We sought to explore the treatment effects and the repair mechanisms of bone marrow derived mesenchymal stem cells (MSCs) during HCl-induced acute lung injury (ALI). MSCs were delivered through the tail veins of rats 24 h after intranasal instillation of HCl. The results showed that MSCs did not ameliorate the histopathologic changes of ALI and pulmonary fibrosis. We found that the activated Wnt/?-catenin signaling may regulate the differentiation of MSCs and is associated with lung fibroblasts activation, pulmonary fibrosis and tissue repair process in ALI rats. Immunofluorescence and histology analysis indicated that activated canonical Wnt/?-catenin signaling induced most MSCs to differentiate into myofibroblasts or fibroblasts in vivo. However, inhibition of Wnt/?-catenin signaling by Dickkopf-1 (DKK1) promotes epithelial differentiation of MSCs induced by native alveolar epithelial cells which are beneficial to repair the injured lung epithelium. Inhibition of Wnt/?-catenin signaling after MSCs transplantation ameliorated pulmonary fibrosis and improved pulmonary function which attenuated the lung injury. In vitro study, activation of the Wnt/?-catenin signaling stimulated MSCs to express myofibroblasts markers, which was attenuated by DKK1. Furthermore, Wnt3? activated Wnt/?-catenin signaling in lung fibroblasts to enhance the expression of collagen I, vimentin and ?-smooth muscle actin, but DKK1 attenuated these proteins expression. These findings demonstrated that canonical Wnt/?-catenin signaling plays a key role in regulating differentiation of MSCs in vivo or in vitro and the pathogenesis of fibrotic diseases. Our study suggested that inhibition of abnormal activated Wnt/?-catenin signaling would promote MSCs epithelial differentiation to repair lung injury and reduce pulmonary fibrosis.
Although gold nanorods (GNRs) have been investigated extensively for optical hyperthermia therapies, the synthesis of rods is far from ideal. In this report, we optimized the synthesis of gold nanorods using hydroquinone as a reducing agent. Compared with the GNRs prepared by traditional ways, the as-synthesized rods have a flexibly tunable size and wider range of longitudinal surface plasmon resonance (LSPR). Furthermore, a series of small-length gold nanorods with length ranging from 30 to 90 nm were synthesized and they are more suitable for in vivo biomedical applications. Finally, we exploited a convenient approach for preparing water-soluble GNRs with less toxicity, better dispersion and flexible functionalization by exchanging hexadecyltrimethylammonium bromide (CTAB) on the surface of the rods with carboxylated bovine serum albumin (BSA) derivative, the BSA modified GNRs showed significant anticancer efficacy through near infrared (NIR) hyperthermia. We believe that the as-prepared gold nanorods will find promising applications in biomedical fields, especially in cancer therapy.
Idiopathic pulmonary fibrosis is a progressive lung disorder of unknown etiology. Previous studies have shown that aberrant activation of the Wnt/?-catenin signaling cascade occurs in lungs of patients with idiopathic pulmonary fibrosis. Given the important roles of the Wnt/?-catenin signaling pathway in the development of pulmonary fibrosis, we targeted this pathway for the intervention of pulmonary fibrosis with XAV939, a small molecule that specifically inhibits Tankyrase 1/2, eventually leading to the degradation of ?-catenin and suppression of the Wnt/?-catenin signaling pathway. Our results demonstrated that XAV939 significantly inhibited the activation of Wnt/?-catenin signaling and attenuated bleomycin-induced lung fibrosis in mice, and thus improved the survival of mice with lung injury. Interestingly, previous investigations have confirmed that endogenous and exogenous mesenchymal stem cells could be recruited to the injured lung, although the exact effects of these cells are debatable. To determine the effect of Wnt/?-catenin signaling in the epithelial differentiation of bone marrow-derived mesenchymal stem cells (BM-MSCs), we established a coculture system that contains BM-MSCs and alveolar type II epithelial cells. The in vitro experiments demonstrated that XAV939 could promote the differentiation of BM-MSCs into an epithelium-like phenotype in the coculture system. We also found that XAV939 could inhibit the proliferation and myofibroblast differentiation of NIH/3T3 fibroblasts. This work supports that inhibition of the Wnt/?-catenin signaling pathway may be exploited for the treatment of idiopathic pulmonary fibrosis for which effective treatment strategies are still lacking.
The estrogenic chemical nonylphenol (NP) and the antiandrogenic agent di-n-butyl phthalate (DBP) are regarded as widespread environmental endocrine disruptors (EEDs) which at high doses in some species of laboratory animals have adverse effects on male reproduction and development. Given the ubiquitous coexistence of various classes of EEDs in the environment, their combined effects warrant investigation. In this study, we attempted to clarify the interactions of NP and DBP on tight junctions (TJs) between rat Sertoli cells. In the in vitro experiment, monobutyl phthalate (MBP), the active metabolite of DBP, was used instead of DBP. Sertoli cells were isolated from Sprague-Dawley rats, and treated with NP and MBP, singly or combined. The morphology of Sertoli cells, and structure and functionality of TJs were measured. In the in vivo experiment, rats were gavaged on postnatal day 23-35 with a single or combined NP and DBP treatment. Testicular weight and morphology of TJs were recorded. These data indicated that NP and DBP/MBP, either in single or in combination, induced the structural and function changes of Sertoli cell tight junctions, both in vivo and in vitro. The combined effect on the regulation of TJ proteins at both the protein and gene levels was correlated to the effect exerted by NP, suggesting that the structure and function of Sertoli cells were more sensitive to exposure to NP than MBP.
