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For MRI, to enhance imaging accuracy, two different imaging modes with specific contrast agents are used。因此，實(shí)驗(yàn)的下一步計(jì)劃是繼續(xù)嘗試合成不同尺寸的Fe3O4納米粒子與PEGGd2O3相連接，并研究它們的性質(zhì)與不同，以確定可激活對(duì)比劑中的Fe3O4納米粒子最佳尺寸。首先配制氫氧化鈉（NaOH）/DEG原液：稱(chēng)取2g NaOH溶于20mL DEG在氮?dú)鈼l件下加熱到120℃、1小時(shí)，然后冷卻至70℃并保持此溫度；稱(chēng)取8g PAA(Polyacrylic Acid) FeCl3溶于17mL DEG在氮?dú)鈼l件下加熱到220℃機(jī)械攪拌30分鐘得到透明黃色液體；然后將1mL NaOH/DEG原液加入到透明黃色液體中（通過(guò)加入不同體積的NaOH/DEG原液以達(dá)到制備不同尺寸Fe3O4納米粒子的目的），將溫度降至210℃，溶液慢慢變黑，繼續(xù)在氮?dú)鈼l件下恒溫加熱1小時(shí)。本文僅僅討論了尺寸為的四氧化三鐵納米粒子應(yīng)用于可激活對(duì)比劑的成像效果研究，對(duì)于不同尺寸的四氧化三鐵的研究還處于制備材料階段。5 總結(jié)與展望本文討論了陽(yáng)性對(duì)比劑氧化釓及陰性對(duì)比劑四氧化三鐵的制備及對(duì)其性質(zhì)進(jìn)行了簡(jiǎn)單的研究。 討論本章節(jié)利用MTT法考察了Fe3O4SSGd2O3納米材料的生物相容性。 不同濃度Fe3O4SSGd2O3納米探針與7860作用24h后的細(xì)胞形態(tài)，細(xì)胞的形態(tài)并未發(fā)生明顯的改變，當(dāng)Fe3O4SSGd2O3納米探針中[Gd3+]，細(xì)胞形態(tài)均未發(fā)生明顯變化，7860腎癌細(xì)胞具有良好的細(xì)胞形態(tài)，由此說(shuō)明Fe3O4SSGd2O3納米探針具有良好的生物相容性，可用于腫瘤細(xì)胞的成像診斷研究。7860腎癌細(xì)胞的加材料實(shí)驗(yàn)過(guò)程與NIH 3T3小鼠成纖維細(xì)胞種板相似，此處不再贅述。 MTT測(cè)試之加材料表序號(hào)培養(yǎng)基Fe3O4SSGd2O3材料濃度A1A12100μL0μL0mM [Gd]B1B6 3mM[Gd3+] [Gd]B7B12 3mM[Gd3+] [Gd]C1C6 3mM[Gd3+] [Gd]C7C1295μL5μL 3mM[Gd3+] [Gd]D1D6 3mM[Gd3+] [Gd]D7D1290μL10μL 3mM[Gd3+] [Gd]NIH 3T3小鼠成纖維細(xì)胞板加材料的實(shí)驗(yàn)步驟為：（1）[Gd3+]，3mM[Gd3+]：首先準(zhǔn)備PEGGd2O3SSNH2：，在37℃電熱恒溫水槽(DK8D)中孵育15min，之后加入與EDC等量的NHS和100μL胱胺，繼續(xù)孵育2小時(shí)。以下各項(xiàng)操作均在無(wú)菌細(xì)胞室進(jìn)行。（3）加入適量的培養(yǎng)液， mL2 mL。 7860腎癌細(xì)胞傳代（1）倒置顯微鏡下觀察細(xì)胞，當(dāng)細(xì)胞處于80％90％匯合階段時(shí)進(jìn)行傳代培養(yǎng)；（2）倒去培養(yǎng)瓶中的培養(yǎng)液，用5—10mL無(wú)菌PBS水洗滌1—2次，然后想瓶?jī)?nèi)加入500μL %的胰蛋白酶消解細(xì)胞；（3）室溫下放置13 min，倒置顯微鏡下觀察培養(yǎng)瓶，當(dāng)細(xì)胞質(zhì)回縮至細(xì)胞形態(tài)近圓形時(shí)，倒去胰蛋白酶，立即加入5—10mL培養(yǎng)基終止消解，輕輕吹打瓶壁細(xì)胞，使之從培養(yǎng)瓶中脫離，形成細(xì)胞懸液；（4）將細(xì)胞懸液轉(zhuǎn)移到10mL的離心管中，2000轉(zhuǎn)離心10min；（5）倒掉上清液，加入1mL培養(yǎng)基使細(xì)胞分散均勻，分裝入數(shù)個(gè)培養(yǎng)瓶中，一般1: 3傳代，但是本實(shí)驗(yàn)由于時(shí)間有限，為了讓細(xì)胞快速生長(zhǎng)，進(jìn)行1:2傳代。（4）加入1/10體積的DMSO，混合均勻。 NIH 3T3小鼠成纖維細(xì)胞計(jì)數(shù)（1）%胰蛋白酶將貼壁的細(xì)胞消化下來(lái)，使之形成細(xì)胞懸液；（2）取計(jì)數(shù)用載玻片，蓋上蓋玻片，吸取少量細(xì)胞懸液滴到載玻片上，使載玻片與蓋玻片之間充滿(mǎn)細(xì)胞懸液；（3）倒置顯微鏡下計(jì)數(shù)載玻片上4個(gè)角的大計(jì)數(shù)格內(nèi)的細(xì)胞總數(shù)。 NIH 3T3小鼠成纖維細(xì)胞培養(yǎng)以下各項(xiàng)操作均在無(wú)菌細(xì)胞室進(jìn)行，NIH 3T3細(xì)胞培養(yǎng)條件為：含10%胎牛血清（FBS）的RPMI1640培養(yǎng)基、37 ℃ 5% CO2培養(yǎng)箱。由于結(jié)晶物形成的量與細(xì)胞數(shù)量及代謝活性成比例，因此吸光度值的大小可反應(yīng)細(xì)胞的數(shù)量及活性。自從1983年美國(guó)學(xué)者M(jìn)osmamn建立了MTT比色法以來(lái)，以其快速、靈敏、簡(jiǎn)便、準(zhǔn)確等優(yōu)點(diǎn)受到人們的普遍重視，現(xiàn)已廣泛應(yīng)用于各個(gè)領(lǐng)域，例如檢測(cè)細(xì)胞對(duì)化學(xué)藥物的敏感性，刺激劑對(duì)免疫細(xì)胞的反應(yīng)、免疫細(xì)胞毒性、細(xì)胞因子活性等[41]。 討論本章節(jié)首先通過(guò)控制單一變量的方法對(duì)制備的可激活磁共振對(duì)比劑Fe3O4SSGd2O