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The elevated concentration of low-density lipoprotein (LDL) is recognized as a leading factor of hyperlipidemia (HLP), and selective adsorption of serum LDL is regarded as a practical therapy. Based on the superior structure-function characteristics of stimuli-responsive materials, a photorenewable nanoadsorbent (SiO

Highly efficient removal of bilirubin from whole blood directly by hemoperfusion for liver failure therapy remains a challenge in the clinical field due to the low adsorption capacity, poor mechanical strength and low biocompatibility of adsorbents. In this work, a new class of nanocomposite adsorbents was constructed through an inorganic-organic co-crosslinked nanocomposite network between vinyltriethoxysilane (VTES)-functionalized hydroxyapatite nanoparticles (V-Hap) and non-ionic styrene-divinylbenzene (PS-DVB) resins (PS-DVB/V-Hap) using suspension polymerization. Notably, our adsorbent demonstrated substantially improved mechanical performance compared to the pure polymer, with the hardness and modulus increasing by nearly 3 and 2.5 times, respectively. Moreover, due to the development of a mesoporous structure, the prepared PS-DVB/V-Hap3 exhibited an ideal adsorption capacity of 40.27 mg g−1. More importantly, the obtained adsorbent beads showed outstanding blood compatibility and biocompatibility. Furthermore, in vivo extracorporeal hemoperfusion verified the efficacy and biosafety of the adsorbent for directly removing bilirubin from whole blood in pig models, and this material could potentially prevent liver damage and improve clinical outcomes. Taken together, the results suggest that PS-DVB/V-Hap3 beads can be used in commercial adsorption columns to threat hyperbilirubinemia patients through hemoperfusion, thus replacing the existing techniques where plasma separation is initially required.

Based on the LDL structural characteristics and intermolecular interactions, a tailored nano-adsorbent (Fe3O4@SiO2@PAA-PE) was prepared aimed at the removal of LDL from hyperlipemia serum with high selectivity.

Immunomagnetic nanoparticles (IMNs) have been widely developed as a detection tool to isolate rare circulating tumor cells (CTCs) from whole blood as a potential method for early cancer diagnosis, metastasis examination, and treatment guidance. However, a spontaneous interaction between nanoparticles and proteins results in the formation of a protein corona that reduces the performance of IMNs when they enter body fluids. To address this issue, the protein corona was precoated onto magnetic nanoparticles (C-MNs), and then their surfaces were conjugated with an immuno-antibody. The adsorption of proteins on C-MNs was decreased 6-fold and non-specific cell binding was reduced 5-fold, compared with magnetic nanoparticles (MNs). Furthermore, the immuno-antibody functionalized C-MNs (IC-MNs) maintained highly specific CTC capture performance when exposed to blood plasma. By using artificial spiked blood samples, IC-MNs exhibited 90.2% CTC isolation efficiency, compared with 60.3% by using IMNs. IC-MNs also successfully captured CTCs with high purity in 24 out of 26 female breast cancer patient blood samples. This work demonstrated that a novel preformed protein corona strategy can provide a useful clinically applicable diagnostic tool.

Lowering of low-density lipoprotein (LDL) levels in blood of patients with hyperlipidaemia can effectively prevent the progression of atherosclerosis and coronary heart disease. The present study demonstrated a facile synthesis strategy to prepare biomembrane-mimetic LDL adsorbent (PVA@COOH-PE) via directional immobilization of phospholipid onto macro-porous cross-linked poly(vinyl alcohol) spheres. The binding between the prepared adsorbent and LDL particles simulates the cytosolic lipid droplets to form a lipid-packing structure. The adsorbent possesses satisfactory removal efficiency for LDL and total cholesterol (TCH) in hyperlipemia serum, while remains high-density lipoprotein (HDL) concentration within the normal range. The adsorption capacities for LDL and TCH are about 1.13 and 1.74 mg/ml respectively, which are nearly three and four times higher than that of HDL (0.42 mg/ml). The adsorbent also possesses satisfactory anticoagulant properties, causes negligible effect on blood cells and produces low hemolysis ratios. The excellent blood compatibility plus LDL removal efficiency of PVA@COOH-PE indicates its good application prospect as hemoperfusion adsorbent in the treatment of hyperlipidaemia.

Elevated levels of low-density lipoproteins (LDL) are recognized as a crucial indicator of hyperlipidemia (HLP) and lowering of LDL levels represents an effective clinical treatment strategy. Inspired by the conjugation of phospholipid monolayers and the lipid content of the LDL particle, the current study describes the preparation of an innovative hemoperfusion adsorbent. The adsorbent was prepared by attachment of phosphatidyl ethanolamine to poly(acrylic acid) modified poly(vinyl alcohol-co-triallyl isocyanurate) beads (PVA@PAA-PE). The interaction between LDL and adsorbent mimics the lipoprotein microemulsion present in the blood and thus promotes efficient binding with high affinity. In vitro adsorption using serum from patients with HLP revealed that the LDL adsorption of PVA@PAA-PE was 4.44 times higher than that of controls and the removal rate of LDL using PVA@PAA-PE was about twice as high as that of the anti-atherogenic high-density lipoprotein (HDL). In vivo whole blood perfusion demonstrated the superior affinity of PVA@PAA-PE for LDL since LDL concentration was significantly reduced from 10.71 ± 2.36 mmol L-1 to 6.21 ± 1.45 mmol L-1, while the HDL level was not severely reduced (from 0.98 ± 0.12 mmol L-1 to 0.56 ± 0.15 mmol L-1). Additionally, PVA@PAA-PE exhibited excellent hemocompatibility and low cytotoxicity. Therefore, PVA@PAA-PE is a potential adsorbent for whole blood perfusion to treat hyperlipidemia.

