Site-specific conjugation of ScFvs antibodies to nanoparticles by bioorthogonal strain-promoted alkyne-nitrone cycloaddition

Particularly suitable: An N-terminal serine mutant of anti-HER2 scFv antibody was conjugated to polymer-coated magnetofluorescent nanoparticles by strain-promoted alkyne-nitrone cycloaddition. The resulting nanoparticles (see scheme) proved effective in targeting and labeling HER2-positive breast cancer cells.     

壳聚糖基阳离子胶束用于共负载两种不同性质药物的研究

以季胺化壳聚糖-O-聚己内酯(TMC-PCL)胶束为载体,用于共负载2种不同亲疏水性质的抗肿瘤物质,阿霉素和吲哚菁绿;并研究了胶束包埋对吲哚菁绿的稳定性和光热效应的影响,以及阿霉素从胶束中的释放行为.结果表明,2种抗肿瘤物质在TMC-PCL胶束中的实际载药量均可达20％,且包封率超过85％.进一步还用MTT法评价了不同载药胶束体系对肿瘤细胞的杀灭作用,发现共负载胶束经近红外激光辐照后,对肿瘤细胞的毒性远高于单载药体系.     

Optimal Halbach permanent magnet designs for maximally pulling and pushing nanoparticles

Optimization methods are presented to design Halbach arrays to maximize the forces applied on magnetic nanoparticles at deep tissue locations. In magnetic drug targeting, where magnets are used to focus therapeutic nanoparticles to disease locations, the sharp fall off of magnetic fields and forces with distances from magnets has limited the depth of targeting. Creating stronger forces at a depth by optimally designed Halbach arrays would allow treatment of a wider class of patients, e.g. patients with deeper tumors. The presented optimization methods are based on semi-definite quadratic programming, yield provably globally optimal Halbach designs in 2 and 3-dimensions, for maximal pull or push magnetic forces (stronger pull forces can collect nanoparticles against blood forces in deeper vessels; push forces can be used to inject particles into precise locations, e.g. into the inner ear). These Halbach designs, here tested in simulations of Maxwell's equations, significantly outperform benchmark magnets of the same size and strength. For example, a 3-dimensional 36 element 2000 cm³ volume optimal Halbach design yields a 5× greater force at a 10 cm depth compared to a uniformly magnetized magnet of the same size and strength. The designed arrays should be feasible to construct, as they have a similar strength (≤1 T), size (≤2000 cm³), and number of elements (≤36) as previously demonstrated arrays, and retain good performance for reasonable manufacturing errors (element magnetization direction errors ≤5°), thus yielding practical designs to improve magnetic drug targeting treatment depths.     

Hepatic lipid composition analysis using 3.0-T MR spectroscopy in a steatotic rat model

Purpose

To investigate the feasibility of in vivo assessment of hepatic lipid composition using 3.0-T proton magnetic resonance spectroscopy (¹H-MRS) in a steatotic rat model and compare it to histopathological and biochemical assessment.

Materials and Methods

Hepatic steatosis was induced by feeding rats with a methionine/choline-deficient (MCD) diet for 1, 2, 3, 5 or 7 weeks (n=5 per group). At the end of the diet period, ¹H-MRS of the liver was performed, and rats were sacrificed for histopathological and biochemical assessment of the liver. Spectra were acquired in a single voxel (1.2 cc) using a point-resolved spectroscopic sequence with TE/TR=35/2000 ms and 64 signal acquisitions. From the MR spectra, peak area ratios were calculated to estimate hepatic lipid composition.

