Preparation and Characterization of pH-Responsive Charge Reversal Nanocomposite for miRNA Delivery

pH-responsive charge reversal loaded miRNA nanocomposite was prepared by electrostatic self-assembly.The morphology,particle size and zeta potential of the nanocomposites were analyzed by transmission electron microscopy and dynamic light scattering.The synthesis of the polymer was analyzed by^(1)H-NMR.The zeta-potential changes and cellular uptake effects of the nanocomplexes under different pH environments were investigated.The experimental results show that the surface morphology of the nanocomposite is spherical,and the average particle size is about 135 nm.As the pH value of the solution gradually decreases,the surface charge of the nanocomposite reverses from negative charge to positive charge(from-9.4 to+17.1 mV).Cellular uptake mediated by pH-responsive nanocomposite is selective for tumor cells,and the cellular uptake effect in tumor cells at pH 6.5 was approximately 3 times higher than that at pH 7.4.This pH responsive charge reversal nanocomposite has promising application prospects for gene delivery in the weak acid environment of tumors.

A high strength pH responsive supramolecular copolymer hydrogel

Constructing high strength pH sensitive supramolecular polymer hydrogel remains very challenging due to the unavoidable network swelling caused by ionization of acid or basic groups at a specified pH.In this work,we proposed a simple and very convenient approach to fabricate high strength pH responsive supramolecular polymer(SP) hydrogels by one-pot copolymerization of N-acryloyl glycinamide(NAGA) and 2-vinyl-4,6-diamino-1,3,5-triazine(VDT),two feature hydrogen bonding monomers.In these PNAGA-PVDT SP hydrogels obtained,the hydrogen bonding of NAGA was shown to play a dominant role in reinforcing strength,while the hydrogen bonding of diaminotriazine served as a pH sensitive moiety.At pH 3,the mechanical properties of PNAGA-PVDT hydrogels decreased to a different extent due to the breakup of hydrogen bonding;in contrast,the hydrogel resumed the original strength while pH was raised to 7.4 because of reconstruction of hydrogen bonding.Over the selected pH range,the PNAGA-PVDT hydrogels exhibited up to 1.25 MPa tensile strength,845% breaking strain,69 kPa Young's modulus and 21 MPa compressive strength.This novel high strength pH-responsive SP hydrogels may find applications in biomedical and industrial fields.

pH responsive Janus polymeric nanosheets

pH responsive polymeric Janus nanosheets with poly（maleic acid） moiety and crosslinked PS onto the corresponding sides have been synthesized by free radical polymerization. The Janus nanosheets can serve as solid emulsifier to stabilize an oil/water emulsion, whose stability is easily triggered by changing pH across pKa of the poly（maleic acid）.

Antibiotic-based small molecular micelles combined with photodynamic therapy for bacterial infections

The appearance of multidrug-resistant bacteria and the formation of bacterial biofilms have necessitated the development of alternative antimicrobial therapeutics.Antibiotics conjugated with or embedded in nano-drug carriers show a great potential and advantage over free drugs,but the mass proportion of carriers generally exceeds 90%of the nano-drug,resulting in low drug loading and limited therapeutic output.Herein,we fabricated a nanocarrier using antibiotics as the building blocks,minimizing the use of carriermaterials,significantly increasing the drug loading content and treatment effect.Firstly,we conjugated betaine carboxylate with ciprofloxacin(CIP)through an ester bond to form the amphiphilic conjugate(CIP-CB),which self-assembled into micelles(CIP-CBMs)in aqueous solutions,with a CIP loading content as high as 65.4%and pH-induced surface charge reversal properties.Secondly,a model photosensitizer(5,10,15,20-tetraphenylporphyrin(TPP))was encapsulated in CIP-CBMs,generating infection-targeted photodynamic/antibiotic combined nanomedicines(denoted as TPP@CIP-CBMs).Upon accumulation at infection sites or in deep bacterial biofilms,the ester bond between the betaine carboxylate and CIP is cleaved to release free TPP and CIP,leading to a synergetic antibacterial and antibiofilm activity in vitro and in vivo.

