Ceria-Containing Hybrid Multilayered Microcapsules for Enhanced Cellular Internalisation with High Radioprotection Efficiency.

Cerium oxide nanoparticles (nanoceria) are believed to be the most versatile nanozyme, showing great promise for biomedical applications. At the same time, the controlled intracellular delivery of nanoceria remains an unresolved problem. Here, we have demonstrated the radioprotective effect of polyelectrolyte microcapsules modified with cerium oxide nanoparticles, which provide controlled loading and intracellular release. The optimal (both safe and uptake efficient) concentrations of ceria-containing microcapsules for human mesenchymal stem cells range from 1:10 to 1:20 cell-to-capsules ratio. We have revealed the molecular mechanisms of nanoceria radioprotective action on mesenchymal stem cells by assessing the level of intracellular reactive oxygen species (ROS), as well as by a detailed 96-genes expression analysis, featuring genes responsible for oxidative stress, mitochondrial metabolism, apoptosis, inflammation etc. Hybrid ceria-containing microcapsules have been shown to provide an indirect genoprotective effect, reducing the number of cytogenetic damages in irradiated cells. These findings give new insight into cerium oxide nanoparticles' protective action for living beings against ionising radiation.

Application of Gamma Radiation on Hard Gelatin Capsules as Sterilization Technique and Its Consequences on the Chemical Structure of the Material.

Hard capsules are made from gelatin, an organic polymer obtained through the hydrolysis of collagen present in animal tissues. Gelatin can be degraded by microorganisms and some strategies can be used to control contaminating micro-organisms. Gamma irradiation is considered as an effective sterilization method; however, its application can alter the chemical structure of the irradiated product. Samples of hard gelatin capsules were irradiated at doses of 5, 15, and 25 kGy at room temperature. The characterizations of the physical and chemical effects were evaluated by scanning electron microscopy, Fourier-transform infrared spectroscopy, X-ray diffractometry, and differential scanning calorimetry techniques. Furthermore, hard gelatin capsule samples were dissolved and inoculated with Bacillus subtilis, a Gram-positive spore-forming bacterium, to evaluate the effect of gamma ray radiation on bacterial counts. The results showed that gamma radiation did not interfere on physical parameters of the capsule, such as moisture content, mass, body and cap length, and disintegration time. Nevertheless, differential scanning calorimetry results demonstrated changes in the glass transition temperature, indicating the formation of crosslinking in irradiated capsules. It was observed that there were significant reductions on the inoculated bacterial population starting from the lowest irradiation dose and there was no detection of bacterial growth from the 15 kGy dose, while in the non-irradiated samples were found with 104 CFU mL-1 of bacteria. Therefore, this work concludes that the gamma radiation is effective on the reduction of the microbial population, cause discrete physical-chemical alterations, and could be used as a hard capsule sterilization technique.

Synthesis and Characterization of Stimuli-Responsive Poly(2-dimethylamino-ethylmethacrylate)-Grafted Chitosan Microcapsule for Controlled Pyraclostrobin Release.

Controllable pesticide release in response to environmental stimuli is highly desirable for better efficacy and fewer adverse effects. Combining the merits of natural and synthetic polymers, pH and temperature dual-responsive chitosan copolymer (CS-g-PDMAEMA) was facilely prepared through free radical graft copolymerization with 2-(dimethylamino) ethyl 2-methacrylate (DMAEMA) as the vinyl monomer. An emulsion chemical cross-linking method was used to expediently fabricate pyraclostrobin microcapsules in situ entrapping the pesticide. The loading content and encapsulation efficiency were 18.79% and 64.51%, respectively. The pyraclostrobin-loaded microcapsules showed pH-and thermo responsive release. Microcapsulation can address the inherent limitation of pyraclostrobin that is photo unstable and highly toxic on aquatic organisms. Compared to free pyraclostrobin, microcapsulation could dramatically improve its photostability under ultraviolet light irradiation. Lower acute toxicity against zebra fish on the first day and gradually similar toxicity over time with that of pyraclostrobin technical concentrate were in accordance with the release profiles of pyraclostrobin microcapsules. This stimuli-responsive pesticide delivery system may find promising application potential in sustainable plant protection.

