Co-Electrospun Silk Fibroin and Gelatin Methacryloyl Sheet Seeded with Mesenchymal Stem Cells for Tendon Regeneration.

Silk fibroin (SF) is a promising biomaterial for tendon repair, but its relatively rigid mechanical properties and low cell affinity have limited its application in regenerative medicine. Meanwhile, gelatin-based polymers have advantages in cell attachment and tissue remodeling but have insufficient mechanical strength to regenerate tough tissue such as tendons. Taking these aspects into account, in this study, gelatin methacryloyl (GelMA) is combined with SF to create a mechanically strong and bioactive nanofibrous scaffold (SG). The mechanical properties of SG nanofibers can be flexibly modulated by varying the ratio of SF and GelMA. Compared to SF nanofibers, mesenchymal stem cells (MSCs) seeded on SG fibers with optimal composition (SG7) exhibit enhanced growth, proliferation, vascular endothelial growth factor production, and tenogenic gene expression behavior. Conditioned media from MSCs cultured on SG7 scaffolds can greatly promote the migration and proliferation of tenocytes. Histological analysis and tenogenesis-related immunofluorescence staining indicate SG7 scaffolds demonstrate enhanced in vivo tendon tissue regeneration compared to other groups. Therefore, rational combinations of SF and GelMA hybrid nanofibers may help to improve therapeutic outcomes and address the challenges of tissue-engineered scaffolds for tendon regeneration.

Natural melanin-loaded nanovesicles for near-infrared mediated tumor ablation by photothermal conversion.

Photothermal therapy requires a biocompatible material to absorb near-infrared (NIR) light and generate sufficient heat. Herein, we suggest natural melanin-loaded nanovesicles (melasicles) as photothermal therapeutic agents (PTA) for NIR mediated cancer therapy in vivo. The mean size of these melasicles was 140 ± 15 nm. They showed excellent colloidal stability. After irradiation from 808 nm NIR laser at 1.5 W cm-2, the melasicles showed good photothermal conversion efficiencies both in vitro and in vivo. In drug release study, laser irradiation increased fluidity of vesicle membrane due to photothermal generation from melanin. Initial drug release in the laser irradiation group was higher than that in the no laser irradiation group. After injecting the melasicles into tail veins of CT-26 bearing mice, tumors were suppressed or eliminated after irradiation at 1.5 W cm-2 for 5 min once or twice. These results suggest that melasicles could be used as attractive PTA for cancer therapy and localized drug release.

Selenium Nanoparticles Formed by Modulation of Carrageenan Enhance Osteogenic Differentiation of Mesenchymal Stem Cells.

Fabrication of nano-sized selenium (Se) particles may help to expend the applications of Se. In this study, we focused on the preparation and characterization of Se nanoparticles (Se NPs) modulated with carrageenan (CA). Furthermore, their influence on osteoblast cell growth was investigated in vitro. Spherical Se-NPs, of 100-200 nm diameter, were prepared simply by adding κ-, ι-, and λ-CA, which has sulfate groups, hydroxyl groups, and carboxyl groups. CA-modulated Se NPs (CA-Se NPs) were readily suspended in liquid medium with no precipitation over long time periods. In particular, it was found through Alizarin Red S staining that the growth of osteoblast D1 cells treated with λ-CA-Se NPs was improved significantly. These results suggest that Se NPs can be prepared simply, using CA, have good suspension stability in liquid medium, and λ-CA-Se NPs may induce the growth of osteoblast cells.

Peptide-loaded nanoparticles and radionuclide imaging for individualized treatment of myocardial ischemia.

