重要的概念：生物玻璃是一种具有优良生物相容性及生物活性的硅酸盐材料。它能够在植入部位迅速发生一系列表面反应,形成能与骨和软组织良好结合碳酸盐羟基磷灰石。Ⅰ型胶原是天然骨基质的主要有机成分,具有良好的生物相容性和生物降解性。透明质酸是一种结构最简单的糖氨聚糖,它的结构疏松,含水及多孔性,特别适于细胞的迁移及增殖。磷酸丝氨酸作为磷蛋白的重要组成成分,是生物矿化的重要调节剂,也是细胞生物膜结构的重要组成成分，为细胞分化、增殖和再生所必需,可维持和促进多种膜结合蛋白和受体的生物活性。

应用要点：通过冷冻干燥和仿生矿化技术，以改性生物活性玻璃粉体和胶原、透明质酸钠、磷酸丝氨酸等天然生物分子复合制备的生物活性玻璃/胶原蛋白/透明质酸/磷酸丝氨酸、生物活性玻璃/胶原蛋白/磷酸丝氨酸、生物活性玻璃/胶原蛋白/透明质酸、生物活性玻璃/胶原蛋白4种仿生复合骨组织工程三维支架材料无毒性，无刺激性，生物安全性能良好。

偏倚或不足：由于骨组织工程材料是长期植入体内，在进入临床试验前，有必要对其遗传毒性、致癌性方面进行相关试验，此项研究工作有待进一步完成。

摘要
背景：制备仿生骨组织工程支架材料材料的原材料生物活性玻璃已被临床广泛应用于软骨、骨组织的修复、替代，已通过生物安全性评价。
目的：利用生物活性玻璃和胶原蛋白、透明质酸、磷酸丝氨酸等天然大分子复合制备4种新型仿生骨组织工程支架材料，观察4种新型仿生骨组织工程支架材料的生物安全性。
设计、时间及地点：随机分组设计、动物对照实验，于2006-03/08在南方医科大学珠江医院血液科实验室完成。
材料：健康昆明系小鼠60只，健康成年家兔11只。采用冷冻干燥和仿生矿化技术，以改性生物活性玻璃粉体和胶原、透明质酸钠、磷酸丝氨酸等天然生物分子复合制备生物活性玻璃/胶原蛋白/透明质酸/磷酸丝氨酸、生物活性玻璃/胶原蛋白/磷酸丝氨酸、生物活性玻璃/胶原蛋白/透明质酸、生物活性玻璃/胶原蛋白4种仿生复合骨组织工程三维支架材料。
方法：对4种新型仿生骨组织工程支架材料进行生物安全性试验：全身急性毒性试验观察小鼠中毒症状，按50 mL/kg尾静脉注射4种材料的浸提液。支架材料加入新鲜兔血进行溶血试验，离心后取上清液，测定吸光度值，计算溶血率。兔脊柱两侧皮内注射浸提液观察皮肤刺激反应。
主要观察指标：注入浸提液后24，48，72 h观察小鼠全身急性毒性反应和皮肤刺激反应，测定溶血率。
结果：全身急性毒性试验结果表明，4组小鼠无死亡，无明显毒性表现、体质量下降等不良反应。溶血试验结果表明，4组支架材料的溶血率均＜5%，满足医用生物材料的应用要求。皮内刺激反应试验结果表明，4组均未见任何刺激反应，无红斑、焦痂、水肿等皮肤反应现象。
结论：仿生复合支架材料生物活性玻璃/胶原蛋白/透明质酸/磷酸丝氨酸、生物活性玻璃/胶原蛋白/磷酸丝氨酸、生物活性玻璃/胶原蛋白/透明质酸、生物活性玻璃/胶原蛋白无毒性，无刺激性，生物安全性能良好。
关键词：仿生；生物活性玻璃；胶原蛋白；透明质酸；磷酸丝氨酸；复合支架；生物安全性；生物材料

张梅霞，姚瑶，李妙，靳安民. 四种新型仿生骨组织工程支架材料生物安全性评价[J].中国组织工程研究与临床康复，2008，12(19):3641-3644 [www.zglckf.com/zglckf/ejournal/upfiles/08-19/19k-3641(ps).pdf]

Biological safety of four kinds of new biomimetic composite scaffolds for bone tissue engineering

Abstract
BACKGROUND: Biomimetic composite scaffolds of bone tissue engineering prepared by natural macromolecules, have achieved wide applications on the repair and substitute of cartilage and bone tissues, moreover, the biological safety has also been verified.
OBJECTIVE: To explore the biological safety on the four kinds of new biomimetic composite scaffolds prepared by bioactive glass (BG), collagen (COL), hyaluronic acid (HYA) and phosphoserine (PS).
DESIGN, TIME AND SETTING: A randomized control animal experiment was conducted in the Hematology Laboratory of Zhujiang Hospital, Southern Medical University, Guangzhou, Guangdong, China from March to August in 2006.
MATERIALS: Sixty healthy Kunming mice, eleven normal adult rabbits, BG-COL-HYA-PS scaffold, BG-COL-PS scaffold, BG-COL-HYA scaffold and BG-COL scaffold.
METHODS: Biological safety detection of the four kinds of scaffold materials was performed. General toxicity test: The extracts from the materials were injected into mouse vein with the dose of 50 mL/kg. The toxic symptoms of mice were observed. Hemolytic test: The scaffold materials with fresh rabbit blood was applied. Subsequent to the centrifugation and the supernatant liquid extraction, the absorbance value and hemolysis rate were detected. Skin stimulation test: The extracts of materials were injected into the bilateral sides of rabbit spine subcutaneously. The skin reaction was observed.
MAIN OUTCOME MEASURES: General toxicity reaction and skin stimulation reaction were observed within 24, 48 and 72 hours after the injection. Hemolysis rate was also detected.
RESULTS: General toxicity test showed that there were no adverse effects in the mice, such as death, toxicity reaction or decreasing weight. Hemolytic test showed that the hemolysis rates of the four scaffold materials were all less than 5%, indicating that the requirements of biomedical material application could be satisfied. Skin stimulation test showed that no erythema, eschar or edema appeared in 4 groups.
CONCLUSION: The biomimetic BG-COL-HYA-PS, BG-COL-PS, BG-COL-HYA and BG-COL composite scaffolds present no toxicity or skin stimulation, and have good biological safety.

Zhang MX, Yao Y, Li M, Jin AM.Biological safety of four kinds of new biomimetic composite scaffolds for bone tissue engineering.Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu 2008;12(19):3641-3644
[www.zglckf.com/zglckf/ejournal/upfiles/08-19/19k-3641(ps).pdf]