中国组织工程研究

中国组织工程研究 ›› 2019, Vol. 23 ›› Issue (30): 4775-4779.doi: 10.3969/j.issn.2095-4344.1403

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

改良低温机械物理研磨法制备纳米脱钙骨基质

王智巍，苑  博，陈雄生，周盛源，贾连顺
  

  1. 解放军第二军医大学上海长征医院骨科医院脊柱外科，上海市  200003
  • 收稿日期:2019-04-21 出版日期:2019-10-28 发布日期:2019-10-28
  • 通讯作者: 陈雄生，博士，博士生导师，主任医师，教授，解放军第二军医大学上海长征医院骨科医院脊柱外科，上海市 200003
  • 作者简介:王智巍，男，1986年生，辽宁省鞍山市人，满族，2018年解放军第二军医大学毕业，博士，解放军第二军医大学流行病与卫生统计学教研室在站博士后，主要从事骨组织工程的基础研究和脊柱韧带骨化病的临床研究。
  • 基金资助:

    上海市科委基础研究重点项目(15JC1491003)，项目负责人：陈雄生

Preparation of nano-demineralized bone matrix by modified low temperature mechanical trituration 

Wang Zhiwei, Yuan Bo, Chen Xiongsheng, Zhou Shengyuan, Jia Lianshun
  

  1. Spine Center, Department of Orthopedics, Shanghai Changzheng Hospital, Second Military Medical University, Shanghai, 200003, China
  • Received:2019-04-21 Online:2019-10-28 Published:2019-10-28
  • Contact: Chen Xiongsheng, MD, Doctoral supervisor, Chief physician, Professor, Spine Center, Department of Orthopedics, Shanghai Changzheng Hospital, Second Military Medical University, Shanghai, 200003, China
  • About author:Wang Zhiwei, MD, Spine Center, Department of Orthopedics, Shanghai Changzheng Hospital, Second Military Medical University, Shanghai, 200003, China
  • Supported by:

    The Major Project of the Science and Technology Commission of Shanghai Municipality, No. 15JC1491003 (to CXS)

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摘要：

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文题释义：
脱钙骨基质：是对异体骨通过化学方法进行脱钙处理后得到的支架，由93%的胶原、5%的可溶性蛋白(诱导成骨的骨形态发生蛋白和转化生长因子、胰岛素样生长因子等协同蛋白)及2%的残余矿化基质组成，它在微结构和组成上与天然骨骼高度相似，可使间充质细胞分化为成软骨细胞，诱导生成的软骨被重吸收之后再诱导骨生成。
低温研磨技术：一些弹性较大、易挥发或热敏感的材料，为达到目标颗粒尺寸，需要在低温条件下进行研磨粉碎处理，材料在低温环境下成分不易被破坏或降解，同时又能使材料变硬，还能避免高挥发性物质的蒸发。
 
 
背景：前期研究证实纳米脱钙骨基质为一种无毒、组织相容性良好、生物利用度高、炎症反应轻、成骨能力强的骨移植替代物，但其制备耗时较长，需多次间断取样行粒度分析，研磨效率低。
目的：采用低温物理研磨法制备纳米脱钙骨基质，分析纳米脱钙骨基质的结构特征及制备效率。
方法：采用改良Urist法制备同种异体脱钙骨基质，通过二次低温物理研磨法，应用CryoMill全自动冷冻研磨仪先将脱钙骨基质初步研磨至微米级别颗粒，再通过E-Max高能水冷球磨仪进行纳米尺度研磨，最后通过扫描电镜观测研磨效果及材料形貌。
结果与结论：采用二次低温物理研磨法制备的纳米脱钙骨基质底层致密，表面充满不规则纳米颗粒，颗粒直径20-50 nm，纳米颗粒相互团聚，表面密布纳米级别凹槽，纳米纤维结构间相互连接，内部形成大量相互连通的微米级别孔隙，其微观结构符合纳米生物材料范畴；二次低温物理研磨法仅需25 min，提高了研磨效率；结果说明，二次低温物理研磨法制备纳米脱钙骨基质的研磨结果更确切、材料粒度分布均匀性更高。

关键词: 纳米材料, 纳米脱钙骨基质, 脱钙骨基质, 纳米化, 低温物理研磨, 显微特征, 研磨

Abstract:

BACKGROUND: Previous studies have confirmed that nano-demineralized bone matrix is an alternative to an autograft with non-toxicity, good tissue compatibility, high bioavailability, mild inflammatory reaction, strong osteogenic ability. But its preparation takes long time. Intermittent sampling is required for particle size analysis.
OBJECTIVE: To prepare nano-demineralized bone matrix by low temperature mechanical trituration and analyze its structure characteristics and preparation efficacy.
METHODS: Allogeneic decalcified bone matrix was prepared by the modified Urist method. The CryoMill automatic freezing grinder was used to initially grind the decalcified bone matrix to the micron-sized particles. Then the E-Max high-energy water-cooled ball mill was used to grind the micro-sized particles into nano-sized particles. Finally, the grinding effect and material appearance were observed by scanning electron microscopy.
RESULTS AND CONCLUSION: After two-step grinding, the prepared decalcified bone matrix had nanostructure composed by irregular 20-50 nm-sized nanoparticles through agglomeration. The surface of nano-particles was densely covered with nano-scale grooves. A large number of interconnected micro-pores formed between nanofiber structures, the microstructure of which conformed to the category of nanobiomaterials. Compared with the Micro superfine mill, the new trituration process only took 25 minutes, which increases grinding efficacy. These results suggest that two steps of low temperature mechanical trituration for preparation of nano-demineralized bone matrix produces more uniform particle size distribution.

Key words: nanomaterials, nano-demineralized bone matrix, demineralized bone matrix, nanocrystallization, low temperature mechanical trituration, micro-characteristic, trituration

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