Controversies and risks continue to be reported about exogenous mesenchymal stem cell-based therapies. In contrast with employing exogenous stem cells, making use of lung resident mesenchymal stem cells (LR-MSCs) could be advantageous. Our study sought to isolate the LR-MSCs and explore their potential to differentiate into alveolar epithelial type II cells (ATII cells). Total lung cells were first precultured, from which the Sca-1(+) CD45(-) CD31(-) population was purified using fluorescence activated cell sorting (FACS). By these methods, it would seem that the Sca-1(+) CD45(-) CD31(-) cells were LR-MSCs. Similar to bone marrow derived mesenchymal stem cells (BM-MSCs), these cells express Sca-1, CD29, CD90, CD44 and CD106, but not CD31 or CD45. They share the same gene expression file with the BM-MSCs and have a similar DNA content during long-term culturing. Furthermore, they could be serially passaged with all these properties being sustained. Above all, LR-MSCs could differentiate into ATII cells when co-cultured with ATII cells in a trans-well system. These findings demonstrated that the Sca-1(+) CD45(-) CD31(-) cells appear to be LR-MSCs that can differentiate into ATII cells. This approach may hold promise for their use in the treatment of lung disease.
In this article, for the first time, a novel, high-yield and template-free method for the synthesis of Ag nanoparticle decorated thionine/infinite coordination polymer (AgNP/THI/ICP) fibres is proposed. The thionine can be adsorbed to the AgNP/THI/ICP fibres by ?-conjugation and act as the redox probe. The AgNP/THI/ICP fibres not only favor the immobilization of antibody but also facilitate the electron transfer. It is found that the AgNP/THI/ICP fibres can be designed to act as a sensitive label-free electrochemical immunosensor for carcinoembryonic antigen (CEA) determination. Under the optimized conditions, the linear range of the proposed immunosensor is estimated to be from 50 fg/mL to 100 ng/mL and the detection limit is estimated to be 0.5 fg/mL at a signal-to-noise ratio of 3, respectively. The prepared immunosensor for detection of CEA shows high sensitivity, reproducibility and stability. Our study demonstrates that the proposed immunosensor has also been used to determine CEA successfully in diluted blood samples.
Acute lung injury may lead to fibrogenesis. However, no treatment is currently available. This study was conducted to determine the effects of bone marrow-derived mesenchymal stem cells (MSCs) in a model of HCl-induced acute lung injury in Sprague-Dawley (SD) rats. Stromal cell-derived factor (SDF)-1 and its receptor CXC chemokine receptor (CXCR)4 have been shown to participate in mobilizing MSCs. Adenovirus carrying the CXCR4 gene was used to transfect MSCs in order to increase the engraftment numbers of MSCs at injured sites. Histological examination data demonstrated that the engraftment of MSCs did not attenuate lung injury and pulmonary fibrosis. The results showed that engraftment of MSCs almost differentiated into myofibroblasts, but rarely differentiated into lung epithelial cells. Additionally, it was demonstrated that activated canonical Wnt/?-catenin signaling in injured lung tissue regulated the myofibroblast differentiation of MSCs in vivo. The in vitro study results demonstrated that activation of the Wnt/?-catenin signaling stimulated MSCs to express myofibroblast markers; however, this process was attenuated by Wnt antagonist DKK1. Therefore, the results demonstrated that the aberrant activation of Wnt signaling induces the myofibroblast differentiation of engrafted MSCs, thus contributing to pulmonary fibrosis following lung injury.
The estrogenic chemical nonylphenol (NP) and the antiandrogenic agent di-n-butyl phthalate (DBP) are regarded as widespread environmental endocrine disruptors (EDCs) which at high doses in some species of laboratory animals, such as mice and rats, have adverse effects on male reproduction and development. Given the ubiquitous coexistence of various classes of EDCs in the environment, their combined effects warrant clarification. In this study, we attempted to determine the mixture effects of NP and DBP on the testicular Sertoli cells and reproductive endocrine hormones in serum in male rats based on quantitative data analysis by a mathematical model. In the in vitro experiment, monobutyl phthalate (MBP), the active metabolite of DBP, was used instead of DBP. Sertoli cells were isolated from 9-day-old Sprague-Dawley rats followed by treatment with NP and MBP, singly or combined. Cell viability, apoptosis, necrosis, membrane integrity and inhibin-B concentration were tested. In the in vivo experiment, rats were gavaged on postnatal days 23-35 with a single or combined NP and DBP treatment. Serum reproductive hormone levels were recorded. Next, Bliss Independence model was employed to analyze the quantitative data obtained from the in vitro and in vivo investigation. Antagonism was identified as the mixture effects of NP and DBP (MBP). In this study, we demonstrate the potential of Bliss Independence model for the prediction of interactions between estrogenic and antiandrogenic agents.
