Danhong Yan

Toxicological Effect of ZnO Nanoparticles Based on Bacteria

Streptococcus agalactiae and Staphylococcus aureus are two pathogenetic agents of several infective diseases in humans. Biocidal effects and cellular internalization of ZnO nanoparticles (NPs) on two bacteria are reported, and ZnO NPs have a good bacteriostasis effect. ZnO NPs were synthesized in the EG aqueous system through the hydrolysis of ionic Zn2+ salts. Particle size and shape were controlled by the addition of the various surfactants. Bactericidal tests were performed in an ordinary broth medium on solid agar plates and in liquid systems with different concentrations of ZnO NPs. The biocidal action of ZnO materials was studied by transmission electron microscopy of bacteria ultrathin sections. The results confirmed that bactericidal cells were damaged after ZnO NPs contacted with them, showing both gram-negative membrane and gram-positive membrane disorganization. The surface modification of ZnO NPs causes an increase in membrane permeability and the cellular internalization of these NPs whereas there is a ZnO NP structure change inside the cells.

Characterization and bacterial response of zinc oxide particles prepared by a biomineralization process.

In this paper, olive-like ZnO particles were successfully synthesized via a facile biomineralization process in the template of silk fibroin (SF) peptide at room temperature. The coat of SF peptide on the surface of ZnO particles had a substantial influence on their morphology during the biomineralization. Room-temperature photoluminescence behavior of ZnO particles indicated that the visible blue emission peak centered at 410 nm was enhanced with the mineralization time. Bacteriological tests revealed that the mineralized ZnO particles with SF peptide were not toxic for Staphylococcus aureus, Escherichia coli, and Streptococcus agalactiae, presenting good cytocompatibility due to the surface coat of peptide. Their potential applications in bio-optical detectors could be envisioned.

Preparation and characterization of the biomineralized zinc oxide particles in spider silk peptides.

In this work, hierarchical ZnO particles were prepared using a biomineralization strategy at room temperature in the presence of peptides acidified from spider silk proteins. A mechanism of the mineralization of the ZnO particles was that the affinity of original ZnO nanoparticles and zinc ions in the peptide chains played an important role in controlling the biocrystallizing formation of the pore ZnO particles. The intensity of their visible green luminescence was enhanced with increases of the mineralization time due to the porous surface defects. The hierarchical ZnO materials with biomolecules will facilitate their photoluminescence spectra applications as biosensors or optoelectronic nanodevices in the future, when covalently coupled with peptides or other biomolecules to achieve patterned growth over large areas of substrate.

Cellular Compatibility of Biomineralized ZnO Nanoparticles Based on Prokaryotic and Eukaryotic Systems

Zinc oxide nanoparticles (NPs) with the size of ∼100 nm were prepared via a facile biomineralization process in the template of silk fibroin (SF) peptide at room temperature. These ZnO NPs have shown the remarkable behavior of low toxicity to gram-positive bacteria (Staphylococcus aureus, Staphylococcus agalactiae), gram-negative bacteria (Escherichia coli), and eukaryotic cells (mouse L929 fibroblasts). Bacteriological testing indicated that ZnO NPs presented a 50% inhibitory effect on Streptococcus agalactiae at the concentrations of >100 mM, whereas at the same concentrations, the growth of Staphylococcus aureus and Escherichia coli were hardly inhibited. On the other hand, a remarkable proliferation of Staphylococcus aureus or Escherichia coli was observed at the concentrations of ZnO NPs <50 mM. Moreover, the cytotoxicity test demonstrated that ZnO NPs mineralized with SF peptide possessed a low toxicity to mouse L929 fibroblasts. The SF peptide coated on the surface of ZnO NPs permitted greater adhesion and consequently greater proliferation of mouse L929 fibroblasts. Besides, from TEM micrographs of the cell ultrastructure, endocytosis of NPs into the cytoplasm can be detected and the ultrastructure of the cell underwent few changes. The cell membrane retained integrity, euchromatin dispersed homogenously inside the cytoplasm, the mitochondrial architecture remained intact, and no intracellular vacuoles were observed. High-resolution transmission electron microscopy images and selected area electron diffraction patterns of ultrathin cell sections indicated that the crystal structure of NPs was not damaged by the organelle or cytoplasm. All these observations indicated that ZnO NPs mineralized with the SF peptide possess good cytocompatibility.

Structure, morphology and fibroblasts adhesion of surface-porous titanium via anodic oxidation

Surface-porous titanium samples were prepared by anodic oxidation in H2SO4, H3PO4 and CH3COOH electrolytes under various electrochemical conditions. X-ray diffraction (XRD), scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDX) were employed to characterize the structure, morphology and chemical composition of the surface layer, respectively. Closer analysis on the effect of the electrochemical conditions on pore configuration was involved. It can be indicated that porous titania was formed on the surface layer, and the pore configuration was influenced by electrolyte composition and crystal structure of the titania. The fibroblast cells experiment showed that anodic oxidation of titanium surface could promote fibroblast adhesion on Ti substrate. The results suggested that anodic oxidation of Ti in CH3COOH was suitable to obtain surface-porous titanium oxides layers, which might be beneficial for better soft tissue ingrowths.