Results

During MCD diet periods, hepatic steatosis significantly increased on histopathology (P<.001). The ¹H-MRS measurements of total hepatic fat content [1.3/(1.3+4.65) ppm] correlated strongly with histological macrovesicular hepatic steatosis (r=0.93, P<.001) and with the biochemical total hepatic fatty acids (r=0.94, P<.001). Total unsaturated fatty acids [TUFA, 5.4/(1.3+4.65) ppm] estimated with ¹H-MRS strongly correlated with the biochemical unsaturated fatty acids (r=0.90, P<.001). Polyunsaturated fatty acids [PUFA, 2.8/(1.3+4.65) ppm] estimated with ¹H-MRS strongly correlated with biochemical PUFA (r=0.91, P<.001). The proportion of total unsaturated fatty acids relative to the amount of total fatty acids (rTUFA, 5.4/1.3ppm) measured with ¹H-MRS strongly correlated with the biochemical amount of unsaturated relative to total hepatic fatty acids (r=0.81, P<.001). The proportion of PUFA relative to the amount of total fatty acids (rPUFA, 2.8/1.3 ppm) measured with ¹H-MRS correlated with the biochemical amount of PUFA relative to total fatty acids (r=0.59, P=0.005,) and with the biochemical amount of omega-6 PUFA relative to total fatty acids (r=0.73, P<.001).PUFA at ¹H-MRS correlated with the histopathologically assessed degree of lobular inflammation in the liver (r=0.57, P=.001).

Conclusion

3.0T ¹H-MRS is able to measure poly- and unsaturated hepatic fatty acids and this strongly correlates with biochemical assessment. This study provides evidence that 3.0-T ¹H-MRS is a noninvasive technique to assess hepatic lipid composition.     

Inclusion of interactions in mathematical modelling of implant assisted magnetic drug targeting

Drug delivery technologies are an important area within biomedicine. Targeted drug delivery aims to reduce the undesired side effects of drug usage by directing or capturing the active agents near a desired site within the body. This is particularly beneficial in, for instance, cancer chemotherapy, where the side effects of general (systemic) drug administration can be severe.One approach to targeted drug delivery uses magnetic nanoparticles as the constituents of carriers for the desired active agent. Once injected into the body, the behaviour of these magnetic carriers can be influenced and controlled by magnetic fields. In implant assisted magnetic drug targeting systems a magnetic implant, typically a stent, wire or spherical seed can be used to target sites deep within the body as the implant acts as a focus for the resulting magnetic force. This can be easily understood as the force depends on the gradient of the magnetic field and the gradient near the implant is large.In designing such a system many factors need to be considered including physical factors such as the size and nature of the implants and carriers, and the fields required. Moreover, the range of applicability of these systems in terms of the regions of the vasculature system, from low blood velocity environments, such as capillary beds to higher velocity arteries, must be considered. Furthermore, assessment criteria for these systems are needed. Mathematical modelling and simulation has a valuable role to play in informing in vitro and in vivo experiments, leading to practical system design.Specifically, the implant assisted magnetic drug targeting systems of Avilés, Ebner and Ritter are considered within this review, and two dimensional mathematical modelling is performed using the open source C++ finite volume library OpenFOAM. In the first system treated, a large ferromagnetic particle is implanted into a capillary bed as a seed to aid collection of single domain nanoparticles (radius 20-100 nm). The Langevin function is used to calculate the magnetic moment of the particles, and the model is further adapted to treat the agglomeration of particles known to occur in these systems. This agglomeration can be attributed to interparticle interactions and here the magnetic dipole-dipole and hydrodynamic interactions for two mutually interacting nanoparticles are modelled, following Mikkelsen et al. who treated two particle interactions in microfluidic systems, with low magnetic field (0.05 T). The resulting predicted performance is found to both increase and decrease significantly depending on initial positions of the particles. Secondly, a ferromagnetic, coiled wire stent is implanted in a large arterial vessel. The magnetic dipole-dipole and hydrodynamic interactions for multiple particles are included. Different initial positions are considered and the system performance is assessed. Inclusion of these interactions yields predictions that are in closer agreement with the experimental results of Avilés et al. We conclude that the discrepancies between the non interacting theoretical predictions and the corresponding experimental results can (as suggested by Avilés et al.) be largely attributed to interparticle interactions and the consequent agglomeration.     

