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Disposable rocking bioreactors facilitate scaling up animal and plant cell biomass propagation and developing specified bioprocesses like manufacturing vaccines or chimeric antigen receptor (CAR) T cells. Future contexts for these bioreactors include supporting regenerative medicine, recognising metabolic responses of biochemically or mechanically stressed cells, continuously performing in vitro bioprocesses, or cell-free protein synthesis systems. [ABSTRACT FROM AUTHOR]

Polymers are of great interest for medical and cosmeceutical applications. The current trend is to combine materials of natural and synthetic origin in order to obtain products with appropriate mechanical strength and good biocompatibility, additionally biodegradable and bioresorbable. Citric acid, being an important metabolite, is an interesting substance for the synthesis of materials for biomedical applications. Due to the high functionality of the molecule, it is commonly used in biomaterials chemistry as a crosslinking agent. Among citric acid‐based biopolyesters, poly(1,8‐octanediol citrate) is the best known. It shows application potential in soft tissue engineering. This work focuses on a much less studied polyester, poly(1,3‐propanediol citrate). Porous and non‐porous materials based on the synthesized polyesters are prepared and characterized, including mechanical, thermal, and surface properties, morphology, and degradation. The main focus is on assessing the biocompatibility and antimicrobial properties of the materials. [ABSTRACT FROM AUTHOR]

Secondary metabolites derived from plants are recognized as valuable products with several successful applications in the pharmaceutical, cosmetic, and food industries. The major limitation to the broader implementation of these compounds is their low manufacturing efficiency. Current efforts to overcome unprofitability depend mainly on biotechnological methods, especially through the application of plant in vitro cultures. This concept allows unprecedented bioengineering opportunities for culture system modifications with in situ product removal. The silica-based xerogels can be used as a novel, porous biomaterial characterized by a large surface area and high affinity to lipophilic secondary metabolites produced by plant tissue. This study aimed to investigate the influence of xerogel-based biomaterials functionalized with methyl, hydroxyl, carboxylic, and amine groups on Rindera graeca transgenic root growth and the production of naphthoquinone derivatives. The application of xerogel-based scaffolds functionalized with the methyl group resulted in more than 1.5 times higher biomass proliferation than for reference untreated culture. The naphthoquinone derivatives' production was noted exclusively in culture systems supplemented with xerogel functionalized with methyl and hydroxyl groups. Applying chemically functionalized xerogels as in situ adsorbents allowed for the enhanced growth and productivity of in vitro cultured R. graeca transgenic roots, facilitating product isolation due to their selective and efficient accumulation. [ABSTRACT FROM AUTHOR]

In situ extraction is a method for separating plant secondary metabolites from in vitro systems of plant biomass cultures. The study aimed to investigate the MTMS-based xerogels morphology effect on the growth kinetics and deoxyshikonin productivity in xerogel-supported in vitro culture systems of Rindera graeca hairy root. Cultures were supplemented with three types of xerogel, i.e., mesoporous gel, microporous gel, and agglomerated precipitate, in the disintegrated or monolithic form. Structure, oil sorption capacity, and SEM analyses for xerogel-based additives were performed. Application of monolithic macroporous xerogel resulted in the highest biomass proliferation, i.e., 5.11-fold fresh biomass increase after four weeks of the screening culture. The highest deoxyshikonin production (i.e., 105.03 µg) was noted when hairy roots were maintained with particles of disintegrated mesoporous xerogel. The detailed kinetics investigations (6-week culture) revealed the highest growth of hairy root biomass and secondary metabolite production, equaling 9.46-fold fresh weight biomass and 204.08 µg deoxyshikonin, respectively. MTMS-based xerogels have been recognized as selective biocompatible scaffolds for boosting the proliferation of transgenic roots or for productivity enhancement of naphthoquinones without detrimental effects on biomass growth, and their successful applicability in in situ removal of secondary plant metabolites has been experimentally confirmed. [ABSTRACT FROM AUTHOR]

