Stability of Engineered Micro or Nanobubbles for Biomedical Applications.

A micro/nanobubble (MNB) refers to a bubble structure sized in a micrometer or nanometer scale, in which the core is separated from the external environment and is normally made of gas. Recently, it has been confirmed that MNBs can be widely used in angiography, drug delivery, and treatment. Thus, MNBs are attracting attention as they are capable of constructing a new contrast agent or drug delivery system. Additionally, in order to effectively use an MNB, the method of securing its stability is also being studied. This review highlights the factors affecting the stability of an MNB and the stability of the MNB within the ultrasonic field. It also discusses the relationship between the stability of the bubble and its applicability in vivo.

Self-Assembling ß-Glucan Nanomedicine for the Delivery of siRNA.

We aimed to design and manufacture a transporter capable of delivering small interfering RNAs (siRNAs) into the skin without causing any damage. ß-glucans are unique chiral polysaccharides with well-defined immunological properties and supramolecular wrapping ability. However, the chiral properties of these polymers have hardly been applied in drug delivery systems. In this study, ß-glucan nanoparticles were designed and manufactured to deliver genetic material to the target cells. The ß-glucan molecules were self-assembled with an siRNA into nanoparticles of 300-400 nm in diameter via a conformational transition process, in order to construct a gene delivery system. The assembled gene nanocarriers were associated with high gene-loading ability. The expression and efficiency of siRNA were verified after its delivery via ß-glucan. Our results provide evidence that ß-glucan nanoparticles can be effectively used to deliver siRNA into the cells.

Study and Evaluation of the Potential of Lipid Nanocarriers for Transdermal Delivery of siRNA.

The topical delivery of siRNA-based therapies has opened new avenues for the treatment of skin disorders. The use of siRNA as a therapeutic, however, is limited due to its rapid degradation and poor cellular uptake. Furthermore, the top layer of skin, the stratum corneum, is a major barrier to the delivery of topical agents. There is an unmet need for efficient topical formulations for delivering siRNA to the site of action. In this study, 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) or lipofectamine is used to prepare a nanocarrier for delivering siRNA against glyceraldehyde 3-phosphate dehydrogenase (GAPDH); GAPDH expression is then evaluated at the cellular level. In addition, a dermal transport assay is designed and implemented to evaluate the penetration and delivery efficacy of siRNA in pig skin using lipid nanocarriers. The delivery of siRNA with the use of a lipid nanocarrier is significantly better than the delivery of siRNA without it. Thus, the findings identify lipid nanocarriers as excellent candidates for the transdermal delivery of siRNA for gene silencing in the skin and thus for applications in related preclinical models.

Development of Antibiofilm Nanocomposites: Ag/Cu Bimetallic Nanoparticles Synthesized on the Surface of Graphene Oxide Nanosheets.

There are numerous issues associated with bacteria, particularly biofilms, which exhibit a strong resistance to antibiotics. This is currently considered an urgent global issue owing to the lack of effective treatments. Graphene oxide (GO) nanosheets are two-dimensional carbon materials that are available as a substrate for metal nanoparticles and have a lower release rate of metal ions than free metal nanoparticles by regulating the oxidation of metal nanoparticles, which is known to reduce the cytotoxicity caused by the free metal nanoparticles. Over centuries, metal particles, including Ag and Cu, have been considered as antibacterial agents. In this study, Ag and Cu bimetallic nanoparticles on a GO surface (Ag/Cu/GO) were synthesized using a chemical reduction method, and their antimicrobial effects against several bacterial species were demonstrated. Ag/Cu/GO nanocomposites were characterized by transmission electron microscopy and energy-dispersive X-ray spectroscopy. The in vitro cytotoxicity of an Ag/Cu/GO nanocomposite was evaluated in human dermal fibroblasts, and its antibacterial activity against Methylobacterium spp., Sphingomonas spp., and Pseudomonas aeruginosa (P. aeruginosa) was also tested. The synthesized Ag/Cu/GO nanocomposite was able to eradicate all three bacterial species at a concentration that was harmless to human cells. In addition, Ag/Cu/GO successfully removed a biofilm originated from the culturing of P. aeruginosa in a microchannel with a dynamic flow. In a small-animal model, a biofilm-infected skin wound was healed quickly and efficiently by the topical application of Ag/Cu/GO. The Ag/Cu/GO nanocomposites reported in this study could be used to effectively remove antibiotic-resistant bacteria and treat diseases in the skin or wound due to bacterial infections and biofilm formation.