Porous nanocomposite materials possess enhanced adsorption properties, but uniformly dispersing nanofillers in polymer matrices is still a challenging technical difficulty. In this study, modification through loading with SiO2 successfully improved the dispersibility of graphene in chitosan suspensions. By employing a phase-inversion method, cross-linked chitosan/SiO2-loaded graphene (CS/graphene-SiO2) composite beads were prepared. The resulting CS/graphene-SiO2 beads were characterized by scanning electron microscopy and Raman spectroscopy, which clearly showed that graphene was well-dispersed in the chitosan matrix. Adsorption results demonstrated that CS/graphene-SiO2 beads possessed much better adsorption capacity for bilirubin (77.87%) compared to pure chitosan (CS) beads (22.47%), mainly due to the synergistic effect between the hydrophobic forces of graphene and the electrostatic interactions provided by the amino groups of CS. The adsorption of bilirubin was found to match a monolayer model, as well as a pseudo-first-order kinetic model. The mechanical strength of composite beads was significantly improved as compared to CS beads, due to the incorporation of graphene. The effects on hemolytic activity and the components of blood were negligible, which indicate an excellent compatibility of the obtained composite beads with blood. Overall, the proposed CS/graphene-SiO2 beads as an efficient adsorbent for bilirubin have high potential in blood purification applications.

A novel nano-CaCO3 (nCaCO3) particle composite-derived polystyrene (PS) resin was successfully synthesized by a suspension polymerization method. The nCaCO3 reinforced PS material (PS/nCaCO3) possessed a structure with abundant mesopores of high porosity, high specific surface area (828.3 m2 g-1) and large pore volume (1.83 cm3 g-1). It was revealed that the incorporation of nCaCO3 into the PS matrix enhanced both the mechanical strength which can prevent the fragmentation and its adsorption capacity for interleukin-6 (IL-6, MW = 24.0 kDa) from human plasma. The adsorption isotherm could be described by the Langmuir model and classified as S-3 type, showing an IL-6 uptake of up to 25.6 ng g-1 at an equilibrium concentration of about 500 ng L-1. The adsorption capacity for IL-6 of PS/nCaCO3 is not only significantly higher than that of PS (without nCaCO3), but also superior to those of currently available adsorbents that are under clinical studies (e.g., CytoSorb™ towards cytokines). In addition, the PS/nCaCO3 adsorbent also had good hemocompatibility and showed no leakage of nCaCO3 in the plasma in a flowing model system. Therefore, the synthesized PS/nCaCO3 nano-composite has a great potential to be used as an efficient adsorbent for the removal of interleukin-6 (IL-6) from blood of inflammatory and auto-immune disease patients through hemoperfusion.

The cytokine network of tumour microenvironment (TME) plays an important role in cancer growth and progression. The current work aims to provide a new strategy for cancer therapy based on the targeted regulation of cytokines in the TME. Here, heparin-coupled polyvinyl alcohol (PVA-H) microspheres have been developed as an adsorbent for selectively remove tumour-induced immunosuppressive cytokines, such as vascular endothelial growth factor (VEGF) and transforming growth factor-beta (TGF-β), but not tumour necrosis factor-alpha (TNF-α) which has an immune-stimulating effect and can inhibit tumour growth. The proliferation and apoptosis of breast cancer cells after perfusion were tested by cell viability assays, flow cytometry analysis and mRNA microarray assays. Results showed that the PVA-H microspheres efficiently absorbed the majority of VEGF (74.39%) and TGF-β (86.39%), but much less TNF-α (4.16%). The regulation of the cytokines had remarkable anti-proliferative and pro-apoptotic effects on breast cancer cells, which was further confirmed from the change of mRNA expression levels. Thus, targeting regulatory pathways within the TME by an affinity adsorbent that selectively depletes immunosuppressive cytokines is potentially a new and promising strategy for cancer therapy.

Tumor necrosis factor (TNF)-α has an important role in the pathogenesis of autoimmune and inflammatory diseases such as rheumatoid and septic arthritis. Removal of excess tumor necrosis factor-α (TNF-α) is a promising treatment. In this study, a series of functionalized carbon nanotube-embedded poly(vinyl alcohol) (PVA) nanocomposite adsorbents were prepared for TNF-α removal for the first time. The resulting nanocomposites were characterized by scanning electron microscopy and Raman spectroscopy, which demonstrated that carbon nanotubes were well-dispersed on the surface of PVA macroporous microspheres. Adsorption tests showed that the carboxylated carbon nanotube-embedded composite microspheres (PVA/MWCNTs-COOH) possessed much better adsorption capacity for TNF-α in both simulated serum solution and rat plasma compared to the aminated (PVA/MWCNTs-NH2) and raw carbon nanotube-embedded microspheres (PVA/MWCNTs-raw). In addition, the effects on hemolytic activity, the anticoagulant property, and the components of blood were negligible, indicating the excellent blood compatibility of composite beads. Our findings suggest that the carboxylated carbon nanotube-embedded composite microspheres may be potentially useful for the treatment of autoimmune and inflammatory diseases by removing TNF-α from the blood.

