第31卷第2期Vol.31 No.22015年2月CHINESE JOURNAL OF INORCANIC CHEMISTRY215-221聚乙二醇-金納米棒介導(dǎo)的近紅外光熱抑菌作用馮曉燕陳瑩*劉玉鵬王春鵬儲(chǔ)富祥(中國(guó)林業(yè)科學(xué)研究院林產(chǎn)化學(xué)工業(yè)研究所;生物質(zhì)化學(xué)利用國(guó)家工程實(shí)驗(yàn)室;國(guó)家林業(yè)局林產(chǎn)化學(xué)工程重點(diǎn)開放性實(shí)驗(yàn)室;江蘇省生物質(zhì)能源與材料重點(diǎn)實(shí)驗(yàn)室,南京210042)摘要:種子生長(zhǎng)法合成縱向表面等離子體共振吸收峰為785nm的金納米棒,并對(duì)其表面進(jìn)行聚乙二醇(PEG)修飾,研究了表面修飾PEG的金納米樺(polyethylene glycol modifed gold nanorods , PEG-GNR)的光熱轉(zhuǎn)化效應(yīng),并測(cè)試了其細(xì)胞毒性。以革蘭氏陽性菌金黃色葡萄球菌蠟狀芽孢桿菌,革蘭氏陰性菌大腸埃希氏菌及銅綠假單胞菌為細(xì)菌模型,詳細(xì)研究了PEC-GNR在808nm波長(zhǎng)近紅外激光照射下金納米棒濃度和照射功率對(duì)抑菌效果的影響。結(jié)果表明,PEG-CNR對(duì)革蘭氏陽性菌和革蘭氏陰性菌在近紅外照射下均有較好的抑菌效果,并且抑菌效果與金納米棒的濃度和照射功率有著密切的關(guān)系;結(jié)合熒光顯微鏡和透射電子顯微鏡對(duì)細(xì)菌壞死狀況的觀察,初步證實(shí)細(xì)菌對(duì)PEG-GNR有效吸收是近紅外光熱殺菌的關(guān)鍵因素。關(guān)鍵詞:金納米棒; PEG修飾;光熱抑菌中圖分類號(hào): TQ131.2文獻(xiàn)標(biāo)識(shí)碼: A文章編號(hào): 1001-4861(2015)02-0215-07DOI: 10.1 1862/CJIC.2015.059Near-IR Photothermal Antibacterial Effects of PolyethyleneGlycol (PEG) Modified Gold NanorodsFENG Xiao-Yan CHEN Ying* LU Yu-Peng WANG Chun-Peng CHU Fu-Xiang(Institute of Chemical Industry of Forestry Products, CAF China; Key Laboratory of Biomass Energy and Material,Jiangsu Province China; National Engineering Laboratory for Biomass Chemical Utilization China;Key and Open Laboratory on Forest Chemical Engineering, SFA, Nanjing 210042, China)Abstract: Gold nanorods (CNR) with longitudinal surface plasma resonance (LSPR) absorption at 785 nm weresynthesized by seed-mediated growth method and their surface was modified with polyethylene glycol (PEG)macromolecular (PEG-GNR). The photothermal conversion effect and cytotoxicity of PEG-GNR were investigated.Different bacteria, including gram-positive bacterium Staphylococcus aureus and Bacillus cereus, gram- negativebacterium Escherichia coli and Pse udomonas aeruginosa were used to analyze the influences of concentration ofPEG-GNR and laser output power on the inhibition effects. The results show that the PEG-GNR has goodantibacterial properties for both Gram positive and negative bacterium under the radiation of near-IR laser. Theconcentration of PEG-GNR and laser output power determined antibacterial effects of the PEG-GNR. Thepreliminary investigation on the antibacterial mechanism was explored by studying of bacteria apoptosis statuswith fluorescence microscope and transmission electronic microscope, suggesting that the effective absorption ofthe PEG GNR by the cells is one of the key factors in the process of photothermal antibiosis.Key words: gold nanorod; PEG modifed; photothermal antibacterial efet收稿日期:2014-04-21。收修改稿日期:2014-12-02。中國(guó)煤化工中央級(jí)公益性科研院所基本科研業(yè)務(wù)費(fèi)專項(xiàng)資金(No.CAFINT2010K04),國(guó)家自然科然科學(xué)基金(No.BK20131071)資 助項(xiàng)目。TYHCNMHG“通訊聯(lián)系人。