Ultrafast Infrared Spectroscopic Study of Microscopic Structural Dynamics in pH Stimulus-Responsive Hydrogels

Hydrogels show versatile properties and are of great interest in the fields of bioelectronics and tissue engineering.Understanding the dynamics of the water molecules trapped in the three-dimensional polymeric networks of the hydrogels is crucial to elucidate their mechanical and swelling properties at the molecular level.In this report,the poly(DMAEMA-co-AA)hydrogels were synthesized and characterized by the macroscopic swelling measurements under different pH conditions.Furthermore,the microscopic structural dynamics of pH stimulus-responsive hydrogels were studied using FTIR and ultrafast IR spectroscopies from the viewpoint of the SCN-anionic solute as the local vibrational reporter.Ultrafast IR spectroscopic measurements showed the time constants of the vibrational population decay of SCN-were increased from 14±1 ps to 20±1 ps when the pH of the hydrogels varied from2.0 to 12.0.Rotational anisotropy measurements further revealed that the rotation of SCNanionic probe was restricted by the three-dimensional network formed in the hydrogels and the rotation of SCN-anionic probe cannot decay to zero especially at the pH of 7.0.These results are expected to provide a molecular-level understanding of the microscopic structure of the cross-linked polymeric network in the pH stimulus-responsive hydrogels.

Molecular origin for pH-responsive wormlike micelles of zwitterionic surfactants triggered by aromatic acid derivatives

Introduction of aromatic acid derivatives(AADs)into zwitterionic surfactants is an efficient method to prepare wormlike micelles with pH-controllable viscosity;however,the coincident molecular origin of AAD/zwitterionic surfactant binary mixtures remains unclear.Herein,the self-assembly of hydroxyl derivatives of benzoic acid(BA)and cetyldimethyl betaine(BS-16)mixtures in water was systematically assessed,and various factors,such as the molecular structure,molar ratio of AAD and BS-16,and solution pH,were investigated.The structure-property relationship of AAD/BS-16 binary mixtures was established,which provided the molecular origin for the effect of AAD on micellar microstructures and the pH-induced morphological transitions.The ortho-substituted hydroxyl moiety in the BA molecule facilitated the formation of larger wormlike micelles,whereas the effect of the meta-substituted moiety was less significant.The para-substituted hydroxyl moiety in BA did not favor micellar growth.This moiety exhibited similar characteristics to the increasing hydroxyl moiety number in the AAD molecules or solution pH where the negative effects of steric hindrance and electrostatic interactions of molecules in micelles aredominant.

PAM-PAA microgel inverse opal photonic crystal and pH response

The colloidal crystal template or opal with a closed-packed face-centered cubic （fcc） lattice was prepared from monodisperse polystyrene （PS） spheres by vertical sedimentation. The template provided void space for infiltration of monomer precursor composed of acrylate acid, acrylamide and ammonium-persulfate, as well as microgel from the subsequent copolymerization. The sample was immersed in dimethylbenzene for completely removing PS spheres to form PAM inverse opal hydrogels （IOHPAM） or PAM/PAA inverse opal hydrogels （IOHPAM/PAA） photonic crystals. The PS spheres were replaced by air spheres, which interconnected each other through the windows. The study of responses to pH show that there are two peaks for both IOHPAM and IOHPAM/PAA films, but the IOHPAM/PAA peaks shift to higher pH, and the peaks are independent with the AA content.

mtDNA-triggered pH response signal-amplified fluorescent probe for multiple cell discrimination

Developing fluorescence probes with multiple responses has vital significance but remains challenging.Herein,for the first time,we present a mitochondrial DNA(mtDNA)-triggered pH response signalamplified fluorescent probe(QCY-DBT)for multiple cell detection.The probe exhibited a large stokes shift(229 nm),excellent DNA selectivity over RNA,and ultrasensitivity of detection limit(DL;74.0 ng/mL).Thus,QCY-DBT was successfully applied to analyze multiple human peripheral blood cells and visualize mtDNA in healthy and apoptotic cells.In the tumor acidic environment(pH 6.0–7.0),the absorbance of QCY-DBT at 436 nm increased,and the fluorescence signal(665 nm)was amplified by mtDNA,which enabled the direct observation of tumor cells.Our study provides help in designing smart probes with multiple responses for efficient abnormal cell detection.