Treatment of Parkinson's disease in rats by Nrf2 transfection using MRI-guided focused ultrasound delivery of nanomicrobubbles.

Parkinson's disease (PD) is a very common neurological disorder. However, effective therapy is lacking. Although the blood-brain-barrier (BBB) protects the brain, it prevents the delivery of about 90% of drugs and nucleotides into the brain, thereby hindering the development of gene therapy for PD. Magnetic resonance imaging (MRI)-guided focused ultrasound delivery of microbubbles enhances the delivery of gene therapy vectors across the BBB and improves transfection efficiency. In the present study, we delivered nuclear factor E2-related factor 2 (Nrf2, NFE2L2) contained in nanomicrobubbles into the substantia nigra of PD rats by MRI-guided focused ultrasound, and we examined the effect of Nrf2 over-expression in this animal model of PD. The rat model of PD was established by injecting 6-OHDA in the right substantia nigra stereotactically. Plasmids (pDC315 or pDC315/Nrf2) were loaded onto nanomicrobubbles, and then injected through the tail vein with the assistance of MRI-guided focused ultrasound. MRI-guided focused ultrasound delivery of nanomicrobubbles increased gene transfection efficiency. Furthermore, Nrf2 gene transfection reduced reactive oxygen species levels, thereby protecting neurons in the target region.

Microcapsules and 3D customizable shelled microenvironments from laser direct-written microbeads.

Microcapsules are shelled 3D microenvironments, with a liquid core. These core-shelled structures enable cell-cell contact, cellular proliferation and aggregation within the capsule, and can be utilized for controlled release of encapsulated contents. Traditional microcapsule fabrication methods provide limited control of capsule size, and are unable to control capsule placement. To overcome these limitations, we demonstrate size and spatial control of poly-l-lysine and chitosan microcapsules, using laser direct-write (LDW) printing, and subsequent processing, of alginate microbeads. Additionally, microbeads were used as volume pixels (voxels) to form continuous 3D hydrogel structures, which were processed like capsules, to form custom shelled aqueous-core 3D structures of prescribed geometry; such as strands, rings, and bifurcations. Heterogeneous structures were also created with controlled initial locations of different cell types, to demonstrate the ability to prescribe cell signaling (heterotypic and homotypic) in co-culture conditions. Herein, we demonstrate LDW's ability to fabricate intricate 3D structures, essentially with "printed macroporosity," and to precisely control structural composition by bottom-up fabrication in a bead-by-bead manner. The structural and compositional control afforded by this process enables the creation of a wide range of new constructs, with many potential applications in tissue engineering and regenerative medicine. Biotechnol. Bioeng. 2016;113: 2264-2274. © 2016 Wiley Periodicals, Inc.

Near-infrared femtosecond laser-triggered nanoperforation of hollow microcapsules.

Fabrication of a nanopore in a hollow microcapsule was demonstrated using near-infrared femtosecond laser irradiation. The shape of the irradiated microcapsules was kept spherical except for a pore in the shell owing to the nonthermal processing by a femtosecond laser. The simulation results for the near-field and far-field scattering around a microcapsule revealed that highly-enhanced optical intensity can be generated at a spot on the shell of a microcapsule, which would in turn contribute to localized ablation. To the best of our knowledge, this is the first demonstration of the nanoperforation of transparent hollow microcapsules by a near-infrared laser without any doping with absorbing metals or dyes that may cause cell toxicity. The presented method is a promising approach for safer drug delivery and the controlled release of therapeutic drugs.

Fabrication of a dual pH-responsive and photothermal microcapsule pesticide delivery system for controlled release of pesticides.

