摘要
目的:通过一步水热法合成葡萄糖酸锌碳点(Zn-CDs),探讨Zn-CDs的细胞成像及其对小鼠前成骨细胞向成骨方向分化的促进作用。方法:一步水热法合成Zn-CDs,采用透射电子显微镜(TEM)、荧光光谱仪和傅里叶变换红外光谱仪(FT-IR)观察检测Zn-CDs的表征。将不同浓度(0.01、0.10、1.00、10.00、100.00和1 000.00mg·L^-1)Zn-CDs浸提液与小鼠前成骨细胞系MC3T3-E1共培养作为实验组,空白对照组仅加入细胞培养液。采用MTT法检测各组MC3T-E1细胞相对增殖率(RGR);采用激光共聚焦成像观察MC3T3-E1细胞的成像特点;采用qRT-PCR法检测各组MC3T3-E1细胞中Runt相关转录因子2基因(Runx2)、碱性磷酸酶基因(ALP)和骨钙素(OC)mRNA相对表达水平;茜素红染色检测各组细胞中钙化结节数。结果:TEM检测,成功合成粒径为5.25nm的Zn-CDs。荧光光谱,Zn-CDs具有360nm紫外光激发和450nm蓝光发射的荧光性质,并表现激发波长依赖特性。FT-IR检测,Zn-CDs表面主要由羧基和羟基基团构成。与空白对照组比较,共培养24h时1 000.00mg·L^-1Zn-CDs组MC3T3-E1细胞的RGR明显降低(P<0.01)。荧光成像,Zn-CDs与MC3T3-E1细胞共培养后,细胞呈现蓝色、绿色和红色的荧光图像,形态轮廓清楚且荧光强度细胞质比细胞核强。qRT-PCR检测,随着Zn-CDs浓度的增加,MC3T3-E1细胞中Runx2、ALP和OCmRNA相对表达水平逐渐升高。茜素红染色,诱导21d后不同浓度Zn-CDs组MC3T3-E1细胞中钙结节数多于空白对照组。结论:Zn-CDs可以有效地进行MC3T3-E1细胞荧光成像,且Zn-CDs具有一定的促MC3T3-E1细胞向成骨方向分化的作用。
Objective:To synthesize the zinc gluconate carbon dots(Zn-CDs)by one-step hydrothermal method,and to investigate their effects on the cell imaging and inducing osteoblastic differentiation of preosteoblasts in the mice.Methods:The Zn-CDs were synthesized by one-step hydrothermal method,and the characteristics were observed and detected by transmission electron microscope(TEM),Fourier transform-infrared spectrum(FT-IR)and fluorescence spectrometer.The MC3T3-E1 cells were divided into blank control group and experimental groups;different concentrations(0.01,0.10,1.00,10.00,100.00,1 000 mg·L^-1)of Zn-CDs were added into the cells in experimental groups,and nothing was added into the cells in blank control group.MTT assay was used to determin the relative growth rate(RGR)of MC3T3-E1 cells in various groups;the imaging characteristics of MC3T3-E1 cells were observed under confocal microscope;the relative expression levels of Runt-relateed transcription factor-2(Runx2),alkaline phosphatase(ALP)and osteocalcin(OC)mRNA in the MC3T3-E1 cells in various groups were detected by qRT-PCR;the number of calcified nodules was detected by alizarin red staining.Results:The TEM results showed that the particle size of Zn-CDs was about 5.25 nm.The fluorescence spectrometer results showed that the Zn-CDs had the fluorescence properties of 360 nm ultraviolet excited light and 450 nm blue emission light,and the Zn-CDs showed the characteristics of excitation wavelength dependence.The FT-IR results showed that the surface of Zn-CDs was mainly composed of carboxyl groups and hydroxyl groups.Compared with blank control group,the RGR of MC T3-E1 cells in 1 000.00 mg·L^-1 Zn-CDs group was decreased significantly at 24 hafter co-culture(P<0.01).The results of fluorescence imaging showed that the blue,green and red fluorescence in the MC3T3-E1 cells,the outline was clear,and the fluorescence intensity of the cytoplasm was stronger than that of the nucleus after co-cultured with Zn-CDs.The qRT-PCR results showed that the relative expression
作者
孟阳
杨明锡
王柳然
刘东宁
于维先
王闻天
武洲
MENG Yang;YANG Mingxi;WANG Liuran;LIU Dongning;YU Weixian;WANG Wentian;WU Zhou(Department of Periodontology, Stomatology Hospital, Jilin University, Changchun 130021, China;State Key Laboratory of Supramolecular Structure and Materials, Jilin University, Changchun 130012, China;Jilin Provincial Key Laboratory of Tooth Development and Jaw Bone Remodeling and Regeneration,Changchun 130021, China;Department of Stomatology, Aerospace Central Hospital, Beijing 100049, China;Department of Dental Medicine,Graduate School, Kyushu University, Fukuoka 812-8581, Japan)
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2019年第3期504-510,I0002,共8页
Journal of Jilin University：Medicine Edition
基金
国家自然科学基金面上项目资助课题(81570983)
吉林省科技厅国际合作项目资助课题(20180414053GH)
吉林省卫计委卫生技术创新项目资助课题(2016J073)
关键词
荧光碳点
生物学成像
前成骨细胞
成骨分化
carbon dots
biological imaging
pro-osteoblast
osteoblastic differentiation
