一种体外流动钙化模型的建立  

Establishment of an in vitro flow calcification assessment model

作　　者：井慧敏 刘婧 李彬寒 冷希岗 王志红 JING Hui-min;LIU Jing;LI Bin-han;LENG Xi-gang;WANG Zhi-hong(Institute of Biomedical Engineering,Chinese Academy of Medical Sciences and Peking Union Medical College,Tianjin 300192,China)

机构地区：[1]中国医学科学院·北京协和医学院生物医学工程研究所,天津300192

出　　处：《生物医学工程与临床》2021年第3期255-263,共9页Biomedical Engineering and Clinical Medicine

基　　金：国家自然科学基金资助项目(31771059;31800815);中国医学科学院医学与健康科技创新工程团队项目(2017-12M-3-002);天津市自然科学基金资助项目(18JCQNJC14200);天津市科技支撑重点项目(19YFZCSY00520);天津市青年人才托举工程(TJSQNTJ-2018-02)。

摘　　要：目的构建一种基于模拟生理流体环境、且可便捷、高效反映心血管植入物钙化特性的体外测试模型,为心血管植入物的快速开发提供新的手段。方法选择新鲜牦牛牛心包(青海裕泰畜产品有限公司,中国);选择3周龄Wistar雄性幼鼠5只,体质量(50±5)g。采用常规戊二醛(GA)与处理牛心包交联方法,制备GA交联的牛心包材料(Yak-GA)。在大鼠皮下埋植14、28、56 d做体内钙化实验。将橡胶管、瓣叶钙化舱、体液模拟液溶液瓶、蠕动泵组成体外流动钙化模拟装置,将Yak-GA置于瓣叶钙化舱,于不同流速(0、2、10 mL/min)下1.5倍模拟体液(SBF)转流14 d,测定钙离子浓度、pH值变化。利用SPSS软件对体内外钙化数据进行拟合,通过拟合优度分析,筛选出最佳的流速并获得体内钙化预估方程。结果Yak-GA体内植入14、28、56 d,组织钙沉积量分别可达到(52.17±11.01)μg/mg、(104.40±21.34)μg/mg、(171.06±15.11)μg/mg。体外钙化模拟实验中,对比0、2、10 mL/min流速下、相同时间点溶液钙损失,2 mL/min流速溶液钙损失量高,材料钙化进展最快;14 d时2 mL/min流速溶液钙损失量与0 mL/min流速比,差异有显著统计学意义[(2.07±0.07)mmol/L vs(1.14±0.10)mmol/L](t=13.41,P<0.01);与10 mL/min流速比,差异也有统计学意义[(2.07±0.07)mmol/L vs(1.89±0.01)mmol/L](t=3.55,P<0.05)。2 mL/min流速体内外钙化数据拟合优度R^(2)=0.941,所得的评估方程为:当1.65<C浓度≤2.05 mmol/L时,公式为Y=-126.71X+407.74;当2.05<C_(浓度)≤3.75 mmol/L时,公式为Y=e^((7.46–1.22X))(X=钙化液中钙离子浓度;Y=体内模型组织中钙沉积量);模型残差在(2.36±11.36)μg/mg之间波动。另外,研究中发现pH值变化与钙离子浓度变化线性相关性可达到0.94,具有极强相关性,相关性方程为Y=2.63×pH+0.37。结论成功开发了一种简单、有效、可实时监测钙化过程的体外流动钙化模型,其中2 m L/min为该系统最佳流速,该流速下所构建�Objective To construct an in vitro calcification test model based on physiological fluid environment, which can reflect the calcification characteristics of implants conveniently and efficiently, and provide novel method for rapid development of cardiovascular implants. Methods Five 3-week-old Wistar male mice with body weight of(50 ± 5) g were selected.The conventional glutaraldehyde(GA) and processed bovine pericardial cross-linking method were adopted to prepare GA crosslinked bovine pericardial material(Yak-GA). In vivo calcification model was subcutaneously implanted in rats at 14, 28 and 56 days. The rubber tube, valve leaflet calcification chamber, body fluid simulation solution bottle and peristaltic pump were composed in vitro to get flow calcification simulation device, which collected the changes of calcium concentration and pH value in calcification solution at different flow rates(0, 2, 10 mL/min) and 1.5 times simulated body fluid(SBF) within 14 days. SPSS software was used to fit the in vitro and in vivo calcification data to obtain optimal flow rate and prediction equation of in vivo calcification. Results The calcium deposition of Yak-GA in vivo reached(52.17 ± 11.01) μg/mg,(104.40 ± 21.34) μg/mg and(171.06 ± 15.11) μg/mg at 14, 28 and 56 days. In vitro calcification results showed that more calcium loss was observed in2 mL/min solution, and material calcification developed fastest. At 14 days, the difference of calcium loss in 2 mL/min[(2.07 ±0.07) mmol/L] and 0 mL/min solution[(1.14 ± 0.10) mmol/L] was statistically significant(t = 13.41, P < 0.01), and the difference of calcium loss in 2 mL/min and in 10 mL/min solution[(1.89 ± 0.01) mmol/L] was statistically significant(t = 3.55, P < 0.05).In 2 mL/min solution, the goodness of fit was R^(2)= 0.941, and evaluation equation: 1.65 < C_(concentration)≤ 2.05 mmol/L, the formula was Y =-126.71 X + 407.74;2.05 < C_(concentration)≤ 3.75 mmol/L, formula was Y = e^((7.46 – 1.22 X))(X = concentration of calcium ions in calcification solu

关 键 词：心血管植入物 钙化模型 钙沉积 曲线拟合 体内外相关性 钙化评价 

分 类 号：R318.08[医药卫生—生物医学工程]