摘要
目的制备半导体聚合物量子点(Pdots)、狂犬病病毒糖蛋白29(RVG29)修饰的Pdots-RVG纳米复合物,体外表征后检测该纳米复合物能否被神经细胞摄取,体内探究其能否穿透小鼠血脑屏障(BBB)。方法纳米共沉淀法制备Pdots,并通过静电吸附连接RVG29制备Pdots-RVG纳米复合物;动态光散射技术检测纳米复合物的平均粒径及Zeta电位;细胞增殖-毒性检测试剂盒(CCK-8)检测Pdots及Pdots-RVG的细胞毒性;纳米颗粒与小鼠中脑多巴胺能神经元细胞系MN9D细胞共孵育,激光扫描共聚焦显微镜下观察纳米颗粒的细胞摄取。分别将磷酸缓冲盐溶液(PBS)、Pdots、Pdots-RVG经腹腔注射进小鼠体内,40 h后制备组织单细胞悬液,并通过流式细胞术分析各组织中Pdots阳性细胞数量及生物富集。结果Pdots平均粒径为105.10 nm,Zeta电位为-33.83 mV,Pdots-RVG纳米复合物平均粒径为339.13 nm,Zeta电位为-18.20 mV;当Pdots浓度为0~50 mg/L时,Pdots与Pdots-RVG均表现出较好的生物相容性;激光共聚焦扫描显微镜下观察到,在MN9D细胞中Pdots-RVG比Pdots摄取更多、更快;流式细胞术分析显示,Pdots在小鼠体内主要分布在肝脏、脾脏、肺等周围脏器中,在脑中无蓄积;Pdots-RVG在小鼠体内主要分布在脑中(如嗅球、皮层、纹状体等),在周围脏器中较少蓄积。结论本实验制备的Pdots-RVG纳米复合物生物相容性较好,能被神经细胞大量且快速摄取,在小鼠体内能穿透BBB,并在嗅球、皮层等部位大量蓄积。
Objective To prepare rabies virus glycoprotein 29(RVG29)modified semiconducting polymer dots(Pdots)nanocomposite(Pdots-RVG),and to explore its ability to be taken up by nerve cells in vitro and whether it can penetrate the blood-brain barrier(BBB)in mice.Methods Pdots were prepared by nano-coprecipitation,and Pdots-RVG nanocomposites were prepared by connecting RVG29 by electrostatic adsorption.The average particle size and Zeta potential of the nanocomposites were detected by dynamic light scattering.The cytotoxicity of Pdots and Pdots-RVG was detected by cell counting kit-8(CCK-8).The nanoparticles were co-incubated with mouse midbrain dopaminergic cells(MN9D),and the cellular uptake of nanoparticles was observed under a laser scanning confocal microscope.Phosphate buffer saline(PBS),Pdots,and Pdots-RVG were injected into mice by intraperitoneal injection,respectively.Tissue single cell suspension was prepared after 40 h and the number of Pdots-positive cells in each tissue was analyzed by flow cytometry to determine the bioaccumulation of Pdots and Pdots-RVG.Results The average particle size of Pdots was 105.10 nm,and the Zeta potential was-33.83 mV.The average particle size of Pdots-RVG nanocomposites was 339.13 nm,and the Zeta potential was-18.20 mV.Both Pdots and Pdots-RVG showed good biocompatibility when the concentration of Pdots was 0-50 mg/L.It was observed under laser confocal scanning microscop that the uptake of Pdots-RVG was more and faster than that of Pdots in MN9D cells.Flow cytometry analysis showed that Pdots was mainly distributed in the liver,spleen,lung and other peripheral organs in mice,and there was no Pdots accumulation in the brain.Pdots-RVG was mainly distributed in the brain(such as olfactory bulb,cortex,striatum,etc.)in mice,but less accumulated in peripheral organs.Conclusion The Pdots-RVG nanocomposite prepared in this study has good biocompatibility and can be taken up by nerve cells in large quantities and quickly.In mice,it can penetrate the BBB and accumulate in the olfactory
作者
尧月
苏炳银
李淑蓉
韩玉萍
Yaoyue;Su Bingyin;Li Shurong;Han Yuping(Key Laboratory of Development and Regeneration of Sichuan Province, Chengdu Medical College, Chengdu 610500, China;Department of Pathology and Pathophysiology, Chengdu Medical College, Chengdu 610500, China;Department of Histology and Embryology, Chengdu Medical College, Chengdu 610500, China)
出处
《成都医学院学报》
CAS
2022年第3期293-299,共7页
Journal of Chengdu Medical College
基金
四川省科技厅应用基础研究重点项目(No:2021YJ0018,No:2019YJ0366)
中国科协青年人才托举项目(No:2019QNRC001)
成都医学院发育与再生四川省重点实验室基金重点项目(No:SYS18-01)
成都医学院发育与再生四川省重点实验室开放基金项目(No:SYS20-04)。