A surfactant-free and template-free method for the high-yield synthesis of biomolecule (serotonin)-based formaldehyde resin (BFR) microspheres is proposed for the first time. The colloidal microspheres loaded with Au nanoparticles (AuNPs) prepared by a convenient in-situ synthesis of AuNPs on BFR (AuNPs/BFR) microsphere surface show good stability. AuNPs/BFR microspheres not only favor the immobilization of antibody but also facilitate the electron transfer. It is found that the resultant AuNPs/BFR microspheres can be designed to act as a sensitive label-free electrochemical immunosensor for carcinoembryonic antigen (CEA) determination. The immunosensor is prepared by immobilizing capture anti-CEA on AuNPs/BFR microspheres assembled on thionine (TH) modified glassy carbon electrode (GCE). TH acts as the redox probe. Under the optimized conditions, the linear range of the proposed immunosensor is estimated to be from 25pg/mL to 2000pg/mL (R=0.998) and the detection limit is estimated to be 3.5pg/mL at a signal-to-noise ratio of 3. The prepared immunosensor for detection of CEA shows high sensitivity, reproducibility and stability. Our study demonstrates that the immunosensor can be used for the CEA detection in humans serum.
The silver-modified gold nanoplate arrays as bimetallic surface-enhanced Raman scattering (SERS) substrates were optimized for the surface-enhanced Raman detection of streptavidin/biotin monolayer assemblies. The bimetallic gold-silver nanoplate arrays were fabricated by coating silver nanoparticles uniformly on the gold nanoplate arrays. Depending on silver nanoparticle coating, the localized surface plasmon resonance (LSPR) peak of the bimetallic gold-silver nanoplate arrays blue-shifted and broadened significantly. The common probe molecule, Niel Blue A sulfate (NBA) was used for testing the SERS activity of the bimetallic gold-silver nanoplate arrays. The SERS intensity increased with the silver nanoparticle coating, due to a large number of hot spots and nanoparticle interfaces. The platforms were tested against a monolayer of streptavidin functionalized over the bimetallic gold-silver nanoplate arrays showing that good quality spectra could be acquired with a short acquisition time. The supramolecular interaction between streptavidin (strep) and biotin showed subsequent modification of Raman spectra that implied a change of the secondary structure of the host biomolecule. And the detection concentration for biotin by this method was as low as 1.0nM. The enhanced SERS performance of such bimetallic gold-silver nanoplate arrays could spur further interest in the integration of highly sensitive biosensors for rapid, nondestructive, and quantitative bioanalysis, particularly in microfluidics.
Despite advances in cancer diagnosis and treatment, ovarian cancer remains one of the most fatal cancer types. The development of targeted nanoparticle imaging probes and therapeutics offers promising approaches for early detection and effective treatment of ovarian cancer. In this study, HER-2 targeted magnetic iron oxide nanoparticles (IONPs) are developed by conjugating a high affinity and small size HER-2 affibody that is labeled with a unique near infrared dye (NIR-830) to the nanoparticles. Using a clinically relevant orthotopic human ovarian tumor xenograft model, it is shown that HER-2 targeted IONPs are selectively delivered into both primary and disseminated ovarian tumors, enabling non-invasive optical and MR imaging of the tumors as small as 1 mm in the peritoneal cavity. It is determined that HER-2 targeted delivery of the IONPs is essential for specific and sensitive imaging of the HER-2 positive tumor since we are unable to detect the imaging signal in the tumors following systemic delivery of non-targeted IONPs into the mice bearing HER-2 positive SKOV3 tumors. Furthermore, imaging signals and the IONPs are not detected in HER-2 low expressing OVCAR3 tumors after systemic delivery of HER-2 targeted-IONPs. Since HER-2 is expressed in a high percentage of ovarian cancers, the HER-2 targeted dual imaging modality IONPs have potential for the development of novel targeted imaging and therapeutic nanoparticles for ovarian cancer detection, targeted drug delivery, and image-guided therapy and surgery.
Surface properties, as well as inherent physicochemical properties, of the engineered nanomaterials play important roles in their interactions with the biological systems, which eventually affect their efficiency in diagnostic and therapeutic applications. Here we report a new class of MRI contrast agent based on milk casein protein-coated iron oxide nanoparticles (CNIOs) with a core size of 15 nm and hydrodynamic diameter ~30 nm. These CNIOs exhibited excellent water-solubility, colloidal stability, and biocompatibility. Importantly, CNIOs exhibited prominent T2 enhancing capability with a transverse relaxivity r2 of 273 mM(-1) s(-1) at 3 tesla. The transverse relaxivity is ~2.5-fold higher than that of iron oxide nanoparticles with the same core but an amphiphilic polymer coating. CNIOs showed pH-responsive properties, formed loose and soluble aggregates near the pI (pH ~4.0). The aggregates could be dissociated reversibly when the solution pH was adjusted away from the pI. The transverse relaxation property and MRI contrast enhancing effect of CNIOs remained unchanged in the pH range of 2.0-8.0. Further functionalization of CNIOs can be achieved via surface modification of the protein coating. Bioaffinitive ligands, such as a single chain fragment from the antibody of epidermal growth factor receptor (ScFvEGFR), could be readily conjugated onto the protein coating, enabling specific targeting to MDA-MB-231 breast cancer cells overexpressing EGFR. T2-weighted MRI of mice intravenously administered with CNIOs demonstrated strong contrast enhancement in the liver and spleen. These favorable properties suggest CNIOs as a class of biomarker targeted magnetic nanoparticles for MRI contrast enhancement and related biomedical applications.