The study aims to approach a coating layer based on alginate functionalized with albumin nanocoated oxygenating factor, constituting a safe and even contact surface with epithelial cells in system components for medical devices to support maintaining the human lung cells' function. The alginate coating layer with incorporated albumin-coated oxygenating element was designed (ALG_Alb-PFD). As an oxygenating factor, perfluorodecalin was applied at three saturation levels with oxygen: 0%, 21%, and 100%. The studies were performed on human adenocarcinoma A549 cell lines derived from human lung epithelial cells cultured in configuration with the designed coatings. Scanning electron microscopy, flow cytometry, and fluorescence microscopy were applied to evaluate cell function and the system morphology. AFM and SEM-EDX characterized the coating layer material. The designed coating ensured the viability of human lung cells cultured on the coating layer during 10 days of culture. An oxygenating factor presence was confirmed by SEM-EDX analysis of the ALG_Alb-PFD. However, no significant discrepancies were noted between the coatings with different saturation levels after 10 days of culture. Moreover, the AFM-analyzed profiles of the coating surface confirmed/indicated the formation of additional elements in the coating due to using an oxygenating element, which increases the effective contact area with cells. The platform based on alginate with the developed surface of contact of oxygenating factor incorporated within the coating layer ensures the sustained growth and morphology of the human lung cells, which can be considered a therapeutic element of system components of medical devices for maintaining human lung cells' function. [Display omitted] [ABSTRACT FROM AUTHOR]

Single-use (i.e. disposable) bioreactors are commonly applied in biotechnology, as well as in biopharmaceutical industry, to efficient scaling-up of cultures of animal cells. Wave-type agitated systems provide sufficient mass transfer conditions in single-use culture bags. Spherical microcarriers made with biocompatible polymers are applying to obtain higher levels of biomass yield in various types of in vitro systems for culture of anchorage-dependent (i.e. adherent) cells. The aim of the study was to determine the applicability of three spherical microcarriers, i.e. Cytodex 1, Cytodex 3 and Cytopore, which differ in surface properties of granules, for submerged batch culture of mice anchorage-dependent L929 fibroblasts under conditions of wave-type agitation. Based on the results of cultures performed in small-scale wave-agitated system (5 cm3, laboratory cradle), the Cytodex 3 microcarrier has been identified as the spherical granules promoting proliferation of L929 cells. Cytodex 3 has been also successfully applied in the ReadyToProcess WAVETM25 bioreactor system (WAVE 25, GE Healthcare Bio-Sciences AB, Sweden) equipped with polymer-based culture bag in volume of 300 cm3, what resulted in enhanced growth of L929 cells characterized by Y*X/S equaled to 1.48·109 cells·gglucose-1. [ABSTRACT FROM AUTHOR]

The physicochemical parameters of the liquid phase have influence significantly on the rate of oxygen absorption in liquid. Feasibility of wave-type agitated single-use bioreactors for bioprocesses involving in vitro cultures makes the study of the influence of liquid properties on values of the volumetric mass transfer coefficient (kLa) interesting and actual research issue. ReadyToProcess WAVE™25 single-use bioreactor system (WAVE 25; GE Healthcare Bio-Sciences AB, Sweden), equipped with 2 dm3 polymer-based culture bag, has been used for studying the oxygen absorption rate in the wave-induced agitation system. The influence of viscosity of liquid on the rate of oxygen absorption under wave-type agitation has been discussed. The increase of liquid phase viscosity negatively influenced on the rate of oxygen absorption, and lead to decreasing in values of the kLa coefficient in studied system. Fetal bovine serum strongly limited significantly the rate of oxygen absorption into the liquid phase, what resulted in a significant decrease in the kLa coefficient. [ABSTRACT FROM AUTHOR]

