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.

Unique biosynthetic abilities revealed by plants determine in vitro cultures of hairy roots as a suitable source of pharmaceutically relevant bioactive compounds. The basic aim of the study was to examine the applicability of aerogel composed of methyltrimethoxysilane (MTMS) for immobilization of Rindera graeca hairy roots by identifying quantitative effects of biomass proliferation and naphthoquinones extracellular secretion in the aerogel-supported culture system. R. graeca hairy roots were simultaneously cultured for 28-days, as (i) nonimmobilized biomass (reference system), (ii) biomass immobilized on macroporous polyurethane foam (PUF), (iii) biomass with disintegrated MTMS aerogel, (iv) biomass immobilized on polypropylene (PP) fibers (as control), and (v) biomass immobilized on monolithic PP-reinforced MTMS aerogel. MTMS aerogel exhibited high level of biocompatibility toward R. graeca hairy roots which grew into the structure of monolithic aerogel-based constructs. Monolithic MTMS-based constructs significantly promoted the proliferation of hairy roots, resulting in 55% higher fresh mass than the reference system. The highest level of naphthoquinones productivity, i.e., 653 µg g_DW⁻¹, was noted for PUF-supported culture system.

Systems of two immiscible liquid phases—aqueous phase (i.e., distilled water (dH₂O) or phosphate-buffered saline (PBS)) and liquid perfluorochemical (i.e., perfluorodecalin (PFD))—were subjected to wave-assisted agitation, i.e., oscillatory rocked, in a disposable bag-like container in a ReadyToProcess WAVE^TM25 bioreactor, to recognize oxygen transfer effects and effectivity of the surface aeration. According to the DoE methodology, values of the volumetric liquid-side mass transfer (k_La) coefficient for dH₂O, PBS, dH₂O-PFD, and PBS-PFD systems were determined for the whole range of operating parameters of the WAVE 25 bioreactor. A significantly higher maximal value of k_La was found for waving dH₂O than for dH₂O-PFD (i.e., 0.00460 s⁻¹ vs. 0.00331 s⁻¹, respectively) compared to more equal maximal values of k_La reached for PBS and PBS-PFD (0.00355 s⁻¹ vs. 0.00341 s⁻¹, respectively). The interface development factor (f) depended on the interfacial area a, and the enhancement factor (E^PFD), depending on k_La, was introduced to quantitatively identify the mass transfer effects in the systems of waving two immiscible liquids. The phase of PFD was identified as the reservoir of oxygen. Dimensional correlations were proposed for the prediction of the k_La coefficient, in addition to the f and E^PFD factors. The presented correlations, and the set of k_La values, can be directly applied to predict oxygen transfer effects reached under continuous oscillatory rocked systems containing aqueous phase and liquid perfluorochemical.