PURPOSE: To determine whether chitosan hydrogel nanoparticles loaded with vascular endothelial growth factor (VEGF) peptides (81-91 fragments) capable of targeting the ischemic myocardium enhance angiogenesis and promote therapeutic effects and whether radionuclide image-guided dosage control is feasible. MATERIALS AND METHODS: Experimental procedures and protocols were approved by the Institutional Animal Care and Use Committee. Rats (n = 32, eight per group) were subjected to myocardial ischemia (control group) and received chitosan hydrogel nanoparticles with VEGF165 proteins (chitosan VEGF) or VEGF81-91 peptides (chitosan peptides) via apical puncture. Ischemic hearts receiving chitosan without angiogenic factors served as the chitosan control. Myocardial perfusion was examined 7 days after surgery by using technetium 99m ((99m)Tc) tetrofosmin (37 MBq) autoradiography, and changes in vascular density with immunohistochemical staining were reviewed. Kruskal-Wallis test and Bonferroni corrected Mann-Whitney U test were used for multiple comparisons. Wilcoxon signed rank test was used to compare myocardial retention of (99m)Tc chitosan. RESULTS: Thirty minutes of myocardial ischemia resulted in perfusion defects (median, 54%; interquartile range [IQR], 41%-62%). Chitosan VEGF decreased perfusion defect extent (median, 68%; IQR, 63%-73%; P = .006 vs control) and increased vascular density (median, 81 vessels per high-power field; IQR, 72-100; P = .009 vs control). Administration of chitosan peptides reduced the degree of perfusion defects (median, 66%; IQR, 62%-73%; P = .006 vs control) and increased vascular density (median, 82 vessels; IQR, 78-92; P = .006 vs control). The effects of chitosan peptides on perfusion and vascular density were comparable to those seen with chitosan VEGF proteins (P = .713 and P = .833, respectively). Chitosan radiolabeled with (99m)Tc was administered twice at reperfusion with a 1-hour interval to determine whether image-guided dosage control is feasible. The hearts initially retained 4.6% (IQR, 4.1%-5.0%) of (99m)Tc chitosan administered and 9.2% (IQR, 6.6%-12.7%; P = .068) with subsequent injection. CONCLUSION: VEGF peptides have angiogenic potential and resulted in therapeutic effectiveness. Adjunct use of single photon emission computed tomography was also demonstrated for individualized treatment of myocardial ischemia by further tailoring the therapeutic dosing. Online supplemental material is available for this article.

Characterization of a spontaneous osteopetrosis model using RANKL-dysfunctional mice.

Tumor necrosis factor superfamily member 11 (TNFSF11), or receptor activator of nuclear factor-κB ligand (RANKL), is a crucial osteoclast-stimulating factor binding to RANK on osteoclast membranes. Mouse models are powerful tools for understanding the genetic mechanisms of related diseases. Here, we examined the utility of Tnfsf11 mutation in mice for understanding the mechanisms of bone remodeling and dysmorphology. The Tnfsf11gum mouse, discovered in 2011 at Jackson Laboratory, was used to study the genetic landscape associated with TNFSF11 inactivation in bone marrow tissues. Tnfsf11gum/+ and Tnfsf11+/+ mice were subjected to Micro-CT observation, ELISA analysis, histological evaluation, and massively-parallel mRNA sequencing (RNA-Seq) analysis. Tnfsf11gum/+ mice exhibited severe osteopetrotic changes in the bone marrow cavity, along with significantly lower serum RANKL levels and a reduced number of tartrate-resistant acid phosphatase (TRAP)-positive osteoclasts in the bone marrow compared to those in Tnfsf11+/+ mice. However, tooth eruption between Tnfsf11gum/+ and Tnfsf11+/+ mice did not differ. Furthermore, genes involved in osteoblast proliferation and differentiation, including Gli1, Slc35b2, Lrrc17, and Junb were differentially expressed. Heterozygous mutation of TNFSF11 was also associated with a slightly increased expression of genes involved in osteoclast proliferation and differentiation, including Tcirg1, Junb, Anxa2, and Atp6ap1. Overall, we demonstrate that single gene mutations in Tnfsf11 cause bone resorption instability without significantly altering the genes related to osteoblast and osteoclast activity in the bone marrow cavity, thus establishing an optimal resource as an experimental animal model for bone resorption in bone biology research.

The effect of mannosylation of liposome-encapsulated indocyanine green on imaging of sentinel lymph node.

