Prevention and treatment of HPV-related cancer through a mRNA vaccine expressing APC-targeting antigen.

Persistent human papillomavirus (HPV) infection is associated with multiple malignancies. Developing therapeutic vaccines to eliminate HPV-infected and malignant cells holds significant value. In this study, we introduced a lipid nanoparticle encapsulated mRNA vaccine expressing tHA-mE7-mE6. Mutations were introduced into E6 and E7 of HPV to eliminate their tumourigenicity. A truncated influenza haemagglutinin protein (tHA), which binds to the CD209 receptor on the surface of dendritic cells (DCs), was fused with mE7-mE6 in order to allow efficient uptake of antigen by antigen presenting cells. The tHA-mE7-mE6 (mRNA) showed higher therapeutic efficacy than mE7-mE6 (mRNA) in an E6 and E7+ tumour model. The treatment resulted in complete tumour regression and prevented tumour formation. Strong CD8+ T-cell immune response was induced, contributing to preventing and curing of E6 and E7+ tumour. Antigen-specific CD8+ T were found in spleens, peripheral blood and in tumours. In addition, the tumour infiltration of DC and NK cells were increased post therapy. In conclusion, this study described a therapeutic mRNA vaccine inducing strong anti-tumour immunity in peripheral and in tumour microenvironment, holding promising potential to treat HPV-induced cancer and to prevent cancer recurrence.

Development of liposome as a novel adsorbent for artificial liver support system in liver failure.

Artificial liver support systems (ALSS), represented by albumin dialysis, are designed to replace the liver detoxification function and to serve as supportive therapy until liver transplantation or liver regeneration. We introduce liposome, which is majorly formed by soybean lecithin as the adsorbent nanomaterial in dialysate for the removal of protein-bound and liver failure-related solutes. The binding rate was detected by ultrafiltration column. In vitro and in vivo dialysis was performed in a recirculation system. Unconjugated bilirubin (52.83-99.87%) and bile salts (50.54-94.75%) were bound by liposomes (5-80 g/L) in a dose-response relationship. The in vitro haemodialysis model showed that the concentration of unconjugated bilirubin (45.64 ± 0.90 µmol/L vs. 54.47 ± 3.48 µmol/L, p < 0.05) and bile salts (153.75 ± 7.72 µmol/L vs. 180.72 ± 7.95 µmol/L, p < 0.05) were significantly decreased in the liposome dialysis group than in the phosphate buffer saline group. The in vivo haemodialysis model showed that 40 g/L liposome-containing dialysate led to a significant higher reduction ratio in total bilirubin (6.56 ± 5.72% vs. -1.86 ± 5.99%, p < 0.05) and more total bile acids (7.63 ± 5.27 µmol vs. 2.13 ± 2.32 µmol, p < 0.05) extracted in the dialysate in comparison with the conventional dialysate. In conclusion, the liposome-added dialysate proved to impose good extraction effects on the unconjugated bilirubin and bile salts. These findings indicate that conventional dialysate supported by this nanomaterial can markedly improve the removal of protein-bound and liver failure-related solutes, thus suggesting a novel and promising liver dialysis system.

Increasing the removal of protein-bound uremic toxins by liposome-supported hemodialysis.

Protein-bound uremic toxins (PBUTs) accumulate at high plasma levels and cause various deleterious effects in end-stage renal disease patients because their removal by conventional hemodialysis is severely limited by their low free-fraction levels in plasma. Here, we assessed the extent to which solute removal can be increased by adding liposomes to the dialysate. The uptake of liposomes by direct incubation in vitro showed an obvious dose-response relationship for p-cresyl sulfate (PCS) and indoxyl sulfate (IS) but not for hippuric acid (HA). The percent removal of both PCS and IS but not of HA was gradually increased with the increased concentration of liposomes in a rapid equilibrium dialysis setup. In vitro closed circulation showed that adding liposomes to the dialysate markedly increased the dialysances of PBUTs without greatly altering that of urea and creatinine. In vivo experiments in uremic rats demonstrated that adding liposomes to the dialysate resulted in higher reduction ratios (RRs) and more total solute removal (TSR) for several PBUTs compared to the conventional dialysate, which was approximately similar to the addition of bovine serum albumin to the dialysate. These findings highlight that as an adjunct to conventional hemodialysis, addition of liposomes to the dialysate could significantly improve the removal of protein-bound uremic solutes without greatly altering the removal of small, water-soluble solutes.

