纳米羟基磷灰石/硫酸钙复合人工骨的生物安全性研究

Biological Security of A New Nano-hydroxyapatite/Calcium Sulphate Composited Bone Substitute

目的研制纳米羟基磷灰石/半水硫酸钙(n-HA/CSH)复合型人工骨,并对其进行体内、外生物安全性测试。方法对n-HA/CSH人工骨进行急性全身毒性试验、皮内刺激试验、致敏试验、MTT细胞毒性试验和遗传毒性实验(Ames试验)并与对照组比较。结果人工骨浸取液静脉及腹腔注射后不引起小鼠呼吸、进食改变或死亡,体重稳定。家兔皮内注射72小时后仅出现红斑或微弱水肿,豚鼠皮内注射后未出现过敏反应。MTT细胞毒性试验显示含HA10%、20%、40%人工骨及纯n-HA、CSH的细胞增殖率均在77%以上,细胞毒性均为0～1级,Ames试验表明含HA40%人工骨的不同浓度生理盐水浸取液引起鼠伤寒沙门氏菌回复突变数均不超过阴性对照组的2倍。结论n-HA/CSH复合材料不引起全身毒性反应、皮内刺激反应和急性过敏反应。且无MTT细胞毒性,细胞相容性良好。同时,复合材料的生理盐水浸取液不引起鼠伤寒沙门氏菌回复突变数增加。

Objective To evaluate the biological security properties of a new nano-hydroxyapatite / calcium sulphate hemidydrate （ n-HA/CSH ） bone substite. Methods The n-HA/CSH were subjected to acute whole body toxity test, endodermal stimulation test, anaphylactic test, MTT cytotoxity assay and Ames test, compared with control groups. Results After intravenous and intra-abdominal injection of soaked solution of material, there were no changes in breath, appetite, body weight of mice and no death. There were only red flecks or mild edema in rabbit skin 72hrs followed soaked solution endodermal injection and no allergic reaction were found in guinea pig after endodermal injection. Test ofMTT cytotoxicity showed that the average cell proliferation rate was over 77% after treated with the solution of 5 bone substitutes （P〈0.05）, with the cytotoxity being 0-1 （non-toxic）. Ames test showed that the number of reversion mutation increased by bone substitute （40%wtHA） with diffrent doses / plate in any 4 strains with or without S9mix didn＇t exceed two times of those of negative controls. Conclusion n-HA/CSH does not cause acute whole body toxic reaction, endodermal stimulation, allergic reaction and no MTT cytotoxity. The material does not increase the number of reversion mutation in Ames test.