Nanobubbles can enhance both the proliferation and metabolic activity of microorganisms (mainly bacteria) and the growth of the whole higher organisms such as mice, fish, or plants. The critical fact is that nanobubbles of different gases can affect given cells differently. As animal cell cultures are used in industry and research studies, investigations of their interactions with nanobubbles should be carried out. This study aims to uncover whether the presence of nanobubbles improves the proliferation rate and metabolic activity of L929 fibroblasts and HL60 leukemia cells as exemplary animal cell lines of adherent and non-adherent cells, respectively. The long-term (8-day) cultures of both L929 and HL-60 cells with nanobubble addition to the appropriate medium were carried out. The medium was not exchanged for the whole duration of the culture. Nanobubbles of two gases - oxygen and nitrogen - were dispersed in the appropriate media and then used to culture cells. The density and viability of cells were assessed microscopically while their metabolic activity was determined using PrestoBlue or XTT assays. Additionally, we have performed the analysis of substrate consumption rate during the growth and activity of lactate dehydrogenase. We have shown that nanodispersion of both gases enhances the proliferation rate and metabolic activity of L929. For HL-60 cultures, reference cultures exhibited better viability, cell density, and metabolic activity than those with either oxygen or nitrogen nanobubbles. Obtained results clearly show that nanobubble dispersions of both oxygen and nitrogen positively affect the cultures of L929 while inhibiting the growth of HL-60 cells. We suspect that a similar positive effect would be visible for other adherent cells, similar to L929. Such results are promising for intensifying the growth of animal or human cells in routine cell cultures.

(1) The generation of nanobubbles by electrolysis is an interesting method of using electrical energy to form bubble nuclei, effectively creating a multiphase system. For every process, the effectiveness of nanobubble generation by electrolysis depends on various process parameters that impact should be determined. (2) In this work, the electrolytic generation of hydrogen and oxygen bubbles was performed in a self-built setup, in which a Nafion membrane separated two chambers. The generation of bubbles of both gases was investigated using Box–Behnken experimental design. Three independent variables were salt concentration, current density, and electrolysis time, while the dependent variables were Sauter diameters of generated bubbles. An ANOVA analysis and multivariate regression were carried out to propose a statistical and power model of nanobubble size as a process parameter function. (3) The generation of bubbles of hydrogen and oxygen by electrolysis showed that different factors or their combinations determine their size. The results presented in this work proved to be complementary to previous works reported in the literature. (4) The Sauter diameter of bubbles increases with salt concentration and stays constant with increasing current density in investigated range. The proposed correlations allow the Sauter diameters of nanobubbles generated during electrolysis to be predicted.

Cleaning of surfaces without complex cleaning agents is an important subject, especially in food, pharmaceutical, and biomedical applications. The subject of microbubble and nanobubble cleaning is considered one of the most promising ways to intensify this process. In this work, we check whether and how the purity of water used for microbubble generation, as well as the gas used, affects the effectiveness of cleaning stainless-steel surfaces. Surfaces contaminated with Pluronic L-121 solution were cleaned by water of three purities: ultrapure water (<0.05 μS/cm), water after reversed osmosis (~6.0 μS/cm), and tap water (~0.8 mS/cm). Similarly, three different gases were supplied to the generation setup for microbubble generation: air, oxygen, and nitrogen. Stainless steel plates were immersed in water during microbubble generation and cleaned for a given time. FTIR (Fourier Transform Infrared Spectroscopy) and contact angle analysis were employed for the analysis of surfaces. The results of cleaning were repeatable between plates and showed different cleaning effects depending on both the purity of water (concentration of ions) and gas composition. We have proposed different mechanisms that are dominant with respect to specific combinations of ion concentration and oxygen content in gas, which are directly connected to the microbubble stability and reactivity of gas.

The repeatable and cost-efficient generation of nanobubbles is still a challenge. In most cases, the hydrodynamic generation of nanobubbles is used at larger scales. Therefore, every cost reduction possible in nanobubble generation is needed. In this work, we decided to check how the generation of nanobubbles changes when the surrounding liquid properties change. The generation of nanobubbles was carried out in a novel setup, designed by us. We investigated the minimum liquid velocity needed for nanobubble generation and propose correlations describing this based on the physicochemical properties of the liquid and gas phases. As carbon dioxide nanobubbles are commonly used for the treatment of ischemia and chronic wounds, the investigation of their stability enhancement is crucial for the wider public. We investigated the minimum rotation rate of the impeller needed for CO₂ nanobubble generation and the influence of a biomedical surfactant (Pluronic P-123) addition and concentration change on the size of nanobubbles and their stability over time. Nanobubbles were stable in the presence of surfactant additions and showed the impact of both changes in generation time and shear stress on their size. We hope that this study will be a step in the direction of the cost-efficient generation of stable carbon dioxide nanobubble dispersions.