Development of NIR-II fluorescence image-guided and pH-responsive nanocapsules for cocktail drug delivery

Nanocapsule-based targeted delivery and stimulus-responsive release can increase drug effectiveness, while reducing the side effects of the drug. However, difficulties in the scale-up synthesis, fast burst release, and low degradability, could hamper the translation of drug nanocapsules from lab to clinic. Here we have controllably functionalized the biodegradable and widely available polysuccinimide, in order to obtain an amphiphilic poly（amino acid）. Using this polymer, we designed nanocapsules （〈 100 nm） for hydrophobic drug delivery, which could facilitate tumor targeting, hydrogen bond-based pH-responsive release, and real-time fluorescence tracking, in the second near-infrared region. This method is versatile, eco-friendly, and easy to scale up at low costs. In addition, this system can carry a cocktail of drugs, obtained by loading multiple anticancer drugs to the same vehicle. Our nanocapsules were observed to be stable in blood vessels （pH = 7.4）, and the pH-responsive release （pH = 5.0 in lysosome） was sustained. The chemotherapy results in tumor-xenografted mice suggested that our nanocapsule was safe and efficient, and may be a useful tool for drug delivery.

Doxorubicin-conjugated pH-responsive gold nanorods for combined photothermal therapy and chemotherapy of cancer

Cancer chemotherapy can be hindered by drug resistance which leads to lower drug efficiency.Here,we have developed a drug delivery system that tethers doxorubicin to the surface of gold nanorods via a pHsensitive linkage(AuNRs@DOX),for a combined photothermal and chemical therapy for cancer.First,AuNRs@DOX is ingested by HepG2 liver cancer cells.After endocytosis,the acidic pH triggers the release of doxorubicin,which leads to chemotherapeutic effects.The gold nanorods are not only carriers of DOX,but also photothermal conversion agents.In the presence of an 808 nm near-infrared laser,AuNRs@DOX significantly enhance the cytotoxicity of doxorubicin via the photothermal effect,which induces elevated apoptosis of hepG2 cancer cells,leading to better therapeutic effects in vitro and in vivo.

Continuous Detection of pH-responsive Drug Delivery System in Cells in situ by Confocal Laser Scanning Microscopy

Mesoporous silica nanoparticles （MSN） were coated by pH-responsive polymer chitosan-poly （methacrylic acid） （CS-PMAA）. This nano drug delivery system showed good application prospects and the polymer-coated microspheres were promising site-specific anticancer drug delivery carriers in biomedical field. A continuous detection of pH-responsive drug delivery system in cells in situ, utilizing MSN/CS-PMAA composite microspheres, was pro- posed. Two kinds of different cell lines, tumor cell line （Hela） and normal somatic cells （293T）, were used to inves- tigate the behaviours of the drug loaded system in the cells. Conclusions could be drawn from the fluorescent im- ages obtained by confocal laser scanning microscopy （CLSM）, modified drug-loaded microspheres （MSN/CS-PMAA） were ingested into cells more easily, the uptake of DOX@FITC-MSN/CS-PMAA by HeLa/293T cells were performed at pH 7.4/pH 6.8, DOX was released during the ingestion process, fluorescence intensity de- creased with time because of efflux transport and photo-bleaching. Fluoresence detection by flow cytometry was performed as comparison. The continuous fluorescent observation in situ could be widely used in the pH-responsive releasing process of drug delivery system in the cells.

Enhanced Viscosity of Poly（acrylamide）Solution in the Presence of Chromium Citrate Triggered by Release of CO2

Partially hydrolyzed polyacrylamide （HPAM） has been widely used for water shut-off and profile control to enhance oil recovery. Herein, we reported a novel technique by which the crosslinking between HPAM and Cr3＋ in aqueous solutions at 60 ℃ can be delayed effectively. Citric acid was selected as an organic complexing agent of Cr3＋ so that the crosslinking between HPAM and Cr3＋ can be prevented completely. Due to the decomposi- tion of the bicarbonate （HCO3-） embedded in solution, CO2 released from solution and the pH value of solution increased gradually. The degree of ionization of HPAM and its ability to complex with Cr3＋ increased accordingly. When the complexation of Cr3＋ with HPAM is stronger than that with citric acid, the viscosity of the HPAM solution increased signif- icantly. Under the closed condition, together with the existence of potassium dihydrogen phosphate （KH2PO4）, the release of CO2 was very slow and the condition was highly con- trolled so that the ionization of HPAM was prevented initially. Furthermore, the hydrogen bonding interactions between HPAM and melamine embedded in solution previously also postponed the ionization of HPAM. As a result, the crosslinking between HPAM and Cr3＋ can be delayed for almost one month, completely meeting the requirements for deep water shut-off and profile control to enhance oil recovery.

