纤维蛋白胶和异种无机骨构建复合支架材料与种子细胞的复合-中国组织工程与临床康复2007年1期目次

纤维蛋白胶和异种无机骨构建复合支架材料与种子细胞的复合*☆

武汉1，2，张春秋3，章培标4，尹飞2，刘景臣2，李玉新1

Combination of compound scaffolds constructed by fibrin
glue and xenogeneic inorganic bone and seed cells*☆

Abstract

AIM： To construct bone combined scaffolds in bone tissue engineering with fibrin glue and xenogeneic inorganic bone and perform three-dimensional culture of rabbit marrow stroma cells (MSCs) as well as probe into the adhesion effect of the scaffold on seed cells.

METHODS： The experiment was co-conducted by the Department of Orthopedics and Trauma of China-Japan Union Hospital and the Department of Mechanical Engineering, College of Mechanical Engineering of Jilin University and the Department of Genetics and Cell of Northeast Normal University between January 2005 and April 2006. The bovine cancellous bone (BCB) was dealt with H2O2, NaOH and alcohol to prepare inorganic bone. After that the BCB was combined with the fibrin glue to make combined scaffolds. Rabbit MSCs were cultured in vitro and transfer, and the MSCs were collected for three-dimension culture with fibrin glue and combined scaffold made of xenogeneic inorganic bone. The cell growth in fibrin glue was examined by HE staining, phase-contrast microscope and transmission electron microscope.

RESULT: ① Observation under the phase contrast microscope showed that the MSCs could be seen evenly mixed with the fibrin glue from inorganic bone gaps. ② At the 4th week of culture, the MSCs formed into dense stereo-net. ③ It could be found under the transmission electron microscope that there were micro-protrusion in local stroma cells at one month after the culture, and mitochondria as well as ribosome were found in the cytoplasm with rough endoplasmic reticulum.

CONCLUSION: The MSCs in scaffold combined by fibrin glue and xenogeneic inorganic bone are better in activity, and they can rapidly proliferate, which indicate that this kind of scaffold fits for the growth of seed cells.

1Postdoctoral Station of Department of Genetics and Cell, Northeast Normal University, Changchun 130032, Jilin Province, China; 2Department of Orthopaedics, China-Japan Union Hospital, Changchun 130033, Jilin Province, China; 3Department of Mechanical Engineering, College of Mechanical Engineering, Jilin University, Changchun 130032, Jilin Province, China; 4Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130032, Jilin Province, China

Wu Han☆, Doctor, Associate professor, Postdoctoral Station of Department of Genetics and Cell, Northeast Normal University, Chanchun 130032, Jilin Province, China; Department of Orthopaedics, China-Japan Union Hospital, Changchun 130033, Jilin Province, China
mdwuhan@163.com

Supported by: the National Natural Science Foundation of China, No. 30570451*

Received: 2006-05-15
Accepted: 2006-08-30

摘要
目的：应用纤维蛋白胶、异种无机骨构建骨组织工程复合支架材料，立体培养兔骨髓基质细胞，探讨这种复合支架材料对种子细胞的黏附作用。
方法：实验于2005－01/2006－04在吉林大学中日联谊医院卫生部创伤骨科研究室、吉林大学机械科学与工程学院工程力学系、东北师范大学遗传与细胞研究所合作完成。牛松质骨经去脂去蛋白等无机化处理后与纤维蛋白胶复合制成复合骨支架材料；将兔骨髓基质细胞作为种子细胞进行传代培养，收集后在与纤维蛋白胶、异种无机骨构成的复合支架中进行立体培养，采用相差显微镜、透射电子显微镜、苏木精-伊红染色等手段观察骨髓基质细胞在纤维蛋白胶中的生长状况。
结果: ①相差显微镜下观察自无机骨孔隙中见骨髓基质细胞均匀混合于纤维蛋白胶中。②细胞培养4周骨髓基质细胞形成密集立体网状。③透射电子显微镜见在复合支架中培养1个月的1个基质细胞局部有细突起，胞质内可见线粒体，核糖体，粗面内质网。
结论:在纤维蛋白胶与异种无机骨构建的复合支架中骨髓基质细胞具有良好活性，可迅速扩增生长，提示这种支架材料具有适合种子细胞生长的环境。
关键词：纤维蛋白；支架；组织工程

武汉张春秋章培标尹飞刘景臣李玉新.纤维蛋白胶和异种无机骨构建复合支架材料与种子细胞的复合[J].中国组织工程研究与临床康复，2006，11(1):52-54 [www.zglckf.com/zglckf/ejournal/upfiles/07-1/1k-52(ps).pdf]

1东北师范大学遗传与细胞研究所博士后流动站，吉林省长春市 130032；2吉林大学中日联谊医院骨科，吉林省长春市 130033；3吉林大学机械科学与工程学院工程力学系，吉林省长春市 130032；4中国科学院长春应用化学研究所，吉林省长春市 130032

武汉☆，男，1967年生，吉林省通化县人，汉族，2003年吉林大学毕业，博士，副教授，主要从事创伤修复与骨组织工程的研究。
mdwuhan@163.com

国家自然科学基金资助项目
（30570451）*

中图分类号:R318.08 文献标识码:A
文章编号:1673-8225
(2007)01-00052-03

收稿日期：2006-05-15
修回日期：2006-08-30
(06-50-5-4144/S·X)

热点资讯：目前新的骨替代产品材料的孔径、孔隙率和生物降解性能、生物相容性等方面还存在诸多问
题。有的骨诱导活性好但机械强度差，有的机械强度好但缺乏骨诱导活性，寻找理想的骨移植替代材料目前仍是骨科领域的热门课题和难题之一，开发两者兼具的理想骨组织工程材料具有重要意义。

创新要点：异种骨具有一定机械强度，但生物活性差；纤维蛋白胶纤维蛋白是一种低抗原的生物大分子材料，植入体内易于降解，可塑性良好，具有合适的孔隙，适合细胞生长，能解决细胞的黏附问题，可作为细胞培养支架材料。去抗原牛松质骨经处理降低了抗原性，复合骨髓基质细胞则具有了诱导成骨活性。骨髓基质细胞/纤维蛋白胶/去抗原牛松质骨复合体形成了既有一定机械强度，又有成骨活性的双支架异种组织工程骨，是一种较好的骨移植材料。

同行评价：论文选题属组织工程骨的热门研究课题，作者尝试应用纤维蛋白胶、异种无机骨构建组织工程骨复合支架，研究结果显示该复合支架异种组织工程骨既具有良好的成骨活性，又具有一定的机械强度，也解决了种子细胞与支架材料不易黏附的问题。论文学术水平较高，是对组织工程骨支架材料研究的有益的探索。