The imaging of sentinel lymph nodes (SLN) has been researched for its role in assessing cancer progression and postsurgical lymphedema. Indocyanine green (ICG) is a near-infrared (NIR) optical dye that has been approved by the Food and Drug Administration. It is known that liposome-encapsulated ICG (LP-ICG) has improved stability and fluorescence signal compared with ICG. We designed mannosylated liposome-encapsulated ICG (M-LP-ICG) as an optical contrast agent for SLN. M-LP-ICG has a higher UV absorbance spectrum and fluorescence intensity than LP-ICG. The stability of M-LP-ICG measured in 50% fetal bovine serum solution by a dialysis method was better than that of LP-ICG. M-LP-ICG demonstrated a high uptake in RAW 264.7 macrophage cell because the density of mannose is high. There were differences between M-LP-ICG and glucosylated liposome-encapsulated ICG (G-LP-ICG), which are geometrical isomers. The result of an inhibition study of M-LP-ICG showed a statistically significant decrease in uptake in RAW 264.7 cells after either co-treatment or pre-treatment with D-(+)-mannose as an inhibitor. Results from an in vitro experiment demonstrated that M-LP-ICG was specifically taken up by macrophage cells through the mannose receptor on its surface. The time-series images acquired from a normal mouse model after subcutaneous injection showed that the signal from M-LP-ICG in SLN and other organs appeared early and disappeared quickly in comparison with signals from LP-ICG. Not only the sentinel but also the draining lymph nodes were observed partly in M-LP-ICG. M-LP-ICG appears to increase the specificity of uptake and retention in macrophages, making it a good candidate contrast agent for an optic imaging system for SLN and the lymphatic system.

Optical imaging of MMP expression and cancer progression in an inflammation-induced colon cancer model.

The purpose of this study was to use a near-infrared (NIR) fluorescent cyclic His-Try-Gly-Phe peptide to characterize and image the expressions of matrix metalloproteinases (MMPs), which are correlated with cancer promotion, in an inflammation-induced colorectal cancer (ICRC) model. We explored the relationship between the development of colon cancer and the expression of MMPs at the same colonic sites in ICRC models. To develop ICRC models, mice were administered a single intraperitoneal dose (10 mg/kg) of azoxymethane (AOM) and exposed orally to 2% dextran sodium sulfate (DSS) for one week. MMP-2 expression and ß-catenin activation in colonic lesions were characterized by immunohistochemical (IHC) staining. After being treated with inducers for some time, cancerous lesions were found to express high ß-catenin and MMP-2. The profiles of MMP expression were correlated with ß-catenin activation in the colonic lesions. c(KAHWGFTLD)NH(2) (C6) peptide was prepared by standard Fmoc peptide synthesis to target MMPs. Molecular weight of Cy5.5-C6 was 1,954.78 g/mol (calculated MW = 1955.23 g/mol). The in vitro characterization of Cy5.5-C6 showed MMP binding specificity in a cell experiment. In vivo NIRF imaging showed high accumulation of Cy5.5-C6 in tumors with associated expression of MMP-2 in colonic lesions after intravenous injection. The MMP-2 specificity of Cy5.5-C6 was confirmed by successful inhibition of probe uptake in the tumor due to the presence of excess C6 peptide. The use of Cy5.5-C6 to target MMP-2 has the potential to be developed into an effective molecular imaging agent to monitor ICRC progress.

The effect of fluorination of zinc oxide nanoparticles on evaluation of their biodistribution after oral administration.

Monitoring of the behavior of metal nanoparticles in the body following exposure is very important for investigation of the physiological fates and safety of these nanoparticles. In this study, we investigated the behavior and accumulation of nano-scaled ZnO (20 nm) and submicro-scaled ZnO (100 nm) particles in organic tissues after oral administration using PET imaging. Both types of ZnO nanoparticle (20 or 100 nm) were labeled with the radionuclide (18)F in high yield via 'click reaction'. (18)F labeling on the ZnO nanoparticles was maintained stably in simulated gastric fluid (pH 1.2) for 7 h. PET images indicated that (18)F and (18)F-ethoxy azide showed radioactivity in the bone and bladder 3 h after oral administration, whereas radioactivity for (18)F-labeled ZnO nanoparticles was seen only in the gastrointestinal (GI) tract. At 5 h post-administration, biodistribution studies demonstrate that (18)F accumulated in the bone (10.19 ± 1.1%ID g(-1)) and (18)F-ethoxy azide showed radioactivity in the bone (7.55 ± 0.6%ID g(-1)), liver, and brain (0.94 ± 0.3%ID g(-1)). Unlike (18)F and (18)F-ethoxy azide, (18)F-labeled ZnO nanoparticles showed radioactivity in the lung, liver and kidney including the GI tract. Submicro-scaled (18)F-labeled ZnO nanoparticles (100 nm) showed stronger radioactivity in the liver and kidney compared to nano-scaled (18)F-labeled ZnO nanoparticles (20 nm). In conclusion, PET imaging has the potential to monitor and evaluate the behavior of ZnO nanoparticles absorbed in organic tissues following oral exposures.

