. Novel Semi-interpenetrating microspheres of dextran-grafted-acrylamide and poly(Vinyl Alcohol) for controlled release of abacavir sulfate[J]. Indu strial & Engineering Chemistry Research, 2011, 50(21): 11778-11784.
[20] Gribova V, Crouzier T, Picart C. A material’s point of view on recent developments of polymeric biomaterials: control of mechanical and biochemical properties[J]. J Mater Chem, 2011, 21(38), 14354-14364.
[21]Lee KY, Mooney DJ. Alginate: properties and biomedical applications[J]. Prog Polym Sci, 2012, 37(1):106-126.
[22] Wang M, Abbah SA, Hu T，et al. Minimizing the severity of rhBMP-2-induced inflammation and heterotopic ossification with a polyelectrolyte carrier incorporating heparin on microbead templates[J]. Spine, 2013, 38(17):1452-1458.
[23]Hudalla GA, Murphy WL. Biomaterials that regulate growth factor activity via bioinspired interactions[J]. Adv Funct Mater, 2011, 21(10):1754-1768 .
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