掺铁TiO2纳米颗粒与恒定磁场对HL60白血病肿瘤细胞的灭活效果

通过研究不同浓度、不同磁场作用下TiO2、掺铁TiO2纳米颗粒对HL60白血病细胞活性的影响,以及在接受光照和不接受光照条件下的细胞活性,探讨基于TiO2、掺铁TiO2纳米颗粒作为光敏剂的光动力疗法(PDT)灭活白血病肿瘤细胞的可行性.实验结果表明,纳米颗粒对细胞具有一定的抑制/毒性作用,纳米浓度越大,抑制/毒性作用越明显;磁场对细胞的毒性/抑制作用跟掺铁的浓度以及磁感应强度有关,掺铁纳米组在强磁场作用下对细胞抑制/毒性作用明显;此外,添加了纳米颗粒的PDT灭杀效率要比不添加纳米颗粒的PDT灭杀效率高.     

纳米级Fe3O4磁性粒子与人肝癌细胞HepG-2及人正常肝细胞L02作用的生物学行为的实验研究

目的:对纳米级Fe3O4磁性粒子与人肝癌细胞HepG-2及人正常肝细胞L02作用的生物学行为进行实验研究。方法:通过化学沉淀法制备粒径为10nm左右的纳米级Fe3O4磁性粒子,观察其表征;将不同浓度纳米级Fe3O4粒子加入培养液分别与HepG-2混合培养检测凋亡坏死率;将相同浓度粒子分别与HepG-2和L02混合培养,对两者作用的差异进行动态观察比较。结果:纳米级Fe3O4磁性粒子能在肝癌细胞HepG-2细胞内稳定存在72小时以上,有良好的生物相容性;透射电镜观察到Fe3O4磁性粒子主要分布于细胞的溶酶体及吞噬泡内。共培养1小时后即有较多的纳米磁性粒子进入HepG-2内,而3小时后才见L02细胞内有少量的磁性粒子进入。结论:此实验结果为磁性纳米粒子与肿瘤细胞微观结构的作用提供了有意义的实验数据,并可能对应用磁性纳米粒子治疗恶性肿瘤提供有价值的依据。     

外加磁场作用下磁性Fe3O4纳米粒子对肝癌细胞增殖和凋亡的影响

目的:观察磁性四氧化三铁(Fe3O4)纳米粒子对肝癌细胞的体外作用,并研究外加稳恒磁场(SMF)或交变磁场(EMF)对Fe3O4纳米粒子作用的影响。方法:光镜下观察CBRH-7919细胞对Fe3O4纳米粒子的吞噬作用;MTT法检测Fe3O4纳米粒子对大鼠肝癌细胞株CBRH-7919的毒性及外加磁场的影响;流式细胞术检测外加磁场作用下Fe3O4纳米粒子对细胞凋亡及线粒体膜电位的影响。结果:光镜下可见CBRH-7919细胞吞噬大量Fe3O4纳米粒子入胞浆,且交变磁场作用下细胞的吞噬量增加。30～100μg/mL Fe3O4纳米粒子作用于CBRH-7919细胞未产生细胞毒性,稳恒磁场对其作用无影响,而交变磁场能增加Fe3O4纳米粒子的毒性,使细胞活性降低、凋亡率增加、线粒体膜电位降低。结论:交变磁场能增加CBRH-7919细胞对Fe3O4纳米粒子的吞噬并产生细胞毒性。     