BACKGROUND: The development of stimulus-responsive and photothermally controlled-release microcapsule pesticide delivery systems is a promising solution to enhance the effective utilization and minimize the excessive use of pesticides in agriculture. RESULTS: In this study, an AVM@CS@TA-Fe microcapsule pesticide delivery system was developed using avermectin as the model drug, chitosan and tannic acid as the wall materials, and tannic acid-Fe complex layer as the photothermal agent. The optical microscope, scanning electron microscope, transmission electron microscope, and Fourier-transform infrared spectroscope were used to characterize the prepared microcapsule. The slow-release, UV-shielding, photothermal performance, and nematicidal activity of the microcapsule were systematically investigated. The results showed that the system exhibited excellent pH-responsive and photothermal-sensitive performances. In addition, the UV-shielding performance of the delivery system was improved. The photothermal conversion efficiency (η) of the system under the irradiation of near-infrared (NIR) light was determined to be 14.18%. Moreover, the nematicidal activities of the system against pine wood nematode and Aphelenchoides besseyi were greatly increased under the irradiation of light-emitting diode (LED) simulated sunlight. CONCLUSION: The release of the pesticide-active substances in such a pesticide delivery system could be effectively regulated with the irradiation of NIR light or LED-simulated sunlight. Thus, the developed pesticide delivery system may have broad application prospects in modern agriculture fields. © 2022 Society of Chemical Industry.

Surface-supported multilayers decorated with bio-active material aimed at light-triggered drug delivery.

In this work, we report on the functionalization of layer-by-layer films with gold nanoparticles, microcapsules, and DNA molecules by spontaneous incorporation into the film. Exponentially growing films from biopolymers, namely, hyaluronic acid (HA) and poly-L-lysine (PLL), and linearly growing films from the synthetic polymers, namely, poly(styrene sulfonate) (PSS) and poly(allylamine hydrochloride) (PAH), were examined for the embedding. The studied (PLL/HA)(24)/PLL and (PAH/PSS)(24)/PAH films are later named HA/PLL and PSS/PAH films, respectively. The HA/PLL film has been found to be more efficient for both particle and DNA embedding than PSS/PAH because of spontaneous PLL transport from the interior of the whole HA/PLL film to the surface in order to make additional contact with embedded particles or DNA. DNA and nanoparticles can be immobilized in HA/PLL films, reaching loading capacities of 1.5 and 100 microg/cm(2), respectively. The capacities of PSS/PAH films are 5 and 12 times lower than that for films made from biopolymers. Polyelectrolyte microcapsules adsorb irreversibly on the HA/PLL film surface as single particles whereas very poor interaction was observed for PSS/PAH. This intrinsic property of the HA/PLL film is due to the high mobility of PLL within the film whereas the structure of the PSS/PAH film is "frozen in". Gold nanoparticles and DNA form micrometer-sized aggregates or patches on the HA/PLL film surface. The diffusion of nanoparticles and DNA into the HA/PLL film is restricted at room temperature, but DNA diffusion is triggered by heating to 70 degrees C, leading to homogeneous filling of the film with DNA. The film has not only a high loading capacity but also can be activated by "biofriendly" near-infrared (IR) laser light, thanks to the gold nanoparticle aggregates on the film surface. Composite HA/PLL films with embedded gold nanoparticles and DNA can be activated by light, resulting in DNA release. We assume that the mechanism of the release is dependent on the disturbance in bonding between "doping" PLL and DNA, which is induced by local thermal decomposition of the HA/PLL network in the film when the film is exposed to IR light. Remote IR-light activation of dextran-filled microcapsules modified by gold nanoparticles and integrated into the HA/PLL film is also demonstrated, revealing an alternative release pathway using immobilized light-sensitive carriers (microcapsules).

Preparation and performance of a BTDA-modified polyurea microcapsule for encapsulating avermectin.

A pesticide microcapsule was prepared by encapsulating avermectin (AVM) in a polyurea microcapsule via interfacial polymerization in acetic ether/water emulsion. The polyurea microcapsule was consisted of chitosan oligomer (CO) as the membrane material and diphenyl methane-4,4'-diisocyanate (MDI) as the crosslinker. A chemical modification was carried out by grafting a UV-absorbent, 3,3',4,4'-benzophenonetetracarboxylic dianhydride (BTDA), to CO before interfacial polymerization to enhance the UV-resistance of the microcapsule. The BTDA grafted CO (CO-BTDA) and the AVM microcapsules were characterized by a variety of instrumental techniques, including NMR, FTIR, UV-vis, GPC-LS, DLS, SEM and TEM. The in vitro release test showed that the polyurea microcapsule maintained the sustained release of AVM for a longer period (up to 120 h) in comparison with the commercial AVM formulations (within 24 h). The photodegradation test revealed that the polyurea microcapsule significantly reduced the AVM degradation and extended the half-life of AVM from 4.16 h to 9.43 h. The AVM degradation was further reduced by using the BTDA-modified polyurea microcapsule. The corresponding half-life was extended up to 17.33 h and can be mediated by changing the mass ratio of BTDA: CO during the synthesis of CO-BTDA. The use of polyurea microcapsule did not raise a concern about pesticide residue as no AVM was detected after the photodegradation test. In addition, the polyurea microcapsule itself was subject to degradation under sunlight exposure, which reduced its residue in the environment.

