Internalization of titanium dioxide nanoparticles by glial cells is given at short times and is mainly mediated by actin reorganization-dependent endocytosis

Many nanoparticles (NPs) have toxic effects on multiple cell lines. This toxicity is assumed to be related to their accumulation within cells. However, the process of internalization of NPs has not yet been fully characterized. In this study, the cellular uptake, accumulation, and localization of titanium dioxide nanoparticles (TiO₂ NPs) in rat (C6) and human (U373) glial cells were analyzed using time-lapse microscopy (TLM) and transmission electron microscopy (TEM). Cytochalasin D (Cyt-D) was used to evaluate whether the internalization process depends of actin reorganization. To determine whether the NP uptake is mediated by phagocytosis or macropinocytosis, nitroblue tetrazolium (NBT) reduction was measured and the 5-(N-ethyl-N-isopropyl)-amiloride was used. Expression of proteins involved with endocytosis and exocytosis such as caveolin-1 (Cav-1) and cysteine string proteins (CSPs) was also determined using flow cytometry.TiO₂ NPs were taken up by both cell types, were bound to cellular membranes and were internalized at very short times after exposure (C6, 30 min; U373, 2 h). During the uptake process, the formation of pseudopodia and intracellular vesicles was observed, indicating that this process was mediated by endocytosis. No specific localization of TiO₂ NPs into particular organelles was found: in contrast, they were primarily localized into large vesicles in the cytoplasm. Internalization of TiO₂ NPs was strongly inhibited by Cyt-D in both cells and by amiloride in U373 cells; besides, the observed endocytosis was not associated with NBT reduction in either cell type, indicating that macropinocytosis is the main process of internalization in U373 cells. In addition, increases in the expression of Cav-1 protein and CSPs were observed.In conclusion, glial cells are able to internalize TiO₂ NPs by a constitutive endocytic mechanism which may be associated with their strong cytotoxic effect in these cells; therefore, TiO₂ NPs internalization and their accumulation in brain cells could be dangerous to human health.     

Generation and preclinical immunogenicity study of dengue type 2 virus-like particles derived from stably transfected mosquito cells

Recent phase IIb/III trials of a tetravalent live attenuated vaccine candidate revealed a need for improvement in the stimulation of protective immunity against diseases caused by dengue type 2 virus (DENV-2). Our attempts to develop particulate antigens for possibly supplementing live attenuated virus preparation involve generation and purification of recombinant DENV-2 virus-like particles (VLPs) derived from stably (prM+E)-expressing mosquito cells. Two VLP preparations generated with either negligible or enhanced prM cleavage exhibited different proportions of spherical particles and tubular particles of variable lengths. In BALB/c mice, VLPs were moderately immunogenic, requiring adjuvants for the induction of strong virus neutralizing antibody responses. VLPs with enhanced prM cleavage induced higher levels of neutralizing antibody than those without, but the stimulatory activity of both VLPs was similar in the presence of adjuvants. Comparison of EDIII-binding antibodies in mice following two adjuvanted doses of these VLPs revealed subtle differences in the stimulation of anti-EDIII binding antibodies. In cynomolgus macaques, VLPs with enhanced prM cleavage augmented strongly neutralizing antibody and EDIII-binding antibody responses in live attenuated virus-primed recipients, suggesting that these DENV-2 VLPs may be useful as the boosting antigen in prime-boost immunization. As the levels of neutralizing antibody induced in macaques with the prime-boost immunization were comparable to those infected with wild type virus, this virus-prime VLP-boost regimen may provide an immunization platform in which a need for robust neutralizing antibody response in the protection against DENV-2-associated illnesses could be tested.     

Influence of Fine Grains on the Bending Fatigue Behavior of Two Implant Titanium Alloys

By means of the ultrasonic surface impact (amplitude of 30 μm, strike number of 48,000 times/mm²), nanograins have been achieved in the surfaces of both Ti6Al4V(TC4) and Ti3Zr2Sn3Mo25Nb(TLM) titanium alloys, mainly because of the dislocation motion. Many mechanical properties are improved, such as hardness, residual stress, and roughness. The rotating–bending fatigue limits of TC4 and TLM subjected to ultrasonic impact are improved by 13.1% and 23.7%, separately. Because of the bending fatigue behavior, which is sensitive to the surface condition, cracks usually initiate from the surface defects under high stress amplitude. By means of an ultrasonic impact tip with the size of 8 mm, most of the inner cracks present at the zone with a depth range of 100~250 μm in the high life region. The inner crack core to TC4 usually appears as a deformed long and narrow α-phase, while the cracks in TLM specimens prefer to initiate at the triple grain boundary junctions. This zone crosses the grain refined layer and the deformed coarse grain layer. With the gradient change of elastic parameters, the model shows an increase of normal stress at this zone. Combined with the loss of plasticity and toughness, it is easy to understand these fatigue behaviors.     