Tumor necrosis factor alpha (TNF-α) is a pro-inflammatory cytokine active in the bodily immune response and serious inflammatory diseases. Traditional ligands targeting TNF-α focus on antibodies and receptors, which always associate with low efficacy and specificity. In the present study, two peptide ligands (T1: Ac-RKEM-NH2 and T2: Ac-RHCLS-NH2) were designed by computer simulation technology considering the weak interactions between TNF-α and its receptor TNFR1. Calculations of binding free energy (BFE) were made by the Molecular Mechanics Poisson-Boltzmann Surface Area (MM-PBSA) method between T1 or T2 and TNF-α (-22.68 and -14.23 kcal mol-1, respectively). To assess the affinity levels, short peptide ligands were fixed on polyvinyl alcohol (PVA) microspheres; adsorption tests showed a stronger affinity of both PVA-T1 and PVA-T2 to TNF-α in PBS buffer than PVA microspheres (79.20 ± 1.32 and 74.27 ± 1.10 vs. 39.03 ± 1.25 pg mg-1, respectively). Moreover, PVA-T1 (74.8%, 17.60 ± 2.98 pg mg-1) and PVA-T2 (63.2%, 15.30 ± 4.81 pg mg-1) exhibit significantly enhanced TNF-α adsorption from the plasma of rats with sepsis to blank PVA and commercial XAD-7 resin. In conclusion, our results show that T1 designed by computer-aided molecular design (CAMD) exhibits a stronger affinity to TNF-α and it can significantly enhance PVA microsphere adsorption efficiency of TNF-α in plasma.

A novel nano-CaCO

Restoration of sufficient blood supply for the treatment of ischemia remains a significant scientific and clinical challenge. Here, a cell-like nanoparticle delivery technology is introduced that is capable of recapitulating multiple cell functions for the spatiotemporal triggering of vascular regeneration. Specifically, a copper-containing protein is successfully prepared using a recombinant protein scaffold based on a de novo design strategy, which facilitates the timely release of nitric oxide and improved accumulation of particles within ischemic tissues. Through closely mimicking physiological cues, the authors demonstrate the benefits of bioactive factors secreted from hypoxic stem cells on promoting angiogenesis. Following this cell-mimicking manner, artificial hybrid nanosized cells (Hynocell) are constructed by integrating the hypoxic stem cell secretome into nanoparticles with surface coatings of cell membranes fused with copper-containing protein. The Hynocell, hybridized with different cell-derived components, provides synergistic effects on targeting ischemic tissues and promoting vascular regeneration in acute hindlimb ischemia and acute myocardial infarction models. This study offers new insights into the utilization of nanotechnology to potentiate the development of cell-free therapeutics.

A novel nano-CaCO3/polystyrene nanocomposite adsorbent (NPS-8) was synthesized for efficient bilirubin removal from human plasma. A comparison with the polystyrene adsorbent (PS-8), which was without the incorporation of nano-CaCO3, revealed that NPS-8 had superior bilirubin adsorption capacity and mechanical strength. The resulting nano-CaCO3 reinforced PS-8 (NPS-8) was tested by transmission electron microscopy (TEM), scanning electron microscopy (SEM), mechanical strength test, and bilirubin adsorption assays. The adsorption results indicated that NPS-8 displayed better adsorption capacity for bilirubin (91%) than that of PS-8 (75.88%). The mechanical strength of NPS-8 was significantly greater than that of PS-8. In addition, both PS-8 and NPS-8 possessed good blood compatibility properties (a negligible hemolytic activity and platelet adhesion). Therefore, a conclusion could be drawn that NPS-8 has a high potential as an efficient bilirubin adsorbent for blood purification in clinical practice. At the same time, the success of organic-inorganic nanocomposite adsorbents might provide a new insight into the improvement of adsorbents in hemoperfusion.

PS-DVB/nano-CaCO3, a novel abundant mesoporous structured polymer nano-composite, was successfully synthesized via suspension polymerization. Characterization of this type of bead nano-composite demonstrated that it exhibits significantly enhanced TNF-α adsorption from blood plasma and possesses good mechanical strength.

A multi-functional polystyrene based adsorbent (NKU-9) with a unique mesoporous and a high surface area was prepared by suspension polymerization for removal of therapeutic toxins in blood purification. The adsorbent produced had an almost equal amount of mesopore distribution in the range from 2 to 50 nm. The adsorption of serum toxins with different molecular weights were examined by in vitro adsorption assays and compared with some clinical currently used adsorbents such as HA-330, Cytosorb and BL-300 which are produced by China, America and Japan, respectively. Test results indicated that the adsorption rate for pentobarbital by NKU-9 was 81.24% which is nearly as high as HA-330 (81.44%). The latter adsorbent is currently used for acute detoxification treatment in China. To reach adsorption equilibrium, NKU-9 was faster than HA-330, which implies short treatment time. For the removal of middle molecular toxins such as β2-microglobulin (98.88%), NKU-9 performed better adsorptive selectivity than Cytosorb (92.80%). In addition, NKU-9 showed high performance for the removal of albumin-bound toxins (e.g., bilirubin), and its adsorption rate for total bilirubin (80.79%) in plasma was 8.4% higher than that of anion exchange resin BL-300 which is currently used to eliminate bilirubin in clinic. Therefore, our results indicate that the newly developed adsorbent with a wide distribution and almost equal amount of mesopores is a multifunctional adsorbent for high efficient removal of serum toxins with different molecular weights which might be an excellent blood purification adsorbent especially to treat diseases that conventional medical methods are low or not efficient.

