羧甲基壳聚糖钙与骨髓基质细胞生物相容性的初步实验

山东大学学报(医学版) ›› 2010, Vol. 48 ›› Issue (6): 57-60.

羧甲基壳聚糖钙与骨髓基质细胞生物相容性的初步实验

崔军1，徐欣1,2，马跃1，董鹏飞3，刘刚利1，文勇1

1.山东大学口腔医学院口腔种植中心, 济南 250012； 2.山东省口腔生物医学重点实验室, 济南 250012；
3.滨州医学院附属医院口腔颌面外科, 山东滨州 256603

收稿日期:2009-12-14 出版日期:2010-06-16 发布日期:2010-06-16
通讯作者: 徐欣，女，教授，主要从事口腔种植的基础与临床研究。 Email:xinxu@sdu.edu.cn
作者简介:崔军（1975- ），男，博士研究生，主治医师，主要从事口腔种植与生物材料学研究。

A preliminary experimental study on bone marrow stromal cell biocompatibility of carboxymethyl chitosan calcium

CUI Jun1, XU Xin1,2, MA Yue1, DONG Pengfei3, LIU Gangli1, WEN Yong1

1. Department of Dental Implant Center, School of Stomatology, Shandong University, Jinan 250012, China;
2. Key Lab of Oral Biomedicine of Shandong Province, Jinan 250012, China;
3. Department of Oral and Maxillofacial Surgery, Affiliated Hospital to Binzhou Medical College,
Binzhou 256603, Shandong, China

Received:2009-12-14 Online:2010-06-16 Published:2010-06-16

摘要/Abstract

摘要：

目的通过体外小鼠骨髓基质细胞培养，从细胞形态和细胞增殖方面评价新型生物材料羧甲基壳聚糖钙的细胞生物相容性。方法合成羧甲基壳聚糖钙，进行红外光谱分析，并用DMEM制备浸提液。分别用含羧甲基壳聚糖钙浸提成分的培养液 (实验组)、DMEM 培养液(阴性对照组)和含6.4g／L苯酚培养液 (阳性对照组)培养骨髓基质细胞，通过倒置相差显微镜观察和MTT比色法评价细胞生长及增殖情况。结果红外光谱分析提示，羧甲基壳聚糖与钙离子发生了络合反应。镜下观察，细胞培养1、2、3d，实验组与阴性对照组细胞数量明显增加，细胞形态正常；阳性对照组细胞数量明显减少，细胞固缩甚至崩解。MTT比色法，实验组与阴性对照组吸光度(A)值间无统计学差异(P>0.05)，实验组各时间点的毒性分级为0级；阳性对照组A值明显低于其他组(P<0.01)，毒性分级为4级。结论羧甲基壳聚糖钙复合材料对小鼠骨髓基质细胞有良好细胞生物相容性，是一种可用于组织工程的新型生物材料。

关键词: 羧甲基壳聚糖钙；骨髓基质细胞；细胞相容性；细胞毒性；生物材料

Abstract:

Objective To evaluate the cellular biocompatibility of carboxymethyl chitosan calcium (CCC)composite by observing the morphology and proliferation of murine bone marrow stromal cells(BMSCs) in vitro. Methods (CCC) was synthesized and then analysed by an infrared spectrometer (IR). CCC extraction prepared with DMEM (experimental group), DMEM culture medium (negative control group) and 6.4g/L phenol culture medium (positive control group) were used for BMSCs culture. The morphology and proliferation of BMSCs were evaluated by inverted phase contrast microscopic observation and MTT assay. Results Infrared spectrum analysis indicated CCC had been formed by complex reaction between carboxymethyl chitosan and calcium ions. On culture day 1, 2 and 3, the cells disaggregated and the cell number decreased noticeably in the positive control group; however, the cell number in the experimental group and negative control group increased remarkably, with normal cell appearance. The absorbance (A) values of the experimental group and the negative control group were increased with time, and there was no statistically significant difference between the two groups (P>0.05). While the A value in the positive control group was significantly decreased compared with the other two groups (P<0.01). At each time point of culture, the cytotoxicity grade was 0 in the experimental group, and 4 in the positive control group. Conclusion (CCC) composite has potential application in tissue engineering owning to good cellular compatibility with BMSCs.

Key words: Carboxymethyl chitosan calcium; Bone marrow stromal cells; Cellular compatibility; Cytotoxicity; Biomaterials

中图分类号:

R318

崔军1，徐欣1,2，马跃1，董鹏飞3，刘刚利1，文勇1. 羧甲基壳聚糖钙与骨髓基质细胞生物相容性的初步实验[J]. 山东大学学报(医学版), 2010, 48(6): 57-60.

CUI Jun1, XU Xin1,2, MA Yue1, DONG Pengfei3, LIU Gangli1, WEN Yong1. A preliminary experimental study on bone marrow stromal cell biocompatibility of carboxymethyl chitosan calcium[J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2010, 48(6): 57-60.

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