Electrodeposition of a biopolymeric hydrogel: potential for one-step protein electroaddressing.

The electrodeposition of hydrogels provides a programmable means to assemble soft matter for various technological applications. We report an anodic method to deposit hydrogel films of the aminopolysaccharide chitosan. Evidence suggests the deposition mechanism involves the electrolysis of chloride to generate reactive chlorine species (e.g., HOCl) that partially oxidize chitosan to generate aldehydes that can couple covalently with amines (presumably through Schiff base linkages). Chitosan's anodic deposition is controllable spatially and temporally. Consistent with a covalent cross-linking mechanism, the deposited chitosan undergoes repeated swelling/deswelling in response to pH changes. Consistent with a covalent conjugation mechanism, proteins could be codeposited and retained within the chitosan film even after detergent washing. As a proof-of-concept, we electroaddressed glucose oxidase to a side-wall electrode of a microfabricated fluidic channel and demonstrated this enzyme could perform electrochemical biosensing functions. Thus, anodic chitosan deposition provides a reagentless, single-step method to electroaddress a stimuli-responsive and biofunctionalized hydrogel film.

Polyethyleneimine-anchored liposomes as scavengers for improving the efficiency of protein-bound uremic toxin clearance during dialysis.

Protein-bound uremic toxins (PBUTs) are significant toxins that are closely related to the prognosis of chronic kidney disease. They cannot be effectively removed by conventional dialysis therapies due to their high albumin binding affinity. Our previous research revealed that cationic liposomes (i.e., polyethyleneimine [PEI]-decorated liposomes) could enhance the clearance of PBUTs via electrostatic interactions. However, the poor biocompatibility (hemolysis) restricted their applications in clinical dialysis treatment. Herein, we produced PEI-anchored, linoleic acid-decorated liposomes (CP-LA liposomes) via the conjugation of PEI to cholesterol chloroformate (Chol-PEI, CP), and linoleic acid (LA) was added to provide liposomal colloidal stability. The CP-LA liposomes outperformed the plain liposomes, demonstrating significantly higher PBUT binding rates and removal rates. In addition, in vitro dialysis simulation verified that the CP-LA liposomes had a better capacity for PBUT clearance than the plain liposomes, especially for PBUTs with a strong negative net charge. Hemolysis and cytotoxicity tests revealed that the biocompatibility of the CP-LA liposomes was better than that of the physically-decorated PEI-liposome. CP-LA liposomes possess great potential for PBUT clearance in clinical dialysis therapy.

Lactate dehydrogenase encapsulated in a metal-organic framework: A novel stable and reusable biocatalyst for the synthesis of D-phenyllactic acid.

In this study, we synthesized a novel biocatalyst by encapsulating lactate dehydrogenase (LDH) in the metal-organic framework ZIF-90 by one-pot embedding. It showed strong biological activity for efficient synthesis of D-phenyllactic acid (D-PLA). The morphology and structure of LDH@ZIF-90 was systematically characterized by powder X-ray diffraction (XRD), scanning electron microscopy (SEM), Fourier transform infrared (FT-IR) spectroscopy, confocal laser scanning microscopy (CLSM) and gas sorption. According to thermogravimetric analysis (TGA), the enzyme loading of the biocatalyst was 3 %. The Michaelis-Menten constant (Km) and maximal reaction rate (Vmax) of LDH@ZIF-90 were similar to those of free LDH, which proved that ZIF-90 had good biocompatibility to encapsulate LDH. At the same time, LDH@ZIF-90 exhibited enhanced tolerance to temperature, pH and organic solvents, and its reusability was greatly improved with 68 % of initial enzyme activity remaining after 7 rounds of recylcing. Overall, LDH encapsulated in ZIF-90 may be an economically competitive and environmentally friendly novel biocatalyst for the synthesis of D-PLA.