Chitosan–carbon nanotube composite beads combines the advantages of chitosan in forming a stable biocompatible framework and carbon nanotube that provide nanometer effects (high strength and high specific surface area etc.). In this study, chitosan/amino multiwalled carbon nanotubes (CS/AMWCNT) composite beads was prepared by phase-inversion method, in which CS and AMWCNT was crosslinked by ethylene glycol diglycidyl ether (EGDE). The CS/AMWCNT nanocomposite beads produced has been characterized by BET, SEM, TGA, and Raman spectroscopy which exhibited enhanced thermal stability due to the incorporation of AMWCNT. Mechanical test results showed that mechanical strength of the CS/AMWCNT composite beads was significantly enhanced when comparing to unmodified chitosan beads, the breakage percentage decreased from 34.1% to 0.67%. The adsorption capacity for bilirubin was measured in PBS and BSA solutions, and the CS/AMWCNT composite beads with 5 wt% AMWCNT showed much higher adsorption capacity (12.7 mg/g in PBS and 7.6 mg/g in BSA) to bilirubin than chitosan beads (8.5 mg/g in PBS and 4.2 mg/g in BSA). Our nanocomposite beads with excellent hemocompatibility has a high potential application in blood purification as an efficient adsorbent for bilirubin. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2016.

Strong base anion exchange resin (e.g., BL-300) is currently used to eliminate bilirubin from blood in clinical hemoperfusion. However, there is a potential weakness of the resin to remove heparin and activation of coagulation. To overcome this weakness, we prepared a novel non-ionic macroporous polystyrene adsorbent (SZ-9) with rich mesopores and high surface area by suspension polymerization. Test results indicated that SZ-9 performed better adsorption capacity for bilirubin than BL-300. In addition, better blood compatibility was observed during whole blood hemoperfusion. Therefore, adsorbent SZ-9 might be an efficient alternative to anion exchange resin for the removal of bilirubin in hemoperfusion.

Mussel-inspired dopamine (DA) self-polymerization over a variety of substrates has become a simple but versatile approach for synthesis of surface protein-imprinted materials. However, relatively high nonspecific binding to the imprinted polydopamine (PDA) coatings has long been an open problem because of their multifunctionalities. We herein propose a facile strategy for reduction of the nonspecific adsorption by covering the imprinted PDA coatings with slightly crosslinked nonlinear poly(ethylene glycol) (PEG) layers via aqueous precipitation polymerization before template removal. Vinyl groups are introduced onto the PDA coatings via Cu2+ mediated metal coordination for facilitating surface polymerization. The Cu2+ and embedded template are removed after polymerization. For proof of hypothesis, the protein imprinted PDA coatings were formed with SiO2 nanoparticles as representative nano-supports and lysozyme as a model protein template. Protein binding tests show that the grafted PEG layers with an optimized feed crosslinking degree can significantly enhance both recognition selectivity and specific binding capacity to the imprinted nanoparticles, typically with the imprinting factor increasing from 2.6 to 6.4. Also, the PEG layers can remarkably improve the stability of the PDA coatings in the acidic template removal solution. The presented strategy represents the first example for PEGylation of protein-imprinted PDA coatings, and may be extended for surface imprinting of other bio/organic molecules over other substrate materials.

Adsorbents are widely used in hemoperfusion for bilirubin removal. However, their performance is often compromised by the presence of plasma proteins. In this study, the bilirubin adsorption capacity of polyvinyl alcohol microspheres (PVAm) functionalized with different amino-alkane ligands has been investigated, with the aim of gaining binding selectivity over albumin. Octylamine-functionalized PVA microspheres (PVAm-8) exhibited an excellent adsorption capacity for bilirubin (75% and 3.95 mg/mL in PBS vs 72% and 3.84 mg/mL in albumin solution) when compared to the clinical adsorbent BPR (92% and 4.84 mg/mL in PBS vs 71%, and 3.80 mg/mL in albumin solution). The bilirubin adsorption capacities of PVAm-8 were largely unaffected by the presence of albumin. Adsorption of bilirubin to PVAm-8 occurs mainly through hydrophobic effects, with adsorption consistent with the monolayer model and the pseudo-first-order model operating in both PBS and albumin solution. The effects of PVAm-8 on hemolytic activity, blood component stability and coagulant activity were negligible, indicating that PVAm-8 has good potential as a high-affinity bilirubin adsorbent for hemoperfusion applications.