外加磁场作用下磁性Fe3O4纳米粒子对肝癌细胞增殖和凋亡的影响

目的：观察磁性四氧化三铁（Fe3O4）纳米粒子对肝癌细胞的体外作用,并研究外加稳恒磁场（SMF）或交变磁场（EMF）对FeID4纳米粒子作用的影响。方法：光镜下观察CBRH-7919细胞对Fe3O4纳米粒子的吞噬作用;MTT法检测Fe304纳米粒子对大鼠肝癌细胞株CBRH-7919的毒性及外加磁场的影响;流式细胞术检测外加磁场作用下Fe3O4纳米粒子对细胞凋亡及线粒体膜电位的影响。结果：光镜下可见CBRH-7919细胞吞噬大量Fe3O4纳米粒子入胞浆,且交变磁场作用下细胞的吞噬量增加。30-100μg／mLFe3O4纳米粒子作用于CBRH-7919细胞未产生细胞毒性,稳恒磁场对其作用无影响,而交变磁场能增加Fe3O4纳米粒子的毒性,使细胞活性降低、凋亡率增加、线粒体膜电位降低。结论：交变磁场能增加CBRH-7919细胞对Fe3O4纳米粒子的吞噬并产生细胞毒性。     

抗人精子蛋白17磁性纳米探针靶向的卵巢癌体内外磁共振成像

用MRI(magnetic resonance imaging)技术探索连接抗人精子蛋白17单克隆抗体(anti-Sp17 mAb)的磁性纳米探针对体外培养及动物体内Sp17+卵巢癌的靶向性。将anti-Sp17mAb连接到表面包覆壳聚糖的超顺磁性氧化铁纳米颗粒上,制成磁性纳米探针anti-Sp17-MNP,用作MRI阴性对比剂。将磁性纳米探针与Sp17+和Sp17-培养的肿瘤细胞共育,进行一系列体外磁共振成像实验。荷瘤小鼠尾静脉注射磁性纳米颗粒,用7T磁共振仪在体成像,观察肿瘤部位的信号变化,并用普鲁士蓝染色肿瘤组织切片,观察有无铁粒子聚集。体外MRI数据显示,anti-Sp17-MNP与细胞靶向结合,并与细胞共育2 h后,Sp17+HO-8910的T2*信号强度比Sp17-HepG2低2倍;anti-Sp17-MNP对肿瘤细胞的靶向作用可被重组人Sp17阻断。7T磁共振仪对动物在体肿瘤成像结果显示,感兴趣区因磁性纳米探针靶向聚集而导致信号降低,并经组织切片普鲁士蓝染色证实。本研究结果表明,用anti-Sp17抗体和新的合成路线制备的纳米探针具有用作MR对比剂进行分子成像的潜能。     

叶酸受体介导的硼中子俘获疗法治疗无功能垂体腺瘤细胞

侵袭性无功能垂体腺瘤手术很难全部切除,且术后经常复发.目前临床上尚无有效药物能控制肿瘤生长.虽然放疗对肿瘤生长有一定抑制作用,但常常导致严重的并发症.硼中子俘获疗法是一种新型的二元靶向放射疗法,能最大程度杀伤肿瘤细胞而对正常组织损伤较小.叶酸受体靶向的含硼碳纳米颗粒是一种新型的硼携带剂,能通过叶酸受体介导的细胞内吞作用被表达叶酸受体的细胞选择性摄取.本实验中,叶酸受体靶向的含硼碳纳米颗粒能选择性地被表达叶酸受体的无功能垂体腺瘤细胞摄取,而无叶酸受体表达的其他类型垂体腺瘤细胞不能摄取这些碳纳米颗粒.叶酸受体介导的硼中子俘获疗法治疗后,无功能垂体腺瘤细胞的细胞活力显著下降,同时凋亡细胞显著增加.而单独的碳纳米颗粒孵育,或单独的热中子照射,并不能抑制肿瘤细胞活力或促进细胞凋亡.同时,叶酸受体介导的硼中子俘获治疗能导致肿瘤细胞中Bcl-2的表达显著下降和Bax的表达显著增强.综上所述,叶酸受体靶向的含硼碳纳米颗粒为无功能垂体腺瘤的BNCT治疗提供了比较理想的硼携带剂.本研究也为侵袭性无功能垂体腺瘤的治疗提供了新的思路,同时拓展了BNCT的应用,尤其是在良性肿瘤中的应用.     