Immune responses against hepatitis C virus genotype 3a virus-like particles in mice: A novel VLP prime-adenovirus boost strategy

Chronic hepatitis C virus (HCV) infection represents a major health threat to global population. In India, approximately 15–20% of cases of chronic liver diseases are caused by HCV infection. Although, new drug treatments hold great promise for HCV eradication in infected individuals, the treatments are highly expensive. A vaccine for preventing or treating HCV infection would be of great value, particularly in developing countries. Several preclinical trials of virus-like particle (VLP) based vaccine strategies are in progress throughout the world. Previously, using baculovirus based system, we have reported the production of hepatitis C virus-like particles (HCV-LPs) encoding structural proteins for genotype 3a, which is prevalent in India. In the present study, we have generated HCV-LPs using adenovirus based system and tried different immunization strategies by using combinations of both kinds of HCV-LPs with other genotype 3a-based immunogens. HCV-LPs and peptides based ELISAs were used to evaluate antibody responses generated by these combinations. Cell-mediated immune responses were measured by using T-cell proliferation assay and intracellular cytokine staining. We observed that administration of recombinant adenoviruses expressing HCV structural proteins as final booster enhances both antibody as well as T-cell responses. Additionally, reduction of binding of VLP and JFH1 virus to human hepatocellular carcinoma cells demonstrated the presence of neutralizing antibodies in immunized sera. Taken together, our results suggest that the combined regimen of VLP followed by recombinant adenovirus could more effectively inhibit HCV infection, endorsing the novel vaccine strategy.     

镍钛形状记忆合金环抱接骨板治疗桡骨近端1/3骨折

目的探讨治疗桡骨近端1/3骨折的理想内固定方法.方法 2001年8月～2004年4月,采用镍钛形状记忆合金环抱接骨板治疗24例桡骨近端1/3骨折.其中男19例,女5例.年龄16～48岁.摔伤9例,压砸伤5例,交通事故伤4例,直接暴力撞击伤3例,高处坠落伤3例.按照形状记忆合金环抱接骨板的安装方法进行骨折复位内固定.术后定期随访,观察骨折愈合及功能恢复情况.结果所有患者均获随访4～21个月,平均10个月.均无术中、术后并发症,切口Ⅰ期愈合;X 线片示骨折均于术后8～12周临床愈合,根据Anderson疗效标准,功能恢复优18例,良5例,可1例,优良率95.83%.结论应用镍钛形状记忆合金环抱接骨板治疗桡骨近端1/3骨折具有创伤小、操作简便安全、固定可靠、组织相容性好、并发症少等优点,有利于促进骨折愈合和肘、腕关节功能康复,是一种治疗桡骨近端1/3骨折的较好方法.     

钛瓷瓷裂的临床研究和原因分析

目的：分析钛-瓷修复体失败的原因。方法：对38例瓷裂的钛-瓷修复体进行分析：①肉眼观察烤瓷崩瓷部铸件的表面颜色并用便携式厚度测试仪测量其厚度，与未发生瓷裂的正常镰铬合金烤瓷比较；②观察烤瓷崩瓷的部位；③X-ray检测铸件内部气孔。结果：本组崩瓷的钛-瓷修复体，其断层或有气泡，或金属表面氧化层明显增厚（P〈0．05）。而且，所有发生崩瓷的部位均在受力点或因技工本身操作不当引起。结论：钛-瓷修复体失败的原因与加工过程中工艺、瓷的受力点以及与钛元素化学性能有关。     

钛对大肠杆菌生长影响的实验研究

以草酸钛钾溶液为受试物，用无菌操作法在不同浓度试验水样及不含钛的对照水样中加入大肠杆菌稀释液1ml（含菌300个），观察6小时和1－7天的细菌生长情况。结果各受试组与对照组比较，10.0mg／L和1．0mg／L组接触6小时至7天大肠杆菌生长均有明显的抑制作用；0．1mg／L组接触6－24小时显示抑制作用，但第4天开始菌落减少的速度比对照组缓慢。提示钛对大肠杆菌的生长有抑制作用，其作用阈浓度为1．0mg／L，阈下浓度为0，1mg／L。     

复合矫治弓丝接头的力学性能及接头组织观察

目的　测试复合矫治弓丝接头的机械性能及分析接头、接头断口变化。方法　测试复合矫治弓丝接头的拉伸强度、剪切强度、扭转强度、弯曲形变残余角,TiNi形状记忆合金丝超弹性,扫描电镜观察及分析接头、断口组织变化。结果　复合矫治弓丝接头抗拉伸强度为32 0～36 0MPa ,剪切强度为2 2 0～2 5 0MPa ,抗扭转强度为30～4 0N·m、断裂时扭转角度不小于1 4 4 0°,弯曲形变残余角为1 2～1 7°;钎焊异质接头中TiNi形状记忆合金丝超弹性几乎没有破坏;接头钎料与TiNi形状记忆合金界面不存在扩散层,与不锈钢丝界面之间有一较宽的扩散层,接头断裂多发生在钎料与TiNi形状记忆合金超弹界面处,部分发生在钎缝中心。结论　复合矫治弓丝机械性能符合正畸临床要求,可应用于临床。     

Effects of neurosurgical titanium mesh on radiation dose

The purpose of this study was to determine the dosimetric impact of a neurosurgical titanium mesh in patients treated with 6- and 18-MV photon beams. The effects of a 0.4-mm-thick titanium mesh on the dose profile at 3 regions within a solid water phantom were measured using extended dose range-2 (EDR2) film for 6- and 18-MV photon beams. All measurements were performed with the titanium mesh placed at a depth of 1.5 cm in the phantom. Films were exposed immediately above the mesh, immediately below the mesh, and at a depth of 5 cm from the surface of the phantom. The films were scanned using a scanning densitometer. In the region directly above the titanium mesh, there was an increase in dose of 7.1% for 6-MV photons and 4.9% for 18-MV photons. Directly below the titanium mesh, there was an average decrease in dose of 1.5% for 6-MV photons and an increase of 1.0% for 18-MV photons. At 5-cm depth, for 6- and 18-MV photons, there was a decrease in dose of 2.2% and 0.6%, respectively. We concluded that for cranial irradiation with high-energy photons, the dosimetric impact of a 0.4-mm titanium mesh is small and does not require modification in treatment parameters.