Calcium alginate (CA) was used as an adsorbent to remove methylene blue (MB) from aqueous solution. The effect of initial dye concentration, contact time, temperature and solution pH on the adsorption of MB onto CA was investigated by batch experiments. The percentage removal of MB decreased with increasing temperature. Comparatively high adsorption capacities were shown over a wide pH range (pH 2–11). More than 93% of MB removal was obtained within 30 min for an initial dye concentration of 1,000 mg/L at an adsorbent dose of 4 g/L. The adsorption equilibrium was investigated by the Langmuir and Freundlich isotherms. The maximum adsorption capacity was 2,355.4 mg/g on the basis of the Langmuir isotherm. Thermodynamic parameters were evaluated, and revealed that the adsorption process was a spontaneous, exothermic and entropy-reduced process. Pseudo-first, pseudo-second and intra-particle diffusion kinetic models were applied to the experimental data, and the results showed that the adsorption was in good agreement with the pseudo-second order kinetic model. Desorption studies implied that CA could be a useful adsorbent for the removal of MB from aqueous solution.

A novel nano-CaCO

PS-DVB/nano-CaCO

Activated carbon (AC) prepared from peanut shell (PS) with microwave treatment was used to remove indigo carmine (IC) from aqueous solution in this study. PS and AC were examined by pore structural analysis, scanning electron microscopy, and elemental analysis. Effect of initial dye concentration, contact time, pH, and temperature on IC removal was investigated by batch experiments. The adsorption capacity increased with the increase of initial concentration, and decreased with increasing pH. Higher temperatures were favorable for the adsorption. The adsorption equilibrium could be reached within 90 min for all studied concentrations. The results showed that the Langmuir isotherm model had a good fit for the equilibrium data. Kinetic studies revealed that the adsorption followed the pseudo-second-order kinetic model. Thermodynamic studies indicated that the adsorption was a spontaneous, endothermic process.

In this study, a heparin-conjugated poly(ε-caprolactone) electrospun fiber was constructed to develop a functional scaffold for controlled release of vascular endothelial growth factors. The immobilization of vascular endothelial growth factor was achieved through affinity binding between heparin and vascular endothelial growth factor molecules. The sustained release of vascular endothelial growth factor from the scaffold was followed for up to 15 days. The endothelial cell adhesion and proliferation assay demonstrated that immobilized vascular endothelial growth factor maintained its activity. The blood compatibility of the scaffold was evaluated by activated partial thromboplastin time, platelet adhesion test, and arteriovenous shunt, and the functionalized scaffold showed improved anticoagulation properties. The biocompatibility was evaluated by subcutaneous implantation. Results showed that this vascular endothelial growth factor–releasing scaffold stimulated neovascularization with minimum immunological rejection compared to the unmodified poly(ε-caprolactone) scaffold. The present study demonstrated a new strategy of building bioactive scaffolds for the development of small-diameter vascular graft.

Background Microenvironment is being increasingly recognized as a critical determinant in tumor progression and metastasis. However, the appropriate regulatory mechanism to maintain the normal balance between differentiation and self-renewal of the cancer cell in microenvironment is not well known. Methods 4T1 breast cancer cells were treated with embryonic stem (ES) cell conditioned medium which was collected from mouse ES cells. Inhibition of tumor cell growth was based on the reduction of cell proliferation and viability, and inhibition of aggressive properties of tumor cells were examined using the wound-healing and mammosphere assays. The expression of stem cell-associated genes was detected by quantitative RT-PCR. Results We used a real-time imaging system to investigate the effect of the mouse ES cell microenvironment on aggressive breast cancer cells in vitro and in vivo. Exposure of breast cancer cells in mouse ES cell conditioned medium resulted in inhibition of growth, migration, metastasis, and angiogenesis of cancer cells. For many tumors, aggressive properties were tightly related to Stat3 signaling activation. We specifically discovered that the ES cell microenvironment sufficiently suppressed Stat3 signaling pathway activation in aggressive tumor cells, leading to a reduction in tumorigenesis and invasiveness. Conclusions We identified important functions of Stat3 and their implications for antitumor effects of ES cell conditioned medium. Some factors secreted by ES cells could efficiently suppress Stat3 pathway activation in breast cancer cells, and were then involved in cancer cell growth, survival, invasion, and migration. This study may act as a platform to understand tumor cell plasticity and may offer new therapeutic strategies to inhibit breast cancer progression.

Supplementary information contains Table S1 presenting primer sequences used for real-time PCR, Figure S1 showing mES cell characterization and reporter gene cells used in the experiment, Figure S2 showing decreased proliferation of 4T1 cells by direct cellâ cell contact of ES cells, Figure S3 showing ES-CM inhibited Stat3 signaling pathway in 4T1 cells (related to Figure 3), and Figure S4 showing ES-CM inhibit tumor growth in vivo (related to Figure 5). (DOCX 4638 kb)

