Enhanced effect of autologous EVs delivering paclitaxel in pancreatic cancer.

We engineered human pancreatic cancer cell (PANC-1)-derived extracellular vesicles (EVs) by conjugating the functional ligand RGD and magnetic nanoparticles (MNPs) onto EV surfaces (rmExo), for pancreatic cancer therapy. Paclitaxel (PTX) loaded into rmExo (rmExo-PTX) was intravenously injected into xenograft mice prepared using PANC-1 cells, which showed a significant reduction in tumor size compared to the free PTX-treated and control groups. The enhanced therapeutic effect was attributed to the modification of the surface of EVs using RGD, which has affinity for αvß3 that is highly expressed in pancreatic cancer cells. Moreover, autologous EVs seemed to have more benefits in delivering PTX due to an unknown homing property to parent tumor cells, as exemplified by the reduced therapeutic effect of RGD-modified PANC-1 EVs on HT29 xenograft mice and RGD-modified U937 EVs on PANC-1 xenograft mice. The RGD-modified autologous EV vehicles were effective at penetrating and internalizing tumor cells, and eventually regressing the tumors, by mediating spontaneous removal of α-smooth muscle actin and collagen type 1 in the extracellular matrix of xenografts. Our results also identified an important molecule involved in the home-driving properties of PANC-1 EVs, integrin ß3, which was expressed both on PANC-1 cells and the EVs derived from them. Additional therapeutic effect by permanent magnet near tumor xenograft was not observed in this study.

Targeted removal of leukemia cells from the circulating system by whole-body magnetic hyperthermia in mice.

Until now, magnetic hyperthermia was used to remove solid tumors by targeting magnetic nanoparticles (MNPs) to tumor sites. In this study, leukemia cells in the bloodstream were directly removed by whole-body hyperthermia, using leukemia cell-specific MNPs. An epithelial cellular adhesion molecule (EpCAM) antibody was immobilized on the surface of MNPs (EpCAM-MNPs) to introduce the specificity of MNPs to leukemia cells. The viability of THP1 cells (human monocytic leukemia cells) was decreased to 40.8% of that in control samples by hyperthermia using EpCAM-MNPs. In AKR mice, an animal model of lymphoblastic leukemia, the number of leukemia cells was measured following the intravenous injection of EpCAM-MNPs and subsequent whole-body hyperthermia treatment. The result showed that the leukemia cell number was also decreased to 43.8% of that without the treatment of hyperthermia, determined by Leishman staining of leukemia cells. To support the results, simulation analysis of heat transfer from MNPs to leukemia cells was performed using COMSOL Multiphysics simulation software. The surface temperature of leukemia cells adhered to EpCAM-MNPs was predicted to be increased to 82 °C, whereas the temperature of free cells without adhered MNPs was predicted to be 38 °C. Taken together, leukemia cells were selectively removed by magnetic hyperthermia from the bloodstream, because EpCAM-modified magnetic particles were specifically attached to leukemia cell surfaces. This approach has the potential to remove metastatic cancer cells, and pathogenic bacteria and viruses floating in the bloodstream.

Immunochromatographic assay to detect α-tubulin in urine for the diagnosis of kidney injury.

BACKGROUNDS: Shortening of primary cilia in kidney epithelial cells is associated with kidney injury and involved with the induced level of α-tubulin in urine. Therefore, rapid detection and quantification of α-tubulin in the urine samples could be used to the preliminary diagnosis of kidney injury. METHODS: Cellulose-based nanobeads modified with α-tubulin were used for the detection probe of competitive immunochromatographic (IC) assay. The concentration of α-tubulin in the urine samples was determined by IC assay and compared with the amount determined by Western blotting analysis. RESULTS: The relationship between α-tubulin concentration and the colorimetric intensity resulted from IC assay was determined by logistic regression, and the correlation coefficient (R2 ) was 0.9948. When compared to the amount determined by Western blotting analysis, there was a linear relationship between the α-tubulin concentrations measured by the two methods and the R2 value was 0.823. CONCLUSIONS: This method is simple, rapid, and adequately sensitive to detect α-tubulin in patient urine samples, which could be used for the clinical diagnosis of kidney injury.

Multimodal endoscopic system based on multispectral and photometric stereo imaging and analysis.

We propose a multimodal endoscopic system based on white light (WL), multispectral (MS), and photometric stereo (PS) imaging for the examination of colorectal cancer (CRC). Recently, the enhancement of the diagnostic accuracy of CRC colonoscopy has been reported; however, tumor diagnosis for a variety of lesion types remains challenging using current endoscopy. In this study, we demonstrate that our developed system can simultaneously discriminate tumor distributions and provide three-dimensional (3D) morphological information about the colon surface using the WL, MS, and PS imaging modalities. The results demonstrate that the proposed system has considerable potential for CRC diagnosis.

