课题背景：已经证实在磷酸钙中加入壳聚糖可使磷酸钙骨水泥的力学强度明显提高，因此作者选择了壳聚糖-磷酸钙支架以加强力学强度，同时利用重组人骨形态发生蛋白2（rhBMP-2）的诱导成骨作用，制备rhBMP-2/壳聚糖-磷酸钙支架复合体，尝试寻找一种新型的骨组织工程材料。

应用要点：实验观察了经诱导的骨髓间充质干细胞在由重组人骨形态发生蛋白2（rhBMP-2）修饰后的壳聚糖-磷酸钙支架材料上的增殖和生长情况，发现rhBMP-2/壳聚糖-磷酸钙支架复合体具有良好的细胞相容性，且材料内部的多孔结构和孔径也能满足于细胞长入和骨的再生，可以作为成骨细胞的支架载体用于构建组织工程化骨。

相关链接：在骨组织工程研究中，为了增强成骨能力，已开始用人工复合材料进行骨诱导。目前有学者提出理想的骨移植材料应是适宜与骨形成蛋白、磷酸钙骨水泥等复合的支架材料，这些理论也已并取得了初步的成功。

摘要
目的: 观察经诱导的骨髓间充质干细胞在由重组人骨形成蛋白2（recombinant human bone morphogenetic protein-2,rhBMP-2）修饰后的壳聚糖-磷酸钙支架材料上的增殖和生长情况，以及其作为成骨细胞的支架载体用于构建组织工程化骨的可行性。
方法：实验于2007-02/08在暨南大学附属第一医院中心实验室及暨南大学理工学院材料科学与工程系实验室完成。①制备壳聚糖-磷酸钙支架，再将其放入rhBMP-2溶液中浸泡30 min，真空抽吸后冷冻干燥。②测量改良后壳聚糖-磷酸钙支架的孔隙率及抗压强度，扫描电镜观察其超微结构；同时，将兔骨髓间充质干细胞接种于复合支架上，用成骨细胞诱导液培养1周，扫描电镜观察成骨细胞在支架上的生长及黏附情况。
结果：①rhBMP-2/壳聚糖-磷酸钙支架复合体的孔径超过100 μm，空隙率为87%，压缩弹性模量为20 MPa。②扫描电镜结果显示支架材料内部为多孔结构，孔内见有细胞生长，细胞表面可见分泌颗粒。
结论：经复合rhBMP-2后的壳聚糖-磷酸钙支架是合适三维立体多孔复合支架，有较好的细胞相容性，可以作为成骨细胞的支架载体，用于构建组织工程化骨。
关键词：壳聚糖-磷酸钙；重组人骨形态发生蛋白2；生物材料；组织工程化骨

谭颖，王晶，李志忠，林永新，查振刚，焦根龙，洪亮，吴波文.重组骨形成蛋白2/壳聚糖-磷酸钙支架载体与经诱导骨髓间充质干细胞共培养的可行性[J].中国组织工程研究与临床康复，2008，12(6):1005-1008
[www.zglckf.com/zglckf/ejournal/upfiles/08-6/6k-1005(ps).pdf]

Feasibility of chitosan-calcium phosphate cement/recombinant human bone morphogenetic protein-2 composite scaffold co-cultured with bone marrow mesenchymal stem cells after induction

Abstract

AIM：To observe the proliferation and growth of the induced bone marrow mesenchymal stem cells on the chitosan-calcium phosphate scaffolds, which are modified with recombinant human bone morphogenetic protein-2 (rhBMP-2), and explore the feasibility of chitosan-calcium phosphate as bone tissue engineering scaffold for chondroblasts.
METHODS: The experiment was done in the Central Laboratory of the First Affiliated Hospital, Jinan University and the Laboratory of Materials Science and Engineering, Jinan University Technology College from February to August in 2007.①The scaffold of chitosan-calcium phosphate cement was produced and taken into rhBMP-2 solution for 30 minutes, followed by vacuum aspiration and freeze-drying.②The porosity and compressive strength of chitosan-calcium phosphate scaffold were measured, and its microstructure was observed via scanning electron microscopy; at the same time, the rabbit bone marrow mesenchymal stem cells were seeded onto the modified scaffold, being cultured in chondroblast culture fluid for one week, and the growth and adhesion of cells on the scaffold were observed with scanning electron microscope.
RESULTS: ①The porosity of rhBMP-2/chitosan-calcium phosphate composite scaffold was 87%, its pore diameter was over 100 μm, and modulus of elasticity was 20 MPa.②Scanning electron microscope showed that the scaffold had an interconnected pore structure. The cells appeared to infiltrate into the scaffold and the extracellular matrix components were observed.
CONCLUSION: The chitosan-calcium phosphate composite with rhBMP-2 is a three-dimensional porous scaffold, with good biocompatibility, and it is a good substrate candidate for bone tissue engineering scaffold for chondroblasts.

Tan Y, Wang J, Li ZZ, Lin YX, Zha ZG, Jiao GL, Hong L, Wu BW.Feasibility of chitosan-calcium phosphate cement/recombinant human bone morphogenetic protein-2 composite scaffold co-cultured with bone marrow mesenchymal stem cells after induction.Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu 2008;12(6):1005-1008(China)
[www.zglckf.com/zglckf/ejournal/upfiles/08-6/6k-1005(ps).pdf]