Aims Parthenogenetic embryonic stem cells (pESCs) derived from artificially activated oocytes without fertilization presumably raise minimal ethical concerns and may serve as attractive candidates for regenerative medicine. Here we investigated whether pESCs could repair myocardial infarction (MI), in comparison to embryonic stem cells (ESCs). Methods and results A total of 89 mice that survived coronary artery ligation randomly received an intramyocardial injection of undifferentiated pESCs, ESCs, or saline. Sham-operated mice ( n = 21) that received no treatment served as control animals. After 7 days, transplantation of pESCs increased expression of pro-angiogenic factors and reduced leucocyte infiltration. By 14 and 30 days post-MI, similar to treatment with ESCs, treatment with pESCs efficiently prevented cardiac remodelling and enhanced angiogenesis, in contrast to saline-treated hearts. Improved heart contractile function was also notable 30 days following transplantation of pESCs. Immunofluorescence staining revealed that tissues regenerated from pESCs in the infarcted myocardium were positive for markers of cardiomyocytes, endothelial cells, and smooth muscle cells. Unlike ESC-treated mice, which exhibited a high incidence of teratoma (6 of 34), the pESC-treated mice showed no teratomas (0 of 30) 30 days following transplantation. Conclusion Transplantation of pESCs could attenuate cardiac dysfunction and adverse ventricular remodelling post-MI, suggesting that pESCs may provide promising therapeutic sources for MI in females.

To investigate whether electroacupuncture (EA) can promote cell survival and enhance heart function of mesenchymal stem cells (MSCs) therapy.MSCs were isolated from bone marrow and expanded in Minimum Essential Medium Alpha (α-MEM). MI was induced in 72 Sprague-Dawley (S-D) rats by ligation of the left anterior descending coronary artery (LAD) for 30 min and reperfusion. MI rats randomly received injection of 1×10(6) DiI-labeled MSCs alone (n =24, MSC group), or plus electroacupuncture (EA) at Neiguan (PC6, n=24, EA+MSC group), or saline (n =24, saline group). EA treatment was performed for 4 days. Another 24 rats were subjected to chest-open surgery without LAD occlusion and treatment (sham group). Three time points, 4, 14 and 28 days (n =8 for each group) were included in this study. The survival of transplanted MSCs and the protective gene expression were analyzed by reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot at day 4 and 14. Left ventricular remodeling, cardiac function, infarction area, fibrosis and capillary density were analyzed at day 28.EA can enhance MSC survival (2.6-fold up) at day 4. Big capillary density was 53% higher in EA+MSC treated group than MSC alone group. Furthermore, the rats treated by EA reduced the fibrosis and had 36% smaller infarct size comparing to MSC alone. EA also attenuated left ventricular remodeling and enhanced the functional recovery of infarcted hearts at week 4.EA at Neiguan acupoint can promote the stem cell survival and improve ischemic heart function. EA could become a useful approach in stem cell therapy for ischemia heart diseases.

Alumina, a support material, was loaded with chitosan to form a chitosan/alumina composite (CA) and was utilized for removing sulfonated azo-dye methyl orange (MO) from aqueous media in this study. The adsorbents were characterized by Fourier transform infrared (FTIR) spectra to confirm that chitosan was successfully immobilized on alumina, and some functional groups were successfully introduced into the surface of alumina after modification. The effects of pH, the amount of adsorbent, initial concentration, time, and temperature were studied in batch experiments. The experimental data were analyzed by Langmuir and Freundlich adsorption isotherms. Thermodynamic parameters such as ΔGo, ΔHo, and ΔSo have also been evaluated, and it has been found that the sorption process was spontaneous and exothermic in nature. Pseudofirst-order and pseudosecond-order kinetic models were used to fit the experimental data. Kinetic parameters, rate constants, equilibrium adsorption capacities, and related correlation coeffi...

Animal models that mimic cardiovascular diseases are indispensable tools for understanding the mechanisms underlying the diseases at the cellular and molecular level. This review focuses on various methods in preclinical research to create small animal models of cardiac diseases, such as myocardial infarction, dilated cardiomyopathy, heart failure, myocarditis and cardiac hypertrophy, and the related stem cell treatment for these diseases.

Transplantation of mesenchymal stem cells (MSCs) derived from adult bone marrow has been proposed as a potential therapeutic approach for post-infarction left ventricular (LV) dysfunction. However, age-related functional decline of stem cells has restricted their clinical benefits after transplantation into the infarcted myocardium. The limitations imposed on patient cells could be addressed by genetic modification of stem cells. This study was designed to improve our understanding of genetic modification of human bone marrow derived mesenchymal stem cells (hMSCs) by polyethylenimine (PEI, branched with Mw 25 kD), one of non-viral vectors that show promise in stem cell genetic modification, in the context of cardiac regeneration for patients. We optimized the PEI-mediated reporter gene transfection into hMSCs, evaluated whether transfection efficiency is associated with gender or age of the cell donors, analysed the influence of cell cycle on transfection and investigated the transfer of therapeutic vascular endothelial growth factor gene (VEGF). hMSCs were isolated from patients with cardiovascular disease aged from 41 to 85 years. Optimization of gene delivery to hMSCs was carried out based on the particle size of the PEI/DNA complexes, N/P ratio of complexes, DNA dosage and cell viability. The highest efficiency with the cell viability near 60% was achieved at N/P ratio 2 and 6.0 μg DNA/cm2. The average transfection efficiency for all tested samples, middle-age group (65 years), female group and male group was 4.32%, 3.85%, 4.52%, 4.14% and 4.38%, respectively. The transfection efficiency did not show any correlation either with the age or the gender of the donors. Statistically, there were two subpopulations in the donors; and transfection efficiency in each subpopulation was linearly related to the cell percentage in S phase. No significant phenotypic differences were observed between these two subpopulations. Furthermore, PEI-mediated therapeutic gene VEGF transfer could significantly enhance the expression level.