Although microcystin-LR (MC-LR) produced by cyanobacteria has been demonstrated with strong reproductive toxicity, the mechanisms remain unclear. This study aimed to probe the effects of MC-LR on induction of autophagy in Sertoli cells, as well as the relationship between autophagy and apoptosis. After exposure to various concentrations of MC-LR for 24 or 48 h, cell viability and membrane integrity were significantly decreased under high MC-LR conditions (50-500 nM). The autophagosome marker protein LC3 was increased at mild MC-LR concentrations (0.5-5 nM). However, autophagosomes accumulated to their peak level under high MC-LR conditions in parallel with significantly up-regulated apoptosis. Treatment with an autophagy inhibitor (3-MA) abrogated autophagosome accumulation and apoptosis. This study demonstrated that MC-LR had toxic effects on Sertoli cells by inducing autophagy and apoptosis. The autophagosome accumulation may be involved in the apoptosis induced by MC-LR.
Development of sensitive and specific imaging approaches for the detection of ovarian cancer holds great promise for improving survival of ovarian cancer patients. Here we describe a dual-modality photoacoustic and fluorescence molecular tomography (PAT/FMT) approach in combination with a targeted imaging probe for three-dimensional imaging of ovarian tumors in mice. We found that the selective accumulation of the HER-2/neu targeted magnetic iron oxide nanoparticles (IONPs) led to about 5-fold contrast enhancements in the tumor for PAT, while near-infrared (NIR) dye labeled nanoparticles emitted strong optical signals for FMT. Both PAT and FMT were demonstrated to be able to detect ovarian tumors located deep in the peritoneal cavity in mice. The targeted nanoprobes allowed mapping tumors in high resolution via PAT, and high sensitivity and specificity via FMT. This study demonstrated the potential of the application of HER-2/neu-targeted PAT/FMT approach for non-invasive or intraoperative imaging of ovarian cancer.
Molecular therapy using a small interfering RNA (siRNA) has shown promise in the development of novel therapeutics. Various formulations have been used for in vivo delivery of siRNAs. However, the stability of short double-stranded RNA molecules in the blood and efficiency of siRNA delivery into target organs or tissues following systemic administration have been the major issues that limit applications of siRNA in human patients. In this study, multifunctional siRNA delivery nanoparticles are developed that combine imaging capability of nanoparticles with urokinase plasminogen activator receptor-targeted delivery of siRNA expressing DNA nanocassettes. This theranostic nanoparticle platform consists of a nanoparticle conjugated with targeting ligands and double-stranded DNA nanocassettes containing a U6 promoter and a shRNA gene for in vivo siRNA expression. Targeted delivery and gene silencing efficiency of firefly luciferase siRNA nanogenerators are demonstrated in tumor cells and in animal tumor models. Delivery of survivin siRNA expressing nanocassettes into tumor cells induces apoptotic cell death and sensitizes cells to chemotherapy drugs. The ability of expression of siRNAs from multiple nanocassettes conjugated to a single nanoparticle following receptor-mediated internalization should enhance the therapeutic effect of the siRNA-mediated cancer therapy.
Antifouling magnetic iron oxide nanoparticles (IONPs) coated with block copolymer poly(ethylene oxide)-block-poly(?-methacryloxypropyltrimethoxysilane) (PEO-b-P?MPS) were investigated for improving cell targeting by reducing nonspecific uptake. Conjugation of a HER2 antibody, Herceptin®, or a single chain fragment (ScFv) of antibody against epidermal growth factor receptor (ScFvEGFR) to PEO-b-P?MPS-coated IONPs resulted in HER2-targeted or EGFR-targeted IONPs (anti-HER2-IONPs or ScFvEGFR-IONPs). The anti-HER2-IONPs bound specifically to SK-BR-3, a HER2-overexpressing breast cancer cell line, but not to MDA-MB-231, a HER2-underexpressing cell line. On the other hand, the ScFvEGFR-IONPs showed strong reactivity with MDA-MB-231, an EGFR-positive human breast cancer cell line, but not with MDA-MB-453, an EGFR-negative human breast cancer cell line. Transmission electron microscopy revealed internalization of the receptor-targeted nanoparticles by the targeted cancer cells. In addition, both antibody-conjugated and non-antibody-conjugated IONPs showed reduced nonspecific uptake by RAW264.7 mouse macrophages in vitro. The developed IONPs showed a long blood circulation time (serum half-life 11.6 hours) in mice and low accumulation in both the liver and spleen. At 24 hours after systemic administration of ScFvEGFR-IONPs into mice bearing EGFR-positive breast cancer 4T1 mouse mammary tumors, magnetic resonance imaging revealed signal reduction in the tumor as a result of the accumulation of the targeted IONPs.
In this study, we present a new method to fabricate large-area two-dimensionally (2D) ordered gold nanobowl arrays based on 3D colloidal crystals by wet chemosynthesis, which combines the advantages of a very simple preparation and an applicability to "real" nanomaterials. By combination of in situ growth of gold nanoshell (GNSs) arrays based on three-dimensional (3D) colloidal silica crystals, a monolayer ordered reversed GNS array (2D ordered GNS array) was conveniently manufactured by an acrylic ester modified biaxial oriented polypropylene (BOPP). 2D ordered gold nanobowl array with adjustable periodic holes, good stability, reproducibility, and repeatability could be obtained when the silica core was etched by HF solution. The surface-enhanced Raman scattering (SERS) enhancement factor (EF) of this 2D ordered gold nanobowl array could reach 1.27 × 10(7), which shows high SERS enhancing activity and can be used as a universal SERS substrate.