Elevated anti-inflammatory effects of eicosapentaenoic acid based self-aggregated glycol chitosan nanoparticles.

The formation of nanoparticles from eicosapentaenoic acid (EPA) is crucial to improving EPA's bioavailability and pharmacological properties, and widening its use in biomedical fields. In this study, we report EPA-conjugated glycol chitosan (GC) that can self-aggregate into core-shell nanoparticles. The EPA-GC nanoparticles were internalized into the cytosol of RAW 264.7 cells by endocytosis, which results in effective delivery of EPA to the cells. There were no differences in the cell viability after the treatment with EPA-GC nanoparticles. In the anti-inflammatory studies, the EPA-GC nanoparticles significantly inhibited lipopolysaccharide (LPS)-stimulated nitric oxide (NO) production and interleukin-1 beta (IL-1 beta) secretion in RAW 264.7 cells. The anti-inflammatory effects of the EPA-GC nanoparticles were far better than those seen for EPA only. Given their excellent bio-physicochemical properties, it is expected that EPA-GC nanoparticles may have a potential for widening the use of EPA in biomedical fields and, in particular, the treatment of inflammatory diseases.

NIR-Mediated drug release and tumor theranostics using melanin-loaded liposomes.

BACKGROUND: Heat generation in a drug delivery carrier by exposure to near-infrared (NIR) light with excellent tissue transmittance is an effective strategy for drug release and tumor therapy. Because liposomes have amphiphilic properties, they are useful as drug carriers. Liposomes are also very suitable for drug delivery strategies using heat generation by NIR laser because lipid bilayers are easily broken by heat. Thermally generated bubbles from liposomes not only induce drug release, but also enable ultrasound imaging. METHODS: Melanin, perfluorohexane (PFH), and 5-fluorouracil (5-FU)-loaded liposomes (melanin@PFH@5-FU-liposomes) that can generate heat and bubble by NIR laser irradiation were prepared by a thin film method. Conversion of light to heat and bubble generation of melanin@PFH@5-FU-liposomes were evaluated using an infrared (IR) thermal imaging camera and an ultrasound imaging system both in vitro and in vivo. To investigate tumor therapeutic effect, NIR laser of 808 nm was used to irradiate tumor site for 10 min after injecting melanin@PFH@5-FU-liposome into tail veins of CT26-bearing mice. RESULTS: Melanin@PFH@5-FU-liposomes showed a spherical shape with a size of 209.6 ± 4.3 nm. Upon NIR laser irradiation, melanin@PFH@5-FU-liposomes exhibited effective temperature increase both in vitro and in vivo. In this regard, temperature increase caused a phase transition of PFH to induce bubble generation dramatically, resulting in effective drug release behavior and ultrasound imaging. The temperature of the tumor site was increased to 52 t and contrast was greatly enhanced during ultrasound imaging due to the generation of bubble. More importantly, tumor growth was effectively inhibited by injection of melanin@PFH@5-FU-liposomes with laser irradiation. CONCLUSIONS: Based on intrinsic photothermal properties of melanin and phase transition properties of PFH, melanin@PFH@5-FU-liposomes exhibited effective heat and bubble generation upon NIR laser irradiation. The elevated temperature induced bubble generation, resulting in contrast enhancement of ultrasound imaging. Melanin@PFH@5-FU-liposomes under NIR laser irradiation induced the death of cancer cells, thereby effectively inhibiting tumor growth. These results suggest that melanin@PFH@5-FU-liposomes can be utilized as a promising agent for photothermal tumor therapy and ultrasound imaging.

Chemo-photothermal therapeutic effect of chitosan-gelatin hydrogels containing methotrexate and melanin on a collagen-induced arthritis mouse model.