葡聚糖包被的纳米磁粒子制备及其在H-22细胞标记中的应用

利用化学共沉淀法制备近微米级葡聚糖T-40包被的超顺磁纳米氧化铁(SPIO)粒子。用X射线衍射(XRD)、透射电镜(TEM)、原子力显微镜(AFM)、红外光谱等手段对其性质进行表征。XRD确定所制备的粒子主要为Fe3O4晶体,具有超顺磁性且饱和磁化强度为64.396emu/g;TEM显示无机核心粒径约为20nm;被葡聚糖包被后,AFM显示粒子呈扁平长方体形貌,三维尺寸为(200～300)nm×(400～600)nm×(50～70)nm。在超声作用下,该类粒子能够对小鼠H-22肝癌细胞进行标记,说明所制备的磁性纳米粒子具有作细胞磁性标记物的特性,同时超声也可促进H-22细胞快速装载磁性微粒。     

荧光磁性纳米粒子标记间充质干细胞靶向胃癌研究

目的:探索一种高效的早期胃癌诊断体系.方法:荧光磁性纳米粒子与间充质干细胞共培养,不同时间点检测干细胞存活率.利用荧光显微镜、普鲁士蓝染色、透射电子显微镜等方法观测干细胞被标记情况;建立裸鼠的胃癌模型,并将标记后的干细胞尾静脉注射入裸鼠体内,14d后用动物成像仪和核磁共振成像仪检测.结果:荧光磁性纳米粒子在低于 100μg/mL浓度下对间充质干细胞没有毒性作用,荧光磁性纳米粒子与干细胞共培养6h后,荧光显微镜、普鲁士蓝染色、透射电子显微镜检测结果显示粒子被内吞后定位于细胞质中,动物实验显示通过检测肿瘤部位的荧光信号和核磁信号,被标记的干细胞能够准确定位到胃癌的发生部位.结论:这种双模式检测体系为胃癌的早期诊断与治疗提供一种新的方法.     

磁性纳米粒子介导的细胞生物学效应

磁性纳米粒子(magnetic nanoparticles, MNPs)由于其独特磁响应性,可将外加磁场的能量转化为机械能和热能。磁性纳米粒子介导的物理信号依赖于MNPs本身的磁学性能及磁场的参数,可定量输出作用于不同类型的细胞,调控细胞命运。MNPs本身Fe~(2+)引发芬顿反应可上调化学信号(reactive oxygen species, ROS),用于肿瘤治疗;在超低频磁场(1 Hz)下产生的机械力可诱导干细胞分化和巨噬细胞极化等过程,用于再生医学领域;在低频磁场(1～100 Hz)下产生的机械力可通过直接物理破坏或间接触发生物信号通路,引起肿瘤细胞死亡;在高频磁场(100 kHz～1 MHz)下产生的热可破坏肿瘤细胞,在神经元信号转导领域也取得一定的突破。研究MNPs介导的化学、物理、生物信号引起的细胞生物学效应对MNPs的设计和磁场的选择具有重要的指导意义。本文就MNPs在不同类型磁场下介导的细胞生物学效应做一概述。     

热处理肿瘤细胞抗原负载树突细胞抗小鼠结肠癌效应

研究了热处理肿瘤细胞抗原负载骨髓来源的树突细胞（dendritic cell,DC）对结肠癌小鼠的治疗作用.将小鼠结肠癌细胞CT26热处理后超声破膜,以其细胞裂解液负载BALB/c小鼠骨髓来源的DC,观察DC诱导肿瘤特异性细胞毒性T淋巴细胞（cytotoxic T lymphocyte,CTL）杀伤活性;并将DC接种于荷瘤小鼠皮下,观察其对肿瘤生长的抑制作用及对荷瘤小鼠生存期的影响.发现致敏DC诱导的CTL对CT26肿瘤细胞具有显著的杀伤作用.用致敏DC免疫小鼠后,对小鼠肿瘤的生长具有显著的抑制作用,并能显著延长荷瘤小鼠的生存时间.热处理肿瘤细胞抗原负载的树突细胞对结肠癌小鼠具有显著的治疗效果.     

