Green Synthesis of Reusable Adsorbents for the Removal of Heavy Metal Ions.

Industrial wastewater often contains heavy metals, like lead, copper, nickel, cadmium, zinc, mercury, arsenic, and chromium. Overdoses of heavy metals will impose a severe threat to human health. Adsorption is the most efficient way of wastewater treatment for eliminating heavy metals. A novel material-reusable hydrogel-based adsorbent was developed in overcoming the regeneration issue. The polyethylene glycol diacrylate-3-sulfopropyl methacrylate potassium salt (PEGDA-SMP) hydrogel performed an ion-exchange rate to remove heavy metals from wastewater in 30-120 min. The adsorption capacity of PEGDA-SMP increases the increasing pH of a solution, in which pH 5 reaches the maximum. Pseudo-second-order adsorption and the Langmuir adsorption model can fully describe the adsorption properties of PEGDA-SMP for heavy metals. PEGDA-SMP prefers to exchange Pb2+ through K+, and its adsorption capacity can achieve 263.158 mg/g. Ag+, Zn2+, Ni2+, and Cu2+ were 227.27, 117.647, 102.041, and 99.010 mg/g, respectively. The hydrated ionic radius of the heavy metal might play an essential role to affect the adsorption preference. The removal efficiency of heavy metals can approach over 95% for each heavy metal. PEGDA-SMP performs rapid desorption and reaches desorption equilibrium in 15 min. After 10 consecutive adsorption-desorption cycles, the adsorption capacity remained over 90%. The hydrogel developed in this study showed reversible heavy metal absorption. Therefore, excellent adsorption-desorption properties of PEGDA-SMP can be potentially extended to industrial wastewater for removing heavy metals.

Control of cell attachment on pH-responsive chitosan surface by precise adjustment of medium pH.

The purpose of this study is to demonstrate pH-responsive chitosan is able to control cell behavior in response to small changes in environmental pH, which is at useful pH suitable for recovering cultured cells without additional enzymatic treatment and extensive washing steps. HeLa cells attached and spread well on chitosan at pH 6.99 and 7.20. When the pH was increased to 7.65, over 90% of cells would rapidly detached from chitosan surface within 1 h. Similarly, fibronectin adsorbed on chitosan at pH 7.20 also rapidly desorbed after increasing the medium pH. Most importantly and interestingly, medium pH adjustment could be facilitated by altering environment pCO(2). It was found over 80% of HeLa cells could be recovered from chitosan surface within 1 h and the viability of detached cells was more than 95% by transferring the culture plate from incubator to atmospheric condition. Additionally, chitosan substrate could effectively control attachment/detachment of various types of cells including cell lines HaCaT, H1299, NIH-3T3, and primary corneal fibroblasts, indicating the technology described here is easily reproducible and should be promising for controlling rapid fibronectin adsorption/desorption and cell attachment/detachment for tissue engineering applications.

The role of adenosine receptor and caveolae-mediated endocytosis in oligonucleotide-mediated gene transfer.

We previously reported the preparation and characterization of ternary nanoparticles with the negative surface charge, which comprises histidine-conjugated polyallylamine (PAA-HIS)/DNA core complex and a single-stranded oligonucleotide outer layer, to transfect various cell lines. As a continued effort, here the investigations on the endocytotic mechanisms involved in the uptake of the oligonucleotide-coated PAA-HIS/DNA complexes are reported. Interestingly, these complexes showed enhanced transfection efficiency only when deoxyadenosine-containing oligonucleotides were deposited on the PAA-HIS/DNA complex surface. The addition of uncomplexed oligonucleotide, free adenosine and adenosine receptor antagonist significantly inhibited the transfection efficiency of oligonucleotide-coated PAA-HIS/DNA complexes. These results indicated that the oligonucleotide-coated PAA-HIS/DNA complexes could specifically recognize adenosine receptors on the cell surface and were taken up by adenosine receptor-mediated process. Uptake and transfection experiments with various endocytic inhibitors suggested that, after receptor/ligand binding, oligonucleotide-coated PAA-HIS/DNA/complexes were mainly internalized via caveolae-mediated pathway to result in effective intracellular processing for gene expression. In conclusion, both adenosine receptor and caveolae-mediated endocytosis play important roles in oligonucleotide-mediated gene transfer.

PEGylated guanidinylated polyallylamine as gene-delivery carrier.

A novel cationic co-polymer was developed by grafting poly(ethylene glycol) (PEG) on guanidinylated polyallylamine (PAA) for gene delivery. Characterization of PEG-g-guanidinylated PAA/DNA complexes demonstrated that particle size increased and surface charge decreased with increasing the amount of PEG. The results of cytotoxicity assay proved that grafted PEG could effectively decrease the cytotoxicity of the complexes. In transfection efficiency assay, HeLa cells treated with PEG(2)-g-guanidinylated PAA (formed with 17.5 µmol guanidinylated PAA and 2 µmol PEG)/DNA (0.2 µg EGFP plasmid) complexes showed a very high level of EGFP expression. In conclusion, combination of guanidinylation and PEGylation could effectively decrease the cytotoxicity and significantly increase the transfection efficiency of PAA.