Effective delivery of mycophenolic acid by oxygen nanobubbles for modulating immunosuppression.

Immunosuppressive drugs are crucial for preventing acute graft rejection or autoimmune diseases. They are generally small molecules that require suitable drug carriers for ensuring stability, bioavailability, and longer half-life. Mycophenolic acid (MPA) is an extensively studied immunosuppressive drug. However, it requires suitable carriers for overcoming clinical limitations. Currently, lipid-shelled micro- and nanobubbles are being thoroughly investigated for diagnostic and therapeutic applications, as they possess essential properties, such as injectability, smaller size, gaseous core, high surface area, higher drug payload, and enhanced cellular penetration. Phospholipids are biocompatible and biodegradable molecules, and can be functionalized according to specific requirements. Methods: In this study, we synthesized oxygen nanobubbles (ONBs) and loaded the hydrophobic MPA within the ONBs to generate ONB/MPA. Peripheral blood mononuclear cells (PBMCs) were treated with ONB/MPA to determine the suppression of immune response by measuring cytokine release. In vivo murine experiments were performed to evaluate the effectiveness of ONB/MPA in the presence of inflammatory stimulants. Results: Our results suggest that ONBs successfully delivered MPA and reduced the release of cytokines, thereby controlling inflammation and significantly increasing the survival rate of animals. Conclusion: This method can be potentially used for implantation and for treating autoimmune diseases, wherein immunosuppression is desired.

Array-Based Screening of Silver Nanoparticle Mineralization Peptides.

The use of biomolecules in nanomaterial synthesis has received increasing attention, because they can function as a medium to produce inorganic materials in ambient conditions. Short peptides are putative ligands that interact with metallic surfaces, as they have the potential to control the synthesis of nanoscale materials. Silver nanoparticle (AgNP) mineralization using peptides has been investigated; however, further comprehensive analysis must be carried out, because the design of peptide mediated-AgNP properties is still highly challenging. Herein, we employed an array comprising 200 spot synthesis-based peptides, which were previously isolated as gold nanoparticle (AuNP)-binding and/or mineralization peptides, and the AgNP mineralization activity of each peptide was broadly evaluated. Among 10 peptides showing the highest AgNP-synthesis activity (TOP10), nine showed the presence of EE and E[X]E (E: glutamic acid, and X: any amino acid), whereas none of these motifs were found in the WORST25 (25 peptides showing the lowest AgNP synthesis activity) peptides. The size and morphology of the particles synthesized by TOP3 peptides were dependent on their sequences. These results suggested not only that array-based techniques are effective for the peptide screening of AgNP mineralization, but also that AgNP mineralization regulated by peptides has the potential for the synthesis of AgNPs, with controlled morphology in environmentally friendly conditions.

NIR Laser-Responsive PNIPAM and Gold Nanorod Composites for the Engineering of Thermally Reactive Drug Delivery Nanomedicine.

When ingesting a drug on its own or injecting it directly into tissue, its concentration increases immediately within the body, which often exacerbates the side effects and increases its toxicity. To solve this problem, we synthesized the thermally reactive polymer poly(N-isopropylacrylamide) (PNIPAM) using reversible addition-fragmentation chain transfer (RAFT) polymerization and prepared nanocarriers by binding PNIPAM to gold nanorods (GRs), with the anticancer agent doxorubicin (DOX) used as a model drug. PNIPAM changes from hydrophilic to hydrophobic at temperatures above its lower critical solution temperature, which represents a coil-to-globule volume phase transition. Because GRs absorb near-infrared (NIR) laser light and emit energy, PNIPAM aggregation occurs when the synthesized PNIPAM/GR are subjected to an NIR laser, and the temperature of the GRs rises. Using this principle, DOX was combined with the PNIPAM/GR complex, and the resulting anticancer effects with and without laser treatment were observed in Hela and MDA-MB-231 cells. In our proposed complex, the GR binding rate of PNIPAM reached 20% and the DOX binding rate reached 15%. The release profile of the drug following laser irradiation was determined using a drug release test and confocal microscopy imaging. It was subsequently confirmed that the release of the drug is higher at higher temperatures, especially with laser treatment. The proposed combination of temperature-reactive polymers and gold nanostructures shows promise for future research into controlled drug release.