Linoleic acid-modified liposomes for the removal of protein-bound toxins: An in vitro study.

INTRODUCTION: Protein-bound uremic toxins (PBUTs) and liver failure-related cholestatic solutes are associated with adverse outcomes in patients with chronic kidney disease (CKD) and liver failure, respectively, and are not easily removed by traditional dialysis therapies. We constructed linoleic acid-modified liposomes (LA-liposomes) as indirect adsorbent in the dialysate, and evaluated their effects on the clearance of the representative PBUTs and cholestatic solutes. METHODS: The LA-liposomes were prepared by the thin-film hydration method. The binding rates of liposomes and protein-bound solutes were detected by the ultrafiltration column. The in vitro dialysis experiments were performed using both non-current and current devices to assay the clearing efficiency of the dialysate supported by LA-liposomes. RESULTS: The LA-liposomes exhibited good binding properties to the PBUTs, bilirubin and bile acids. The LA-liposome dialysate showed higher solute reduction rates of the representative PBUTs and cholestatic solutes than the traditional dialysate or dialysate supported by the unmodified plain liposomes. Also, albumin binding of the PBUTs was significantly inhibited by the addition of linoleic acid (LA), and the removal efficiency of PBUTs was greatly enhanced by the combination of indirect adsorbent LA-liposomes and LA as the competitive displacer. CONCLUSION: LA-liposomes were efficient in the clearance of the representative PBUTs and liver failure-related solutes. Moreover, the combination of indirect adsorbent LA-liposomes and competitive displacer suggested a potential application for the extremely highly-bound solutes.

Improving the clearance of protein-bound uremic toxins using cationic liposomes as an adsorbent in dialysate.

Anionic and protein-bound uremic toxins, represented by indoxyl sulfate (IS), may be associated with cardiovascular outcomes and the progression of chronic kidney disease in cases of injured kidney function and are not easily cleared by traditional dialysis therapy. We fabricated cationic liposomes that were modified with polyethyleneimine (PEI), octadecylamine (Oct), and hexadecyl trimethyl ammonium bromide (CTAB), and evaluated the effects on the clearance of the representative protein-bound uremic toxins (PBUTs). The binding rate was obtained by ultrafiltration and in vitro dialysis was performed in a Rapid Equilibrium Dialysis (RED) device to assay the clearing efficiency of the dialysate supported by three types of cationic liposomes. The cationic liposomes showed a higher binding rate with IS (1.24-1.38 fold higher) and p-cresol (1.07-1.09 fold higher) than in the unmodified plain liposomes. The dialysate supported by cationic liposomes also exhibited better clearing efficiency for IS (PEI-20: 57.65 ± 1.74 %; Oct-5: 62.80 ± 0.69 %; CTAB-10: 66.54 ± 0.91 %; p < 0.05) and p-cresol (PEI-20: 67.05 ± 3.09 %; Oct-5: 79.26 ± 0.43 %; CTAB-5: 68.45 ± 1.72 %; p < 0.05) than for phosphate buffer saline (IS: 29.70 ± 2.38 %; p-cresol: 33.59 ± 3.44 %) or dialysate supported by bovine serum albumin (IS: 50.00 ± 4.01 %; p-cresol: 53.06 ± 0.97 %). In conclusion, cationic liposomes are efficient in the clearance of anionic PBUTs, and these modified liposomes suggest a potential application in blood purification.

Removal of Protein-Bound Uremic Toxins by Liposome-Supported Peritoneal Dialysis.

