课题背景：课题属2005广东省科技三项经费计划项目(No. 2005B34001008)，项目名称：组织工程骨复合骨唾液酸蛋白的基础与临床研究。课题已取得的成果：成功将人骨唾液酸蛋白基因克隆到毕赤酵母表达载体pPICZαA上，以融合蛋白的形式表达重组骨唾液酸蛋白。

相关链接：骨唾液酸蛋白是骨发生过程中的关键因子之一，广泛参与骨形成和骨重建的诸多环节。从目前研究情况看，骨唾液酸蛋白有良好的成骨效应，在组织工程骨成为研究热点的今天，完全可以成为一种新的骨生长因子应用于组织工程骨修复骨缺损。目前关于骨唾液酸蛋白成骨的动物实验较少，且缺乏系统性，尚有许多工作要完成。

应用要点：实验证实了骨唾液酸蛋白在骨缺损内的成骨活性。与目前惟一应用于临床的骨生长因子骨形态发生蛋白对比，骨唾液酸蛋白来源方便，造价低廉，且由于其具多重成骨效应，有可能成为一种新型的与组织工程骨复合的生长因子，组织工程骨与骨唾液酸蛋白复合体的产业化将具有很好的应用前景。

摘要
目的：研制骨唾液酸蛋白-β-磷酸三钙（BSP-β-TCP）人工骨并观察BSP-β-TCP中骨唾液酸蛋白的释放规律。
方法：实验于2006-12/2007-07在解放军广州军区广州总医院医学实验科完成。①BSP-β-TCP人工骨制备：采用低温负压冻干的方法将骨唾液酸蛋白复合于β-磷酸三钙中，按骨唾液酸蛋白含量不同制备两种样品：A样品含骨唾液酸蛋白1﹪， B样品含骨唾液酸蛋白2﹪。②观察指标：两种样品分别在恒温空气浴振荡器内(37±1)℃持续振荡。试验开始2 周内每24 h，以后每72 h时分别吸出浸泡液100 μL / 次,共观察4 周。用考马斯亮蓝法测定各个时间点溶液中蛋白质含量；绘制出蛋白质释放量和时间关系的散点图，再将散点图的曲线直线化，分析BSP-β-TCP人工骨中骨唾液酸蛋白的释放动力学。
结果：①骨唾液酸蛋白释放率：72 h内有一个爆发性的释放，释放率A样品为25.44﹪，B样品为35.11﹪；10 d内的释放速度较快，A样品为39.76﹪，B样品为44.13﹪；持续4周仍有缓慢释放，A样品为55.43﹪，B样品为56.82﹪。②骨唾液酸蛋白的释放动力学：将散点图曲线进行拟和后可见释放量与时间（h）平方根成线性关系，两者的关系符合Higuchi方程。
结论：骨唾液酸蛋白以扩散方式释放，β-磷酸三钙可作为良好的载体与BSP共同形成缓释系统。
关键词：骨唾液酸蛋白；β-磷酸三钙；释放动力学；组织工程骨

王玮，章莹，吴文，骨唾液酸蛋白-β-磷酸三钙人工骨的研制及释放实验[J].中国组织工程研究与临床康复，2008，12(1):27-30
[www.zglckf.com/zglckf/ejournal/upfiles/08-1/1k-27(ps).pdf]

Preparation and protein release of bone sialoprotein-beta-tricalcium phosphate artificial bone

Abstract

AIM：To prepare bone sialoprotein-β-tricalcium phosphate (BSP-β-TCP) artificial bone and observe releasing rule of BSP in the artificial bone.
METHODS: The experiment was carried out in the Medical Laboratory of Guangzhou General Hospital of Guangzhou Military Area Command of Chinese PLA between December 2006 and July 2007.①BSP in combination with β-TCP was conducted by using low temperature, negative pressure and freeze-dried device, to prepare BSP-β-TCP artificial bone. Two kinds of BSP-β-TCP artificial bone were produced according to the content of BSP: A sample (containing 1% BSP) and B sample (containing 2% BSP).②The two samples were persis0
tently oscillated in constant temperature air bath (37±1) ℃. The soak was inhausted 100 μL per time every 24 hours within two weeks after experiment, and then every 72 hours later, the observation lasted 4 weeks. Coomassie brilliant blue method was used to determine the protein content in solution at every time point. The scatterplot of relation between protein releasing amount and time was drawn, then lined curve of scatterplot was conducted to analyze releasing kinetics of BSP in BSP-β-TCP artificial bone.
RESULTS: ①BSP had an breaking out delivery in 72 hours, and the BSP release rate of A sample was 25.44% while that of B sample was 35.11%; BSP kept a high releasing rate in 10 days, 39.76% in A sample and 44.13% in B sample; slow releasing still remained for 4 weeks, the BSP release rate of A sample was 55.43%, and that of B sample was 56.82%.②After making the curve fitting of scatterplot, the linear correlation relationship between the release amount of BSP and the square root of time (hour) was found, in agreement with Higuchi equation.
CONCLUSION: BSP release from BSP-β-TCP artificial bone in a diffuse manner. As a carrier, β-TCP can form slow-release system with BSP.

Wang W, Zhang Y, Wu W.Preparation and protein release of bone sialoprotein-beta-tricalcium phosphate artificial bone.Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu 2008;12(1):27-30(China)
[www.zglckf.com/zglckf/ejournal/upfiles/08-1/1k-27(ps).pdf]