Immunogenicity of the capsid precursor and a nine-amino-acid site-directed mutant of the 3C protease of foot-and-mouth disease virus coexpressed by a recombinant goatpox virus.

The myristoylated capsid precursor mP1-2A of foot-and-mouth disease virus (FMDV), when expressed in mammalian cells and processed by the FMDV 3C protease, can self-assemble into virus-like particles (VLPs). In the present study, nine amino acids of the 3C protease were replaced by site-directed mutagenesis to create a mutant 3C protease, 9m3C. To coexpress mP1-2A and 9m3C and test the resulting proteolytic processing and VLP assembly, two recombinant goatpox viruses (rGTPVs) were constructed by the insertion of two coding regions, one for mP1-2A and the other for either 9m3C (rGTPV-mP1-2A-9m3C) or Theileria protective antigen (TPA) as a control (rGTPV-mP1-2A-TPA). The two exogenous genes were inserted into an intergenic region between loci gp_24 and gp_24.5 of the rGTPV genome. Western blotting of cells infected with rGTPV-mP1-2A-9m3C showed that proteins VP0, VP1, and VP3 from the mP1-2A processed by the 9m3C protease could be detected by polyclonal FMDV sera. As observed by electron microscopy, the infected cells produced VLPs with a diameter of about 25 ± 2 nm. Titers of neutralizing antibody against FMDV were significantly higher in mice inoculated with rGTPV-mP1-2A-9m3C, which expresses the 9m3C protease together with mP1-2A, than mice inoculated with the control rGTPV-mP1-2A-TPA, which does not express the protease. An ovine immunization test determined that sheep inoculated intramuscularly with rGTPV-mP1-2A-9m3C produced FMDV-specific neutralizing antibody, but its titers did not meet the requirement of the World Organization for Animal Health. The result indicates that further modifications of rGTPV-mP1-2A-9m3C are necessary to produce an effective vaccine.

Photodynamic antibacterial research on hypericin-loaded PEGylated mesoporous silica delivery system.

In this study, a novel drug delivery system (MSN-PEG-Hypericin) was successfully fabricated using tetraethyl orthosilicate and 3-aminopropyltriethoxysilane as raw materials, and the PEGylation of the prepared aminated mesoporous silica and grafting of hypericin onto the carrier were further conducted to obtain MSN-PEG-Hypericin. The successful preparation of MSN-PEG-Hypericin was characterized by several physical-chemical techniques. Furthermore, the MSN-PEG-Hypericin system increased the ability of hypericin to generate reactive oxygen species (ROS) in vitro. The cytotoxicity assay and hemolysis analysis showed that MSN-PEG-Hypericin had good biocompatibility. For antibacterial studies, the irradiation time and incubation time of photodynamic therapy (PDT) for S. aureus and E. coli were respectively 8 min and 8 h, and the concentrations of hypericin were 2.5 and 5 µg/mL. The result of triphenyl tetrazolium chloride assay indicated that MSN-PEG-Hypericin had stronger photodynamic antibacterial activity than free hypericin, and S. aureus was more sensitive to PDT than E. coli, which was related to their cell structural differences. The antibacterial mechanism study indicated that the generated ROS could destroy the bacterial structures and cause bacterial death due to the leakage of the contents. The MSN-PEG-Hypericin system prepared in this study had potential application prospects in the antibacterial field.

NIR-Activated Thermosensitive Liposome-Gold Nanorod Hybrids for Enhanced Drug Delivery and Stimulus Sensitivity.