Background:Protein-bound uremic toxins (PBUTs) are poorly cleared by peritoneal dialysis (PD). This study aimed to enhance PBUT removal in PD by adding a binder to the peritoneal dialysate and to evaluate the feasibility and efficacy of liposome-supported PD (LSPD) to increase the removal of PBUTs compared with albumin PD.Methods:Removal of p-cresyl sulfate (PCS), indoxyl sulfate (IS), and indole-3-acetic acid (3-IAA) was first evaluated in an in vitro PD model using artificial plasma preloaded with test solutes. Male Sprague-Dawley rats (n = 24) were then subjected to 5/6 nephrectomy and fed for 16 weeks to establish end-stage renal failure, after which they were treated with either conventional glucose-based PD, albumin-based PD, or liposome-based PD. Removal of PBUTs and small water-soluble solutes was determined during a 6-hour PD dwell.Results:In vitro experiments showed that adding albumin as a toxin binder to the dialysate markedly increased the removal of PCS, IS, and 3-IAA compared with the control. The uptake capacity of liposomes was comparable with that of albumin for PCS and 3-IAA, though slightly inferior for IS. In vivo PD in uremic rats demonstrated that LSPD resulted in higher intraperitoneal concentrations and more total mass removal for PBUTs than the conventional glucose-based PD, which was comparable with albumin PD.Conclusions:Supplementing conventional glucose-based PD solutions with a binder could efficiently increase the removal of PBUTs. This preliminary study suggested that LSPD may be a promising alternative to albumin PD for increasing PBUT removal in the development of next-generation PD solutions for PD patients.

Bile acids regulate cysteine catabolism and glutathione regeneration to modulate hepatic sensitivity to oxidative injury.

Bile acids are signaling molecules that critically control hepatocellular function. Disrupted bile acid homeostasis may be implicated in the pathogenesis of chronic liver diseases. Glutathione is an important antioxidant that protects the liver against oxidative injury. Various forms of liver disease share the common characteristics of reduced cellular glutathione and elevated oxidative stress. This study reports a potentially novel physiological function of bile acids in regulating hepatic sulfur amino acid and glutathione metabolism. We found that bile acids strongly inhibited the cysteine dioxygenase type-1-mediated (CDO1-mediated) cysteine catabolic pathway via a farnesoid X receptor-dependent mechanism. Attenuating this bile acid repressive effect depleted the free cysteine pool and reduced the glutathione concentration in mouse liver. Upon acetaminophen challenge, cholestyramine-fed mice showed impaired hepatic glutathione regeneration capacity and markedly worsened liver injury, which was fully prevented by N-acetylcysteine administration. These effects were recapitulated in CDO1-overexpressing hepatocytes. Findings from this study support the importance of maintaining bile acid homeostasis under physiological and pathophysiological conditions, as altered hepatic bile acid signaling may negatively impact the antioxidant defense mechanism and sensitivity to oxidative injury. Furthermore, this finding provides a possible explanation for the reported mild hepatotoxicity associated with the clinical use of bile acid sequestrants in human patients.

Study on ß-cyclodextrin-complexed nanogels with improved thermal response for anticancer drug delivery.

For achievement of controllability in drug delivery, development of nanocarriers with thermal response is one of the most investigated stimulative strategies for oncological treatment. How to improve the thermosensitivity of the nanocarriers is an important factor for their successful drug delivery applications. In this study, a kind of complexed nanogels (PNACD) was developed by incorporating ß-cyclodextrin (ß-CD) into the nanogels of copolymers of N-isopropylacrylamide (NIPAM) and acrylic acid (AA) during their polymerization via in situ crosslinking of N,N'-methylenebisacrylamide (MBA) as a crosslinker. The complexed PNACD nanogels displayed a significantly enhanced thermosensitivity near body temperature compared to the ß-CD-free nanogels (PNA), which is probably associated with the rapid volumetric transformation during heating/cooling process due to the formation of complexed (decomplexed) structure between ß-CD and PNIPAM element. The PNACD nanogels can be used for loading of an anticancer drug (doxorubicin, DOX) with an encapsulation efficiency of 54±5%. The DOX-loaded nanogels displayed pH-/thermo-dual responsivenesses in drug release, which can be effectively internalized into KB cells (a human epithelial carcinoma cell line) to exert good anticancer bioactivity. This approach may enlighten design of novel nanocarriers for delivery of drugs beyond anticancer chemotherapeutic reagents.

