新型双层骨引导再生壳聚糖膜修复大鼠颅骨缺损的实验研究

中国口腔颌面外科杂志 ›› 2015, Vol. 13 ›› Issue (2): 97-104.

• 基础研究 • 下一篇

新型双层骨引导再生壳聚糖膜修复大鼠颅骨缺损的实验研究

陈琦¹, 万光勇^{2, *}, 张善勇^{3, *}, 焦子先³

1.潍坊医学院口腔医学院,山东潍坊 261000;
2.泰安市中心医院口腔科,山东泰安 271000;
3.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:2014-10-13 修回日期:2014-11-24 出版日期:2015-03-20 发布日期:2015-04-23
通讯作者: 万光勇,E-mail: wgykq@163.com;张善勇,E-mail: zhangshanyong@126.com。*共同通信作者
作者简介:陈琦（1989-）,男,在读硕士研究生,E-mail：chenqill@163.com
基金资助:
国家自然科学基金（81371168）; 上海市科学技术委员会西医引导类项目(134119a5400)

An experimental study of a novel double-layer chitosan membrane for guided bone regeneration in rat calvarial defects

CHEN Qi¹, WAN Guang-yong², ZHANG Shan-yong³, JIAO Zi-xian³

1. College of Stomatology, Weifang Medical University. Weifang 26100, Shandong Province;
2. Department of Stomatology, Tai’an Central Hospital. Tai’an 271000, Shandong Province;
3. 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:2014-10-13 Revised:2014-11-24 Online:2015-03-20 Published:2015-04-23
Supported by:
Supported by National Natural Science Foundation of China (81371168) and Western Medicine Guide Project of Science and Technology Committee of Shanghai Municipality (134119a5400)

摘要/Abstract

摘要： 目的: 通过比较新型双层壳聚糖膜和Bio-Gide膜对大鼠颅骨缺损的修复效果,评估其作为引导骨再生膜的可行性。方法: 双层壳聚糖膜与MC3T3-E1细胞共培养,应用扫描电子显微镜（scanning electron microscope,SEM）观察细胞对膜的黏附效果;采用cell counting kit-8（CCK-8）法在3、7和10 d检测膜的细胞毒性;选择18只Sprague-Dawley（SD）大鼠,随机分为A、B、C组,在其颅骨中线两侧制作5 mm的极量骨缺损,左侧为实验侧,覆盖双层壳聚糖膜,右侧为对照侧覆盖Bio-Gide膜,分别于术后第2、4及8周时处死各组大鼠,通过大体观察、微计算机体层扫描技术（micro-computed tomography,micro-CT）及组织学方法,评价2种膜修复骨缺损的效果。采用SPSS20.0软件包对结果进行统计学分析。结果: 通过MC3T3-E1细胞在双层壳聚糖膜上培养2 d后的扫描电镜照片,观察到细胞较好地粘附于多孔层上。第3、7及10天时,实验组细胞相对增殖率分别为114.49%、107.17%和98.73%,细胞毒性级别为0或1级,表明膜的细胞毒性轻微。2周时,Bio-Gide膜组的骨形成量（BV）和骨体积分数（BVF、BV/TV）与实验组差异显著（P<0.05）;但在第4和8周时,2组的BV和BVF无显著差异（P>0.05）。结论: 新型双层壳聚糖的体外和体内生物学性能符合GBR技术的要求,具有作为GBR膜的潜力。

关键词: 壳聚糖膜, 胶原膜, 生物材料, 骨缺损, 引导骨再生, 微计算机断层扫描技术, 颅骨缺损, 大鼠

Abstract: PURPOSE: To compare new bone formation in rat calvarial defects using novel double-layer chitosan membranes (DLCM) or collagen (Bio-Gide) membranes, and evaluate the feasibility as guided bone regeneration (GBR) membrane. METHODS: The MC3T3-E1 cells were cultured on the DLCM for 2 days, then SEM was used to observe the attachment of the cell line to the membrane. The cytotoxicity of the DLCM was tested by CCK-8 assay at day 3, 7 and 10. Eighteen (18)SD rats were selected, and then divided into 3 groups (A,B and C). Two calvarial critical-sized defects (CSDs), 5 mm in diameter, were created symmetrically on the bilateral sides of the midline using a trephine bur. Right defect was covered with a resorbable collagen membrane (Bio-Gide) as a control and the left side was covered with DLCM. The rats were sacrificed 2, 4 and 8 weeks after surgery. The specimens were then analysed using micro-CT and histological methods,to compare new bone formation in rat calvarial defects using the two membranes. The data was analyzed for Student’s t test with SPSS20.0 software package. RESULTS: From the SEM image of MC3T3-E1 cells on the double-layer chitosan membrane after being cultured for 2 days. It was found that the cells adhered well to the porous layer. The results of cytotoxicity showed that cell proliferation rates of the experimental groups were 114.49%, 107.17% and 98.73% at day 3,7 and10, respectively, while the cytotoxicity was in grade 0 or 1, indicating that the chitosan membrane had low cytotoxicity. At 2 week, there were significant differences in bone volume (BV) and bone volume/total volume (BVF, BV/TV) (P<0.05) between the collagen group and the chitosan membrane group. But at 4 and 8 week, there was no significant difference in BV and BVF (P>0.05) between the 2 groups. CONCLUSIONS: This pilot study indicated that the in vitro and in vivo bioactivities of the novel chitosan membrane fulfilled the requirements for GBR, and has significant potential as a GBR membrane.

Key words: Chitosan membrane, Collagen membrane, Biomaterials, Bone defect, Guided bone regeneration, Micro-computed tomography, Calvarial defect, Rat

中图分类号:

R782.1

陈琦, 万光勇, 张善勇, 焦子先. 新型双层骨引导再生壳聚糖膜修复大鼠颅骨缺损的实验研究[J]. 中国口腔颌面外科杂志, 2015, 13(2): 97-104.

CHEN Qi, WAN Guang-yong, ZHANG Shan-yong, JIAO Zi-xian. An experimental study of a novel double-layer chitosan membrane for guided bone regeneration in rat calvarial defects[J]. China J Oral Maxillofac Surg, 2015, 13(2): 97-104.

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新型双层骨引导再生壳聚糖膜修复大鼠颅骨缺损的实验研究

An experimental study of a novel double-layer chitosan membrane for guided bone regeneration in rat calvarial defects

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