涡旋磁纳米颗粒——崭新的生物医用磁性纳米平台

相比于超顺磁性纳米颗粒,具有涡旋磁畴的磁性纳米颗粒,由于独特的磁化闭合分布、较大的粒径尺寸及外加磁场中的磁化翻转特性,使得其兼具弱的颗粒间磁相互作用和更优异的磁学性能,在生物医学领域展现出了更好的应用优势和潜力.本综述结合近年来国内外对涡旋磁畴的研究及涡旋磁纳米颗粒在生物医学领域的报道,提出了一类新型的生物医用涡旋磁溶胶体系,并以涡旋磁氧化铁纳米盘和纳米环为例,介绍了涡旋磁纳米颗粒的化学合成,并着重论述了这类具有独特涡旋畴结构的纳米颗粒在磁共振成像、抗肿瘤治疗等生物医学应用上的最新研究进展.     

Synthesis of Immunotargeted Magneto-plasmonic Nanoclusters

Magnetic and plasmonic properties combined in a single nanoparticle provide a synergy that is advantageous in a number of biomedical applications including contrast enhancement in novel magnetomotive imaging modalities, simultaneous capture and detection of circulating tumor cells (CTCs), and multimodal molecular imaging combined with photothermal therapy of cancer cells. These applications have stimulated significant interest in development of protocols for synthesis of magneto-plasmonic nanoparticles with optical absorbance in the near-infrared (NIR) region and a strong magnetic moment. Here, we present a novel protocol for synthesis of such hybrid nanoparticles that is based on an oil-in-water microemulsion method. The unique feature of the protocol described herein is synthesis of magneto-plasmonic nanoparticles of various sizes from primary blocks which also have magneto-plasmonic characteristics. This approach yields nanoparticles with a high density of magnetic and plasmonic functionalities which are uniformly distributed throughout the nanoparticle volume. The hybrid nanoparticles can be easily functionalized by attaching antibodies through the Fc moiety leaving the Fab portion that is responsible for antigen binding available for targeting.     

Cell-specific targeting of nanoparticles by multivalent attachment of small molecules

Nanomaterials with precise biological functions have considerable potential for use in biomedical applications. Here we investigate whether multivalent attachment of small molecules can increase specific binding affinity and reveal new biological properties of such nanomaterials. We describe the parallel synthesis of a library comprising 146 nanoparticles decorated with different synthetic small molecules. Using fluorescent magnetic nanoparticles, we rapidly screened the library against different cell lines and discovered a series of nanoparticles with high specificity for endothelial cells, activated human macrophages or pancreatic cancer cells. Hits from the last-mentioned screen were shown to target pancreatic cancer in vivo. The method and described materials could facilitate development of functional nanomaterials for applications such as differentiating cell lines, detecting distinct cellular states and targeting specific cell types.     

Effect of nanoheat stimulation mediated by magnetic nanocomposite hydrogel on the osteogenic differentiation of mesenchymal stem cells

Hyperthermia has been considered as a promising healing treatment in bone regeneration. We designed a tissue engineering hydrogel based on magnetic nanoparticles to explore the characteristics of hyperthermia for osteogenic regeneration. This nanocomposite hydrogel was successfully fabricated by incorporating magnetic Fe₃O₄ nanoparticles into chitosan/polyethylene glycol (PEG) hydrogel, which showed excellent biocompatibility and were able to easily achieve increasing temperatures under an alternative magnetic field (AMF). With uniformly dispersed nanoparticles, the composite hydrogel resulted in high viability of mesenchymal stem cells (MSCs), and the elevated temperature contributed to the highest osteogenic differentiation ability compared with direct heat treatment applied under equal temperatures. Therefore, the nanoheat stimulation method using the magnetic nanocomposite hydrogel under an AMF may be considered as an alternative candidate in bone tissue engineering regenerative applications.     