Heat stimulation can promote osteoblast differentiation and bone formation. Combining photothermal therapy and chemotherapy is an effective strategy for treating rheumatoid arthritis (RA). Herein, we prepared chitosan/gelatin/ß-glycerophosphate-melanin-methotrexate (CMM) hydrogel that could be used to perform simultaneous chemotherapy and photothermal therapy for patients with RA. The CMM solution was successfully converted to a gel state at body temperature. Due to intrinsic photothermal properties of melanin, CMM hydrogel exhibited effective temperature increase both in vitro and in vivo with increasing time of near-infrared (NIR) laser irradiation. After NIR laser irradiation, 50 % of methotrexate was rapidly released from the hydrogel within 3 h. Its release rate showed an instantaneous increase with additional NIR laser irradiation. After CMM hydrogel was injected directly into the paw joint of each collagen-induced arthritis (CIA) mouse followed by irradiation with a NIR laser (808 nm, 0.5 W/cm2, 3 min), swelling and redness at the inflamed area were significantly alleviated at 14 days after treatment. Micro-CT analysis confirmed that treated joints of mice were similar to normal joints. Hence, CMM hydrogel could be used as an attractive RA therapeutic agent for simultaneous chemo-photothermal therapy.

Synthesis and drug release properties of melanin added functional allopurinol incorporated starch-based biomaterials.

The main objective of this study was to prepare functional allopurinol (ALP) incorporated biomaterials using mungbean starch, polyvinyl alcohol, melanin (MEL), and plasticizers. Prepared biomaterials were characterized by FE-SEM and FT-IR analysis. Photothermal conversion efficiencies and ALP release properties of biomaterials were evaluated with NIR laser irradiation. When biomaterials were irradiated with the NIR laser, temperatures increase of MEL-added biomaterials were higher than those of MEL-non-added biomaterials. After NIR laser irradiation, ALP release rates of MEL-added biomaterials were 1.62 times faster than those of MEL-non-added biomaterials. In addition, ALP release using an artificial skin was increased by NIR laser irradiation. ALP release from biomaterials followed Fickian diffusion mechanism, while ALP release using an artificial skin followed a non-Fickian diffusion mechanism. Xanthine oxidase inhibitory (%) for MEL-added biomaterials with/without the addition of GL and XL were 47.5%, 61.7%, and 65.1%, respectively.

Biomedical Membrane of Fish Collagen/Gellan Gum Containing Bone Graft Materials.

The development of a guided bone regeneration (GBR) membrane with non-mammalian fish collagen has the advantage of low risk for transmission of infectious diseases in tissue regeneration. In this work, a fish collagen/gellan gum and bone graft material (FC/GG-BGM) composite GBR membrane were fabricated through solution blending and casting procedures in a vacuum. The membranes were characterized using Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), scanning electron microscopy observation (SEM), and atomic force microscope (AFM) analyses. FT-IR results suggested that ionic interactions were formed between FC and GG both in composite powder and membranes. In vivo experiments showed that these FC/GG-BGM composite membranes could generate osteoblast minerals and promote loose bone calcification, thus accelerating bone regeneration. At 2 weeks, the defected site of rats treated with FC/GG-BGM membrane (0.377 ± 0.012 mm3) showed higher regeneration than that of rats treated with the bovine collagen membrane (0.290 ± 0.015 mm3) and control rats without membrane (0.160 ± 0.008 mm3). Compared with bovine collagen membrane, the FC/GG-BGM composite membrane displays better bone regeneration ability. Therefore, FC/GG-BGM composite membrane is suitable as a GBR membrane for bone regeneration.

Alginate nanocapsules by water-in-oil emulsification and external gelation for drug delivery to fine dust stimulated keratinocytes.

Methodologies for synthesizing drug-loaded alginate nanocapsules were optimized and indomethacin and phloroglucinol loading capacities were studied. Their biological effects were studied for ameliorating fine dust (FD) induced detrimental effects in keratinocytes. The 1 % alginate to oil phase ratio of 1:20 was the optimal parameter for water in oil emulsification. The oil phase was optimized to contain sunflower oil: span 80 ratios of 17:3. Nanocapsule drug encapsulation efficiencies were 36.91 ± 5.56 and 32.41 ± 4.05 % respectively for phloroglucinol (EG2P) and indomethacin (EG2I) while the loading capacities were 25.28 ± 3.36 and 23.15 ± 2.84 %. Dried nanocapsules indicated a 40-140 nm diameter range while their hydrodynamic diameter was 989.69 nm at pH 7.0. Nanocapsules swelling was pH-dependent and in releasing media of pH values 4.5, 7.4, and 8.5, the drug release indicated a complex mechanism of swelling, diffusion, and erosion while at pH 2.0 the drug release followed the non-Fickian release. EG2P and EG2I treatment dose-dependently lowered FD-induced intracellular ROS production, apoptosis and inflammatory responses mediated through the NF-κB pathway in FD stimulated HaCaT keratinocytes and reduced epidermal barrier degradation. Further research could investigate the use of this technique in formulating cosmeceuticals containing drug-loaded alginate nanocapsules for achieving controlled release.

