摘要
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。
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 sh
作者
彭晖
张学慧
Peng Hui;Zhang Xue-hui(The 3rd Dental Center,Peking University Hospital of Stomatology,Beijing 100191,China;Department of Dental Materials,Peking University Hospital of Stomatology,Beijing 100081,China)
出处
《中国组织工程研究》
CAS
北大核心
2018年第18期2807-2812,共6页
Chinese Journal of Tissue Engineering Research
基金
国家高技术研究发展(863)计划课题(2015AA033601)~~