Agar/gelatin bilayer gel matrix fabricated by simple thermo-responsive sol-gel transition method.

We present a simple and environmentally-friendly method to generate an agar/gelatin bilayer gel matrix for further biomedical applications. In this method, the thermally responsive sol-gel transitions of agar and gelatin combined with the different transition temperatures are exquisitely employed to fabricate the agar/gelatin bilayer gel matrix and achieve separate loading for various materials (e.g., drugs, fluorescent materials, and nanoparticles). Importantly, the resulting bilayer gel matrix provides two different biopolymer environments (a polysaccharide environment vs a protein environment) with a well-defined border, which allows the loaded materials in different layers to retain their original properties (e.g., magnetism and fluorescence) and reduce mutual interference. In addition, the loaded materials in the bilayer gel matrix exhibit an interesting release behavior under the control of thermal stimuli. Consequently, the resulting agar/gelatin bilayer gel matrix is a promising candidate for biomedical applications in drug delivery, controlled release, fluorescence labeling, and bio-imaging.

Layer-by-layer assembled biopolymer microcapsule with separate layer cavities generated by gas-liquid microfluidic approach.

In this work, a layer-by-layer (LbL) assembled biopolymer microcapsule with separate layer cavities is generated by a novel and convenient gas-liquid microfluidic approach. This approach exhibits combined advantages of microfluidic approach and LbL assembly method, and it can straightforwardly build LbL-assembled capsules in mild aqueous environments at room temperature. In particular, using this approach we can build the polyelectrolyte multilayer capsule with favorable cavities in each layer, and without the need for organic solvent, emulsifying agent, or sacrificial template. Various components (e.g., drugs, proteins, fluorescent dyes, and nanoparticles) can be respectively encapsulated in the separate layer cavities of the LbL-assembled capsules. Moreover, the encapsulated capsules present the ability as colorimetric sensors, and they also exhibit the interesting release behavior. Therefore, the LbL-assembled biopolymer capsule is a promising candidate for biomedical applications in targeted delivery, controlled release, and bio-detection.

Multistimulative Nanogels with Enhanced Thermosensitivity for Intracellular Therapeutic Delivery.

The flexibility and hydrophilicity of nanogels suggest their potential for the creation of nanocarriers with good colloidal stability and stimulative ability. In the present study, biocompatible AGP and AGPA nanogels with triple-stimulative properties (thermosensitivity, pH sensitivity, and redox sensitivity) were prepared by incorporating poly(N-isopropylacrylamide) (PNIPAM) or poly(N-isopropylacrylamide-co-acrylic acid) (P(NIPAM-AA)) into alginate (AG) emulsion nanodrops, followed by fixation with a disulfide-containing molecule (cystamine dihydrochloride (Cys)). Compared to AG/PNIPAM(AGP) nanogels, AG/P(NIPAM-AA) (AGPA) nanogels exhibited more sensitive volumetric expansion by switching the temperature from 40 to 25 °C under physiological medium. This expansion occurs because P(NIPAM-AA) with -COOH groups can be fixed inside the nanogels via chemical bonding with Cys, whereas PNIPAM was encapsulated in the nanogels through simple physical interactions with the AG matrix. AGPA nanogels carrying an anticancer drug tend to easily enter cells upon heating, thereby exerting toxicity through a cold shock and reverse thermally induced release of an anticancer drug. Upon internalization inside cells, the nanogels use the reducible and acidic intracellular environments to effectively release the drug to the nucleus to impart anticancer activity. These results demonstrate that multifunctional nanogels may be used as a general platform for therapeutic delivery.

In situ biodegradable crosslinking of cationic oligomer coating on mesoporous silica nanoparticles for drug delivery.