Orthogonal near-infrared upconversion co-regulated site-specific O2 delivery and photodynamic therapy for hypoxia tumor by using red blood cell microcarriers.

Pre-existing hypoxia in tumors can result in an inadequate oxygen supply during photodynamic therapy (PDT), which in turn hampers photodynamic efficacy. To overcome this problem, we developed an orthogonal near-infrared upconversion controlled red blood cell (RBC) microcarriers to selectively deliver O2 in hypoxia area. Moreover, this RBC microcarriers are able to overcome a series of complex biological barriers which include transporting across the inflamed endothelium, evading mononuclear phagocyte system, reducing reticuloendothelial system uptake. Based on these abilities, RBC microcarriers have efficient tumors accumulation and are capable of delivery a large amount of O2 for PDT under near-infrared (NIR) irradiation to realize effective solid tumor eradication.

Stability studies on colloidal suspensions of polyurethane nanocapsules.

Generally nanocapsules suspensions are a colloidal system in a metastable state, there is aggregation due to attraction and repulsion forces between particles. The objective of this work was to bring the role of the polymeric membrane in the protection of the active drug against damaging caused by external agents and to select the monomer which leads to obtain stable formulation with the highest possible payload of the active drug. The stability testing involving visual aspect, particle size measurement, transmission electron microscopy (TEM) examination, and drug loss was conduced after 6 months of storage at different temperatures (4, 25, and 45 degrees C). The colloidal suspensions of nanocapsules were obtained using the combined interfacial polycondensation and spontaneous emulsification, the technique was used to encapsulate alpha-tocopherol using polyurethanes polymers. It is a one step procedure: An organic phase composed of a water miscible solvent (acetone), lipophilic monomer (Isophorone diisocyanate IPDI), oil, and a lipophilic surfactant, is injected in an aqueous phase containing hydrophilic monomer (diol with various molecular weight: 1,2-ethanediol (ED), 1,4-butanediol (BD), and 1,6-hexanediol (HD)) and hydrophilic emulsifying agent. The water miscible solvent diffuses to the aqueous phase, the oil precipitates as nano-droplets, and the two monomers react at the interface, forming a membrane around the nanoemulsion leading to nanocapsules. A good physical stability of suspensions corresponds to absence of symptoms such as sedimentation or agglomeration, significant size change and alpha-tocopherol degradation due to external agents such as oxygen, temperature, and ultraviolet (UV) irradiation. The size of nanocapsules before storage was about 232 +/- 3, 258 +/- 29, and 312 +/- 4 nm for ED, BD, and HD, respectively. After 6 months of storage, polyurethanes nanocapsules possess good stability against aggregation at 4 and 25 degrees C. Comparing results obtained using different monomers, it reveals that the polyurethane based on HD offers good protection of alpha-tocopherol against damaging caused by the temperature and UV irradiation.

Acoustic destruction of a microcapsule having a hard plastic shell.

Acoustic destruction of a microcapsule having a hard plastic shell is discussed. In an ultrasonic drug delivery system, microcapsules having thin elastic shells release drugs that are contained therein when the shell is destroyed. In this paper, two subjects related to capsule destruction are discussed: the driving pulse duration for capsule destruction and the frequency dependence of capsule destruction. Optical observation of microcapsule destruction is performed with a high-speed video camera. In the case of capsule destruction by a pulse wave, the internal gas of the microcapsule cannot be ejected completely, and a portion of the internal gas remains inside the broken shell. It is found that capsule destruction by pulse waves depends on both the amplitude of the driving pressure and the pulse duration. The frequency dependence of microcapsule destruction also is investigated. In the case of capsule destruction by a low-amplitude acoustic wave, the destruction rate under the resonance condition is higher than under nonresonance conditions. By controlling the driving frequency, selective capsule destruction can be achieved.