Combinatorial photothermal therapy and chemotherapy is an extremely promising tumor therapeutic modality. However, such systems still remain challenges in stimulus sensitivity, avoiding drug leakage, and therapeutic safety. To solve these problems, we engineered actively loaded doxorubicin (DOX) and gold nanorod (GNR) liposomes through embedding stiff hollow mesoporous silica nanoparticles (HMSNs) in the liposomal water cavity (HMLGDB) to resist the influence of shear force of GNRs to prevent drug leakage. Under 808 nm laser irradiation, the ambient temperature was raised greatly because of the photothermal conversion of GNRs, thereby rupturing the lipid layer and then triggering the DOX release. The results of in vitro experiments showed that the low concentration of HMLGDB (15 µg/mL) could effectively overcome the MCF-7 cells (human breast cancer cell line) by the increase of DOX concentration intracellularly and the good photothermal effect of GNRs. After intravenous injection, HMLGDB exhibited intratumor aggregation and PTT capacity. Furthermore, the combined chemo-photothermal antitumor strategy demonstrated a high inhibition of tumor growth and low damage to normal tissues. The developed hybrids provide a paradigm for efficient combinatorial photothermal therapy (PTT) and chemotherapy (CT).

Enzyme functionalized PEOz modified magnetic polydopamine with enhanced penetration for cascade-augmented synergistic tumor therapy.

In recent years, reactive oxygen species (ROS)-mediated cancer therapies have been widely recognized for their high selectivity and good biological safety. However, due to the difficulties of endogenous tumor microenvironment (TME), penetration of tumor tissues and integration of multimodal tumor ablation, the treatment with traditional therapies could not achieve satisfactory tumor inhibition effects. Here, a doxorubicin (DOX)-glucose oxidase (GOx) dual-loaded and poly (2-ethyl-2-oxazoline) (PEOz) decorated magnetic polydopamine nanoparticles (Fe3O4-DOX@PDA-GOx@PEOz, FDPGP) were constructed for tumor ablation. GOx-mediated cascade enzyme reactions could amplify oxidative stress damage and further synergistically inhibit breast cancer. Its pH-responsive charge reversal, drug-controlled release, photothermal, and cascade reactions were evaluated through extracellular experiments. Cellular uptake, cell cytotoxicity, tumor penetration and therapeutic efficacy of FDPGP were investigated through intracellular experiments. Finally, in vivo distribution, photothermal, synergistic antitumor therapeutic effect and biosafety were evaluated comprehensively by in vivo experiments. Excitingly, outstanding tumor enrichment and penetration, superior anticancer effects and biosafety were achieved by the combination of photothermal therapy (PTT)/starvation therapy (ST)/chemodynamic therapy (CDT)/chemotherapy (CT). As such, the FDPGP nanoplatform provides a new insight into the development of collaboratively multimodal enhanced tumor therapy.

High-efficiency Ni2+-NTA/PAA magnetic beads with specific separation on His-tagged protein.

To effective capture and universal enrichment of His-tagged protein, polyacrylic acid (PAA) brushes were used to encapsulate Fe3O4 nanoparticles, connect NTA, and Ni2+ to prepare magnetic beads. These materials provide many advantages, such as excellent stability, tuneable particle size, and a surface for further functionalisation with biomolecules. His-tagged green fluorescence protein (GFP) was separated efficiently, and the binding capacity of Fe3O4/MPS@PAA/NTA-Ni2+ was 93.4 mg/g. Compared with High-Affinity Ni-NTA Resin and Ni-NTA Magnetic Agarose Beads, Fe3O4/MPS@PAA/NTA-Ni2+ nanocomposites exhibited higher separation efficiency and binding capacity towards His-tagged GFP. Moreover, the selectivity and recyclability of them for the target proteins were maintained well after six cycles. This study would widen the application of PAA in constructing multifunctional nanocomposites for biomedical fields.

Silver triethanolamine-loaded PVB/CO films for a potential liquid bandage application.

Many studies have reported that silver has excellent antibacterial properties. However, silver ions can easily react with oxygen to form Ag2O, thus leading to a color change and a reduction in its anti-microbial characteristics. In this study, silver triethanolamine- (ST) loaded PVB/CO solution was prepared as a potential candidate liquid bandage. PVB/CO/ST retained high transparency after exposure to light for 12 months, which allowed convenient inspection of the wound bed without removal of the dressing. The PVB/CO/ST film exhibited favorable properties, such as speed of drying, excellent tensile strength and elongation characteristics and water vapor transmission rate (WVTR). It was comfortable and waterproof, and therefore effective at preventing bacterial invasion, providing effective biosafety. PVB/CO/ST solution-treated wounds exhibited accelerated healing and reduced inflammation in a nude mouse mode. Our data suggested that PVB/CO/ST solution could serve as a promising liquid bandage for treatment of minor trauma.