Oleyl-chitosan nanoparticles based on a dual probe for optical/MR imaging in vivo.

Oleic acid-conjugated chitosan (oleyl-chitosan) is a powerful platform for encapsulating oleic acid-decorated iron oxide nanoparticles (ION), resulting in a good magnetic resonance imaging (MRI) probe. Oleyl-chitosan could self-assemble into core-shell structures in aqueous solution and provide the effective core compartment for loading ION. ION-loaded oleyl-chitosan nanoparticles showed good enhanced MRI sensitivity in a MR scanner. Cy5.5 dye was accessed to the oleyl-chitosan conjugate for near-infrared (NIR) in vivo optical imaging. After intravenous injection of ION-loaded Cy5.5-conjugated oleyl-chitosan (ION-Cy5.5-oleyl-chitosan) nanoparticles in tumor-bearing mice, both NIRF and MR imaging showed the detectable signal intensity and enhancement in tumor tissues via enhanced permeability and retention (EPR) effect. Tumor accumulation of the nanoparticles was confirmed through ex vivo fluorescence images and Prussian blue staining images in tumor tissues. It is concluded that ION-Cy5.5-oleyl-chitosan nanoparticle is highly an effective imaging probe for detecting tumor in vivo.

In vivo tumor accumulation of nanoparticles formed by ionic interaction of glycol chitosan and fatty acid ethyl ester.

In this study, novel fatty acid ethyl ester (FAEE)-based glycol chitosan (GC) nanoparticles were prepared through an electrostatic interaction. Additionally, the tumor accumulation of the FAEE-GC nanoparticles was evaluated in order to determine the enhanced permeability and retention (EPR) effect in a breast tumor model. FAEE, including eicosapentaenoic acid ethyl ester (EPAEE) and docosahexanoic acid ethyl ester (DHAEE), successfully formed ionic complexes with GC. The FAEE-GC complexes were self-aggregated with a spherical shape and a mean diameter of 110-333 nm in aqueous media. Cy7 was labeled to the FAEE-GC complexes for the in vivo optical imaging. A tumor animal model was developed by inoculating MDA-MB231 breast tumor cells into the right flanks of mice. The Cy7-labeled FAEE-GC nanoparticles were injected into the tail vein of the tumor-bearing mice. Fluorescence signals were strongly observed in the tumor because of the EPR effect. In the ex vivo imaging of the mice, the highest fluorescence intensity, except for the kidney, was observed in the tumor. Therefore, the FAEE-GC nanoparticles could be a useful vehicle for FAEE formulation as well as an in vivo tumor-selective drug delivery carrier.

Poly (l-lactic acid) membrane crosslinked with Genipin for guided bone regeneration.

In this study, we chemically modified poly(L-lactic acid) (PLLA) with functional amine groups and fabricated a PLLA membrane crosslinked with genipin as a biomembrane for inducing guided bone regeneration (GBR). The mechanical strength of the PLLA-amine membrane was improved by crosslinking with genipin compared to pure PLLA membrane. The surface of the PLLA-amine membrane crosslinked with genipin had many more uniform pores. Attachment and proliferation of MC3T3-E1 cells were increased and improved on the PLLA-amine membrane crosslinked with genipin. In an in vitro osteogenesis study, MC3T3-E1 cells on the PLLA membrane showed higher alkaline phosphatase (ALP) activity and calcification ability evaluated by alizarin red S staining than those on the pure PLLA membrane. When a skull defect hole of a rat was covered with the PLLA-amine membrane crosslinked with genipin, vigorous new bone regeneration determined by computed tomography at 8 weeks post operation was superior to that when the skull defect was covered with the pure PLLA membrane. Taken together, these results demonstrate that the PLLA-amine membrane crosslinked with genipin has a promising therapeutic application to GBR as a barrier membrane for covering the defect site.