Although layer-by-layer assembly using anionic and cationic polymer has been a popular way to develop core-shell nanoparticles, the strong electrostatic interactions may limit shell degradability, thus hampering their application as a platform for controlled therapeutic delivery. In this study, we demonstrate a simple approach to developing mesoporous nanohybrids via a process of pre-drug loading (using doxorubicin (DOX) as a model drug) into mesoporous silica nanoparticles (MSN), followed by surface functionalization with a kind of cationic oligomer (low molecular weight polyethylene imine, LPEI) and in situ crosslinking by degradable N,N'-bis(acryloyl)cystamine (BAC). The presence of LPEI shell affords the nanohybrids with charge-reversal ability, which means that the acidic tumor extracellular microenvironment can transform the negative surface charge at neutral conditions into positive-charged ones. The nanohybrids displayed a pH- and redox-dual sensitivity in DOX release under conditions that mimic intracellular reductive conditions and acidic tumor microenvironments. The nanohybrids can be effectively internalized into A549 cells (a carcinomic human alveolar basal epithelial cell line), resulting in a high DOX intracellular accumulation and an improved anticancer cytotoxicity when compared with free DOX, suggesting their high potential as a new platform for therapeutic delivery.

Preferential tumor accumulation and desirable interstitial penetration of poly(lactic-co-glycolic acid) nanoparticles with dual coating of chitosan oligosaccharide and polyethylene glycol-poly(D,L-lactic acid).

Despite advances in polymeric nanoparticles (NPs) as effective delivery systems for anticancer drugs, rapid clearance from blood and poor penetration capacity in heterogeneous tumors still remain to be addressed. Here, a dual coating of poly (ethylene glycol)-poly (d,l-lactic acid) (PEG-PDLLA) and water-soluble chitosan oligosaccharide (CO) was used to develop PLGA-based NPs (PCPNPs) with colloidal stability for delivery of paclitaxel (PTX). The PCPNPs were prepared by a modified nanoprecipitation process and exhibited homogeneous size of 165.5nm, and slight positive charge (+3.54mV). The single PEG-PDLLA-coated PLGA NPs (PPNPs) with negative charge (-13.42mV) were prepared as control. Human breast cancer MDA-MB-231 cell and mice MDA-MB-231 xenograft model were used for in vitro and in vivo evaluation. Compared to Taxol®, both PCPNPs and PPNPs increased the intracellular uptake and exerted stronger inhibitory effect on tumor cells in vitro, especially for PCPNPs. Particularly, due to the near neutral surface charge and shielding by the dual coating, the blank cationic NP presented low cytotoxicity. With the synergistic action of PEG-PDLLA and CO, PCPNPs not only strongly inhibited macrophage uptake and extended the blood circulation time, but also improved the selective accumulation and interstitial penetration capacity to/in tumor site. Consequently, a significantly enhanced antitumor efficacy was observed for the cationic PCPNPs. Our findings suggest that, the dual PEG-PDLLA/CO coating can effective improve the tumor accumulation and interstitial penetration of NPs and, therefore may have great potential for tumor treatment. STATEMENT OF SIGNIFICANCE: Rapid clearance from blood and poor penetration capacity in heterogeneous tumors represent great challenge for polymeric nanoparticles (NPs) as effective delivery systems for anticancer drugs. This study provides a promising cationic nanoparticle (PCPNPs) with dual coating of chitosan oligosaccharide (CO) and PEG-PDLLA to address the above problem. The PCPNPs prepared with 165.5nm and slight positive charge (+3.54mV) showed an improved accumulation and interstitial penetration capacity to/in tumor site, and thus led to an enhanced antitumor efficacy. This is the first time to report the cooperative effect of PEG-PDLLA and CO on PLGA NPs in this field. This work can arouse broad interests among researchers in the fields of nanomedicine, nanotechnology, and drug delivery system.

Polymer fractionation using chromatographic column packed with novel regenerated cellulose beads modified with silane.

