中国组织工程研究 ›› 2018, Vol. 22 ›› Issue (18): 2807-2812.doi: 10.3969/j.issn.2095-4344.0865

• 组织工程骨及软骨材料 tissue-engineered bone and cartilage materials • 上一篇    下一篇

煅烧鹿角松质骨在骨缺损修复过程中的早期血管化

彭 晖1,张学慧2   

  1. 1北京大学口腔医院第三门诊部,北京市 100191;2北京大学口腔医院口腔材料研究室,北京市 100081
  • 收稿日期:2018-04-21 出版日期:2018-06-28 发布日期:2018-06-28
  • 通讯作者: 张学慧,副研究员,北京大学口腔医院口腔材料研究室,北京市 100081
  • 作者简介:彭晖,女,1982年生,汉族,河北省蔚县人,2012年北京大学医学部口腔医学院毕业,硕士,医师,主要从事骨组织工程方面的研究。
  • 基金资助:

    国家高技术研究发展(863)计划课题(2015AA033601)

Early vascularization of calcined antler cancellous bone in repairing bone defects

Peng Hui1, Zhang Xue-hui2   

  1. 1The 3rd Dental Center, Peking University Hospital of Stomatology, Beijing 100191, China; 2Department of Dental Materials, Peking University Hospital of Stomatology, Beijing 100081, China
  • Received:2018-04-21 Online:2018-06-28 Published:2018-06-28
  • Contact: Zhang Xue-hui, Associate researcher, Department of Dental Materials, Peking University Hospital of Stomatology, Beijing 100081, China
  • About author:Peng Hui, Master, Physician, the 3rd Dental Center, Peking University Hospital of Stomatology, Beijing 100191, China
  • Supported by:

    the National High-Technology Research and Development Research of China (863 Program), No. 2015AA033601

摘要:

文章快速阅读:

 

文题释义:
煅烧鹿角松质骨:是去除鹿角皮质骨后,经过化学制剂对鹿角松质骨进行脱脂脱蛋白处理,采用600 ℃高温煅烧方法获得的一种颗粒状异种无机骨填充材料,前期研究已经证实其具有良好的成骨性能。
早期血管化:血管化对骨缺损修复至关重要,尤其对于大范围骨缺损,尽早建立血管化可保证缺损区内细胞有充足的营养和氧分,加速代谢物质的转运,建立良好的动态微环境,从而促进骨缺损修复。
 
 
背景:前期研究成功制备了煅烧鹿角松质骨材料,并证实其可明显促进骨缺损的修复。
目的:考察煅烧鹿角松质骨修复骨缺损的早期血管化作用。
方法:去除鹿角皮质骨,将鹿角松质骨经脱脂脱蛋白和高温煅烧制备成颗粒状骨填充材料煅烧鹿角松质骨,采用扫描电镜和X射线衍射仪对其进行表面形貌观察和晶相结构检测。将小鼠前成骨细胞MC3T3-E1接种于煅烧鹿角松质骨支架材料上,共培养1 d后扫描电镜观察细胞铺展情况。制备兔双侧下颌骨临界尺寸骨缺损模型,一侧植入煅烧鹿角松质骨材料,另一侧植入Bio-Oss®颗粒,空白对照组不植入任何材料,术后4,8周取下颌骨标本,进行苏木精-伊红染色、Masson染色和CD31免疫组织化学染色,观察成骨和成血管情况。

结果与结论:①扫描电镜观察可见煅烧鹿角松质骨为互通多孔结构,棱柱状规则磷灰石晶体,X射线衍射图谱显示煅烧鹿角松质骨各特征峰与人骨一致,为结晶度良好的羟基磷灰石相;②成骨细胞在材料表面铺展良好,并与羟基磷灰石晶体紧密结合;③材料植入兔下颌骨缺损4周后,苏木精-伊红染色结果显示煅烧鹿角松质骨组和Bio-Oss®组植入材料周围有明显新生血管;Masson染色和CD31免疫组织化学结果显示,两组材料周围有新生成的胶原纤维和新生血管,且煅烧鹿角松质骨组较Bio-Oss®组血管化效果更为明显;④材料植入8周后苏木精-伊红染色结果显示,煅烧鹿角松质骨组、Bio-Oss®组骨缺损处均有大量新骨形成;⑤结果表明,煅烧鹿角松质骨在早期骨缺损修复过程中具有良好的血管化作用,这可能是其促进骨缺损修复的关键因素。

ORCID: 0000-0001-9595-1295(彭晖) 

关键词: 鹿角, 骨再生, 骨组织工程, 骨充填材料, 骨缺损修复, 血管化, 骨生成, 血管生成, 支架材料, 生物材料

Abstract:

BACKGROUND: Calcinated antler cancellous bone (CACB) was successfully prepared in the early stage, and was confirmed that it could significantly promote bone defect repair.

OBJECTIVE: To evaluate the early neovascularization of CACB in a bone defect model.
METHODS: CACB was prepared by defatted and deproteinized process and combined with calcination after removal of the cortical bone. The surface morphology and phase composition of CACB granules were evaluated by using scanning electron microscope (SEM) and X-ray diffraction spectrum (XRD). Mouse osteoblasts MC3T3-E1 were cultured with CACB for 1 day to assess the cellular affinity by SEM. Models of critical bone defects were prepared on the bilateral mandibles of 16 New Zealand rabbits. CACB was implanted into the defects on the one side, and Bio-Oss® was implanted into the defects on the other side. Nothing was implanted into the defects as blank control. All the rabbit mandibles after 4 and 8 weeks post-surgery were subjected to tissue processing and sectioning. Hematoxylin-eosin, Masson’s trichrome staining and CD31 immunohistochemical method were performed separately on consecutive tissue sections to observe the vascularization and osteogenesis.
RESULTS AND CONCLUSION: (1) SEM images showed that CACB exhibited a well-interconnected porous structure and was present as regularly prismatic crystals of hydroxyapatite. The XRD spectrograph revealed that the characteristic peaks of CACB corresponded to the peaks of calcinated human cancellous bone, indicating that the hydroxyapatite phase with good crystallinity is the same as that of human bone. (2) The cellular experiment findings showed that CACB could be conducive for attachment and spreading of MC3T3-E1 cells integrated with hydroxyapatite closely. (3) At 4 weeks post-implantation, hematoxylin-eosin staining results showed a lot of newly formed small blood vessels at the periphery of the scaffold of CACB and Bio-Oss®. Masson and CD31 immunohistochemical staining results showed the newly formed collagen fibers and blood vessels around the bone filling materials. Moreover, the ability of vascularization of CACB was more effective than that of Bio-Oss®. (4) At 8 weeks post-implantation, hematoxylin-eosin staining results showed there was a large number of newly formed bones in the bone defects and the remaining scaffold was surrounded by newly formed bones. To conclude, our findings reveal that CACB possesses good vascularization during the bone defect repair, which may have a potentially positive effect on promoting osteogenesis. 

Key words: Bone Regeneration, Osteogenesis, Tissue Engineering

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