Prostate cancer-targeted imaging using magnetofluorescent polymeric nanoparticles functionalized with bombesin.

PURPOSE: In this work, the aim was to prepare and characterize a magnetofluorescent polymeric nanoparticle for prostate cancer imaging in vivo. METHODS: Glycol chitosan (GC) was chemically modified with N-acetyl histidine (NAHis) as a hydrophobic moiety, and bombesin (BBN) was conjugated to the hydrophobically modified GC for use in targeting gastric-releasing peptide receptors (GRPR) overexpressed in prostate cancer cells. NAHis-GC conjugates were labeled with the near-infrared (NIR) fluorophore Cy5.5 (C-NAHis-GC conjugate). RESULTS: BBN-conjugated C-NAHis-GC nanoparticles (BC-NAHis-GC nanoparticles) showed significantly higher binding to the PC3 cell surface than nanoparticles without BBN, and the cellular binding was clearly inhibited by BBN. The tumor-to-muscle ratios of C- and BC-NAHis-GC nanoparticles were 2.26 +/- 0.66 and 5.37 +/- 0.43, respectively. The tumor accumulation of BC-NAHis-GC nanoparticles was clearly reduced by co-injection of BBN. Further, iron oxide nanoparticles (IO) were loaded into BC-NAHis-GC nanoparticles to investigate the possibility of use as a probe for MRI. IO-BC-NAHis-GC nanoparticles were well observed in the PC3 cells, and the blocking with BBN significantly reduced the cellular binding of the nanoparticles. CONCLUSION: These results demonstrate that the BBN conjugation to NAHis-GC nanoparticles improves their tumor accumulation in PC3-bearing mice in comparison to nanoparticles without BBN, suggesting that BC-NAHis-GC nanoparticles may be useful for prostate cancer imaging.

Synthesis of Tc-99m labeled 1,2,3-triazole-4-yl c-met binding peptide as a potential c-met receptor kinase positive tumor imaging agent.

The mesenchymal-epithelial transition factor (c-Met), which is related to tumor cell growth, angiogenesis and metastases, is known to be overexpressed in several tumor types. In this study, we synthesized technetium-99m labeled 1,2,3-triazole-4-yl c-Met binding peptide (cMBP) derivatives, prepared by solid phase peptide synthesis and the 'click-to-chelate' protocol for the introduction of tricarbonyl technetium-99m, as a potential c-Met receptor kinase positive tumor imaging agent, and evaluated their in vitro c-Met binding affinity, cellular uptake, and stability. The (99m)Tc labeled cMBP derivatives ([(99m)Tc(CO)(3)]12, [(99m)Tc(CO)(3)]13, and [(99m)Tc(CO)(3)]14) were prepared in 85-90% radiochemical yields. The cold surrogate cMBP derivatives, [Re(CO)(3)]12, [Re(CO)(3)]13, and [Re(CO)(3)]14, were shown to have high binding affinities (0.13 microM, 0.06 microM, and 0.16 microM, respectively) to a purified cMet/Fc chimeric recombinant protein. In addition, the in vitro cellular uptake and inhibition studies demonstrated the high specific binding of these (99m)Tc labeled cMBP derivatives ([(99m)Tc(CO)(3)]12-14) to c-Met receptor positive U87MG cells.

Surface engineering of quantum dots for in vivo imaging.

The aim of this study was to investigate the effect of gluconic acid (GA) conjugation on the biodistribution of cysteamine-capped quantum dots (amino-QDots) in vivo. Cadmium selenide/zinc sulfide (CdSe/ZnS) was capped with cysteamine through a thiol exchange method, and different amounts of GA were conjugated to the amine groups of cysteamine via the formation of an amide bond. The emission maxima of the synthesized QDots, the amino-QDots and the GA-conjugated amine-QDots (GA-QDots) were located at 720, 600 and 610 nm, respectively. In the cell viability studies, the GA-QDots showed very low toxicity against CHO cells as compared to the cytotoxicity of the amino-QDots. The QDots were next intravenously injected into normal mice and then we performed ex vivo optical imaging. The majority of the amino-QDots were accumulated in the lung. In contrast, the GA-QDots were cleared out of the body through the kidney. Therefore, we expect that the conjugation of GA onto the amino-QDots can create opportunities for using amino-QDots for in vivo imaging.