无溶剂法荧光碳点的制备及卵巢癌细胞荧光成像研究

Fluorescent carbon dots prepared by solvent-free method and their ovarian cancer cells fluorescence imaging application

目的:构建基于碳点(carbon dots,CDs)的荧光纳米探针CDs@Tf,探讨其材料制备及应用于卵巢癌细胞荧光成像的潜能。方法:以二水合柠檬酸钠和尿素为原料,在高温下经无溶剂法制备一种高效荧光碳点,用新型四唑化合物(MTS)法检测其对人卵巢癌细胞HO-8910及人脐静脉血管内皮细胞EA.hy926的细胞毒性。昆明小鼠经尾静脉注射CDs 7 d和21 d后采集主要器官(心、肺、肾、肝、脾)进行组织学分析,评价CDs的活体毒性。利用1-(3-二甲基氨基丙基)-3-乙基碳化二亚胺盐酸(EDC·HCl)及N-羟基琥珀酰亚胺(NHS)作为化学耦合剂,将制备的CDs与转铁蛋白(Tf)共价结合成荧光分子探针CDs@Tf。对其表征分析并应用于细胞成像。结果:制得一种平均粒径为3.85 nm的球形荧光CDs,最佳激发波长为400 nm,最大发射波长为520 nm,量子产率高达93%。所制备的CDs具有较高的荧光强度和低细胞毒性等性质。建立的CDs@Tf分子探针能靶向识别H0-8910卵巢癌细胞。结论:成功构建一种光学性能优良、毒性低的靶向性新型荧光纳米探针CDs@Tf,有助于应用于卵巢癌的早期筛查。

Objective:To construct CDs@Tf fluorescent nanoprobe based on carbon dots,and to explore the material preparation and the potential application in fluorescence imaging of ovarian cancer cells.Methods:Using sodium citrate dihydrate and urea as raw materials,a high efficiency fluorescent carbon dots(CDs)were prepared by solvent-free method at high temperature.The cell cytotoxicity of CDs was determined by MTS assay against HO-8910 ovarian cancer cells and EA.hy926 umbilical vein endothelial cells.Major organs(heart,lung,kidney,liver,spleen)were collected from Kunming mice after 7 days and 21 days of CDs injection via tail vein for histological analysis to evaluate the toxicity of CDs in vivo.Using 1-(3-dimethylaminopropyl)-3-ethylcarbonide diimine hydrochloride(EDC·HCl)and N-hydroxysuccinimide(NHS)as chemical coupling agents,the prepared CDs were covalently combined with transferrin(Tf)to form fluorescent molecular probe CDs@Tf.The probe was characterized and applied to cell imaging.Results:The spherical fluorescent CDs with an average diameter of 3.85 nm were prepared.The CDs exhibited an optimal excitation wavelength of 400 nm and a maximum emission wavelength of 520 nm,and a quantum yield of 93%.The CDs had excellent properties such as high fluorescence intensity and low cytotoxicity.The established CDs@Tf molecular probe could target and identify HO-8910 ovarian cancer cells.Conclusion:A novel targeted fluorescent nanoprobe CDs@Tf with excellent optical performance and low toxicity is successfully constructed,which is helpful for the early screening of ovarian cancer.