Preparation, characterization, in vitro bioactivity, and osteoblast adhesion of multi-level porous titania layer on titanium by two-step anodization treatment

To combine the advantages of different electrolytes in anodic oxidation, pure titanium samples were anodized in CH(3) COOH electrolyte according to a novel anodizing treatment regime and then in H(2) SO(4) electrolyte in potentialstatic mode. The in vitro bioactivity of the as-prepared titanium samples was evaluated by simulated body fluid (SBF) test. In addition, MG63 osteoblast-like cells were cultured on surfaces of the as-prepared titanium samples to evaluate osteoblast adhesion ability. The titanium samples after the two-step anodization treatment were covered by titania layers of anatase and/or rutile with several micrometres thickness and presented a multi-level porous surface morphology consisting of interlaced grooves about 20-μm wide overlaid with submicron scale pores. The SBF test results showed that the crystal titania layers prepared at appropriate conditions were able to induce apatite-forming in 7 days, indicating that the abundance of surface Ti-OH groups and (101)-oriented rutile structure both played important roles in in vitro bioactivity of titania layers. The cell experiment results showed that the macroscopic grooves could effectively promote osteoblast adhesion and growth and submicron scale pores might be beneficial to osteoblast adhesion. The two-step anodization treatment might be a promising candidate for surface modification of titanium implant.

Biomineralization of Uniform Gallium Oxide Rods with Cellular Compatibility

Monodispersed single crystalline alpha-GaOOH rods coated by silk fibroin (SF) have been prepared via a facile biomineralization process in the template of SF peptide. The carbon-coated alpha-Ga(2)O(3) and beta-Ga(2)O(3) rods are obtained by thermal treatment of the alpha-GaOOH rods at 600 and 800 degrees C, respectively. In vitro cytotoxicity studies of these gallium oxide rods showed no significant effect leading to restraint of cell proliferation of L929, Hela, and HaCat cells in less than 0.1 mg/mL prepared rods. On the basis of their excellent luminescence emission properties and cellular compatibilities, possible applications for bio-optoelectronic devices can be envisioned.

In vitro screening of ovarian tumor specific peptides from a phage display peptide library

To develop more biomarkers for diagnosis and therapy of ovarian cancer, a 12-mer phage display library was used to isolate peptides that bound specifically to the human ovarian tumor cell line SK-OV-3. After five rounds of in vitro screening, the recovery rate of phages showed a 69-fold increase over the first round of washings and a group of phage clones capable of binding to SK-OV-3 cells were obtained. A phage clone named Z1 with high affinity and specificity to SK-OV-3 cells was identified in vitro. More importantly, the synthetic biotin-labeled peptide, ZP1 (=SVSVGMKPSPRP), which corresponded to the sequence of the inserted fragment of Z1, demonstrated a high specificity to SK-OV-3 cells especially when compared to other cell lines (A2780 and 3T3). ZP1 might therefore be a biomarker for targeting drug delivery in ovarian cancer therapy.

A specific cell-penetrating peptide induces apoptosis in SKOV3 cells by down-regulation of Bcl-2

Peptides are emerging as pharmaceutical agents in cancer therapy. The peptide, TLSGAFELSRDK (TLS) is a targeting ligand that can specifically triggers cellular uptake by binding to SKOV3 cells. Cell surface proteins and the C-terminal basic residues of the TLS are required for effective cell penetration, and the uptake process is energy-dependent. It inhibited the proliferation of SKOV3 cells and induced early-stage apoptosis by down-regulation of Bcl-2 expression mediated through a caspase-dependent pathway. The synergistic anti-proliferative effects of the peptide TLS and doxorubicin on SKOV3 cells were further investigated. Taken together, TLS, acting as a combination of a targeted ligand and a therapeutic agent, was a promising candidate for the development of peptide-based therapies in ovarian cancer.

A novel peptide specifically targeting ovarian cancer identified by in vivo phage display

Discovery of peptide ligands that can target human ovarian cancer and deliver chemotherapeutics offers new opportunity for cancer therapy. The advent of phage‐displayed peptide library facilitated the screening of such peptides. In vivo screening that set in a microanatomic and functional context was applied in our study, and a novel peptide WSGPGVWGASVK targeting ovarian cancer was isolated. The phage clone PC3‐1 displaying peptide WSGPGVWGASVK can gain effective access to accumulate in the tumor sites after intravenous injection while reducing its accumulation in normal organs. Positive immunostaining of PC3‐1 was located in both sites of tumor cells and tumor blood vessels, which resulted in a diffuse binding pattern through the tumor. In vitro study results confirmed the capability of peptide WSGPGVWGASVK binding to and being internalized by both tumor cells and angiogenic endothelial cells. Flow cytometry analysis revealed that the peptide bound to SKOV3 cells with Kd value of 5.43 ± 0.4 μM. Taken together, it suggested that peptide WSGPGVWGASVK is a lead candidate for delivering therapeutics to penetrate into tumors. Copyright