Yibing Wang

A Structural Study of Escherichia coli Cells Using an In Situ Liquid Chamber TEM Technology

Studying cell microstructures and their behaviors under living conditions has been a challenging subject in microbiology. In this work, in situ liquid chamber TEM was used to study structures of Escherichia coli cells in aqueous solutions at a nanometer-scale resolution. Most of the cells remained intact under electron beam irradiation, and nanoscale structures were observed during the TEM imaging. The analysis revealed structures of pili surrounding the E. coli cells; the movements of the pili in the liquid were also observed during the in situ tests. This technology also allowed the observation of features of the nucleoid in the E. coli cells. Overall, in situ TEM can be applied as a valuable tool to study real-time microscopic structures and processes in microbial cells residing in native aqueous solutions.

One-step surface modification of polyurethane using affinity binding peptides for enhanced fouling resistance

Affinity binding peptides were examined for surface fabrication of synthetic polymeric materials. Peptides possessing strong binding affinities toward polyurethane (PU) were discovered via biopanning of M13 phage peptide library. The apparent binding constant (Kapp) was as high as 2.68 × 109 M−1 with surface peptide density exceeded 1.8 μg/cm2. Structural analysis showed that the ideal peptide had a high content (75%) of H-donor amino acid residues, and that intensified hydrogen bond interaction was the key driving force for the highly stable binding of peptides on PU. PU treated with such affinity peptides promises applications as low-fouling materials, as peptides increased its wettability and substantially reduced protein adsorption and cell adhesion. These results demonstrated a facile but highly efficient one-step strategy for surface property modification of polymeric materials for biotechnological applications.

A hyperbranched amphiphilic acetal polymer for pH-sensitive drug delivery

Nanoparticles are appealing drug delivery systems since they promise prolonged circulation time and predictable release behaviors. The current work reports a novel hyperbranched amphiphilic block copolymer synthesized using deactivation-enhanced atom transfer radical polymerization (DE-ATRP) for smart drug delivery. PEG2000-Br was applied as a macroinitiator to initiate acid-cleavable divinyl (ACD) monomers linked by acetal groups, formulating the hydrophilic (PEG) and hydrophobic (ACD) segments. The polymer appeared to self-assemble into micelles with diameters in the range of 70–100 nm, and was examined for the controlled release of doxorubicin (DOX). Results showed that DOX-loaded micelles (∼90 nm) could achieve drug loading as high as 8.2 wt%, with interesting pH-dependent release behaviors. Studies with flow cytometry (FCM) and confocal laser scanning microscopy (CLSM) showed that DOX-loaded micelles exhibited a high cellular uptake performance by HeLa cells, which indicated promising anti-tumor efficacy for such a drug delivery system. Additionally, such DOX-loaded micelles exhibited remarkable cytotoxicity against HeLa cells in a dose- and time-dependent manner due to the enhanced cell uptake behavior of micelles. These results indicated that the polymeric micelles might be used as a promising candidate for a pH-responsive drug delivery for cancer therapy.

Rational design of curcumin loaded multifunctional mesoporous silica nanoparticles to enhance the cytotoxicity for targeted and controlled drug release

Curcumin has attracted increasing attentions in recent years due to its promising anticancer activities. However, the hydrophobicity of curcumin has limited greatly its efficacy in clinical trials. In this study, folate (FA)-receptor targeting mesoporous silica nanoparticles that promise high loadings of curcumin via pH-sensitive Schiff base reactions were constructed and examined for targeted delivery of curcumin. Such nano-delivery system showed significantly improved stability and biocompatibility of curcumin under physiological conditions. Further investigations demonstrated that this nanocarrier had high values of drug loading efficiency (9.5%) and pH-responsive drug release property. Moreover, the particles could be efficiently internalized by FA-receptor-rich MCF-7 cells through the receptor-mediated endocytosis, whereas FA-receptor-poor HEK-293T normal cells showed much lower endocytosis of the nanoparticles under the same conditions. The in vitro cytotoxicity assay indicated that the curcumin-loaded nanoparticles exhibited significantly enhanced cytotoxicity against MCF-7 cell than HEK-293T cells because of the higher cellular uptake efficiency of nanocarriers. More broadly, this work demonstrates a new type of mesoporous silica nanocarrier particularly useful for targeted and controlled drug release applications.

Manipulation of pH-Sensitive interactions between podophyllotoxin-chitosan for enhanced controlled drug release

Podophyllotoxin (PPT) offers a broad-spectrum of anticancer activities, but little has been reported for its controlled release. This work shows that by manipulating molecular interactions between PPT and Chitosan, efficient nanoscale capsulation of PPT can be realized. The drug encapsulation efficiency is as high as 52%, with a final particle drug loading in the order of 10% (wt/wt). It further demonstrates that changes in pH can also significantly affect the rate of drug release from the Chitosan nanoparticles. Upon contact with cancer cells, chitosan nanoparticles enable efficient internalization and drug release. In vitro evaluations with HepG-2 and MCF-7 cells indicate that the chitosan nanoparticle carriers can improve drug efficacy in comparison to free PPT, most likely by regulating the intrinsic apoptotic signaling pathway to induce apoptosis. Overall, PPT chitosan nanoparticles promise a safe and efficient drug delivery system for PPT.

