Fluorocarbon Nanodroplets: Their Formation and Stability in Complex Solution Systems.

Perfluorocarbon (PFC) nanodroplets (NDs) are expanding in a wide range of applications in biotechnology and nanotechnology. Their efficacy in biological systems is significantly influenced by their size uniformity and stability within bioelectrolyte contexts. Presently, methods for creating monodisperse, highly concentrated, and well-stabilized PFC NDs under harsh conditions using low energy consumption methods have not been thoroughly developed, and their stability has not been sufficiently explored. This gap restricts their applicability for advanced medical interventions in tissues with high pH levels and various electrolytic conditions. To tackle these challenges and to circumvent potential toxicity from surface stabilizers, we have conducted an in-depth investigation into the formation and stability of uncoated perfluorohexane (PFH) NDs, which were synthesized by using a low-energy consumption solvent exchange technique, across complex electrolyte compositions or a broad spectrum of pH levels. The results indicated that low concentrations of low-valent electrolyte ions facilitate the nucleation of NDs and consistently accelerate Ostwald ripening over an extended period. Conversely, high concentrations of highly valent electrolyte ions inhibit nucleation and decelerate the ripening process over time. Given the similarities between the properties of NDs and nanobubbles, we propose a potential stabilization mechanism. Electrolytes influence the Ostwald ripening of NDs by adjusting the adsorption and distribution of ions on the NDs' surface, modifying the thickness of the electric double layer, and fine-tuning the energy barrier between droplets. These insights enable precise control over the stability of PFC NDs through the meticulous adjustment of the surrounding electrolyte composition. This offers an effective preparation method and a theoretical foundation for employing bare PFC NDs in physiological settings.

Correction: Gram-selective antibacterial peptide hydrogels.

Correction for 'Gram-selective antibacterial peptide hydrogels' by Yangqian Hou et al., Biomater. Sci., 2022, 10, 3831-3844, https://doi.org/10.1039/D2BM00558A.

Gram-selective antibacterial peptide hydrogels.

The human microbiome plays fundamental roles in human health and disease. However, widely used broad-spectrum antibiotics severely disrupt human-related microbial communities, eventually leading to resistant bacteria, posing a growing threat to global medical health. Antimicrobial peptides (AMPs) are promising antimicrobial agents that barely cause bacterial resistance. Excellent broad-spectrum antimicrobial activities have been achieved using hydrogels self-assembled from AMPs, but there is still a lack of AMP hydrogels that can target Gram-positive and Gram-negative bacteria. Herein, several hydrogels self-assembled from AMPs, termed IK1, IK3, and IK4, were designed and synthesized. In vitro antibacterial results indicated that the IK1 and IK4 hydrogels specifically targeted Gram-positive and Gram-negative bacteria, respectively, while the IK3 hydrogel targeted both Gram-positive and Gram-negative bacteria. The desired broad-spectrum or Gram-selective AMP hydrogels are believed to be obtained through the rational design of the hydrophilicity, hydrophobicity, and charge properties of the peptide molecules. Good in vivo Gram-selective antibacterial properties and the ability to promote wound healing have been demonstrated via treating mouse wound models with these AMP hydrogels. We believe that these Gram-selective AMP hydrogels could potentially have important applications in treating common recurring infections.

Controllable formation of bulk perfluorohexane nanodroplets by solvent exchange.

Perfluorocarbon (PFC) nanodroplets have rapidly developed into useful ultrasound imaging agents in modern medicine due to their non-toxic and stable chemical properties that facilitate disease diagnosis and targeted therapy. In addition, with the good capacity for carrying breathing gases and the anti-infection ability, they are employed as blood substitutes and are the most ideal liquid respirators. However, it is still a challenge to prepare stable PFC nanodroplets of uniform size and high concentration for their efficient use. Herein, we developed a simple and highly reproducible method, i.e., propanol-water exchange, to prepare highly homogeneous and stable perfluorohexane (PFH) bulk nanodroplets. Interestingly, the size distribution and concentration of formed nanodroplets could be regulated by controlling the volume fraction of PFH and percentage of propanol in the propanol-water mixture. We demonstrated good reproducibility in the formation of bulk nanodroplets with PFH volume fractions of 1/2000-1/200 and propanol percentage of 5-40%, with uniform particle size distribution and high droplet concentration. Also, the prepared nanodroplets were very stable and could survive for several hours. We constructed a ternary phase diagram to describe the relationship between the PFH volume ratio, propanol concentration, and the size distribution and concentration of the formed PFH nanodroplets. This study provides a very useful method to prepare uniform size, high concentration and stable PFC nanodroplets for their medical applications.

Gelation of a Pentapeptide in Alcohols.