LIV-1, a zinc transporter, is an effector molecule downstream from soluble growth factors. This protein has been shown to promote epithelial-to-mesenchymal transition (EMT) in human pancreatic, breast, and prostate cancer cells. Despite the implication of LIV-1 in cancer growth and metastasis, there has been no study to determine the role of LIV-1 in prostate cancer progression. Moreover, there was no clear delineation of the molecular mechanism underlying LIV-1 function in cancer cells. In the present communication, we found increased LIV-1 expression in benign, PIN, primary and bone metastatic human prostate cancer. We characterized the mechanism by which LIV-1 drives human prostate cancer EMT in an androgen-refractory prostate cancer cells (ARCaP) prostate cancer bone metastasis model. LIV-1, when overexpressed in ARCaP(E) (derivative cells of ARCaP with epithelial phenotype) cells, promoted EMT irreversibly. LIV-1 overexpressed ARCaP(E) cells had elevated levels of HB-EGF and matrix metalloproteinase (MMP) 2 and MMP 9 proteolytic enzyme activities, without affecting intracellular zinc concentration. The activation of MMPs resulted in the shedding of heparin binding-epidermal growth factor (HB-EGF) from ARCaP(E) cells that elicited constitutive epidermal growth factor receptor (EGFR) phosphorylation and its downstream extracellular signal regulated kinase (ERK) signaling. These results suggest that LIV-1 is involved in prostate cancer progression as an intracellular target of growth factor receptor signaling which promoted EMT and cancer metastasis. LIV-1 could be an attractive therapeutic target for the eradication of pre-existing human prostate cancer and bone and soft tissue metastases.
A SERS active gold nanostar layer on the surface of ITO glass slip has been prepared by a low-cost electrostatically assisted APTES-functionalized surface-assembly method for SERS analysis. The two-dimensional morphology of the SERS substrate was examined by scanning electron microscopy. Comparative analysis revealed that the optical characteristics and SERS efficiency of these substrates varied as a function of nanostar morphology. It was found that the substrate assembled with the longest branches of nanostars generated the best SERS efficiency, whether the excitation source is 785 or 633 nm. The potential use of these substrates in detection applications was also investigated by using Nile blue A and Rhodamine 6G. The detection limits are 5 × 10(-11) M and 1 × 10(-9) M, respectively, when using the 785 nm excitation source. Apart from this high enhancement effect, the substrate here also shows extremely good reproducibility at the same time. All of these indicate that gold nanostars are a very good structure for SERS substrate assembly.
A novel method based on surface-enhanced Raman scattering (SERS) was developed to estimate the antioxidant activity of antioxidants by using self-assembled three-dimensionally (3D) ordered gold nanoparticles (GNPs) precursor composite (SiO(2)/GNPs) arrays as nanoprobes. H(2)O(2) could reduce AuCl(4)(-) to Au(0) which deposited onto the surface of the SiO(2)/GNPs arrays and enlarged the GNPs. As the concentration of H(2)O(2) increased, the surface coverage of the resultant gold on the silica cores increased accordingly until continuous gold nanoshells (GNSs) were formed. The change of the intensities of the SERS spectra correlated well with H(2)O(2) concentration which indicated that this SiO(2)/GNPs array was a potential SERS nanoprobe for H(2)O(2). The presence of antioxidant will prevent the growth of GNPs on the surface of the silica arrays from forming the structure which has strongest SERS-activity and the corresponding change in SERS intensity correlated well with the H(2)O(2) scavenging activity of the antioxidants. The H(2)O(2) scavenging activities of four plant-based antioxidants, tannic acid, citric acid, ferulic acid, and tartaric acid were studied. Our results showed the H(2)O(2) scavenging activities (SA(HP) values) of these four compounds were: tannic acid > ferulic acid > citric acid > tartaric acid.
Novel core-shell structures were presented here which consist of Poly(N-isopropyl acrylamide) (PNIPAM) microspheres as cores and gold as shells. The fabrication of these structures was convenient because the modifications to PNIPAM or gold nanoparticles (GNPs) were omitted. GNPs were attached to the surface of PNIPAM microspheres by means of electrostatic adsorption, and then acted as seeds to grow quickly into complete shells by optimizing the pH of the HAuCl4 solution to control the growth rate of gold nanoshells (GNSs). These structures combine the thermo-responsive behavior of PNIPAM microspheres with optical property of GNSs, and the localized surface plasmon resonance (LSPR) of the GNSs can be changed by adjusting the temperature of the PNIPAM microspheres which make them have great prospects for drug release. Their good biological stability in bovine serum albumin (BSA) and the LSPR located near 700 nm are expected for optical biosensors and optical analysis of whole-blood.