Novel microporous beads with the particle size of about 90 microm were prepared, for the first time, from cellulose and konjac glucomannan (RC/KGM3) in 1.5 M NaOH/0.65 M thiourea aqueous solution by emulsification method. The microporous beads were then modified with silane to avoid the adsorption of polymers containing hydroxyl groups, coded as RC/KGM3-Si. A preparative size-exclusion chromatographic (SEC) column (500 mm x 20 mm) was packed with RC/KGM3-Si, and its exclusion limit and fractionation range of the stationary phase were, respectively, weight-average molecular masses (Mw) of 4.8 x 10(5) g/mol and 5.3 x 10(3)-4.8 x 10(5) g/mol for polystyrene in tetrahydrofuran. The preparative SEC column was used to fractionate poly(epsilon-caprolactone) (PCL, Mw = 8.31 x 10(4) g/mol polydispersity index d= 1.55) in tetrahydrofuran and a polysaccharide PC3-2 (Mw = 1.21 x 10(5) g/mol, d= 1.70) in 0.05 M NaOH aqueous solution, respectively. The Mw values of the fractions determined by analytical SEC combined with laser light scattering were from 1.2 x 10(4) to 1.84 x 10(5) for PCL and from 8.5 x 10(4) to 2.13 x 10(5) for PC3-2, as well as d from 1.2 to 1.5. The results indicated that the preparative SEC has good fractionation efficiency in both organic solvent and alkaline aqueous solution for the various polymers.

Surface properties of polyurethanes modified by bioactive polysaccharide-based polyelectrolyte multilayers.

Lentinan, a mushroom polysaccharide, isolated from Lentinus edodes (Shiitake mushroom) was sulfated in dimethylsulfoxide to obtain a water-soluble derivative coded as LS. Then, two polysaccharide-based polyelectrolytes, polyanionic lentinan sulfate (LS) and polycationic chitosan (CS), were alternatively deposited onto the surfaces of polyurethane (PU) via layer-by-layer (LbL) assembly technique. The surfaces modified by polysaccharide-based multilayers were investigated by X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM) and contact angle measurements. The fibrinogen adsorption and platelet adhesion to the surfaces, cytocompatibility to L-929 cells, and antibacterial activity against Pseudomonas aeruginosa of unmodified PU and LbL-modified PU were tested in vitro, respectively. The results showed that the water contact angle decreased gradually during the successive buildup of the polysaccharide-based multilayers, and decreased slowly after four bilayers were assembled. The surface roughness of PU modified by five bilayers (LS as topmost layer) increased compared with that of unmodified PU. The fibrinogen adsorption on the surface decreased 81% after assembly of five bilayers (LS as topmost layer). The number of adherent platelets on the surface modified by five bilayers (LS as topmost layer) is reduced, in comparison with that of the unmodified PU. The tests of L-929 cells indicated that LbL-modified PU surfaces had better cytocompatibility than unmodified PU. In addition, PU modified by polysaccharide-based multilayers showed antibacterial activity against P. aeruginosa.

Coupling electrodeposition with layer-by-layer assembly to address proteins within microfluidic channels.

Two thin-film assembly methods are coupled to address proteins. Electrodeposition confers programmability and generates a template for layer-by-layer (LbL) assembly. LbL enables precise control of film thickness and the incorporation of labile biological components. The capabilities are demonstrated using glucose oxidase (GOx) based electrochemical biosensing within a microfabricated fluidic device.

Surface modification on polyurethanes by using bioactive carboxymethylated fungal glucan from Poria cocos.

In this work, a water-insoluble ß-D-glucan (PCSG), isolated from Poria cocos, was carboxymethylated to create a water-soluble derivative named as CP. After free amino groups have been introduced, CP was covalently immobilized onto PU surface. The hydrophilicity and the concentration of carboxyl group on the modified PU surface were determined. The fibrinogen and albumin adsorption to the surface, in vitro blood compatibility, and antibacterial activity of the surface against Pseudomonas aeruginosa were evaluated. The water contact angle measurement indicated that the hydrophilicity of PU surface increased after modification. The fibrinogen adsorption of the modified PU surface decreased 51.5%, compared with control PU. CP immobilization could prolong the blood coagulation time was suggested by APTT experiment. Antibacterial activity experiments indicated that CP modified surface obviously suppressed the growth of P. aeruginosa. Thereby, CP immobilization improves blood compatibility of PU surface and introduces special antibacterial bioactivity.