Fluid Viscosity Affects the Fragmentation and Inertial Cavitation Threshold of Lipid-Encapsulated Microbubbles.

Ultrasound and microbubble optimization studies for therapeutic applications are often conducted in water/saline, with a fluid viscosity of 1 cP. In an in vivo context, microbubbles are situated in blood, a more viscous fluid (∼4 cP). In this study, ultrahigh-speed microscopy and passive cavitation approaches were employed to investigate the effect of fluid viscosity on microbubble behavior at 1 MHz subject to high pressures (0.25-2 MPa). The propensity for individual microbubble (n = 220) fragmentation was found to significantly decrease in 4-cP fluid compared with 1-cP fluid, despite achieving similar maximum radial excursions. Microbubble populations diluted in 4-cP fluid exhibited decreased wideband emissions (up to 10.2 times), and increasingly distinct harmonic emission peaks (e.g., ultraharmonic) with increasing pressure, compared with those in 1-cP fluid. These results suggest that in vitro studies should consider an evaluation using physiologic viscosity perfusate before transitioning to in vivo evaluations.

Conducting polymers with defined micro- or nanostructures for drug delivery.

Conducting polymers (CPs) are redox active materials with tunable electronic and physical properties. The charge of the CP backbone can be manipulated through redox processes, with accompanied movement of ions into and out of the polymer to maintain electrostatic neutrality. CPs with defined micro- or nanostructures have greatly enhanced surface areas, compared to conventionally prepared CPs. The resulting high surface area interface between polymer and liquid media facilities ion exchange and can lead to larger and more rapid responses to redox cycling. CP systems are maturing as platforms for electrically tunable drug delivery. CPs with defined micro- or nanostructures offer the ability to increase the amount of drug that can be delivered whilst enabling systems to be finely tuned to control the extent and rate of drug release. In this review, fabrication approaches to achieve CPs with micro- or nanostructure are outlined followed by a detailed review and discussion of recent advances in the application of the materials for drug delivery.

Interfacial Rheological Properties of Contrast Microbubble Targestar P as a Function of Ambient Pressure.

In this Technical Note, we determine the interfacial rheological parameters of the encapsulation of the contrast agent Targestar P using ultrasound attenuation. The characteristic parameters are obtained according to two interfacial rheological models. The properties-surface dilatational elasticity (0.09 ± 0.01 N/m) and surface dilatational viscosity (8 ± 0.1E-9 N·s/m)-are found to be of similar magnitude for both models. Contrast microbubbles experience different ambient pressure in different organs. We also measure these parameters as functions of ambient pressure using attenuation measured at different overpressures (0, 100 and 200 mm Hg). For each value of ambient hydrostatic pressure, we determine the rheological properties, accounting for changes in the size distribution caused by the pressure change. We discuss different models of size distribution change under overpressure: pure adiabatic compression or gas exchange with surrounding medium. The dilatational surface elasticity and viscosity are found to increase with increasing ambient pressure.

Folate-conjugated gene-carrying microbubbles with focused ultrasound for concurrent blood-brain barrier opening and local gene delivery.

Previous studies have demonstrated that circulating DNA-encapsulated microbubbles (MBs) combined with focused ultrasound (FUS) can be used for local blood-brain barrier (BBB) opening and gene delivery. However, few studies focused on how to increase the efficiency of gene delivery to brain tumors after the released gene penetrating the BBB. Here, we proposed the use of folate-conjugated DNA-loaded cationic MBs (FCMBs). When combined with FUS as a trigger for BBB opening, FCMBs were converted into nanometer-sized vesicles that were transported to the brain parenchyma. The FCMBs can selectively aggregate around tumor cells that overexpressed the folate receptor, thus enhancing gene delivery via folate-stimulated endocytosis. Our results confirmed that FCMBs can carry DNA on the surface of the MB shell and have good targeting ability on C6 glioma cells. In addition, the optimized FUS parameters for FCMBs-enhanced gene delivery were confirmed by cell experiments (center frequency = 1 MHz; acoustic pressure = 700 kPa; pulse repetition frequency = 5 Hz; cycle number = 10000; exposure time = 1 min; FCMBs concentration = 4 × 10(7) MB/mL). In vivo data also indicated that FCMBs show better gene transfection efficiency than MBs without folate conjugation and the traditional approach of directly injecting the gene. This study described our novel development of multifunctional MBs for FUS-triggered gene delivery/therapy.

