新型壳聚糖基温敏凝胶膜引导骨再生性能的体外实验研究

上海口腔医学 ›› 2015, Vol. 24 ›› Issue (4): 442-446.

新型壳聚糖基温敏凝胶膜引导骨再生性能的体外实验研究

梁杰¹, 崔军², 许胜¹, 周倜¹, 徐伟成¹, 徐欣^{3, 4}

1.烟台市口腔医院,山东烟台 264001; 2.济南市口腔医院,山东济南 250001; 3.山东大学口腔医学院口腔种植中心,山东济南 250012; 4.山东省口腔生物医学重点实验室,山东济南 250012

收稿日期:2014-07-24 出版日期:2015-08-20 发布日期:2015-09-10
通讯作者: 徐欣,Tel:0531-88382636,E-mail: xinxu@sdu.edu.cn E-mail:xinxu@sdu.edu.cn
作者简介:梁杰(1986-),女,硕士研究生,医师,E-mail：kathyliangjie@163.com
基金资助:
烟台市科技发展计划项目（2013ws248）; 山东省中青年科学家科研奖励基金计划项目（BS2013YY056）

In vitro study on the properties of guided bone regeneration of new type chitosan-based thermosensitive hydrogel membranes

LIANG Jie¹, CUI Jun², XU Sheng¹, ZHOU Ti¹, XU Wei-cheng¹, XU Xin^{3, 4}

(1.Yantai Stomatological Hospital. Yantai 264001; 2. Jinan Stomatological Hospital. Jinan 250001; 3. Department of Implantology, School of Stomatology Shandong University. Jinan 250012; 4. Key Lab of Oral Biomedicine of Shandong Province. Jinan 250012, Shandong Province, China

Received:2014-07-24 Online:2015-08-20 Published:2015-09-10
Supported by:
Supported by Science and Technology Development Planning Project of Yantai City(2013WS248) and Science and Technology Award Program for Young and Middle-aged Scientists of Shandong Province(BS2013YY056)

摘要/Abstract

摘要： 目的制备负载釉基质蛋白（enamel matrix proteins,EMPs）的壳聚糖/β甘油磷酸钠(chitosan/β-glycerophosphate salt,CS/β-GP)温敏凝胶膜,探讨其体外引导骨再生的能力。方法利用CS/β-GP体系的温敏相转变特性,制备新型生物膜并添加牛血清白蛋白（bovine serum albumin,BSA）,利用蛋白浓度测定试剂盒（加强型）检测不同时点的蛋白释放浓度,绘制蛋白缓释曲线。添加EMPs的CS/β-GP（1.0 g）复合膜作为A组; 单纯CS/β-GP（1.0 g）复合膜作为B组;同时,设空白对照组即C组,分别与ST2细胞共培养。检测膜的理化性能, MTT法检测复合膜的生物相容性, PNPP法检测碱性磷酸酶（alkaline phosphatase, ALP）的活性。采用SPSS16.0软件包对数据进行统计学分析。结果不同浓度的复合膜可缓慢释放蛋白12 d以上,并且随着β-GP浓度的升高,蛋白缓释总量增大。负载蛋白后,复合膜的理化性能均未发生显著变化（P>0.05）。MTT检测A、B、C 3组的吸光度（OD）值,组间差异具有显著性,A、B组OD值均高于C组,A组OD值高于B组（P<0.05）。A、B组ALP表达高于空白对照组（P<0.05）,A、B 2组之间ALP活性表达亦具有显著差异,A组高于B组（P<0.05）。结论负载EMPs的新型壳聚糖基温敏凝胶膜在体外实验中表现出一定的引导骨再生活性,是一种具有应用潜力的新型生物膜材料。

关键词: 壳聚糖, β, 甘油磷酸钠, 温敏凝胶, 引导骨再生, 牛血清白蛋白, 釉基质蛋白

Abstract: PURPOSE：To prepare chitosan/β-glycerophosphate salt (CS/β-GP) thermosensitive hydrogel membranes loaded enamel matrix proteins and detect the guided bone regeneration properties. METHODS：A newly membrane was synthesized using thermal phase inversion property of the CS/β-GP system. The membrane was synthesized and added with protein BSA. The concentration of protein was detected at different time points by enhanced protein assay kit, and the protein release curve was drawn. CS/β -GP membrane added EMPs (1.0 g) as group A, CS/β -GP (1.0 g) membrane as group B and nothing as blank control group (group C). They were co-cultured with ST2 cells. The mechanical properties of the membranes were tested in vitro, and the compatibility properties were detected by MTT method. The activity of ALP was assayed by PNPP method. The data was analyzed by SPSS 16.0 software package. RESULTS：Membranes with different concentration of CS/β -GP could release protein slowly more than 12 days, and the total quantity of the released protein increased with the concentration of the β-GP. The changes of mechanical properties of the membranes were not significant（P>0.05）. The OD value of group A, B and C had statistically significant difference in the MTT test. The values of group A and B were higher than that of group C, while the value of group A was higher than that of group B (P<0.05). The activities of ALP were different in the three groups .The activities of group A and B were higher than that of the blank control group(P<0.05）.The difference in expression of ALP between group A and B was also statistically significant. The expression in group A was higher than that in group B (P<0.05). CONCLUSIONS: The new type CS/β-GP membrane shows property of guided bone regeneration in vitro, which have the potentials for clinical use.

Key words: Chitosan, β-Glycerophosphate salt, Thermosensitive hydrogel, Guided bone regeneration, Bovine serum albumin, Enamel matrix proteins Shanghai J Stomatol, 2015, 24(4):442-446.

中图分类号:

R781.34

梁杰, 崔军, 许胜, 周倜, 徐伟成, 徐欣. 新型壳聚糖基温敏凝胶膜引导骨再生性能的体外实验研究[J]. 上海口腔医学, 2015, 24(4): 442-446.

LIANG Jie, CUI Jun, XU Sheng, ZHOU Ti, XU Wei-cheng, XU Xin. In vitro study on the properties of guided bone regeneration of new type chitosan-based thermosensitive hydrogel membranes[J]. Shanghai Journal of Stomatology, 2015, 24(4): 442-446.

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