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可生物降解镁及其合金的溶血和细胞毒性机制。

Hemolysis and cytotoxicity mechanisms of biodegradable magnesium and its alloys.

机构信息

Center for Biomedical Materials and Tissue Engineering, Academy for Advanced Interdisciplinary Studies, Peking University, Beijing 100871, China.

School of Material Science and Engineering, University of Science and Technology Beijing, Beijing 100083, China.

出版信息

Mater Sci Eng C Mater Biol Appl. 2015 Jan;46:202-6. doi: 10.1016/j.msec.2014.08.038. Epub 2014 Aug 19.

Abstract

Good hemocompatibility and cell compatibility are essential requirements for coronary stents, especially for biodegradable magnesium alloy stents, which could change the in situ environment after implanted. In this work, the effects of magnesium ion concentration and pH value on the hemolysis and cytotoxicity have been evaluated. Solution with different Mg(2+) concentration gradients and pH values of normal saline and cell culture media DMEM adjusted by MgCl2 and NaOH respectively were tested for the hemolysis and cell viability. Results show that even when the concentration of Mg(2+) reaches 1000 μg/mL, it has little destructive effect on erythrocyte, and the high pH value over 11 caused by the degradation is the real reason for the high hemolysis ratio. Low concentrations of Mg(2+) (<100 μg/mL) cause no cytotoxicity to L929 cells, of which the cell viability is above 80%, while high concentrations of Mg(2+) (>300 μg/mL) could induce obvious death of the L929 cells. The pH of the extract plays a synergetic effect on cytotoxicity, due to the buffer action of the cell culture medium. To validate this conclusion, commercial pure Mg using normal saline and PBS as extract was tested with the measurement of pH and Mg(2+) concentration. Pure Mg leads to a higher hemolysis ratio in normal saline (47.76%) than in buffered solution (4.38%) with different pH values and low concentration of Mg(2+). The Mg extract culture media caused no cytotoxicity, with pH=8.44 and 47.80 μg/mL Mg(2+). It is suggested that buffered solution and dynamic condition should be adopted in the hemolysis evaluation.

摘要

良好的血液相容性和细胞相容性是冠状动脉支架的基本要求,尤其对于可生物降解的镁合金支架,在植入后会改变局部环境。在这项工作中,评估了镁离子浓度和 pH 值对溶血和细胞毒性的影响。用不同 Mg(2+)浓度梯度的溶液和生理盐水及细胞培养液 DMEM(分别用 MgCl2 和 NaOH 调至相应 pH 值)进行了溶血和细胞活力测试。结果表明,即使 Mg(2+)浓度达到 1000 μg/mL 时,对红细胞也几乎没有破坏作用,降解导致的高 pH 值(超过 11)才是溶血率高的真正原因。低浓度的 Mg(2+)(<100 μg/mL)对 L929 细胞没有细胞毒性,细胞活力超过 80%,而高浓度的 Mg(2+)(>300 μg/mL)会导致 L929 细胞明显死亡。提取液的 pH 值对细胞毒性具有协同作用,这是由于细胞培养液的缓冲作用。为了验证这一结论,使用生理盐水和 PBS 作为提取液对商用纯镁进行了测试,测量了 pH 值和 Mg(2+)浓度。纯镁在生理盐水(47.76%)中的溶血率高于缓冲溶液(4.38%),且缓冲溶液的 pH 值和 Mg(2+)浓度较低。Mg 提取培养液无细胞毒性,pH 值为 8.44 和 47.80 μg/mL 的 Mg(2+)。建议在溶血评价中采用缓冲溶液和动态条件。

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