脱细胞脱钙人牙用于大鼠颅盖骨缺损修复的实验研究

doi:10.19438/j.cjoms.2018.05.003

中国口腔颌面外科杂志 ›› 2018, Vol. 16 ›› Issue (5): 397-401.doi: 10.19438/j.cjoms.2018.05.003

脱细胞脱钙人牙用于大鼠颅盖骨缺损修复的实验研究

刘宗霖^1,*, 郭克^2,*, 王文超^1,*, 张耀升¹, 徐伟峰², 张善勇^2#, 刘秀明^2#, 金佳梅^2#

1.潍坊医学院口腔医学院,山东潍坊 261053;
2.上海交通大学医学院附属第九人民医院·口腔医学院口腔外科,上海市口腔医学重点实验室,上海 200011;

收稿日期:2018-06-08 出版日期:2018-09-20 发布日期:2018-11-06
通讯作者: 张善勇,E-mail: zhangshanyong@126.com;刘秀明,E-mail：sniperr@126.com;金佳敏,E-mail：pterion84@126.com。^#共同通信作者
作者简介:刘宗霖（1990-）,男,在读硕士研究生,E-mail: miaomiao332777020@qq.com;王文超（1990-）,男,在读硕士研究生,E-mail:16382358@qq.com;郭克（1991-）,男,在读硕士研究生,E-mail: 814233948@qq.com。并列第一作者
基金资助:
浦东新区卫计委联合攻关项目（PW2016D-1）; 上海市科委西医引导类项目（134119a5400 ）; 上海交通大学医学院大学生科创项目（20160508）; 2017年上海市卫计委面上项目（M20170232）; 2018年上海市科委生物医药支撑项目（18441900100）

Experimental study of acellular and demineralized human tooth in repair of calvarial bone defect in rats

LIU Zong-lin¹, GUO Ke², WANG Wen-chao¹, ZHANG Yao-sheng¹, XU Wei-feng², ZHANG Shan-yong², LIU Xiu-ming², JIN Jia-min²

1.College of Stomatology, Weifang Medical University. Weifang 261053, Shandong Province;
2.Department of Oral Surgery, Shanghai Ninth People's Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine; Shanghai Key Laboratory of Stomatology. Shanghai 200011, China

Received:2018-06-08 Online:2018-09-20 Published:2018-11-06

摘要/Abstract

摘要： 目的: 探讨脱细胞脱钙牙作为骨支架材料的可行性和诱导大鼠颅盖骨缺损成骨能力。方法: 离体人牙24颗,分别进行脱钙和脱钙脱细胞处理。实验1—取6只10~12周龄雄性SD大鼠,在大鼠腹部皮下同时包埋脱钙牙、脱细胞脱钙牙,4周后处死大鼠,进行H-E染色,观察2种材料周围炎症反应情况。实验2—取12只10~12周龄雄性SD大鼠,制备双侧颅盖骨区域骨缺损,在左侧骨缺损处植入脱细胞脱钙牙,以未植入任何材料的右侧作为空白对照,4周、8周时分别处死6只,获取完整标本,进行Micro-CT扫描,从大体形态学上比较成骨量。取4周大鼠进行H-E染色,观察周围炎症反应,免疫组织化学观察新生成骨程度。采用SPSS 23.0软件包进行重复测量方差分析。结果: 实验1—4周时H-E染色显示,脱细胞脱钙牙与周围组织炎症反应程度显著小于脱钙牙。实验2—取4周和8周的大鼠颅盖骨进行Micro-CT扫描和骨缺损处新生骨量定量分析,显示植入脱细胞脱钙牙的骨缺损处成骨量显著多于空白对照组量（P<0.05）。4周时H-E染色结果显示,脱细胞脱钙牙周围的炎症反应程度与空白对照处相似。免疫组织化学染色显示,脱细胞脱钙牙周围新生的成骨细胞显著多于空白对照组。结论: 脱细胞脱钙人牙是一种具有良好生物相容性、成骨诱导能力和可塑性的生物支架材料。

关键词: 颅盖骨缺损, 脱钙骨基质, 脱细胞骨基质, 离体人牙, 大鼠

Abstract: PURPOSE: To investigate the feasibility of acellular decalcified human teeth as bone scaffold materials and their ability to induce bone defect and osteogenesis in rats. METHODS: Twenty-four isolated human teeth were treated with decalcification and decellularization respectively. In experiment 1, 6 male SD rats aged 10-12 weeks were used to embed decalcified teeth and acellular and demineralized teeth under the skin of the abdomen of the rats. After 4 weeks, the rats were sacrificed and H-E staining was performed to observe the inflammatory reaction around the two materials. In experiment 2, 12 male SD rats of 10-12 weeks old were selected to prepare bilateral calvarial bone defects,the left side of the bone defect was implanted with decalcified and acellular teeth, while the right side of the bone defect without any material was used as a blank control. Six rats were randomly selected for 4 weeks and 8 weeks respectively to be sacrificed. Complete specimens were obtained and Micro- CT scan was performed to compare bone mass from the general morphology. Rats at 4 weeks were taken for H-E staining and immunohistochemistry to observe the peripheral inflammatory response and the extent of newly formed bone. SPSS 23.0 software package was used for ANOVA of repeated measurement. RESULTS: After 4 weeks, H-E staining showed that the inflammatory reaction between acellular and demineralized teeth and surrounding tissues was significantly less than that of decalcified teeth. Micro-CT scanning and quantitative analysis of 4 and 8 weeks of rat calvarial bone showed that the bone mass in the bone defect implanted with acellular and demineralized teeth was significantly higher than that in the blank control (P<0.05). The results of H-E staining of rats at 4 weeks showed that the inflammatory response around the acellular and demineralized teeth was similar to the blank control. Immunohistochemical results showed that the number of new osteoblasts around the acellular and demineralized teeth was significantly higher than that of the blank control. CONCLUSIONS: Decalcified and decellularized teeth are a kind of biological scaffold material with good biocompatibility,osteogenic induction ability and plasticity.

Key words: Calvarial bone defect, Demineralized bone matrix, Acellular bone extracellular matrix, Extracted human teeth, Rat

中图分类号:

R318.08

刘宗霖, 郭克, 王文超, 张耀升, 徐伟峰, 张善勇, 刘秀明, 金佳梅. 脱细胞脱钙人牙用于大鼠颅盖骨缺损修复的实验研究[J]. 中国口腔颌面外科杂志, 2018, 16(5): 397-401.

LIU Zong-lin, GUO Ke, WANG Wen-chao, ZHANG Yao-sheng, XU Wei-feng, ZHANG Shan-yong, LIU Xiu-ming, JIN Jia-min. Experimental study of acellular and demineralized human tooth in repair of calvarial bone defect in rats[J]. China J Oral Maxillofac Surg, 2018, 16(5): 397-401.

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脱细胞脱钙人牙用于大鼠颅盖骨缺损修复的实验研究

Experimental study of acellular and demineralized human tooth in repair of calvarial bone defect in rats

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