Release of Anticancer Drug 5-Fluorouracil from Different Ionically Crosslinked Alginate Beads

In this research;the release of 5-Fluorouracil (5-FU) from different ionically crosslinked alginate (Alg) beads was investigated by using Fe3+, Al3+, Zn2+, and Ca2+, ions as crosslinking agent. The prepared beads were characterized by Fourier Transform Infrared Spectroscopy (FTIR) Differential Scanning Calorimetry (DSC) and Scanning Electron Micros-copy (SEM). The drug release studies were carried out at three pH values 1.2, 6.8 and 7.4 respectively each for two hours. The effects of the preparation conditions as crosslinker type, drug/polymer (w/w) ratio, crosslinker concentration and time of exposure to crosslinker on the release of 5-FU were investigated for 6 hours at 37℃. It was observed that 5-FU release from the beads followed the order of Fe > Zn > Al > Ca-Alg and increased with increasing drug/polymer ratio. At the end of 6 hours, the highest 5-FU release was found to be 90% (w/w) for Fe-Alg beads at the drug/polymer ratio of 1/8 (w/w), crosslinker concentration of 0.05 M, exposure time of 10 minutes respectively. The swelling measurements of the beads supported the release results. Release kinetics was described by Fickian and non-Fickian approaches.

Smart Polymers and Coatings Obtained by Ionizing Radiation: Synthesis and Biomedical Applications

Gamma radiation has been shown particularly useful for the functionalization of surfaces with stimuli-responsive polymers. This method involves the formation of active sites (free radicals) onto the polymeric backbone as a result of the exposition to high-energy radiation, in which a proper microenvironment for the reaction among monomer and/or polymer and the active sites takes place, thus leading to propagation which forms side chain grafts. The modification of polymers using high-energy irradiation may be performed by the following methods: direct or simultaneous, pre-irradiation oxidative and pre-irradiation. The most frequent ones correspond to the pre-irradiation oxidative method and the direct one. Radiation-grafting has many advantages over conventional methods considering that it does not require catalyst nor additives to initiate the reaction, and in general, no changes on the mechanical properties with respect to the pristine polymeric matrix are observed. This chapter focused on the synthesis of smart polymers and coatings obtained by the use of gamma radiation. In addition, diverse applications of these materials in the biomedical field are also reported.

Antibacterial and Alkali-responsive Cationic Waterborne Polyurethane Based on Modification of Aloe Emodin

Cationic water-based polyurethane(CWPU)was synthesized to explore aloe-emodin modifies to obtain CWPU materials with better comprehensive performance.It provides a simple way to synthesize antibacterial waterborne polyurethane,which is to introduce the end-blocking group of herbal extracts into the structure.It contains synergistic antibacterial effect of herbal antibacterial and quaternary ammonium ion on Escherichia coli.It makes the material resist the erosion of bacterial,and increase the service life of materials.When the pH value of the environment changes,the UV absorbance of the aloe-emodin modified cationic water-based polyurethane(AE-CWPU)also changes.Therefore,within a certain detection range,AE-CWPU has great applications in the field of smart response materials.The modified thermodynamic properties have been improved,and the mechanical properties basically maintained the maximum stress,and the elongation at break was reduced.

Synthesis of pH-responsive triazine skeleton nano-polymer composite containing AIE group for drug delivery

We exploited a unique porous structure of the nano-covalent triazinepolymer(NCTP)containing aggregation-induced emission(AlE)group to achievecontrolled release and drug tracking in tumor acidic microenvironment.NCTP wassynthesized by the Friedel-Crafts alkylation and the McMurry coupling reaction.It notonly had strong doxorubicin(DOX)-loading capacity due to its high specific surface areaand large pore volume,but also showed the significant cumulative drug release as aresult of the pH response of triazine polymers.NCTP was induced luminescence aftermass accumulation near tumor cells.Besides,it had excellent biocompatibility andobvious antineoplastic toxicity.The results demonstrate that NCTP as a utility-type drugcarrier provides a new route for designing the multi-functional drug delivery platform.