Magnetic Nanoparticles from Magnetospirillum gryphiswaldense Increase the Efficacy of Thermotherapy in a Model of Colon Carcinoma

Magnetic nanoparticles (MNPs) are capable of generate heating power under the influence of alternating magnetic fields (AMF); this behaviour recently opened new scenarios for advanced biomedical applications, mainly as new promising tumor therapies. In this paper we have tested magnetic nanoparticles called magnetosomes (MNs): a class of MNPs naturally produced by magnetotactic bacteria. We extracted MNs from Magnetospirillum gryphiswaldense strain MSR-1 and tested the interaction with cellular elements and anti-neoplastic activity both in vitro and in vivo, with the aim of developing new therapeutic approaches for neoplastic diseases. In vitro experiments performed on Human Colon Carcinoma HT-29 cell cultures demonstrated a strong uptake of MNs with no evident signs of cytotoxicity and revealed three phases in the interaction: adherence, transport and accumulation in Golgi vesicles. In vivo studies were performed on subcutaneous tumors in mice; in this model MNs are administered by direct injection in the tumor volume, then a protocol consisting of three exposures to an AMF rated at 187 kHz and 23kA/m is carried out on alternate days, over a week. Tumors were monitored by Magnetic Resonance Imaging (MRI) to obtain information about MNs distribution and possible tissue modifications induced by hyperthermia. Histological analysis showed fibrous and necrotic areas close to MNs injection sites in mice subjected to a complete thermotherapy protocol. These results, although concerning a specific tumor model, could be useful to further investigate the feasibility and efficacy of protocols based on MFH. Magnetic nanoparticles naturally produced and extracted from bacteria seem to be promising candidates for theranostic applications in cancer therapy.     

Recent advances in synthesis and surface modification of lanthanide-doped upconversion nanoparticles for biomedical applications

Lanthanide (Ln)-doped upconversion nanoparticles (UCNPs) with appropriate surface modification can be used for a wide range of biomedical applications such as bio-detection, cancer therapy, bio-labeling, fluorescence imaging, magnetic resonance imaging and drug delivery. The upconversion phenomenon exhibited by Ln-doped UCNPs renders them tremendous advantages in biological applications over other types of fluorescent materials (e.g., organic dyes, fluorescent proteins, gold nanoparticles, quantum dots, and luminescent transition metal complexes) for: (i) enhanced tissue penetration depths achieved by near-infrared (NIR) excitation; (ii) improved stability against photobleaching, photoblinking and photochemical degradation; (iii) non-photodamaging to DNA/RNA due to lower excitation light energy; (iv) lower cytotoxicity; and (v) higher detection sensitivity. Ln-doped UCNPs are therefore attracting increasing attentions in recent years. In this review, we present recent advances in the synthesis of Ln-doped UCNPs and their surface modification, as well as their emerging applications in biomedicine. The future prospects of Ln-doped UCNPs for biomedical applications are also discussed.     

Intrinsically superparamagnetic Fe-hydroxyapatite nanoparticles positively influence osteoblast-like cell behaviour

ABSTRACT: BACKGROUND: Superparamagnetic nanoparticles (MNPs) have been progressively explored for their potential in biomedical applications and in particular as a contrast agent for diagnostic imaging, for magnetic drug delivery and more recently for tissue engineering applications. Considering the importance of having safe MNPs for such applications, and the essential role of iron in bone remodelling, this study developed and analysed novel biocompatible and bioreabsorbable superparamagnetic nanoparticles, that avoid the use of poorly tolerated magnetite based nanoparticles, for bone tissue engineering applications. RESULTS: MNPs were obtained by doping hydroxyapatite (HA) with Fe ions, by directly substituting Fe2+ and Fe3+ into the HA structure yielding superparamagnetic bioactive phase. In the current study, we have investigated the effects of increasing concentrations (2000 mug/ml; 1000 mug/ml; 500 mug/ml; 200 mug/ml) of FeHA MNPs in vitro using Saos-2 human osteoblast-like cells cultured for 1, 3 and 7 days with and without the exposure to a static magnetic field of 320 mT. Results demonstrated not only a comparable osteoblast viability and morphology, but increased in cell proliferation, when compared to a commercially available Ha nanoparticles, even with the highest dose used. Furthermore, FeHA MNPs exposure to the static magnetic field resulted in a significant increase in cell proliferation throughout the experimental period, and higher osteoblast activity. In vivo preliminary results demonstrated good biocompatibility of FeHA superparamagnetic material four weeks after implantation into a critical size lesion of the rabbit condyle. CONCLUSIONS: The results of the current study suggest that these novel FeHA MNPs may be particularly relevant for strategies of bone tissue regeneration and open new perspectives for the application of a static magnetic field in a clinical setting of bone replacement, either for diagnostic imaging or magnetic drug delivery.     