Erythropoietin (EPO) protects the myocardium from ischaemic injury and promotes beneficial remodelling. We assessed the therapeutic efficacy of intracardiac EPO injection and EPO-mediated stem cell homing in a rat myocardial infarction (MI) model. Following MI, EPO (3000 U/kg) or saline was delivered by intracardiac injection. Compared to myocardial infarction control group (MIC), EPO significantly improved left ventricular function (n= 11–14, P< 0.05) and decreased right ventricular wall stress (n= 8, P< 0.05) assessed by pressure-volume loops after 6 weeks. MI-EPO hearts exhibited smaller infarction size (20.1 ± 1.1%versus 27.8 ± 1.2%; n= 6–8, P< 0.001) and greater capillary density (338.5 ± 14.7 versus 259.8 ± 9.2 vessels per mm; n= 6–8, P< 0.001) than MIC hearts. Direct EPO injection reduced post-MI myocardial apoptosis by approximately 41% (0.27 ± 0.03%versus 0.42 ± 0.03%; n= 6, P= 0.005). The chemoattractant SDF-1 was up-regulated significantly assessed by quantitative realtime PCR and immunohistology. c-Kit+ and CD34+ stem cells were significantly more numerous in MI-EPO than in MIC at 24 hrs in peripheral blood (n= 7, P< 0.05) and 48 hrs in the infarcted hearts (n= 6, P< 0.001). Further, the mRNAs of Akt, eNOS and EPO receptor were significantly enhanced in MI-EPO hearts (n= 7, P< 0.05). Intracardiac EPO injection restores myocardial functions following MI, which may attribute to the improved early recruitment of c-Kit+ and CD34+ stem cells via the enhanced expression of chemoattractant SDF-1.

The aim of the present study was to investigated the construction of polycaprolactone-lecithin (PCL-L) electrospun fibers as a novel scaffold material for a tissue-engineered ureter. The effect of bone marrow mesenchymal stem cells (BM-MSCs) on the neovascularization of the scaffolds and the viability of planted urothelial cells (UCs) on PCL-L were also studied. UCs were obtained from New Zealand rabbit bladders, cultured and then seeded onto the lumen of the tubular scaffolds before being subcutaneously transplanted into the space of nude mice. The cultured UCs showed vacuolar degeneration after 7 days of transplantation and they gradually degraded thereafter. To facilitate the regeneration of the tissue-engineered ureter and the survival of UCs in the implant, MSCs were seeded into the tubular grafts by rolling up the nanofibrous membrane, followed by the seeding of UCs. This facilitated the survival of the UCs, which formed several cellular layers after 30 days. The mean microvessel density was significantly increased in tissues seeded with MSCs. Cell-tracking experiments revealed that the transplanted MSCs did not integrate directly into capillaries for angiogenesis. Our results demonstrated that the PCL-L electrospun fibrous scaffold has a high potential for a tissue-engineered ureter, especially when seeded with BM-MSCs, which enhanced angiogenesis.

Aims Controversy exists regarding the effects of granulocyte colony-stimulating factor (G-CSF) on post-infarction remodelling, which is regulated by matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs). The aim of this study was to investigate the impact of G-CSF administration on cardiac MMP/TIMP ratios and long-term remodelling in a rat model of acute myocardial infarction (MI). Methods and results Sprague–Dawley rats underwent coronary ligation to produce MI. Rats surviving the MI for 3 h were randomized to receive G-CSF (50 µg/kg/day for 5 consecutive days, n = 16) or saline ( n = 10). Sham-operated animals received no treatment ( n = 10). G-CSF injection significantly increased circulating white blood cells, neutrophils, and monocytes. Western blotting revealed that the ratios of MMP-2/TIMP-1 and MMP-9/TIMP-1 were significantly decreased in the infarcted myocardium. At 3 months, echocardiographic and haemodynamic examinations showed that the G-CSF treatment induced left ventricular (LV) enlargement and dysfunction. Histological analysis revealed that the extent of myocardial fibrosis and infarct size were larger in the G-CSF group than in the Saline group. Furthermore, G-CSF treated animals showed a significantly lower post-MI survival during the study period. Conclusion Decrease of cardiac MMP/TIMP ratios by G-CSF after infarction may be important as a mechanism in promotion of myocardial fibrosis, which further facilitates infarct expansion and LV dysfunction.

The adsorption effects of several macroporous adsorption resins for gibberellic acid (GA3) were investigated. The dynamic adsorption capacity is 58.38 mg/g dry beads for resin R4 and 96.46 mg/g dry beads for resin R5 which is consistent with the surface area. Aqueous methanol (50%, V/V) is a good eluent and the yield of GA3 is above 95%. The concentration of GA3 could increase five-fold after an adsorption-elution cycle and this is important when considering further crystallization of GA3 in an industrial process.