E-mail:yingchencaf@gmail.com216無機(jī)化學(xué)學(xué)報(bào)第31卷細(xì)菌的耐藥性已成為全球醫(yī)療領(lǐng)域中倍受關(guān)血酸、巰基化的聚乙二醇(mPEGoorSH)為國(guó)產(chǎn)分析注的問題,多藥耐藥性細(xì)菌的出現(xiàn)嚴(yán)重威脅著人類純?cè)噭?活死細(xì)胞試劑盒(LIVE/DEAD BacLight kit,的健康12。人們?yōu)E用抗生素導(dǎo)致多藥耐藥細(xì)菌的種美國(guó));金黃色葡萄球菌(Staphylococcus aureus ,S.類越來越多,現(xiàn)有抗菌藥物的療效越來越小，這是aureus ,CICC10384)、蠟狀芽孢桿菌(Bacillus cereus ,人類公共保健領(lǐng)域面臨的最大挑戰(zhàn)之---。激光光熱B. cereus ,CICC10352)、大腸埃希氏菌(Escherichia coli,療法是利用外加能量在含有病變的組織中產(chǎn)生一E. coli ,CICC10354)和銅綠假單胞菌(Pseudomonas定范圍的高溫,達(dá)到殺死病原體細(xì)胞而不傷害正常aeruginosa,P. aeruginosa,CICC10351)購(gòu)自 中國(guó)工業(yè)細(xì)胞的目的30,由于沒有特異性的靶目標(biāo),因而不.微生物菌種保藏管理中心。易產(chǎn)生耐藥性。用于光熱療法的激光主要使用可見儀器:紫外可見分光光度計(jì)(UV-1800SPECT-光激光，但由于該波段激光對(duì)組織的穿透有限,因ROPHOTOMETER,上海美譜達(dá))、透射電子顯微鏡此激光熱療法具有一定的局限性[。近年來研究報(bào)(TEM JEOL-1010, 日本)、酶標(biāo)儀(ChroMa4300,美道,利用光熱轉(zhuǎn)換材料,如有機(jī)光熱染料吲哚菁綠國(guó))、Zeta電位分析儀(Zetasizer nano zS，MALVERN,或者貴金屬和碳納米材料,能夠選擇性地增強(qiáng)病變英國(guó))、近紅外光激光器(LE-LS-808-3000TFCA,深圳部位的熱損傷從而提高治療效果811。眾所周知,金理歐光電公司)、熒光顯微鏡(OlympusX81,日本)。納米棒是一類經(jīng)典的光熱轉(zhuǎn)換材料,具有合適比率1.2 聚乙二醇(PEG)修飾金納米棒(PEG-GNR)的的金納米棒在近紅外區(qū)域?qū)す饽芰坑袕?qiáng)烈的吸制備收效應(yīng)，選擇與金納米棒縱向表面等離子共振金納米棒溶液的制備7:6.25mL二次蒸餾水(LSPR)波長(zhǎng)相匹配的近紅外激光作為光源,能夠誘中分別加入100 μL HAuCL(0.01 mol.L~)溶液,1.88導(dǎo)皮下深層組織的金納米棒顆粒產(chǎn)生熱效應(yīng),促使mL CTAB(0.2 mol.L)溶液,混合均勻后,再加入0.5細(xì)胞致死1-1。由于生物組織在近紅外區(qū)的光吸收mL NaBH(0.01 mol.L)溶液,加完后劇烈搖晃,得很弱,金納米棒因其獨(dú)特的近紅外光吸收性和光穩(wěn)到棕黃色的溶液,即為金納米棒的種子溶液,將它定性,能有效地代替光吸收染料在激光熱療中的應(yīng)置于30 C恒溫條件下靜置2h,待用。在另一容器用。近年來,金納米棒在腫瘤治療光熱療法等方面中分別加106.9mL的二次蒸餾水,7.425mL得到了廣泛的關(guān)注和研究[1416相比之下,金納米棒HAuCl4 (0.01 mol.L-) 溶液和106.4 mL CTAB(0.2在抗菌感染光熱療法的報(bào)道卻很少見。由于激光光mol.L")溶液,混合均勻后加入1.35 mL AgNO3(0.01熱療法的抑菌機(jī)理與普通小分子抗生素的抑菌機(jī)mol.L)和1.35 mL抗壞血酸(0.1 mol.L )溶液,該溶理有著顯著的不同,因此研究金納米棒的近紅外光液為生長(zhǎng)液,待此溶液變?yōu)闊o色后,加人1.8 mL上熱抑菌對(duì)于探索和解決細(xì)菌耐藥性的問題有著十述金納米棒的種子溶液，混合均勻,30 C恒溫條件分重要的意義。鑒于此,本研究通過種子生長(zhǎng)法制下靜置過夜,即得金納米棒溶液(CTAB-GNR)。將所備獲得了縱向等離子體峰為785 nm的金納米棒,得的金納米棒溶液置于4 C條件下靜置4h,析出并用mPEG0o-SH對(duì)其進(jìn)行了表面修飾，制備在生溶液中多余的CTAB分子,然后通過高速離心處理理?xiàng)l件下更穩(wěn)定并有較長(zhǎng)的體內(nèi)循環(huán)時(shí)間的聚乙.去除溶液中殘余的自由CTAB分子,并將溶液濃縮二醇修飾的金納米棒(PEG-CNR),研究了在近紅外30倍(原子吸收光譜測(cè)得金溶液濃度最終為160.5光照射下PEG修飾的金納米棒對(duì)各種測(cè)試菌(包括μg.mL-)，超聲分散后與巰基化的聚乙二醇革蘭氏陽性菌金黃色葡萄球菌(Staphylococcus (mEG0rSH)溶 液混合,mPEG0SH的最終濃度為aureus)、蠟狀芽孢桿菌(Bacillus cereus) 、革蘭氏陰性2.0mg*mL-',靜置,過夜。最終所得的棕紅色溶液即菌大腸埃希氏菌(Escherichiacoli)及銅綠假單胞菌為聚乙二醇(PEG)修飾的金納米棒(PEG-GNR)溶液,(Pseudomonas aeruginosa)的抑 菌效果。備用。1.3 PEG-GNR 的光熱轉(zhuǎn)換效應(yīng)的測(cè)定1實(shí)驗(yàn)部分將制得的PEG_GNR溶液用二次蒸餾水稀釋到1.1實(shí)驗(yàn)試劑與儀器金濃度分別中國(guó)煤化工ug.mL*,各實(shí)驗(yàn)試劑:氣金酸(HAuCl4)、硼氫化鈉(NaBH4)、取1 mL溶液| YHCN M H G紅外光激光照十六烷基三甲基溴化銨(CTAB)、硝酸銀(AgNO,)抗壞射.上述各個(gè)濃度的PEG-GNR溶液22 min,定時(shí)記第2期馮曉燕等:聚乙二醇-金納米棒介導(dǎo)的近紅外光熱抑菌作用221的關(guān)系;細(xì)菌壞死實(shí)驗(yàn)證實(shí)細(xì)菌對(duì)PEG-CNR有效Chin. 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