Biological properties of photocrosslinked alginate microcapsules.

An alternative form of gene therapy using recombinant cell lines delivering therapeutic products encapsulated in alginate hydrogel has proven effective in treating many murine models. The lack of long-term capsule stability has led to a new strategy to reinforce the microcapsules with a photopolymerized interpenetrating covalent network of N-vinylpyrrolidone (NVP) and sodium acrylate. Here the properties for potential application in gene therapy are reported. In assessing potential toxicity of the unpolymerized residues, HPLC showed that even after 1 week of washing, no toxic monomers could be detected. Their ability to sustain cell growth was monitored with growth of the encapsulated cells in vitro and in vivo. Although the initial photopolymerization caused significant cell damage, the cells were able to recover normal growth rates thereafter. After implanting into mice, the NVP-modified capsules showed a high level of biocompatibility as measured by hematological and biochemical functional tests. There was also no difference in the amount and type of plasma proteins adsorbing to the NVP-modified and the classical alginate capsules, thus indicating their similar biological compatibility. Both in vitro and in vivo tests confirmed that the NVP-modified capsules were more resistant to osmotic stress than the alginate microcapsules. Furthermore, when applied to the treatment of a murine model of human cancer by delivering encapsulated cells secreting angiostatin, the NVP-modified microcapsules suppressed tumor growth as successfully as the regular alginate microcapsules. In conclusion, the covalently modified microcapsules have shown a high level of biocompatibility, safety, increase in stability, and clinical efficacy for use as immunoisolation devices in gene therapy.

Delivery of molecules to the lymph node via lymphatic vessels using ultrasound and nano/microbubbles.

Lymph node (LN) dissection is the primary option for head and neck cancer when imaging modalities and biopsy confirm metastasis to the sentinel LN. However, there are no effective alternative treatments to dissection for LN metastasis. Here, we describe a novel drug delivery system combining nano/microbubbles (NMBs) with ultrasound (US) that exhibits considerable potential for the delivery of exogenous molecules into LNs through the lymphatic vessels. A solution containing fluorophores (as a model of a therapeutic molecule) and NMBs was injected into the subiliac LNs of MXH10/Mo-lpr/lpr mice, which develop systemic swelling of LNs (up to 13 mm in diameter, similar to human LNs). It was found that the NMBs were delivered to the entire area of the proper axillary LN (proper-ALN) via the lymphatic channels and that these were retained there for more than 8 min. Furthermore, exposure to US in the presence of NMBs enhanced the delivery of fluorophores into the lymphocytes near the lymphatic channels, compared with exposure to US in the absence of NMBs. It is proposed that a system using US and NMBs to deliver therapeutic drugs via lymphatic vessels can serve as a new treatment method for LN metastasis.

Evaluation of the potential of doxorubicin loaded microbubbles as a theranostic modality using a murine tumor model.

In this study, a novel phospholipid-based microbubble formulation containing doxorubicin and perfluoropropane gas (DLMB) was developed. The DLMBs were prepared by mechanical agitation of a phospholipid dispersion in the presence of perfluoropropane (PFP) gas. An anionic phospholipid, distearoyl phosphatidylglycerol (DSPG) was selected to load doxorubicin in the microbubbles by means of electrostatic interaction. The particle size, zeta potential, echogenicity and stability of the DLMBs were measured. Drug loading was ⩾ 92%. The potential of the DLMBs for use as a theranostic modality was evaluated in tumor bearing mice. Gas chromatography analysis of PFP showed significant enhancement of PFP retention when doxorubicin was used at concentrations of 10-82% equivalent to DSPG. The inhibitory effects on the proliferation of B16BL6 melanoma murine cells in vitro were enhanced using a combination of ultrasound (US) irradiation and DLMBs. Moreover, in vivo DLMBs in combination with (US) irradiation significantly inhibited the growth of B16BL6 melanoma tumor in mice. Additionally, US echo imaging showed high contrast enhancement of the DLMBs in the tumor vasculature. These results suggest that DLMBs could serve as US triggered carriers of doxorubicin as well as tumor imaging agents in cancer therapy.