Properties of solvents such as polarity and H-bond-forming ability are critical for the formation of an organogel and have a significant impact on the gel behavior, as solvents are the majority of organogel systems. However, so far, there is still a lack of systematic studies regarding the effects of molecular structures of solvents on the characteristics of organogels. Motivated by revealing such a relationship, in this paper, we studied the morphologies of assemblies, gelation behaviors, and secondary structures of a pentapeptide termed EAF-5 in a wide variety of alcohols. The side chains and lengths of carbon chains of the solvent molecules were found to play a critical role in the self-assembly and gelation of EAF-5. EAF-5 was capable of self-assembling into fibers and entangling into a network in alcohols including ethanol, propanol, butanol, n-pentanol, and n-hexanol, which further immobilized the corresponding alcohols to form gels. In these organogels, increasing ß-sheet secondary structures of the peptides were formed by introducing side chains and extending the length of primary alcohol molecules. We hypothesized that alcohol molecules with extended lengths and side chains reduced the gelator-solvent interactions and promoted the gelator-gelator interactions, resulting in the self-assembly of EAF-5 into fibril structures and development of gels. These findings provide a new sight into the interactions between gelators and solvents and are helpful for designing peptide-based organogelators.

Macrochirality of Self-Assembled and Co-assembled Supramolecular Structures of a Pair of Enantiomeric Peptides.

Although macrochirality of peptides' supramolecular structures has been found to play important roles in biological activities, how macrochirality is determined by the molecular chirality of the constituted amino acids is still unclear. Here, two chiral peptides, Ac-LKLHLHLQLKLLLVLFLFLALK-NH2 (KK-11) and Ac-DKDHDHDQDKDL DVDFDFDADK-NH2 (KKd-11), which were composed entirely of either L- or D-amino acids, were designed for studying the chiral characteristics of the supramolecular microstructures. It was found that monocomponent KK-11 or KKd-11 self-assembled into right- or left-handed helical nanofibrils, respectively. However, when they co-assembled with concentration ratios varied from 1:9 to 9:1, achiral nanowire-like structures were formed. Both circular dichroism and Fourier transform infrared spectra indicated that the secondary structures changed when the peptides co-assembled. MD simulations indicated that KK-11 or KKd-11 exhibited a strong propensity to self-assemble into right-handed or left-handed nanofibrils, respectively. However, when KK-11 and KKd-11 were both presented in a solution, they had a higher probability to co-assemble instead of self-sort. MD simulations indicated that, in their mixtures, they formed nanowires without handedness feature, a good agreement with experimental observation. Our results shed light on the molecular mechanisms of the macrochirality of peptide supramolecular microstructures.

Antimicrobial d-Peptide Hydrogels.

Biofilms are widely involved in human lives, such as in medical infection, environmental remediation, and industrial processes. However, the control of the biofilm has still been a challenge because of its strong drug resistance. Here, we designed and synthesized an amphipathic antimicrobial peptide (Ac-DKDHDHDQDKDLDVDFDFDADK-NH2 (KKd-11)) that was composed of d-amino acids (DAAs). KKd-11 was found to self-assemble into a hydrogel with an improved long-term antimicrobial ability and a better antiprotease activity as compared to the hydrogel formed by Ac-LKLHLHLQLKLLLVLFLFLALK-NH2 (KK-11). Our results indicated that KKd-11 was not only able to inhibit the formation of biofilms but also could effectively damage preformed mature biofilms and kill the bacteria within the biofilms. Besides, cell viability assays indicated that the KKd-11 peptide had very good biocompatibility. We think d-peptide hydrogels may have great potential in the treatment of biofilm-induced infections.

The effects of nanobubbles on the assembly of glucagon amyloid fibrils.

Some recent studies have shown that the surface and interface play an important role in the assembly and aggregation of amyloid proteins. However, it is unclear how the gas-liquid interface affects the protein assembly at the nanometer scale although the presence of gas-liquid interfaces is very common in in vitro experiments. Nanobubbles have a large specific surface area, which provides a stage for interactions with various proteins and peptides on the nanometer scale. In this work, nanobubbles produced in solution were employed for studying the effects of the gas-liquid interface on the assembly of glucagon proteins. Atomic force microscopy (AFM) studies showed that nanobubble-treated glucagon solution formed fibrils with an apparent height of 4.02 ± 0.71 nm, in contrast to the fibrils formed with a height of 2.14 ± 0.53 nm in the control. Transmission electron microscopy (TEM) results also showed that nanobubbles promoted the assembly of glucagon to form more fibrils. Thioflavin T (ThT) fluorescence and Fourier transform infrared (FTIR) analyses indicated that the nanobubbles induced the change of the glucagon conformation to a ß-sheet structure. A mechanism that explains how nanobubbles affect the assembly of glucagon amyloid fibrils was proposed based on the above-mentioned experimental results. Given the fact that there are a considerable amount of nanobubbles existing in protein solutions, our results indicate that nanobubbles should be considered for fully understanding the protein aggregation events in vitro.

Generating Bulk Nanobubbles in Alcohol Systems.

Bulk nanobubbles (NBs) have attracted wide attention due to their peculiar physicochemical properties and great potential in applications in various fields. However, so far there are no reports on bulk NBs generated in pure organic systems, which we think is very important as NBs would largely improve the efficiency of gas-liquid mass transfer and facilitate chemical reactions to take place. In this paper, we verified that air and N2 NBs could be generated in a series of alcohol solutions by using various methods including acoustical cavitation, pressurization-depressurization, and vibration. The experiments proved that NBs existed in alcohol solutions, with a highest density of 5.8 × 107 bubble/mL in propanol. Our results also indicated that bulk NBs could stably exist for at least hours in alcohol systems. The parameters in generating NBs in alcohols were optimized. Our findings open up an opportunity for improving gas-liquid mass transfer efficiency in the field of the chemical industry.