Stepwise inhibition of T cell recruitment at post-capillary venules by orally active desulfated heparins in inflammatory arthritis.

Identification of the structure-function relationship of heparin, particularly between 2-O-, 6-O-, and N-sulfation and its anticoagulant or anti-inflammatory activities, is critical in order to evaluate the biological effects of heparin, especially in conjunction with modifications for oral formulation. In this study, we demonstrated that removal of 2-O, 6-O, or N-desulfation and their hydrophobic modifications have differential effects on the blocking of interactions between sLeX and P-and L-selectins, with highest inhibition by 6-O desulfation, which was consistent with their in vivo therapeutic efficacies on CIA mice. The 6-O desulfation of lower molecular weight heparin (LMWH) retained the ability of LMWH to interfere with T cell adhesion via selectin-sLeX interactions. Furthermore, 6DSHbD coated on the apical surface of inflamed endothelium directly blocked the adhesive interactions of circulating T cells, which was confirmed in vivo by suppressing T cell adhesion at post-capillary venular endothelium. Thus, in series with our previous study demonstrating inhibition of transendothelial migration, oral delivery of low anticoagulant LMWH to venular endothelium of inflamed joint tissues ameliorated arthritis by the stepwise inhibition of T cell recruitment and provides a rationale for the development of modified oral heparins as innovative agents for the treatment of chronic inflammatory arthritis.

Endogenous inspired biomineral-installed hyaluronan nanoparticles as pH-responsive carrier of methotrexate for rheumatoid arthritis.

Methotrexate (MTX), an anchor drug for rheumatoid arthritis (RA), has been suffered from refractoriness and high toxicity limiting effective dosage. To mitigate these challenges, the ability to selectively deliver MTX to arthritis tissue is a much sought-after modality for the treatment of RA. In this study, we prepared mineralized nanoparticles (MP-HANPs), composed of PEGylated hyaluronic acid (P-HA) as the hydrophilic shell, 5ß-cholanic acid as the hydrophobic core, and calcium phosphate (CaP) as the pH-responsive mineral. Owing to the presence of CaP as the diffusion barrier, mineralized HANPs revealed the pH-responsiveness of release kinetics of MTX across neutral to acidic conditions. HANPs were internalized via receptor-mediated endocytosis in macrophages which involved molecular redundancy among major hyaladherins, including CD44, stabilin-2, and RHAMM. Following endocytosis, MP-HANPs loaded with doxorubicin revealed pH-dependent demineralization followed by dramatic acceleration of drug release into the cytosol compared to other HANPs. Furthermore, an in vivo study showed a significantly high paw-to-liver ratio of fluorescent intensity after systemic administration of MP-HANP-Cy5.5, indicating improved biodistribution of nanoparticles into arthritic paws in collagen-induced arthritis mice. Treatment with MTX-loaded MP-HANPs ameliorated inflammatory arthritis with remarkable safety at high dose of MTX. We highlight the distinct advantages of combining key benefits of biomineralization and PEGylation with HA-based nanoparticles for arthritis-selective targeting, thus suggesting MP-HANPs as a promising carrier of MTX for treatment of RA.

Volume-normalized transfer constant as an imaging biomarker for chronic inflammatory arthritis: A dynamic contrast enhanced MRI study.

PURPOSE: Proper diagnosis and treatment of rheumatoid arthritis are extremely important to optimize treatment outcomes. Dynamic contrast-enhanced MRI (DCE-MRI) may be used as a biomarker to detect inflammatory changes in synovial joints and to discriminate active and inactive stages of the disease. The aim of this study was to investigate vascular permeability changes associated with inflammatory arthritis progression and its treatment. METHODS: Arthritis was induced in DBA/1J mice by immunization with type-II collagen emulsified in complete Freund's adjuvant. Severity of arthritis was monitored using the clinical arthritis index. MR images of mice were obtained at different stages of arthritis progression and at 3 weeks after methotrexate treatment. Immunohistochemical staining using an anti-CD31 antibody was used to assess vessel density. RESULTS: Volume-normalized transfer constant increased progressively until the active stage of arthritis was reached, and thereafter declined gradually. The pattern of volume-normalized transfer constant changes quantified using DCE-MRI correlated with vascular densities and immunohistochemical findings. Furthermore, volume-normalized transfer constant and densities decreased significantly in a dose-dependent manner after treatment with methotrexate. CONCLUSION: Volume-normalized transfer constant assessed by DCE-MRI can be used as an imaging biomarker for tracking disease progression and for monitoring therapeutic efficacy in inflammatory arthritis. Magn Reson Med 76:926-934, 2016. © 2016 Wiley Periodicals, Inc.