Xiaodong Ye

Smart Superstructures with Ultrahigh pH-Sensitivity for Targeting Acidic Tumor Microenvironment: Instantaneous Size Switching and Improved Tumor Penetration.

The currently low delivery efficiency and limited tumor penetration of nanoparticles remain two major challenges of cancer nanomedicine. Here, we report a class of pH-responsive nanoparticle superstructures with ultrasensitive size switching in the acidic tumor microenvironment for improved tumor penetration and effective in vivo drug delivery. The superstructures were constructed from amphiphilic polymer directed assembly of platinum-prodrug conjugated polyamidoamine (PAMAM) dendrimers, in which the amphiphilic polymer contains ionizable tertiary amine groups for rapid pH-responsiveness. These superstructures had an initial size of ∼80 nm at neutral pH (e.g., in blood circulation), but once deposited in the slightly acidic tumor microenvironment (pH ∼6.5-7.0), they underwent a dramatic and sharp size transition within a very narrow range of acidity (less than 0.1-0.2 pH units) and dissociated instantaneously into the dendrimer building blocks (less than 10 nm in diameter). This rapid size-switching feature not only can facilitate nanoparticle extravasation and accumulation via the enhanced permeability and retention effect but also allows faster nanoparticle diffusion and more efficient tumor penetration. We have further carried out comparative studies of pH-sensitive and insensitive nanostructures with similar size, surface charge, and chemical composition in both multicellular spheroids and poorly permeable BxPC-3 pancreatic tumor models, whose results demonstrate that the pH-triggered size switching is a viable strategy for improving drug penetration and therapeutic efficacy.

pH Dependence of Structure and Surface Properties of Microbial EPS

The flocculation of microorganisms plays a crucial role in bioreactors, and is substantially affected by pH. However, the mechanism for such an effect remains unclear. In this work, with an integrated approach, the pH dependence of structure and surface property of microbial extracellular polymeric substances (EPS), excreted from Bacillus megaterium TF10, and accordingly its flocculation is elucidated. From the Fourier transform infrared spectra and acid-base titration test results, the main functional groups and buffering zones in the EPS responsible for the microbial flocculation are indentified. The laser light scattering analysis reveals that the deprotonated or protonated states of these functional groups in EPS result in more dense and compact structure at a lower pH because of hydrophobicity and intermolecular hydrogen bonds. The zeta potential measurements identify the isoelectric point and indicate that the electrostatic repulsion action of EPS is controlled by pH. The highest flocculation efficiency is achieved near the isoelectric point (pH 4.8). These results clearly demonstrate that the EPS structure, surface properties, and accordingly the microbial flocculation are dependent heavily on pH in solution.

Coagulation kinetics of humic aggregates in mono- and di-valent electrolyte solutions.

Coagulation behaviors of humic acids (HAs) aggregates in electrolyte solutions at different pHs, valences and concentrations of electrolyte cations were investigated using dynamic light scattering technique in combination of other analytical tools. For monovalent electrolyte sodium chloride (NaCl) solution, at its low concentrations the average hydrodynamic radius () of aggregates kept nearly constant. However, at high NaCl concentrations, could be scaled to the time t as ∝ t(a), suggesting a diffusion-limited colloid aggregation (DLCA). The coagulation value of NaCl in a buffer at pH 7.1 was calculated to be in a range of 61.3-84.4 mM. Divalent cation Mg(2+) was far more effective in enhancing the HA coagulation, as evidenced by a lower coagulation value (between 1.0 and 1.7 mM) and a more rapid coagulation rate. Such an enhancement could be explained by the combined effects of electrostatic repulsion, complexation and bridging. The highest coagulation rate (d/dt) and coagulation value at different pHs followed the order of: acidic > neutral > alkaline, and alkaline > neutral > acidic, respectively. Such a difference was associated with the extent of hydrogen bond and electrostatic repulsion at different protonation/deprotonation states of carboxyl and phenolic -OH groups. Transmission electron microscopic imaging reveals that HA was predominantly globular aggregates with a rough periphery at pH 5.26, and was changed to smooth spherical particles at pH 10.00. These results are useful for better understanding the coagulation behaviors of HAs in both natural and engineered aqueous systems.