A variable gene delivery carrier--biotinylated chitosan/polyethyleneimine.

A variable gene delivery system has been developed based on conjugating chitosan to biotin through a functionalized poly(ethylene glycol) (PEG) spacer, which can be used to further bind different molecules on the outer layer of a polymer/DNA complex by streptavidin (SA)-biotin linkage. In this study, TAT-conjugated SA was used as the model molecule to prove the conjugation function of the prepared complex. In addition, low-molecular-weight poly(ethyleneimine) (PEI) was added into the polymer/DNA complex to increase the transfection efficiency. The results of the luciferase assay show that the transfection efficiency of the prepared complex was significantly correlated with the amount of PEI and was further enhanced when TAT was conjugated to the complex by SA-biotin linkage. Considered to have negligible cytotoxic effects, the variable gene delivery complex prepared in this study would be of considerable potential as carriers for in vitro applications.

Polycation/DNA complexes coated with oligonucleotides for gene delivery.

Ternary nanoparticles with negatively charged surface were prepared by coating single-stranded oligonucleotides (5'-C(10)A(20)-3') on histidine-conjugated polyallylamine (PAA-HIS)/DNA complexes for gene delivery. Characterization of PAA-HIS/DNA/oligonucleotide complexes demonstrated that nanoparticles possessed the negative surface charge -27 mV and size of around 100 nm when the molar ratio of oligonucleotide/PAA-HIS exceeded 1.5. The negatively charged oligonucleotide-coated PAA-HIS/DNA complexes could be entirely internalized by the living HeLa cells to exhibit high gene expression with low cytotoxicity and the resistance against erythrocyte agglutination and serum inhibition. Since the gene expression of PAA-HIS/DNA complexes was significantly inhibited by coating other polyanions and oligonucleotides, the ternary PAA-HIS/DNA/deoxyadenosine-rich oligonucleotide complexes were uptaken by specific receptor-mediated process. Additionally, the deposition of a layer of oligonucleotides onto the binary PAA-HIS/DNA complexes could effectively transfect various types of cells including HEK-293, HepG2 and Hs68 cells, indicating the technology of coating specific oligonucleotides on PAA-HIS/DNA complexes or other cationic binary DNA complexes might facilitate the use of nanoparticles for safe and efficient gene delivery and eventual therapy.

Change in neuron aggregation and neurite fasciculation on EVAL membranes modified with different diamines.

In this study, we modified poly(ethylene-co-vinyl alcohol) (EVAL) membranes by the covalent bonding of diamines via epoxidation of surface hydroxyl groups of EVAL to analyze the effect of immobilized diamines with different carbon chain length on the cultured cerebellar granule neurons. Morphological studies showed that neurons seeded on the diamine-immobilized EVAL membrane were able to survive and regenerate with formation of an extensive neuritic network. Furthermore, cultured neurons showed that the presence of diamine with different carbon chain length was able to effectively regulate the neuron adhesion, migration, aggregation, and neurite growth pattern, but mediated neuronal activity with equal efficacy. The short-chain amine stimulated neuron migration, aggregation, and neurite fasciculation, whereas the long carbon chain diamine maintained single neuron distribution with the defasciculated feature of the neurite. Although it is known that positively charged amine molecules can interact directly with cell surface proteoglycans to mediate cell attachment, this study further demonstrated that the terminal primary amine with different carbon chain length is involved in mediating cell-substrate interaction to further regulate neuron aggregation and neurite fasciculation. This indicates a delicate interaction of neuron with the immobilized diamine molecules on the EVAL membrane surface. This work is encouraging because the diamine- immobilized EVAL membranes can be applied for the establishment of different neural culture systems useful for future investigations of neuron biology under in vitro conditions.

PMMA particle-mediated DNA vaccine for cervical cancer.

DNA vaccination is a novel immunization strategy that possesses many potential advantages over other vaccine strategies. One of the major difficulties hindering the clinical application of DNA vaccination is the relative poor immunogenicity of DNA vaccines. Poly(methyl methacrylate) (PMMA) is a synthetic polymer approved by the Food and Drug Administration for certain human clinical applications such as the bone cement. In vivo, PMMA particles are phagocytosable and have the potential to initiate strong immune responses by stimulating the production of inflammatory cytokines. In this study, we synthesized a series of PMMA particles (PMMA 1-5) with different particle sizes and surface charges to test the feasibility of implementing such polymer particles for DNA vaccination. To our knowledge, this is the first report to show that the gene gun can deliver DNA vaccine by propelling PMMA particles mixed with plasmid DNA for cervical cancer. It was found that PMMA 4 particles (particle size: 460 +/- 160 nm, surface charge: +11.5 +/- 1.8 mV) stimulated the highest level of TNF-alpha production by macrophages in vitro and yielded the best result of antitumor protection in vivo. Therefore, our results possess the potential for translation and implementation of polymer particles in gene gun delivering DNA vaccination.