Functional polymeric dialdehyde dextrin network capped mesoporous silica nanoparticles for pH/GSH dual-controlled drug release

Multi-stimulation responsive nanomaterial-based drug delivery systems promise enhanced therapeutic efficacy in cancer therapy. This work examines a smart pH/GSH dual-responsive drug delivery system by using dialdehyde dextrin (DAD) end-capped mesoporous silica nanoparticles (MSNs). Specifically, DAD was applied as a “gatekeeper polymer” agent to seal drug loads inside the mesoporous of MSNs via a pH-sensitive Schiff bond, whereas the formed DAD polymer shells were further cross-linked by GSH-sensitive disulfide bonds. Results revealed that the DAD gatekeeper polymer could tightly close the mesopores of MSNs to control premature drug release under physiological conditions and respond to acidic and GSH conditions to release the trapped drugs. Significantly, fluorescent microscopy observation and cytotoxicity studies indicated that drug-loaded nanoparticles could be rapidly internalized through a passive targeting effect to inhibit cancer growth. Taken together, these polymer-modified pH/GSH dual-responsive MSNs could be used as promising candidates for “on-demand” anticancer drug delivery applications.

A self-targeting and controllable drug delivery system constituting mesoporous silica nanoparticles fabricated with a multi-stimuli responsive chitosan-based thin film layer

Surface modification and functionalization of nanomaterials have been adopted widely in devising smart drug delivery systems. This work examines the fabrication of multi-stimuli responsive surfaces on mesoporous silica nanoparticles (MSN) for environmentally sensitive site specific drug delivery with reduced risk of premature drug leakage. Chitosan cross-linked via disulfide bonds was applied to form a thin film on drug-loaded MSN, realizing a capsulation and stimuli-sensitive regulating gate membrane; that was further conjugated with folate for site specific targeting toward cancer cells. The chitosan thin film was very stable under neutral conditions and could effectively prevent drug leakage, but was sensitive to both pH and GSH stimulations to reach rapid drug release. Thus, drug release could be triggered by changes in such factors that are common to cancer cells. However, complete and accelerated release could only be realized when triggered simultaneously by both acidic pH and GSH. Moreover, tests with HepG-2 cells confirmed that folate-receptor mediated endocytosis successfully enhanced the cellular uptake of the nanoparticle and antitumor activity toward cancer cells. It is expected that this surface chemical modification strategy promises a powerful approach constructing smart drug delivery systems for efficient and safe chemotherapy.

pH-responsive nanoreservoirs based on hyaluronic acid end-capped mesoporous silica nanoparticles for targeted drug delivery

Mesoporous silica nanoparticles (MSNs) are greatly appealing for efficient drug delivery due to their excellent drug loading capacities. However, it remains as a major challenge to realize site-specific controlled release with MSNs. This work examines a smart pH-responsive drug release system using MSNs for CD44-targeting drug delivery. Specifically, hyaluronic acid (HA) was applied as an end-capping agent to seal drug loads inside the mesoporous of MSNs through the acid labile hydrazine bonds. HA exposed on the surface of the particles can also serve as a targeting agent at the same time, enable site specific targeting toward CD-44 overexpressing cells. The system showed a good stability at physiological pHs, yet drug release could be triggered in response to changes in pH. Further studies showed that the HA-fabricated particles could achieve much enhanced cellular uptake via CD44 receptor-mediated endocytosis by Hela cells (CD44 receptor-positive), and as a result, doxorubicin-loaded MSNs exhibited significantly enhanced drug efficacy toward cancer cells overexpressing CD44 receptor (IC50 = 0.56 μg/mL), whereas the normal cells showed weakly cytotoxicity (IC50 = 1.03 μg/mL). Such a fabrication strategy may provide a new platform for preparation of high performance drug delivery systems for cancer therapy.

Novel surfactant peptide for removal of biofilms

Conventional chemical surfactants attach on blots randomly, accompanied with health and environmental issues. To address this, a surfactant peptide was designed to mimic chemical surfactants with an affinity binding peptide as a hydrophobic tail for the cleanup of biofilm contaminations. The micelle forming and structural changes of the peptide in aqueous solution were systematically investigated. More importantly, the biofilm removal efficiency toward Escherichia coli O157:H7 biofilm reached 75% in neutral aqueous solutions at the concentration of 125 mg/L (critical micelle concentration 91 mg/L), a significant improvement in comparison to conventional surfactants and random surfactant peptide. The dynamic removal process reported by confocal laser scanning microscope (CLSM) also displayed the different constituents of biofilm blots, which associated with surfactant peptide binding efficiency. Hopefully, this surfactant strategy will eventually provide new scopes in the design of surface active biological agents.