Effect of urea on phase transition of poly(N-isopropylacrylamide) investigated by differential scanning calorimetry.

The effect of urea on the phase transition of PNIPAM was studied using differential scanning calorimetry (DSC). For a certain urea concentration, the enthalpy change of phase transition of poly(N-isopropylacrylamide) (PNIPAM) aqueous solution increases with the number of DSC cycles, presumably due to the displacement of water molecules bound to the amide groups of PNIPAM by urea molecules at the temperature higher than the lower critical solution temperature (LCST) of PNIPAM and causes the decrease in the absolute value of the exothermic heat related to the dehydration of hydrophilic groups and interactions of hydrophilic residues to around 0. Moreover, the enthalpy change decreases with the urea concentration during the heating process of the first DSC cycle, indicating the replacement of water molecules around the apolar isopropyl groups by urea molecules at the temperature lower than LCST, and the endothermic heat caused by the dehydration of apolar groups decreases. Furthermore, the urea molecules which replace the water molecules at high temperature can be replaced again by water molecules at the temperature lower than LCST, but this process needs several days to complete.

Can Coil-to-Globule Transition of a Single Chain Be Treated as a Phase Transition?

The effects of the concentration (C) and heating rate on the collapse and association of poly(N-isopropylacrylamide) chains in water have been investigated by use of ultrasensitive differential scanning calorimetry. In the dilute solutions, both the phase transition temperature (Tp) and enthalpy change (DeltaH) increase with the heating rate but decrease with concentration. By extrapolation to zero heating rate and zero concentration, Tp and DeltaH for coil-to-globule transition of a single chain in thermodynamic equilibrium can be obtained. In semidilute solutions, both Tp and DeltaH increase with the heating rate but slightly vary with the concentration. Tp and DeltaH for pure interchain association in equilibrium are obtained by extrapolation to zero heating rate. Our experiments reveal that only intrachain contraction occurs when the concentration is infinitely close to zero. When the concentration is above the overlap concentration (C*), only interchain association exists. In the range 0

Hydration interactions and stability of soluble microbial products in aqueous solutions

Soluble microbial products (SMP) are organic compounds excreted by microorganisms in their metabolism and decay and the main constituents in effluent from biological wastewater treatment systems. They also have an important contribution to the dissolved organic matters in natural aqueous systems. So far the interactions between SMP colloids have not been well explored. In this work, the interactions between SMP colloids in water and salt solutions were studied by using a combination of static and dynamic light scattering, Fourier transform infrared spectra, Zeta potential and acid-base titration techniques. The second osmotic virial coefficient had a larger value in a 750-mM salt solution than that in a 50-mM solution, indicating that repulsion between SMP colloids increased with an increase in salt concentration, which is contrary with the classic Derjaguin, Landau, Verwey, and Overbeek (DLVO) theory. Such a repulsion was attributed to water structuring and enhanced by the accumulation of hydrophilic counter ions around SMP colloids and the formed hydration force. The repulsion and hydration effect led to the dispersing and deeper draining structure, accompanied by a decreased hydrodynamic radius and increased diffusion coefficient. This hydration force was related to so-called ion specific effect, and electrolyte sodium chloride had a more substantial effect on hydration force than KCl, CsCl, NaBr and NaI. Our results provide an experimental approach to explore the SMP structures, inter-colloid interactions and confirm the non-DLVO forces.

A comparative study of urea-induced aggregation of collapsed poly(N-isopropylacrylamide) and poly(N,N-diethylacrylamide) chains in aqueous solutions.

