Chinese Journal of Tissue Engineering Research ›› 2019, Vol. 23 ›› Issue (18): 2871-2879.doi: 10.3969/j.issn.2095-4344.1653
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Received:
2018-11-08
Online:
2019-06-28
Published:
2019-06-28
Contact:
Quan Renfu, Chief physician, Jiangnan Hospital, Zhejiang Chinese Medical University, Hangzhou 311200, Zhejiang Province, China
About author:
Chai Le, Master candidate, Zhejiang Chinese Medical University, Hangzhou 310053, Zhejiang Province, China
Supported by:
the Medical and Health Scientific Program of Zhejiang Province, No. 2014KYA191 (to QRF); the Major Scientific Research Fund of Zhejiang Province, No. 2014C03031 (to QRF)
CLC Number:
Chai Le, Quan Renfu, Lü Jianlan, Huang Xiaolong, Zhang Can, Ren Weifan, Qian Jiansheng. Physicochemical properties and cellularization of hydroxyapatite/zirconium dioxide foam ceramics[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(18): 2871-2879.
2.4 鼠伤寒沙门氏菌回复性突变和细胞毒性分析 受试物的致突变作用(-S9):在不存在代谢活化体系的情况下,不同浓度羟基磷灰石/ZrO2泡沫陶瓷材料对鼠伤寒沙门氏菌TA97a、TA98、TA100、TA102、TA1535的致突变实验结果,见表2。二甲亚砜对照组的自发回复突变个数分别为(15±1),(6±1),(18±4),(48±8),(4±2)个克隆/孔;TA97a菌株阳性对照化合物(9-氨基吖啶缘酸盐一水合物组)、TA98菌株阳性对照化合物(2-硝基芴组)、TA100菌株阳性对照化合物(叠氮钠组)、TA102菌株阳性对照化合物(过氧化氢异丙苯组)、A1535菌株阳性对照化合物(叠氮钠组)的回复突变个数分别为(56±4),(50±10),(106±13),(193±13),(50±10)个克隆/孔,分别为DMSO对照组的3.7,8.4,5.9,4.0,16.7倍。不同浓度羟基磷灰石/ZrO2泡沫陶瓷材料组TA97a、TA98、TA100、TA102、TA1535的回复突变克隆数均与DMSO对照组相当、略高或略低,都未达到2倍,克隆形态正常,提示受试物无遗传毒性。 受试物代谢产物的致突变作用(+S9):在存在代谢活化体系的情况下,不同浓度羟基磷灰石/ZrO2泡沫陶瓷材料对鼠伤寒沙门氏菌TA97a、TA98、TA100、TA102、TA1535的致突变实验结果,见表3。在加入S9代谢活化体系的情况下,DMSO对照组的自发回复突变个数分别为(17±2),(5±1),(8±1),(45±2),(2±1)个克隆/孔;阳性对照化合物2-氨基蒽组的回复突变个数分别为(143±10),(224± 19),(114±5),(208±15),(63±5)个克隆/孔,分别为DMSO组的8.6,48.1,14.3,4.6,27倍。羟基磷灰石/ZrO2泡沫陶瓷材料组TA97a、TA98、TA100、TA102、TA1535的回复突变克隆数均与DMSO对照组相当、略高或略低,都未达到2倍,且克隆形态正常,提示受试物的代谢产物无遗传毒性。 细胞毒性:背景菌落由依赖组氨酸的细菌微小克隆形成,可通过10倍显微镜观察。受试物羟基磷灰石/ZrO2泡沫陶瓷材料处理后的背景菌落情况,见表4。羟基磷灰石/ZrO2泡沫陶瓷材料对背景菌落无明显影响,提示受试物在所试剂量下没有表现出细胞毒性。在加入S9代谢活化体系的情况下,该材料对背景菌落也无明显影响,提示该材料的代谢物在所试剂量下没有表现出细胞毒性,见表5。 "
2.6 羟基磷灰石/ZrO2泡沫陶瓷材料与间充质干细胞共培养实验结果 碱性磷酸酶分泌量:随着培养时间的延长,碱性磷酸酶的分泌量增加,其中3 d分泌量为52.55 U/L,7 d分泌量为57.34 U/L,10 d分泌量为61.58 U/L,14 d分泌量为 72.83 U/L,见图5;χ2检验显示,5个时间点间碱性磷酸酶分泌量比较差异有显著性意义(P < 0.05),说明能随着时间的延长,间充质干细胞放入成骨分化能力增强。 Ⅰ型胶原表达:共培养第14天,免疫组织化学显示细胞较为均匀的分布于材料之上,并且可见大量Ⅰ型胶原,见图6。 细胞爬行及黏附状态:扫面电镜观察可见材料表面均匀分布着间充质干细胞,细胞黏附良好且数量较多,见图7A,其中红色部分圆球状为间充质干细胞的细胞核,细小灰色部分(条索样状)为该细胞的树突与轴突;细胞生长正常,形态良好,见图7B。"
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