Biocompatibility of helicoidal multilamellar arginine-glycine-aspartic acid-functionalized silk biomaterials in a rabbit corneal model.

Silk proteins represent a unique choice in the selection of biomaterials that can be used for corneal tissue engineering and regenerative medical applications. We implanted helicoidal multilamellar arginine-glycine-aspartic acid-functionalized silk biomaterials into the corneal stroma of rabbits, and evaluated its biocompatibility. The corneal tissue was examined after routine hematoxylin-eosin staining, immunofluorescence for collagen I and III, and fibronectin, and scanning electron microscopy. The silk films maintained their integrity and transparency over the 180-day experimental period without causing immunogenic and neovascular responses or degradation of the rabbit corneal stroma. Collagen I increased, whereas Collagen III and fibronectin initially increased and then gradually decreased. The extracellular matrix deposited on the surface of the silk films, tightly adhered to the biomaterial. We have shown this kind of silk film graft has suitable biocompatibility with the corneal stroma and is an initial step for clinical trials to evaluate this material as a transplant biomaterial for keratoplasty tissue constructs.

Star-shaped PCL/PLLA blended fiber membrane via electrospinning.

Electrospun fiber mesh has been a candidate for guided bone regeneration membrane. However, its poor mechanics property has been limited in clinical application. In this study, various star-shaped poly(ε-caprolactones) (PCLs) are successfully synthesized by ring-opening polymerization and mixed with poly(l-lactide) (PLLA) to be made into blended membranes through electrospinning. Their corresponding properties are evaluated including morphology, thermodynamics, mechanics, and cytotoxicity. The blended fibers show smooth surface and well-distributed structure, which have slight differences in morphology with the change of arm number of star-shaped PCL. Crystallization of the fibrous membrane is influenced by star-shaped PCLs. Glass temperature drops from 64.23 °C for pure PLLA membrane to 53.62-49 °C for the blended membranes. The membranous tensile strength is depended strongly on star-shaped PCLs. The tensile strength goes up with arm number increasing; on the contrary, at the same arm number, the mechanics strength decreases with molecular weight increasing. And the fibrous membrane containing 20 wt.% star-shaped PCL shows better mechanics property compared to the other membranes. The star-shaped PCL/PLLA fiber membrane is not cytotoxicity.

Biofilm formation characteristics of bacterial isolates retrieved from a reverse osmosis membrane.

High-quality water purification systems using reverse osmosis (RO) membrane separation have faced a major challenge related to biofilm formation on the membrane surface, or biofouling. To understand this issue, the biofilm formation characteristics of four bacterial isolates previously retrieved from an RO membrane treating potable water were investigated. Biofilm formation of all four isolates occurred to different extents in microtiter plates and could be related to one or more cell properties (hydrophobicity, surface charge, and motility). For Dermacoccus sp. strain RO12 and Microbacterium sp. strain RO18, bacterial adhesion was facilitated by cell surface hydrophobicity, and for Rhodopseudomonas sp. strain RO3, adhesion was assisted by its low surface charge. Sphingomonas sp. strain RO2 possessed both twitching and swarming motilities, which could be important in mediating surface colonization. Further, strains RO2, RO3, and RO12 did not exhibit swimming motility, suggesting that they could be transported to RO membrane surfaces by other mechanisms such as convective permeate flow. The biofilm formation of RO2 was further tested on different RO membranes made of cellulose acetate, polyamide, and thin film composite in continuous flow cell systems. The resultant RO2 biofilms were independent of membrane surface properties and this was probably related to the ex-opolysaccharides secreted bythe biofilm cells. These results suggested that RO2 could colonize RO membranes effectively and could be a potential fouling organism in RO membranes for freshwater purification.