论文部分内容阅读
背景:有报道指出石墨碳纳米颗粒具有强大的吸附能力,只要尽可能将其控制在有效浓度范围内,石墨碳纳米颗粒会具有很好的细胞相容性及增敏效应。目的:了解石墨碳纳米颗粒的形态特征,观察石墨碳纳米颗粒其对体外培养细胞增殖与超微结构的影响。方法:取石墨碳纳米颗粒0.5g,加100mL三蒸水,振荡后微孔过滤,即为石墨碳纳米颗粒母液。取处于对数生长期的HepG2细胞、L02细胞、Hl7702细胞、3T3细胞,调整密度为5×107L-1接种于6孔板,0.5mL/孔,加入含胎牛血清、青霉素、链霉素的RPMI-1640培养基1.5mL,培养24h后弃去旧液,设1～5号孔为实验组,分别加入质量浓度为25,10,7.5,5,0.25mg/L石墨碳纳米颗粒培养液2.0mL,设6号孔为空白对照组,不加石墨碳纳米颗粒溶液,继续培养24h后终止培养。用原子力显微镜测量石墨碳纳米颗粒的粒径,电子显微镜观察石墨碳纳米颗粒的形态特征;用细胞计数板在光学显微镜下计数不同浓度石墨碳纳米颗粒对细胞数量的影响;透射电镜观察7.5mg/L石墨碳纳米颗粒对细胞超微结构的影响。结果与结论:石墨碳纳米颗粒呈球形微粒,粒径约20nm。与空白对照组比较,各浓度石墨碳纳米颗粒培养液组除HepG2细胞外,其余3种细胞数量基本都有所增加,其中7.5mg/L石墨碳纳米颗粒培养液对L02细胞、Hl7702细胞、3T3细胞、HepG2细胞数量的影响最为显著(P<0.05)。对7.5mg/L石墨碳纳米颗粒作用后的细胞进行透射电镜观察,可见石墨碳纳米颗粒分布于细胞内部,如细胞质、细胞核、线粒体中,未见亚细胞结构受损及细胞凋亡坏死现象发生。证实石墨碳纳米颗粒对体外培养的细胞无损伤不良反应,且能够促进细胞生长增殖,其作用强度与质量浓度有关,7.5mg/L为较佳质量浓度。
BACKGROUND: It has been reported that graphite carbon nanoparticles have strong adsorption capacity. As long as they are controlled within the effective concentration range, graphite carbon nanoparticles have good cell compatibility and sensitization effects. OBJECTIVE: To understand the morphological characteristics of graphite carbon nanoparticles and observe the effect of graphite carbon nanoparticles on cell proliferation and ultrastructure in vitro. Methods: Take graphite carbon nanoparticles 0.5g, add 100mL three steam water, after oscillation microporous filtration, that is graphite carbon nanoparticles mother liquor. Take logarithmic growth phase of HepG2 cells, L02 cells, Hl7702 cells, 3T3 cells, the density of 5 × 107L-1 inoculated on a 6-well plate, 0.5mL / well, adding fetal bovine serum, penicillin, streptomycin RPMI-1640 culture medium 1.5mL, the old liquid was discarded after culturing for 24h, and holes No. 1 to No. 5 were set as the experimental group, respectively, adding 25,10,7.5,5,0.25mg / L graphite carbon nanoparticle culture solution 2.0 mL, set hole No. 6 as a blank control group, without adding graphite carbon nanoparticles solution, continue to culture after 24h termination of culture. The particle size of graphite carbon nanoparticles was measured by atomic force microscopy. The morphological characteristics of graphite carbon nanoparticles were observed by electron microscopy. The effects of different concentrations of graphite carbon nanoparticles on cell number were counted under a light microscope with a cell counting plate. Transmission electron microscope (TEM) L graphite carbon nanoparticles on the ultrastructure of cells. RESULTS AND CONCLUSION: Graphite carbon nanoparticles were spherical particles with particle size of about 20nm. Compared with the blank control group, except for HepG2 cells, the concentration of all three kinds of cells in the graphitized carbon nano-particle culture medium basically increased, of which 7.5 mg / L graphite carbon nanoparticle culture medium had no effect on L02 cells, H17702 cells, 3T3 The number of cells and HepG2 cells had the most significant effect (P <0.05). After treated with 7.5 mg / L graphitic carbon nanoparticles, TEM images showed that the graphite carbon nanoparticles were distributed in the cytoplasm, such as cytoplasm, nucleus and mitochondria, and no sub-cellular structure was found and apoptosis necrosis occurred . It was confirmed that graphite carbon nanoparticles had no adverse reaction to cells cultured in vitro and could promote the proliferation of cells. The intensity of the effect was related to the mass concentration, and 7.5 mg / L was the best mass concentration.