Vascularization and bone regeneration in a critical sized defect using 2-N,6-O-sulfated chitosan nanoparticles incorporating BMP-2
Vascularization and bone regeneration in a critical sized defect using 2-N,6-O-sulfated chitosan nanoparticles incorporating BMP-2
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DOI：
10.1016/j.biomaterials.2013.10.005
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发表时间：
2014-01-01
期刊：
影响因子：
14
通讯作者：
Liu, Changsheng
中科院分区：
文献类型：
--
作者：
Cao, Lingyan;Wang, Jing;Liu, Changsheng
理想的骨组织工程移植物应该同时具有良好的促成骨和促血管生成能力，才能在体内快速实现骨再生。为了实现这一目标，成功地制备了基于2-N，6-O-磺化壳聚糖(26SCS)的纳米粒(S-NP)，并显示出剂量依赖的体外血管生成促进作用。为了修复兔桡骨临界大小的缺损，我们研制了蛋白质负载率为1.4+/-0.2%的骨形态发生蛋白-2/S-NP(BMP-2/S-NP)，并制备了骨形态发生蛋白-2/S-NP(BMP-2/S-NP/G)明胶海绵种植体。该植入物具有骨形态发生蛋白-2的释放，最初24 h突释为15.3+/-4.1%，21天后逐渐释放至77.8+/-3.6%。体外碱性磷酸酶检测显示，BMP-2/S-NP/G细胞在给药后3d和7d释放的BMP-2活性保持不变，14d后比原来的BMP-2释放的活性进一步增强。采用同步辐射显微计算机断层成像、三维显微计算机断层成像、组织学分析、免疫组织化学和生物力学测量等方法，研究骨形态发生蛋白-2/S-纳米粒/G对兔桡骨临界大小缺损区(18 Mm)骨再生和血管形成的影响。结果表明，骨形态发生蛋白-2/S-NP/G组的周围血管和新生血管的形成均显著增加，最早2周达到桥接，8周达到愈合，12周达到骨髓腔的愈合。结果表明，BMP-2/S-NP/G对活性骨形态发生蛋白-2的控制释放和在缺损区良好的血管生成在促进和促进成骨方面起着至关重要的作用。本研究提示，骨形态发生蛋白-2/S-NP/G在大面积骨缺损的临床病例中显示出良好的血管化和骨再生前景。(C)2013爱思唯尔有限公司。保留所有权利。
An ideal bone tissue engineering graft should have both excellent pro-osteogenesis and pro-angiogenesis to rapidly realize the bone regeneration in vivo. To meet this goal, 2-N,6-O-sulfated chitosan (26SCS) based nanoparticle (S-NP) was successfully developed and showed a dose-dependent enhancement on angiogenesis in vitro. For the repair of a critical sized defect in rabbit radius, we developed BMP-2 loaded S-NP (BMP-2/S-NP) with protein loading efficiency of 1.4 +/- 0.2% and fabricated a gelatin sponge (G) based implant loaded with BMP-2/S-NP (BMP-2/S-NP/G). This implant exerted a delivery of BMP-2 with an initial burst release of 15.3 +/- 4.1% in first 24 h and a gradual release for 21 days to 77.8 +/- 3.6%. The in vitro ALP assay revealed that the activity of released BMP-2 from BMP-2/S-NP/G was maintained after 3-d and 7-d delivery and further enhanced after 14-d delivery compared with the original BMP-2. Furthermore, the in vivo effects of BMP-2/S-NP/G on the bone regeneration and vessel formation in the critical sized defect (18 mm) of rabbit radius were investigated by synchrotron radiation-based micro-computed tomography (SR mu CT) imaging, three dimensional micro-computed tomographic (ACT) imaging, histological analysis, immunohistochemistry and biomechanical measurement. Based on the results, both peripheral vessel and new vessel formation were significantly increased by the BMP-2/S-NP/G treatment, along with the bridged defects at as early as 2 weeks, the healed defects at 8 weeks and the reunion of bone marrow cavity at 12 weeks. The results indicated that both controlled release of active BMP-2 and favorable vascularization at the defect site contributed by BMP-2/S-NP/G played a crucial role in accelerating and promoting bone augmentation. This study suggests that BMP-2/S-NP/G demonstrates promise for vascularization and bone regeneration in clinical case of large defect. (C) 2013 Elsevier Ltd. All rights reserved.