Effective Delivery of Anti-Cancer Drug Molecules with Shape Transforming Liquid Metal Particles.

Liquid metals are being studied intensively because of their potential as a drug delivery system. Eutectic gallium-indium (EGaIn) alloy liquid metals have a low melting point, low toxicity, and excellent tissue permeability. These properties may enable them to be vascular embolic agents that can be deformed by light or heat. In this study, we developed EGaIn particles that can deliver anticancer drugs to tumor cells in vitro and change their shapes in response to external stimuli. These particles were prepared by sonicating a solution containing EGaIn and amphiphilic lipids. The liquid metal (LM)/amphiphilic lipid (DSPC, 1,2-distearoyl-sn-glycero-3-phosphocholin) particles formed a vehicle for doxorubicin, an anticancer drug, which was released (up to 50%) when the shape of the particles was deformed by light or heat treatment. LM/DSPC particles are non-toxic and LM/DSPC/doxorubicin particles have anticancer effects (resulting in a cell viability of less than 50%). LM/DSPC/doxorubicin particles were also able to mimic blood vessel embolisms by modifying their shape using precisely controlled light and heat in engineered microchannels. The purpose of this study was to examine the potential of EGaIn materials to treat tumor tissues that cannot be removed by surgery.

Biological Responses of Onion-Shaped Carbon Nanoparticles.

Nanodiamonds are emerging as new nanoscale materials because of their chemical stability, excellent crystallinity, and unique optical properties. In this study, the structure of nanodiamonds was engineered to produce carbon nano-onion particles (CNOs) with multiple layers. Following a series of physicochemical characterizations of the CNOs, various evaluations for biological responses were conducted for potential biotechnological applications of the CNOs. The possibility of biological applications was first confirmed by assessment of toxicity to animal cells, evaluation of hemolysis reactions, and evaluation of reactive oxygen species. In addition, human immune cells were evaluated for any possible induction of an immune response by CNOs. Finally, the toxicity of CNOs to Escherichia coli present in the human colon was evaluated. CNOs have the chemical and physical properties to be a unique variety of carbon nanomaterials, and their toxicity to animal and human cells is sufficiently low that their biotechnological applications in the future are expected.

Surface Composition and Preparation Method for Oxygen Nanobubbles for Drug Delivery and Ultrasound Imaging Applications.

Phospholipids have been widely investigated for the preparation of liposomes, and micro and nanobubbles. They comprise biocompatible and biodegradable molecules and offer simple preparation with a variety of functions in diagnostic and therapeutic applications. Phospholipids require emulsifiers and surfactants to assemble in the form of bubbles. These surfactants determine the size, zeta potential, and other characteristics of particles. Polyethylene glycol (PEG) and its various derivatives have been employed by researchers to synthesize micro and nanobubbles. The stability of phospholipid-shelled nanobubbles has been reported by various researchers owing to the reduction of surface tension by surfactants in the shell. Nanobubbles have been employed to deliver oxygen to tissues and hypoxic cells. In this study, we investigated the effects of different ratios of phospholipids to PEG on the size, distribution, and characterization of oxygen nanobubbles (ONBs). ONBs were synthesized using a sonication technique. We analyzed and compared the sizes, numbers of generated particles, and zeta potentials of different compositions of ONBs using dynamic light scattering and nanoparticle tracking analysis. Then, we employed these oxygen nanobubbles to enhance the cellular microenvironment and cell viability. ONBs were also investigated for ultrasound imaging.

A theoretical model to predict the low-frequency sound absorption of a helmholtz resonator array.

A theoretical method based on mutual radiation impedance is proposed to compute the sound absorption performance of a Helmholtz resonator array in the low-frequency range. Any configuration of resonator arrangement can be allowed in the method, while all the resonators may or may not be identical. Comparisons of the theoretical predictions with those done by the past studies or experiments show that the present method can accurately predict the absorption performance in more general cases.