The urea-induced aggregation of poly(N-isopropylacrylamide) (PNIPAM) and poly(N,N-diethylacrylamide) (PDEAM) globules was studied by using a combination of static and dynamic light scattering. Our results have revealed that urea acting as a cross-linker via formation of two hydrogen bonds with the amide groups of PNIPAM and PDEAM in different globules causes the aggregation, and the aggregation of PNIPAM and PDEAM globules is a reaction-limited cluster-cluster aggregation (RLCA) process. The aggregates have a uniform sphere structure that may be due to the restructuring of the aggregates. The aggregation rate of PNIPAM globules is slower than that of PDEAM, which might mainly contribute to the reasons that the amides groups of PNIPAM have more chance to be inside the globules because of the formation of intra- and inter-hydrogen bonds and the smaller number density of the PNIPAM aggregates at the original time. When the aqueous urea solutions were cooled and reheated to 40 °C, the aggregation became faster than the first heating process, indicating that the urea molecules have replaced some water molecules binding to the amide groups at high temperature and some of the urea molecules remain interacting with the polymers even at the temperature lower than the cloud point temperature.

Spatial configuration of extracellular polymeric substances of Bacillus megaterium TF10 in aqueous solution

Configuration of extracellular polymeric substances (EPS) excreted by microorganisms is related greatly to the inherent properties of EPS, and has a significant effect on the physicochemical characteristics of microbial aggregates, such as activated sludge for wastewater treatment, as well as their interaction with other substances in aqueous systems. In this work, the spatial configuration of microbial EPS is characterized using laser light scattering (LLS) technique, with EPS extracted from Bacillus megaterium TF10 as an example. The combined utilization of static light scanning (SLS) and dynamic light scanning (DLS) offers an effective avenue to explore the EPS configuration in aqueous solution, thus enables a better understanding about the physicochemical properties of EPS. The results show that EPS exist in the form of colloids in neutral aqueous solution (pH 7.0) and that their shape is random coil with incompletely extending chains. The attraction interaction between EPS colloids is related with the high flocculability of B. megaterium TF10. The cryo-electron microscopy image further confirms the spherical shape of EPS colloids. The LLS approach offers a powerful and convenient tool for characterizing microbial EPS configuration and understanding their behaviors in biological wastewater treatment systems.

A facile one-pot strategy for preparation of small polymer nanoparticles by self-crosslinking of amphiphilic block copolymers containing acyl azide groups in aqueous media

In this paper, novel amphiphilic block copolymers, poly(N-isopropylacrylamide-co-4-(acyl azido) phenyl methacrylate)-block-poly(ethylene oxide), abbreviated as PEO-b-P(NIPAM-co-AAPMA), have been designed and synthesized by reversible addition-fragmentation chain transfer (RAFT) polymerization at room temperature. Then self-assembly of the block copolymers has been studied and small cross-linked polymer nanoparticles have been successfully prepared. The units of acyl azido methacrylate act as hydrophobic moieties and crosslinking sites, PEO chains act as stabilizers for the formed particles, and PNIPAM chains make the particles sensitive to temperature. It was found that the size of the polymer particles was related to the unit ratio of NIPAM and AAPAM and by tuning the unit ratio we could easily prepare small cross-linked polymer nanoparticles in aqueous media without using any additional crosslinking agents. The size of polymer nanoparticles was characterized to be about 17 nm by atomic force microscopy (AFM) and transmission electron microscopy (TEM). And the distribution of the particles is very narrow from the Laser light scattering (LLS) characterization.

pH-Induced Conformational Change and Dimerization of DNA Chains Investigated by Analytical Ultracentrifugation

pH-induced conformational change of i-motif DNA has been studied by analytical ultracentrifugation. As pH increases, the hydrodynamic radius of individual DNA chains in aqueous solutions prepared by being heat-treated suddenly increases while the molar mass is constant, indicating that the conformation changes from an i-motif to a random coil. When DNA concentrations are higher than 1.0 μM, relatively stable dimers are formed as pH sharply decreases from 7.5 to 4.5. Moreover, the weight percentage of the dimers increases with the initial DNA concentration. The study can help to understand the functions of the telomeres containing repeated cytosine-rich sequences and to develop DNA-based devices.