中华放射学杂志
中華放射學雜誌
중화방사학잡지
Chinese Journal of Radiology
2013年
7期
654-658
,共5页
肝肿瘤%金属纳米粒子%二氧化硅%叶酸
肝腫瘤%金屬納米粒子%二氧化硅%葉痠
간종류%금속납미입자%이양화규%협산
Liver neoplasms%Metal nanoparticles%Silicon dioxide%Folic acid
目的 探讨叶酸偶联的二氧化硅包覆的金纳米棒(GNRs@SiO2-FA)探针制备方法和其光学性质,检测探针靶向性,并观察其进入细胞过程.方法 采用种子生长法制备出金纳米棒,以SiO2作为壳材,制备出GNRs@SiO2,最后偶联叶酸得到GNRs@SiO2-FA.(1)利用透射电镜、紫外光谱对制备出的GNRs@SiO2-FA探针进行检测.(2)取对数生长期肝癌HepG2细胞随机分为2组,分别加入金元素浓度为40.0×10-6、20.0×10-6、10.0×10-6、5.0×10-6、2.5 ×10-6的GNRs@SiO2-FA和GNRs溶液,采用四甲基偶氮唑盐(MTT)微量酶反应比色法测得吸光度值(A值),以此判断其细胞毒性.(3)将细胞随机分为2组,分别加入相同金元素浓度的GNRs@SiO2-FA和GNRs@SiO2溶液,然后孵育2、4、8、16、24 h时,应用电感耦合等离子体质谱法(ICP-MS)监测GNRs@SiO2-FA细胞摄入的靶向性.(4)用透射电镜观察GNRs@SiO2-FA进入细胞过程.所得数据用(-x)±s表示,单因素方差分析进行两两组间比较,P ＜0.05为差异有统计学意义.结果 紫外图谱证实成功制备GNRs@SiO2-FA.对2组金浓度相同细胞的A值进行方差分析比较,差异均有统计学意义,金元素浓度从高到低(40.0×10-6、20.0×10-6、10.0×10-6、5.0×10-6、2.5×10-6),2组比较F值分别为191.876、265.419、77.987、52.061、18.745,P值均＜0.01.ICP-MS监测结果显示,GNRs@SiO2组细胞在孵育2、4、8、16、24 h时细胞固体金元素含量分别为(256.7±3.3)、(602.8±2.4)、(1067.1±3.6)、(1998.5±4.3)、(2078.5±1.3) mg/kg,GNRs@SiO2-FA组分别为(693.1 ±2.0)、(1432.0±2.6)、(2331.3±3.5)、(2484.5 ±5.0)、(2589.7±2.1)mg/kg,组间两两比较差异有统计学意义(F值分别为3278.070、34287.199、85434.870、18333.454、42412.973,P值均＜0.01),证明叶酸偶联的探针具有很强的靶向性.透射电镜观察到,GNRs@SiO2-FA与细胞共同培养1h后,细胞质内见少量纳米探针；4 ～24 h,细胞质内见大量探针,但细胞核内未见.结论 制备出的GNRs@SiO2-FA探针具有很好的生物安全性和靶向性.
目的 探討葉痠偶聯的二氧化硅包覆的金納米棒(GNRs@SiO2-FA)探針製備方法和其光學性質,檢測探針靶嚮性,併觀察其進入細胞過程.方法 採用種子生長法製備齣金納米棒,以SiO2作為殼材,製備齣GNRs@SiO2,最後偶聯葉痠得到GNRs@SiO2-FA.(1)利用透射電鏡、紫外光譜對製備齣的GNRs@SiO2-FA探針進行檢測.(2)取對數生長期肝癌HepG2細胞隨機分為2組,分彆加入金元素濃度為40.0×10-6、20.0×10-6、10.0×10-6、5.0×10-6、2.5 ×10-6的GNRs@SiO2-FA和GNRs溶液,採用四甲基偶氮唑鹽(MTT)微量酶反應比色法測得吸光度值(A值),以此判斷其細胞毒性.(3)將細胞隨機分為2組,分彆加入相同金元素濃度的GNRs@SiO2-FA和GNRs@SiO2溶液,然後孵育2、4、8、16、24 h時,應用電感耦閤等離子體質譜法(ICP-MS)鑑測GNRs@SiO2-FA細胞攝入的靶嚮性.(4)用透射電鏡觀察GNRs@SiO2-FA進入細胞過程.所得數據用(-x)±s錶示,單因素方差分析進行兩兩組間比較,P ＜0.05為差異有統計學意義.結果 紫外圖譜證實成功製備GNRs@SiO2-FA.對2組金濃度相同細胞的A值進行方差分析比較,差異均有統計學意義,金元素濃度從高到低(40.0×10-6、20.0×10-6、10.0×10-6、5.0×10-6、2.5×10-6),2組比較F值分彆為191.876、265.419、77.987、52.061、18.745,P值均＜0.01.ICP-MS鑑測結果顯示,GNRs@SiO2組細胞在孵育2、4、8、16、24 h時細胞固體金元素含量分彆為(256.7±3.3)、(602.8±2.4)、(1067.1±3.6)、(1998.5±4.3)、(2078.5±1.3) mg/kg,GNRs@SiO2-FA組分彆為(693.1 ±2.0)、(1432.0±2.6)、(2331.3±3.5)、(2484.5 ±5.0)、(2589.7±2.1)mg/kg,組間兩兩比較差異有統計學意義(F值分彆為3278.070、34287.199、85434.870、18333.454、42412.973,P值均＜0.01),證明葉痠偶聯的探針具有很彊的靶嚮性.透射電鏡觀察到,GNRs@SiO2-FA與細胞共同培養1h後,細胞質內見少量納米探針；4 ～24 h,細胞質內見大量探針,但細胞覈內未見.結論 製備齣的GNRs@SiO2-FA探針具有很好的生物安全性和靶嚮性.
