本文選題：四氧化三鐵　切入點：納米復合物　出處：《中國組織工程研究》2016年38期

【摘要】：背景:將磁性納米顆粒作為骨組織工程領(lǐng)域新興材料并植入生物體時,其毒性機制基礎(chǔ)研究及安全性評價就顯得格外重要。目的:探索磁性四氧化三鐵納米顆粒作用于前成骨細胞的生物相容性。方法:將含有0,200,400,800 mg/L磁性四氧化三鐵納米顆粒的細胞培養(yǎng)液作用于小鼠前成骨細胞24 h后,采用堿性磷酸酶活性測試試劑盒、骨鈣素酶聯(lián)免疫試劑盒、CCK-8試劑盒、倒置顯微鏡、細胞骨架熒光染色法及實時熒光定量PCR,觀察處理后小鼠前成骨細胞的堿性磷酸酶活性/總蛋白比值、骨鈣素濃度、細胞增殖率、細胞形態(tài)和骨架改變、細胞凋亡以及自噬相關(guān)半胱氨酸蛋白酶3、LC3A、LC3B基因m RNA表達情況。結(jié)果與結(jié)論:(1)磁性四氧化三鐵納米顆粒作用小鼠前成骨細胞24 h后,200 mg/L組與對照組(0 mg/L)相比,各項指標差異無顯著性意義;(2)作用24 h后,400,800 mg/L組細胞內(nèi)堿性磷酸酶活性/總蛋白比值、骨鈣素濃度上升,細胞增殖率明顯下降,細胞形態(tài)及骨架結(jié)構(gòu)改變明顯,LC3B基因轉(zhuǎn)錄水平升高,而半胱氨酸蛋白酶3、LC3A基因轉(zhuǎn)錄水平與對照組相比差異無顯著性意義;(3)結(jié)果提示,高質(zhì)量濃度的磁性四氧化三鐵納米顆粒作用于小鼠前成骨細胞24 h后,雖具有一定的促成骨作用,但同時可造成細胞毒性損傷,促進細胞自噬相關(guān)基因LC3B表達上調(diào),影響細胞形態(tài)、骨架結(jié)構(gòu)及細胞增殖率。
[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.
【作者單位】： 中山大學光華口腔醫(yī)學院·附屬口腔醫(yī)院口腔頜面外科廣東省口腔醫(yī)學重點實驗室;
【基金】：國家自然科學基金(81070818)~~
【分類號】：R318.08

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