Distinguishable Targeting of Non-Small Cell Lung Cancer Using Hyaluronan Functionalized Platinum Nanoclusters and Their Inhibition Behaviors of Proliferation,Invasion, Migration

Lung cancer is the leading cause of cancer deaths worldwide and most cancer patients receiving conventional chemotherapy suffer from severe side effects due to the non-selective effects of chemotherapeutic drugs on normal cells. Targeted nanomaterials can obtain excellent accumulation at the tumor site through their active or passive targeting mechanisms, thereby reducing the toxicity of the drugs in various ways. In this study, hyaluronic acid (HA) which could specifically bind to CD44 on the surface of tumor cells, was used to modify amine-caged platinum nanoclusters (Pt NCs-NH₂) to obtain targeting HA-Pt NCs-NH₂. Based on the differential expression of CD44 on the surface of three lung cells (non-small cell lung cancer cell H1299, small cell lung cancer cell H446, and embryonic lung fibroblast HFL1), HA-Pt NCs-NH₂ can differentially enter the three cells and achieve their targeting of non-small cell lung cancer cell (NSCLC) cells. Pt NCs significantly inhibited the proliferation, migration and invasion of NSCLC cells and induced their apoptosis in comparison of classical cisplatin and carboplatin, showing a bright future in early diagnosis and treatment of NSCLC.     

The versatility of carbohydrates in antimicrobial applications

The worsening situation of global drug resistance is urgently demanding for novel antimicrobial agents. Considerable efforts have been concentrated on developing new antibacterial therapies with new mode of actions, exerting no selective pressure on bacterial mutation, and minimizing toxicity to host cells. In this context, active targeting can greatly contribute to selectivity between pathogens and mammalian cells in which carbohydrates, playing important roles in numerous biological processes, can be employed as targeting ligands or “trojan horse.” This short account has discussed the recent results of carbohydrate-based antimicrobial agents developed by our group. Other excellent works by other scientists and possible directions in the future are also discussed.     

Targeted destruction of murine macrophage cells with bioconjugated gold nanorods

Gold nanorods manifest a readily tunable longitudinal plasmon resonance with light and consequently have potential for use in photothermal therapeutics. Recent work by others has shown how gold nanoshells and rods can be used to target cancer cells, which can then be destroyed using relatively high power laser radiation (∼1×10⁵ to 1×10¹⁰ W/m²). Here we extend this concept to demonstrate how gold nanorods can be modified to bind to target macrophage cells, and show that high intensity laser radiation is not necessary, with even 5×10² W/m² being sufficient, provided that a total fluence of ∼30 J/cm² is delivered. We used the murine cell line RAW 264.7 and the monoclonal antibody CD11b, raised against murine macrophages, as our model system and a 5 mW solid state diode laser as our energy source. Exposure of the cells labeled with gold nanorods to a laser fluence of 30 J/cm² resulted in 81% cell death compared to only 0.9% in the control, non-labeled cells.     

三明治结构rGO/Fe3O4@mSiO2载体的构筑及协同抗肿瘤性能

采用温和的反应条件,制备出三明治结构rGO/Fe_3O_4@mSiO_2,利用SEM、TEM、FTIR、XRD和N_2吸附-脱附等对其形貌和性能进行表征,考查了其对Hela细胞的毒性和细胞荧光成像效果,并探讨了其形成机理。实验结果表明:rGO/Fe_3O_4@mSiO_2具有较高的比表面积(217 m~2·g~(-1)),对抗癌药物五氟尿嘧啶(5-FU)的载药率达到57.34%;它还具有较好的磁性,磁饱和强度为32 emu·g-1;而且rGO/Fe_3O_4@mSiO_2纳米复合物在光照条件下具有优异的光热转换性能,对He La细胞表现出明显的杀伤效果。