磁性四氧化三铁纳米颗粒作用于前成骨细胞的生物相容性

发布时间：2018-03-23 21:25

本文选题：四氧化三铁　切入点：纳米复合物　出处：《中国组织工程研究》2016年38期

【摘要】：背景:将磁性纳米颗粒作为骨组织工程领域新兴材料并植入生物体时,其毒性机制基础研究及安全性评价就显得格外重要。目的:探索磁性四氧化三铁纳米颗粒作用于前成骨细胞的生物相容性。方法:将含有0,200,400,800 mg/L磁性四氧化三铁纳米颗粒的细胞培养液作用于小鼠前成骨细胞24 h后,采用碱性磷酸酶活性测试试剂盒、骨钙素酶联免疫试剂盒、CCK-8试剂盒、倒置显微镜、细胞骨架荧光染色法及实时荧光定量PCR,观察处理后小鼠前成骨细胞的碱性磷酸酶活性/总蛋白比值、骨钙素浓度、细胞增殖率、细胞形态和骨架改变、细胞凋亡以及自噬相关半胱氨酸蛋白酶3、LC3A、LC3B基因m RNA表达情况。结果与结论:(1)磁性四氧化三铁纳米颗粒作用小鼠前成骨细胞24 h后,200 mg/L组与对照组(0 mg/L)相比,各项指标差异无显著性意义;(2)作用24 h后,400,800 mg/L组细胞内碱性磷酸酶活性/总蛋白比值、骨钙素浓度上升,细胞增殖率明显下降,细胞形态及骨架结构改变明显,LC3B基因转录水平升高,而半胱氨酸蛋白酶3、LC3A基因转录水平与对照组相比差异无显著性意义;(3)结果提示,高质量浓度的磁性四氧化三铁纳米颗粒作用于小鼠前成骨细胞24 h后,虽具有一定的促成骨作用,但同时可造成细胞毒性损伤,促进细胞自噬相关基因LC3B表达上调,影响细胞形态、骨架结构及细胞增殖率。
[Abstract]:Background: magnetic nanoparticles are used as emerging materials in bone tissue engineering and implanted into organisms. It is particularly important to study the mechanism of toxicity and evaluate its safety. Objective: to explore the biocompatibility of magnetic ferric oxide nanoparticles acting on preosteoblasts. Methods: 0200400800 mg/L magnetic ferrite tetroxide was added to the preosteoblasts. The cell culture medium of rice granules was applied to mouse preosteoblasts for 24 h. Alkaline phosphatase activity test kit, osteocalcin enzyme linked immunoassay kit and CCK-8 kit were used. The ratio of alkaline phosphatase activity to total protein, osteocalcin concentration, cell proliferation rate, morphology and cytoskeleton of preosteoblast were observed by fluorescence staining of cytoskeleton and real-time fluorescence quantitative PCR. Cell apoptosis and m RNA expression of autophagy associated cysteine protease 3 (LC3A) LC3B gene were compared with those of control group (0 mg / L) after treatment with magnetic ferric oxide nanoparticles for 24 h. The ratio of alkaline phosphatase activity to total protein, osteocalcin concentration and proliferation rate of the cells in the 400 800 mg/L group were significantly decreased after 24 h treatment. The changes of cell morphology and cytoskeleton structure showed that the transcriptional level of LC3B gene increased, but the transcription level of cysteine protease 3 + LC3A gene was not significantly different from that of the control group. After high concentration of magnetic ferric oxide nanoparticles acted on mouse preosteoblast for 24 h, although it promoted bone formation, it also caused cytotoxic injury and up-regulated the expression of autophagy related gene LC3B. It affects cell morphology, cytoskeleton structure and cell proliferation rate.
【作者单位】：中山大学光华口腔医学院·附属口腔医院口腔颌面外科广东省口腔医学重点实验室;
【基金】：国家自然科学基金(81070818)~~
【分类号】：R318.08

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