목적 탐토협산우련적이양화규포복적금납미봉(GNRs@SiO2-FA)탐침제비방법화기광학성질,검측탐침파향성,병관찰기진입세포과정.방법 채용충자생장법제비출금납미봉,이SiO2작위각재,제비출GNRs@SiO2,최후우련협산득도GNRs@SiO2-FA.(1)이용투사전경、자외광보대제비출적GNRs@SiO2-FA탐침진행검측.(2)취대수생장기간암HepG2세포수궤분위2조,분별가입금원소농도위40.0×10-6、20.0×10-6、10.0×10-6、5.0×10-6、2.5 ×10-6적GNRs@SiO2-FA화GNRs용액,채용사갑기우담서염(MTT)미량매반응비색법측득흡광도치(A치),이차판단기세포독성.(3)장세포수궤분위2조,분별가입상동금원소농도적GNRs@SiO2-FA화GNRs@SiO2용액,연후부육2、4、8、16、24 h시,응용전감우합등리자체질보법(ICP-MS)감측GNRs@SiO2-FA세포섭입적파향성.(4)용투사전경관찰GNRs@SiO2-FA진입세포과정.소득수거용(-x)±s표시,단인소방차분석진행량량조간비교,P ＜0.05위차이유통계학의의.결과 자외도보증실성공제비GNRs@SiO2-FA.대2조금농도상동세포적A치진행방차분석비교,차이균유통계학의의,금원소농도종고도저(40.0×10-6、20.0×10-6、10.0×10-6、5.0×10-6、2.5×10-6),2조비교F치분별위191.876、265.419、77.987、52.061、18.745,P치균＜0.01.ICP-MS감측결과현시,GNRs@SiO2조세포재부육2、4、8、16、24 h시세포고체금원소함량분별위(256.7±3.3)、(602.8±2.4)、(1067.1±3.6)、(1998.5±4.3)、(2078.5±1.3) mg/kg,GNRs@SiO2-FA조분별위(693.1 ±2.0)、(1432.0±2.6)、(2331.3±3.5)、(2484.5 ±5.0)、(2589.7±2.1)mg/kg,조간량량비교차이유통계학의의(F치분별위3278.070、34287.199、85434.870、18333.454、42412.973,P치균＜0.01),증명협산우련적탐침구유흔강적파향성.투사전경관찰도,GNRs@SiO2-FA여세포공동배양1h후,세포질내견소량납미탐침；4 ～24 h,세포질내견대량탐침,단세포핵내미견.결론 제비출적GNRs@SiO2-FA탐침구유흔호적생물안전성화파향성.
Objective To develop a cancer cell targeting probe based on silica-coated gold nanorods and investigate its optics properties and its targeting effect on human hepatocellular carcinoma HepG2 cells in vitro.Methods Preparation of gold nanorods (GNRs) by seeded growth method,and then the spherical core-shell silica-coated gold nanorods were successfully prepared by a sol-gel method,finally the GNRs@SiO2 was conjugated with folate (GNRs@SiO2-FA).The characteristics of GNRs@SiO2-FA were studied using transmission electron microscopy,and UV spectra.The cells were divided into 2 groups randomly,adding GNRs@SiO2-FA and GNRs solution respectively at the gold concentration of 40.0 × 10-6,20.0 ×10-6,10.0 × 10-6,5.0 × 10-6,2.5 × 10-6,and the MTT method was applied to detect the absorbance (A value) and study the cytotoxicity of GNRs@SiO2-FA and GNRs.The cells were divided into 2 groups randomly,and incubated with the same concentration of GNRs@SiO2-FA and GNRs@SiO2 solution respectively,at 2,4,8,16,24 h,and the targeting of GNRs@SiO2-FA cellular uptake were detected by ICPMS by observing the process of GNRs@SiO2-FA into the cells by transmission electron microscopy (TEM).The data represented by (-x) ± s ; single factor analysis of variance were compared between the 2 groups ; and the differences were significant when P ＜ 0.05.Results UV spectrum confirmed the successful preparation of GNRs@SiO2-FA.The A value of the same concentration group was variance analysised,and the differences between the 2 groups was statistically significant (all P ＜ 0.01) with the gold element concentration from high to low:F =191.876,265.419,77.987,52.061,18.745.The ICP-MS confirmed GNRs@SiO2-FA could specifically bind with HepG2 cells.GNRs@SiO2 group gold element content at 2,4,8,16,24 h was (256.7±3.3),(602.8±2.4),(1067.1±3.6),(1998.5±4.3),(2078.5±1.3) mg/kg and GNRs@SiO2-FA group was(693.1 ±2.0),(1432.0 ±2.6),(2331.3 ±3.5),(2484.5 ±5.0),(2589.7 ±2.1)mg/kg,and the 2 groups was statistically significant (F =3278.070,34287.199,85434.870,18333.454,42412.973,P ＜0.01).TEM results showed that a small amount of nano probes were in the cytoplasm after cultured with GNRs@SiO2-FA cells 1 h,and that,a lot of probe were in the cytoplasm,4-24 h later,but there was no probe in the nucleus.Conclusion The prepared Folate-conjugated gold nanorods has good performance on biocompatibility and targeting.