In the present work, the gold nanoshells (GNSs) precursor composites were preadsorbed onto the surface of ITO substrates. With the treatment of modified electrodes immersed in the gold nanoparticles (GNPs) growth solution containing different phenolic acids, the GNSs precursor composites were enlarged to varying degrees. Phenolic acids with one or more phenolic hydroxyl groups served as reductants for the growth of GNPs. The enlargement conditions varied with the different reducing capacity of phenolic acids, exhibiting specific morphologies differ from the complete GNSs. Consequently, the UV-vis-NIR spectra and cyclic voltammetry curves for the phenolic acid-treated ITO electrode were gradually changed. Results showed that the higher reducing capacity for phenolic acid to reduce AuCl(4)(-) to Au(0) resulted in the intensified localized surface plasmon resonance features and reduced cathodic currents. The spectral wavelength peaks red shifted hundreds of nanometers across the visible region. Moreover, the antioxidant capacity of phenolic acids correlates well with their reducing activity, both of which reflect their tendency to donate electrons. Thus, the optical and electrochemical results could be used to evaluate the antioxidant capacity of phenolic acids by utilizing GNSs precursor composites as nanoprobes. The method is simple, rapid and could be used in visual analysis to a certain extent.
Near-IR fluorescence imaging has great potential for noninvasive in vivo imaging of tumors. In this study, we show the preferential uptake and retention of two hepatamethine cyanine dyes, IR-783 and MHI-148, in tumor cells and tissues.
Prostate stromal cells may play binary roles in the process of prostate cancer development. As the first to be encountered by infiltrating prostate cancer cells, prostate stromal cells form the first defense line against prostate cancer progression and metastasis. However, interaction between prostate cancer and stromal cells may facilitate the formation of a tumor microenvironment favoring cancer cell growth and survival. To establish an experimental system for studying the interaction between cancer and stromal cells, we isolated three matched pairs of normal and cancer-associated human prostate stromal clones. In this report, we describe the morphologic and behavioral characteristics of these cells and their effect on LNCaP prostate cancer cells in co-culture. Unlike LNCaP prostate cancer cells, the isolated prostate stromal clones are large fibroblast-like cells with a slow proliferation rate. Growth and survival of these clones are not affected by androgens. The stromal cells display high resistance to serum starvation, while cancer-associated stromal clones have differentiated survival ability. In co-culture experiments, the stromal cells protected some LNCaP prostate cancer cells from death by serum starvation, and cancer-associated stromal clones showed more protection. This work thus established a panel of valuable human prostate stromal cell lines, which could be used in co-culture to study the interaction between prostate cancer and prostate stromal cells.
In the present work, we have developed a novel nanocomposite-based method for the detection of L-DOPA and tyrosinase (TR) activity. This was accomplished by growth-sensitive gold nanoshells (GNSs) precursor nanocomposites (SiO(2)/GNPs II) and the formation process of GNSs. L-DOPA can reduce AuCl(4)(-) to Au(0), depositing on the surface of SiO(2)/GNPs II and mediating the enlargement of gold nanoparticles (GNPs). Here, the preadsorbed GNPs on SiO(2)/GNPs II serve as nucleation sites for Au(0) deposition. As the concentration of L-DOPA increases, the surface coverage of resultant gold on silica cores increases accordingly until continuous GNSs are formed. In this growth procedure, the spectra changes in wavelength correlate well with the concentration of L-DOPA, which indicates that this nanocomposite is a good nanoprobe for detecting L-DOPA. Because TR can catalyze the hydroxylation of L-tyrosine to form L-DOPA, this approach can also be employed to analyze the activity of TR, which possesses vast clinical and food industrial importance.
Chitosan has natural abundance, unique bioactivity and attractive physicochemical properties. Recent years, the synthesis of chitosan-based metal nanomaterials has attracted increasing attention. The synthesis of metal nanoparticles utilizing biomolecular or organism offers a mild medium, and thus a greater degree of control over the nanoparticles produced, along with higher reproducibility. In particular, preparation of metal nanoparticles based on biomolecular or organism has its unique facility in integrating "minimum feature sizes" into labile biological components to an excellent synergy and bifunctional effect and consequently a more broad application. Herein, we review the new development of chitosan, chitosan-based synthesis of metal nanomaterials, and their application.
In the present work, we have developed a novel nanocomposite-based method for estimating antioxidant activity. The assay implements a new enzyme-free optical nanoprobe for assessing hydrogen peroxide (H(2)O(2)) scavenging activity based on the formation process of gold nanoshells (GNSs). H(2)O(2) could enlarge the gold nanoparticles (GNPs) on the surface of GNSs precursor nanocomposites (SiO(2)/GNPs), and the preadsorbed GNPs served as nucleation sites for Au deposition. As the concentration of H(2)O(2) increases, more GNPs on the SiO(2) cores are enlarged until continuous GNSs are formed. During the growth procedure, the spectra changes correlate well with H(2)O(2) concentrations which indicate that this nanocomposite is a good nanoprobe for detecting H(2)O(2). H(2)O(2) scavenging activities of several antioxidants were determined by restraining the H(2)O(2)-mediated formation of GNSs from SiO(2)/GNPs, and the changes of the corresponding plasmon absorption bands correlated well with H(2)O(2) scavenging activity of antioxidants. The spectra were monitored by a UV-vis-near-infrared (NIR) spectrophotometer, and the wavelength changes were adopted as detection signal. The results obtained expressed the difference of H(2)O(2) scavenging activity between various tested compounds, and the relationship between function and structure of antioxidants was also discussed in this article. The new method based on the formation process of GNSs is simple, rapid, and sensitive and, additionally, can be used in visual analysis to a certain extent for antioxidant functional evaluation.
