中国口腔颌面外科杂志 ›› 2018, Vol. 16 ›› Issue (3): 222-226.doi: 10.19438/j.cjoms.2018.03.006

• 论著 • 上一篇    下一篇

生物陶瓷材料负载冻干BMP-2对牙槽突裂骨改建的影响

欧阳婉璐,钱玉芬   

  1. 上海交通大学医学院附属第九人民医院·口腔医学院口腔正畸科,国家口腔疾病临床研究中心,上海市口腔医学重点实验室,上海市口腔医学研究所,上海200011
  • 收稿日期:2018-03-05 修回日期:2018-04-05 出版日期:2018-05-20 发布日期:2018-06-12
  • 通讯作者: 钱玉芬,E-mail:qianyf1960@163.com
  • 作者简介:欧阳婉璐(1992-),女,在读硕士研究生,E-mail:wanlu_oy@sina.com

Bone remolding in alveolar cleft site repaired by inorganic ceramic scaffolds loaded with BMP-2 via lyophilization

OUYANG Wan-lu, QIAN Yu-fen.   

  1. Department of Orthodontics, Shanghai Ninth People's Hospital, College of Stomatology, Shanghai Jiaotong University School of Medicine; National Clinical Research Center of Stomatology; Shanghai Key Laboratory of Stomatology & Shanghai Research Institute of Stomatology. Shanghai 200011, China
  • Received:2018-03-05 Revised:2018-04-05 Online:2018-05-20 Published:2018-06-12

摘要: 目的: 负载冻干BMP-2到一新型无机生物陶瓷类支架材料OsteoBoneTM,验证其对组织工程骨修复牙槽突裂成骨作用的影响。方法: 通过冻干法并添加海藻糖作为冻干保护剂,将BMP-2与OsteoBoneTM支架材料相结合;之后置于PBS溶液中并收集缓释液,BCA法检测随时间变化的蛋白浓度,ALP检测成骨诱导活性。选取18只8周龄雄性SD大鼠,随机分为3组,每组6只,在大鼠左右侧牙槽骨上人为制作4 mm×3 mm×3 mm牙槽突裂,将单纯材料(A组)、BMSC-材料复合物(B组)与构建出的BMP-2-材料复合物(C组),分别植入大鼠牙槽骨骨缺损区。分别于术后3周与6周取材,H-E染色进行组织学观察,比较3组之间的成骨差异,并进行micro-CT影像学分析。采用SPSS19.0软件包对实验数据进行统计学分析。结果: 通过冻干法并添加海藻糖作为冻干保护剂,将BMP-2与OsteoBoneTM支架材料结合后,BMP-2-材料复合物具备持续的随时间渐增的缓释BMP-2的能力,且表现出明显的ALP活性。体内实验术后3周,A、B、C组H-E染色观察可见较稀疏编织骨,3组间无显著差异;micro-CT分析BV/TV和骨小梁厚度,3组间无显著差异。术后6周,组织学观察B组与C组新骨形成趋势比A组更明显;micro-CT分析BV/TV和骨小梁厚度,B、C组显著大于A组,B组与C组间无显著差异。结论: 负载冻干BMP-2到OsteoBoneTM支架材料能显著提高其成骨能力,达到与间充质干细胞相近的效果。

关键词: 组织工程骨, 牙槽突裂, 支架材料, BMP-2, 冻干法

Abstract: PURPOSE: To load BMP-2 on a novel inorganic ceramic scaffold material, OsteoBoneTM, through lyophilization, and explore its effect on bone formation after bone graft in vivo. METHODS: BMP-2 was loaded onto OsteoBoneTM scaffolds by coating it via lyophilization with trehalose added as stabalizer. The samples were incubated in PBS in vitro and the release rate was examined with BCA method and alkaline phosphatase (ALP) activity was detected. Eighteen SD rats were used as experimental animals. Critical size defects (CSD) of 4 mm×3 mm×3 mm were established on maxillary alveolar bone in SD rats. Scaffolds alone (group A), BMSC-scaffolds (group B)and BMP-2-scaffolds (group C) were used to be grafted into CSDs. Micro-CT and H-E staining were performed respectively 3,6 weeks post-surgery. SPSS 19.0 software package was used for statistical analysis. RESULTS: BMP-2 performed sustained release and maintained bioactivity for enhanced osteogenesis from lyophilized OsteoBoneTM scaffolds in vitro. Three weeks after surgery, H-E staining showed sparse braided bone, and there was no significant difference between the three groups; there was no significant difference in micro-CT analysis of bone volume/tissue volume (BV/TV) and trabecular thickness. Six weeks after surgery, H-E staining revealed abundant bone regeneration and osseous tissue was mature in group B and group C, and new bone formation trend was more apparent than in group A. Micro-CT analysis showed that BV/TV and trabecular thickness in group B and C was significantly greater than that in group A, and there was no significant difference between group B and group C. CONCLUSIONS: Loading BMP-2 on OsteoBoneTM scaffolds through lyophilization can significantly enhance the osteogenic capacity and achieve a similar effect to mesenchymal stem cells.

Key words: Tissue engineering bone, Alveolar cleft, Scaffold, BMP-2, Lyophilization

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