Cell Labeling and Targeting with Superparamagnetic Iron Oxide Nanoparticles

Targeted delivery of cells and therapeutic agents would benefit a wide range of biomedical applications by concentrating the therapeutic effect at the target site while minimizing deleterious effects to off-target sites. Magnetic cell targeting is an efficient, safe, and straightforward delivery technique. Superparamagnetic iron oxide nanoparticles (SPION) are biodegradable, biocompatible, and can be endocytosed into cells to render them responsive to magnetic fields. The synthesis process involves creating magnetite (Fe₃O₄) nanoparticles followed by high-speed emulsification to form a poly(lactic-co-glycolic acid) (PLGA) coating. The PLGA-magnetite SPIONs are approximately 120 nm in diameter including the approximately 10 nm diameter magnetite core. When placed in culture medium, SPIONs are naturally endocytosed by cells and stored as small clusters within cytoplasmic endosomes. These particles impart sufficient magnetic mass to the cells to allow for targeting within magnetic fields. Numerous cell sorting and targeting applications are enabled by rendering various cell types responsive to magnetic fields. SPIONs have a variety of other biomedical applications as well including use as a medical imaging contrast agent, targeted drug or gene delivery, diagnostic assays, and generation of local hyperthermia for tumor therapy or tissue soldering.     

综述——新型纳米生物材料的研发：医学纳米颗粒对类细胞膜作用的仿真研究进展

细胞膜是包围细胞质、维持细胞内部组分动态平衡的一个半透膜,参与细胞黏附、离子传导、信号传导等分子生物学过程．类细胞膜提供了有效的模型研究这些生物学过程,故而分子层面上研究医学纳米颗粒对类细胞膜的作用有助于评估纳米颗粒的生物安全性以及促进纳米颗粒的生物医学应用．本文初步探讨了医学纳米颗粒对类细胞膜作用的仿真研究进展,并在此基础上结合膜生物物理学的研究热点对后续的研究进行了展望．     

Modulating pharmacokinetics, tumor uptake and biodistribution by engineered nanoparticles

BACKGROUND: Inorganic nanoparticles provide promising tools for biomedical applications including detection, diagnosis and therapy. While surface properties such as charge are expected to play an important role in their in vivo behavior, very little is known how the surface chemistry of nanoparticles influences their pharmacokinetics, tumor uptake, and biodistribution. METHOD/PRINCIPAL FINDINGS: Using a family of structurally homologous nanoparticles we have investigated how pharmacological properties including tumor uptake and biodistribution are influenced by surface charge using neutral (TEGOH), zwitterionic (Tzwit), negative (TCOOH) and positive (TTMA) nanoparticles. Nanoparticles were injected into mice (normal and athymic) either in the tail vein or into the peritoneum. CONCLUSION: Neutral and zwitterionic nanoparticles demonstrated longer circulation time via both i.p. and i.v. administration, whereas negatively and positively charged nanoparticles possessed relatively short half-lives. These pharmacological characteristics were reflected on the tumor uptake and biodistribution of the respective nanoparticles, with enhanced tumor uptake by neutral and zwitterionic nanoparticles via passive targeting.