Metal nanoparticle-chitosan (NPs-chitosan) bioconjugates were formed by exposure of chitosan to an aqueous solution of metal salts under thermal treatment. The metal nanoparticles that are formed strongly bound to chitosan, which encouraged us to investigate their catalytic performance. It was demonstrated that the metal NPs-chitosan bioconjugates functioned as effective catalysts for the reduction of 4-nitrophenol in the presence of NaBH(4), which was monitored by means of spectrophotometry as a function of reaction time. The silver NPs-chitosan bioconjugates exhibited excellent catalytic activity and were reusable for up to seven cycles. In contrast, the gold NPs-chitosan catalyst displayed poor catalytic activity, even in the second cycle. A highlight of our approach is that chitosan simultaneously acts as an active support for the synthesis and assembly of nanoparticles, and the resultant bioconjugates bear the advantage of easy separation from the reaction medium.
Reflectometry interference spectroscopy (RIfS) is known as a highly sensitive and robust technique for direct, label-free detection of the interaction of biomacromolecules in real time and in situ. The aim of the present study was to investigate the competitive protein adsorption on the surface of fluorocarbon end-capped poly(carbonate) urethane (PCUF) and polystyrene (PS) based on the RIfS method. The surface energy and microstructures of PCUF and PS were characterized by contact angle measurement and atomic force microscopy. Interfacial energies between these surfaces and the proteins were then calculated. The protein adsorption experiments were carried out with both single solution and ternary solutions composed of albumin, fibrinogen and immunoglobulin-G (IgG). The results of surface characterization showed that PCUF was more hydrophilic than PS with a smaller surface energy, and micro-phases separation of PCUF was observed. RIfS analysis results revealed that more albumins, less fibrinogen and IgG were detected on the PCUF surface compared with PS after simplex and competitive protein adsorption, which indicated that PCUF had a preferential adsorption for albumin. The special morphology, smaller surface energy and calculated interfacial energies between PCUF and proteins may be responsible for the better blood compatibility of PCUF compared to PS. The results suggest that RIfS could serve as a novel, effective method for studying the competitive protein adsorption on biomaterial surfaces.
Chitosan-based silver nanoparticles were synthesized by reducing silver nitrate salts with nontoxic and biodegradable chitosan. The silver nanoparticles thus obtained showed highly potent antibacterial activity toward both Gram-positive and Gram-negative bacteria, comparable with the highly active precursor silver salts. Silver-impregnated chitosan films were formed from the starting materials composed of silver nitrate and chitosan via thermal treatment. Compared with pure chitosan films, chitosan films with silver showed both fast and long-lasting antibacterial effectiveness against Escherichia coli. The silver antibacterial materials prepared in our present system are promising candidates for a wide range of biomedical and general applications.
We present here a facile route to in-situsynthesis of free-standing metal (gold or silver) nanoparticles-embedded chitosan films by thermal treatment. The produced nanoparticles were confirmed by UV-vis spectroscopy and transmission electron microscopy (TEM). Interestingly, an exquisite dendritic structure was observed in the resulting silver-chitosan film, which was not present in pure chitosan and gold-chitosan samples. We speculated that the formation of dendritic structures may be due to the presence of nano-sized silver particles which has an influence on the crystallization behavior and thereby the morphology of biopolyer chitosan. The application of the as-prepared metal-chitosan films in surface-enhanced Raman spectroscopy (SERS) was investigated by using Rhodamine 6G (R6G) as probe molecules. It was found that the resultant metal-chitosan samples, especially silver-chitosan one, could be used as SERS substrates exhibiting excellent enhancement ability.
A simple biosensor for hydrogen peroxide (H(2)O(2)) and glucose was fabricated by incorporating gold nanoparticles (GNPs) onto a cuttlebone-derived matrix substrate (CDMS). Such a three-dimensional chamber-like structure naturally bears abundant amino groups for the direct immobilization of GNPs without a series of modifications. And preferably, the framework endows CDMS with a very high surface area for the attachment of GNPs, resulting in effective optical signal transduction and improved sensitivity of the detection system. The principle behind this biosensor is that the localized surface plasmon resonance (LSPR) of the immobilized GNPs changes with the enlargement of GNPs by H(2)O(2)-mediated chemical reduction of chloroauric acid. Using this approach, we demonstrate the proof of an optical biosensor to quantify the concentration of H(2)O(2) as well as glucose. UV-vis absorption spectra were recorded to obtain quantitative information about the H(2)O(2) or glucose concentration. The detection range of our biosensor to H(2)O(2) concentration was from 2 x 10(-6) to 1.5 x 10(-4)M, while the linear response range of glucose concentration was from 5 x 10(-6) to 5 x 10(-5)M. Inspiringly and interestingly, the growth of GNPs on CDMS gives rise to color changes, this phenomenon shows that the rapid detection by our sensor has the superiority in visual detection to a certian extent, which has been a potential application in qualitative or semiquantitative analysis for medicine and biotechnology.