Stem cell therapy has been proved to be an effective approach to ameliorate the heart remodeling post myocardial infarction (MI). However, poor cell engraftment and survival in ischemic myocardium limits the successful use of cellular therapy for treating MI. Here, we sought to transplant adipose derived-mesenchymal stem cells (AD-MSCs) with a hydrogel (NapFF-NO), naphthalene covalently conjugated a short peptide, FFGGG, and β-galactose caged nitric oxide (NO) donor, which can release NO molecule in response to β-galactosidase. AD-MSCs, either from transgenic mice that constitutively express GFP and firefly luciferase (Fluc), or express Fluc under the control of VEGFR2 promoter, were co-transplanted with NapFF-NO hydrogel into murine MI models. Improved cell survival and enhanced cardiac function were confirmed by bioluminescence imaging (BLI) and echocardiogram respectively. Moreover, increasing VEGFR2-luc expression was also tracked in real-time in vivo, indicating NapFF-NO hydrogel stimulated VEGF secretion of AD-MSCs. To investigate the therapeutic mechanism of NapFF-NO hydrogel, cell migration assay, paracrine action of AD-MSCs, and histology analysis were carried out. Our results revealed that condition medium from AD-MSCs cultured with NapFF-NO hydrogel could promote endothelial cell migration. Additionally, AD-MSCs showed significant improvement secretion of angiogenic factors VEGF and SDF-1α in the presence of NapFF-NO hydrogel. Finally, postmortem analysis confirmed that transplanted AD-MSCs with NapFF-NO hydrogel could ameliorate heart function by promoting angiogenesis and attenuating ventricular remodeling. In conclusion, NapFF-NO hydrogel can obviously improve therapeutic efficacy of AD-MSCs for MI by increasing cell engraftment and angiogenic paracrine action.

Bone marrow mesenchymal stem cells (MSCs) have demonstrated their pluripotency to differentiate into different cell lineages and may be an alternative cell source for vascular tissue engineering. The objective of this study is to create small diameter vessels by seeding and culture of genetically modified MSCs onto a synthetic polymer scaffold produced by an electrospinning technique. A tubular scaffold (2 mm in diameter) with a microstructure of nonwoven fibers was produced by electrospinning of poly (propylene carbonate) (PPC). Rat MSCs obtained from bone marrow were expanded in culture and modified with vasculoprotective gene endothelial nitric oxide synthase (eNOS) or marker gene green fluorescent protein (GFP). These MSCs were seeded onto the electrospun fibrous grafts (internal diameter = 2 mm), and cultured in 5% CO(2) at 37 degrees C. The growth of MSCs in the scaffold was analyzed with scanning electron microscopy (SEM) and hematoxylin and eosin (H&E) staining. The gene transfer and transgenic gene expression were examined with fluorescence-activated cell sorting (FACS), immunochemical staining, reverse transcriptase-polymerase chain reaction (RT-PCR), and western blot. The production of nitric oxide (NO) by the engineered vessels was measured with an NO detection kit. Our data showed that the seeded cells integrated with the microfibers of the scaffold to form a three-dimensional cellular network, indicating a favorable interaction between this synthetic PPC scaffold with MSCs. High transduction efficiency was obtained with the use of concentrated retrovirus in the gene transfection of MSCs. The eNOS gene transcripts and protein were detected in the grafts seeded with eNOS-modified MSCs by RT-PCR and immunochemical staining. The amount of NO produced by grafts seeded with eNOS-modified MSCs was comparable to that produced by native blood vessels, and it was significantly higher than that in the grafts seeded with nonmodified MSCs. In summary, the vascular graft produced by culture of eNOS gene-modified MSCs onto the electrospun tubular scaffolds shows promising results in terms of function. The use of MSCs and therapeutic genes in tissue engineering of blood vessels could be helpful in improving vessel regeneration and patency.

Fibrin glue has been used for tissue adhesives and wound sealing. This study aimed to establish an approach to treat rabbit urethral injuries using fibrin glue and mesenchymal stem cells (MSCs).Rabbit urethral injury was performed by excising the subcutaneous tissue from the ventral aspect of the penile urethra. Thirty-six New Zealand rabbits were divided into four groups: injured without treatment (control), glue treatment (glue), glue and MSCs treatment (glue + MSCs), and without injury (normal). Animals were sacrificed and the penises were excised for analysis after treatment for 10 days (n=4) and 30 days (n=5).The fibrin glue had a porous structure with cellular compatibility to MSCs. Two rabbits developed urethral fistulae in the control group, but none did in either the glue or the glue + MSCs groups at day 10 and 30. Hematoxylin and eosin staining showed that the distance between the skin and the epithelial layer at the ventral aspect of the penis increased in the glue and glue + MSCs groups compared to the control group at day 10 and 30. The density of blood vessels in the injured area increased in the glue and glue + MSCs groups compared to the control at day 10 and 30. The average thickness of the epithelial layer in the glue + MSCs group showed the same increasing trend among the three groups at day 10 and 30. The thickness and the area of the smooth muscle that surrounds the urethral mucosa at the operative region increased compared to the thickness in the control and glue groups at day 10 and 30. Pancytokeratins (AE1/AE3) staining showed weak signals in the urethral fistulae in the control group compared with the glue and glue + MSCs groups. The transplanted MSCs have differentiated into endothelial and smooth muscle cells, contributing to the formation of blood vessels and the recovery of urethral smooth muscle.MSCs combined with fibrin glue showed improved neovascularization and smooth muscle formation in repairing urethral injury, resulting in effective prevention of the development of urethral fistulae. Our data indicate that fibrin glue combined with MSCs may be a feasible approach for repairing urethral injury.