Charge-guided masking of a membrane-destabilizing peptide enables efficient endosomal escape for targeted intracellular delivery of proteins

Intracellular delivery of biologicals such as peptides,proteins,and nucleic acids presents a great opportunity for innovative therapeutics.However,the endosome entrapment remains a major bottleneck in the intracellular delivery of biomacromolecules,largely limiting their therapeutic potential.Here,we converted a cell-penetrating peptide(CPP),low molecular weight protamine(LMWP),to endosomal escape peptides(EEPs)by masking LMWP with a pH-responsive counter-ionic peptide.The resulting masked CPPs(mLMWP and mLMWP2)effectively promoted the escape of peptide/protein cargoes from endosomes into the cytoplasm.Consequential lysosome repair and lysophagy were initiated upon the endolysosomal leakage.Minimal reactive oxygen species(ROS)elevation or cell death was observed.Based on mLMWP2,we constructed an intracellular protein delivery system containing an antibody as a targeting module,mLMWP2 as an endosomal escape module,and the desired protein cargo.With the HER2-targeting delivery system,we efficiently translocated cyclization recombination enzyme(Cre)and BH3-interacting domain death agonist(BID)into the cytosol of HER2^(+)cells to exert their biological activity.Thereby,the modular delivery system shows its potential as a promising tool for scientific studies and therapeutic applications.

Biomimetic inorganic-organic hybrid nanoparticles from magnesium-substituted amorphous calcium phosphate clusters and polyacrylic acid molecules

Amorphous calcium phosphate(ACP)has been widely found during bone and tooth biomineralization,but the meta-stability and labile nature limit further biomedical applications.The present study found that the chelation of polyacrylic acid(PAA)molecules with Ca^(2+)ions in Mg-ACP clusters(~2.1±0.5 nm)using a biomineralization strategy produced inorganic-organic Mg-ACP/PAA hybrid nanoparticles with better thermal stability.Mg-ACP/PAA hybrid nanoparticles(~24.0±4.8 nm)were pH-responsive and could be efficiently digested under weak acidic conditions(pH 5.0-5.5).The internalization of assembled Mg-ACP/PAA nanoparticles by MC3T3-E1 cells occurred through endocytosis,indicated by laser scanning confocal microscopy and cryo-soft X-ray tomography.Our results showed that cellular lipid membranes remained intact without pore formation after Mg-ACP/PAA particle penetration.The assembled Mg-ACP/PAA particles could be digested in cell lysosomes within 24 h under weak acidic conditions,thereby indicating the potential to efficiently deliver encapsulated functional molecules.Both the in vitro and in vivo results preliminarily demonstrated good biosafety of the inorganic-organic Mg-ACP/PAA hybrid nanoparticles,which may have potential for biomedical applications.

Reversibly size-switchable polyion complex micelles for antiangiogenic cancer therapy

Size is one of the most important characteristics of nanoparticles to influence their biodistribution and antitumoral efficacy.Particles with large sizes have difficulty in deep tumor penetration,while small particles are easily removed from tumor tissues due to the high tumor interstitial fluid pressure.To address these issues,an intelligent core-crosslinked polyion complex micelle(cPCM)with a reversibly sizeswitchable feature was engineered in this study.The micelles are consisting of methoxy poly(ethylene glycol)-poly(D,L-lactide)copolymer(mPEG-PLA),mPEG-PLA-(HE)6CC,and mPEG-PLA-(RG)6CC at an optimal mass ratio of 6:1:1 with an antiangiogenic compound,dabigatran etexilate(DE),encapsulated.The net charge inside the micelles is switchable when exposed to different pH conditions,thereby leading to revisable size-change of micelles.DE-loaded micelles(DE@cPCM)can swell and release drugs at the tumor sites with a mildly acidic pH,while they shrink and protect the cargo from leaking into the blood circulation with a neutral pH.Results indicated that DE@cPCM can inhibit tumor angiogenesis in vitro and in vivo,thereby efficiently restraining tumor growth in a 4T1-bearing mouse model.Collectively,the sizeswitchable cPCM is a promising nanoplatform for targeting delivery of anticarcinogens into the matrix of tumor tissues.

Responsive Composite Janus Cages

pH responsive composite Janus cages of PS-silica-PEO are prepared using silica nanoparticles to crosslink two PAA contained di-block copolymers at an emulsion interface sequentially. Transverse channels are in situ created across the shell, which facilitates mass transfer. Shell is pH responsive owing to the presence of residual PAA chains tethered onto the silica nanoparticles. The Janus cages are capable to preferentially capture oil from their aqueous surroundings. The saturation absorption capacity is determined by pH and oil property. Other composite Janus cages are expected by extension the method using other copolymers and functional nanoparticles.