Biologically derived materials provide a rich variety of approaches toward new functional materials because of their fascinating structures and environment-friendly features, which is currently a topic of research interest. In this paper, we show that the cuttlebone-derived organic matrix (CDOM) is an excellent scaffold for the one-step synthesis and assembly of silver nanoparticles (AgNPs), which can be further used as substrate for surface-enhanced Raman scattering (SERS). Formation of AgNPs-CDOM composite was accomplished by the reaction of CDOM with AgNO(3) and NH(3).H(2)O solution at 80 degrees C without using any other stabilizer and reducing agents. UV-vis spectra and TEM were utilized to characterize the AgNPs and investigate their formation process. Results demonstrate that the size and distribution of AgNPs can be partly regulated by changing incubation time; the concentration of NH(3).H(2)O is critical to the formation rate of AgNPs. As a proof of principle, we show that the AgNPs-CDOM composite can be employed in trace analysis using SERS.
To visualize human prostate cancer cells in mouse bone with bioconjugated near-infrared quantum dot (QD) probes. Near-infrared fluorescent probes using QDs can visualize tumors in deep tissues in vivo.
We introduce a simple but robust label-free method to detect DNA based on large-scale gold nanoplate (GNP) films with tunable localized surface plasmon resonance (LSPR) and highly surface-enhanced Raman scattering (SERS) activity. The common probe molecule, Neil Blue A sulfate (NBA) is used for testing the SERS activity of the GNP films at very low concentrations. It is found that the SERS properties are highly dependent on the edge lengths of gold nanoplate and gold nanoplate density in the films. Multiple-layer GNP films which are constructed by gold nanoplate with an edge length of 134±6nm have the density of 916±40GNPsGNPs/spot. It shows the highest signal intensity with SERS enhancement factor (EF) as high as 5.4×10(7) and also has excellent stability, reproducibility and repeatability. The optimized SERS-active substrate with the largest enhancement ability could be used to detect double-strand DNA without a dye label, and the detection limit is down to 10(-6)mg/mL.
In recent years, more and more controversies and risks have been reported about the exogenous mesenchymal stem cells-based therapies. In contrast with employing exogenous stem cell, making use of lung resident mesenchymal stem cells (LR-MSCs) should be advantageous. Therefore, the purpose of our study is to isolate the LR-MSCs and explore their potential to differentiate into alveolar epithelial type II cells (ATII cells). In this work, the total lung cells were first precultured, then the Sca-1+CD45-CD31- population was purified using fluorescence activated cell sorting (FACS). With this methods, we suggested that the Sca-1+CD45-CD31- cells were LR-MSCs. Similar to bone marrow derived mesenchymal stem cells (BM-MSCs), these cells expressed Sca-1, CD29, CD90, CD44 and CD106, but not CD31 or CD45; shared the same gene expression file with the BM-MSCs; had a similar DNA content to the BM-MSCs during a long-term culturing; and could be serially passaged with all the properties maintained. Above all, the LR-MSCs could differentiate into ATII cells when co-cultured with ATII cells in the trans-well system. Conclusively, these findings demonstrated that the Sca-1+CD45-CD31- cells appeared to be LR-MSCs and could differentiate into ATII cells. This approach may hold promise in using LR-MSC for treatment in lung disease.
This study focuses on understanding the growth and control of the gold nanoplates by seed-mediated growth approach. These monodispersive size-controlled gold nanoplates have the average thickness of 8-10 nm and average size tunable from 70 to 150 nm, exhibiting strong surface plasmon absorption in the near infrared (NIR) region. For the gold nanoplates formation, the seeds serve as nucleation sites, ascorbic acid (AA) serves as a new reductant to reduce hydrogen tetrachloroaurate (HAuCl4), surface activity system cetyltrimethylammonium bromide (CTAB) and potassium iodide (KI) are critical factors. X-ray diffraction (XRD) and selected-area electron diffraction (SAED) analyses reveal that gold nanoplates with the (111) lattice plane as the basal plane are single crystals. CTAB are absorbed on the surface of the (111) lattice plane of the single crystals, accounting for self-assembled monolayer and head-to-head arrays. The two arrays have been shown to serve as effective surface-enhanced Raman scattering substrates using Niel blue A (NBA) sulfate as Raman report molecule.
In the present work, the enzymatically controlled growth process of gold nanoshells (GNSs) in the presence of O2/glucose/glucose oxidase (GOx) and its chloroaurate ion electron acceptor is described. The biocatalytically stimulated growth process is one of the bio-inspired synthetic procedures directed by biological molecules which occur under ambient conditions. It is found that hydrogen peroxide (H2O2) could enlarge the gold nanoparticles (GNPs) on the surface of GNSs precursor composites, of which the preadsorbed GNPs serve as nucleation sites for further gold deposition. Here, GOx is harnessed for its unparalled level of catalytic activity and substrate specificity while H2O2 is produced as a by-product during the oxidation of D-glucose to gluconic acid by GOx. Then the bio-generated H2O2 is used as the reducing agent in the catalytic deposition process of GNSs formation. During the procedure, the localized surface plasmon resonance peaks range across hundreds of nanometers from visible to near infrared region accompanying by the resultant formation of uniform and continuous core-shell nanostructures. The corresponding optical, morphological and enzyme kinetic properties are all well investigated. The novel protocol offers a new perspective for the bio-directed synthesis method in nanotechnology.
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