[Display omitted] • Study on mixing time in disposable bioreactor by sensor method and DoE-methodology. • Mixing time in a 2-litre disposable container ranges from 3 to 1350 s. • Mixing intensity influenced by rocking angle, frequency, and the volume of liquid. • Correlation based on dimensionless terms predicts t 95% for vessels of various sizes. • A step towards the better systematization of liquid flow inside rocking bioreactors. Rocking single-use bioreactors (SUBs) adopt an effective mixing mechanism based on the rocking movement of a platform on which a disposable bag-like container is placed. Non-typical liquid flow patterns make the hydrodynamical properties of rocking SUBs not yet well systematized. The aim of the study was to determine a quantitative characteristic of mixing time in a ReadyToProcess WAVE 25 system equipped with a 2-litre Cellbag disposable container, with the use of a sensor method and the Design of Experiment (DoE) methodology. The DoE-based approach has been used to screen significant operational parameters and evaluate their impact on the values of mixing time reached in the studied rocking SUB system. To quantify relations between operational parameters and important process quantities in rocking disposable bag-like containers of different sizes, a set of original formulas for generalization of experimental data and predicting mixing time values has been proposed. The set includes exponential correlations and a correlation involving a dimensionless filling level term and a modified Reynolds number expression. Comparisons between estimations and experimental data have shown satisfactory levels of accuracy within ± 30% of experimental values. The correlation involving dimensionless terms can be applied for disposable bag-like containers offered by different manufacturers and of various sizes. [ABSTRACT FROM AUTHOR]

• Microcarrier-supported intensification of CP5 cells growth in disposable bioreactor. • Screening microcarrier for the intensified proliferation of CP5 chondrocytes. • Scaling-up of disposable culture system from 5 cm3 to 300 cm3. • Wave-assisted agitation intensifies chondrocytes proliferation on microcarriers. The growth of CP5 chondrocytes on microcarriers beads in a disposable bioreactor under various conditions of wave-assisted agitation has been compared and discussed. Influences of Reynolds number for liquid (Re L) and the k L a coefficient on intensification of the growth rate (μ) and the specific yield of biomass (Y* X/S) have been identified. Culture system has been established in ReadyToProcess WAVETM25 with a 2-litre flexible bag-like container. Cytodex 3 beads have been identified as the most suitable microcarrier fulfilled CP5 chondrocytes requirements. Based on simplified dimensionless correlations, the values of Re L (i.e., 510–10,208), and k L a coefficient (i.e., 2.83–13.55 h–1), have been both determined for various presets of the WAVE 25 system. The highest values of μ = 0.0129 h–1 and Y * X/S = 10.9 × 108 cells g glc –1 have been noted for culture performed under wave-induced agitation characterised by Re L equalled to 5104. The results might be highly applicable in scaling-up, and intensifying microcarrier-engineered bioprocesses focused on adherent mammalian or human cells maintained in disposable bioreactors. [Display omitted] [ABSTRACT FROM AUTHOR]

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 WAVETM25 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 (kLa) 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 kLa 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 kLa 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 (EPFD), depending on kLa, 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 kLa coefficient, in addition to the f and EPFD factors. The presented correlations, and the set of kLa values, can be directly applied to predict oxygen transfer effects reached under continuous oscillatory rocked systems containing aqueous phase and liquid perfluorochemical. [ABSTRACT FROM AUTHOR]

The knowledge of a volumetric liquid-side mass transfer coefficient (kLa) characterizing the oxygen transfer in bioreactor working at defined operating parameters, is a fundamental principle for establishing the aeration strategy for aerobic bioprocesses. The design of experiments (DoE) methodology has been applied for distinguishing relevant from irrelevant operating parameters, and for prediction of characteristics of oxygen mass transfer effects, in the whole range of values of operating parameters accessible in setup of ReadyToProcess WAVE™ 25 (WAVE 25; GE Healthcare) bioreactor equipped with 2 dm³ disposable culture bag. Due to DoE-aided analysis, rocking speed (ω), rocking angle (α) and volumetric flow of gas phase through the culture bag (QG) have been indicated as the operating parameters robustly impacting on the value of the k[sub L]a coefficient. All relevant operational parameters, i.e. ω, α a and QG, exerted monotonically increasing influence on kLa . Otherwise, the influence of volume of liquid poured into culture bag (VL) and oxygen partial pressure in applied gas phase (p1) on kLa proved to be negligible. Two original correlations have been proposed to generalize the experimental results and to estimate the kLa coefficient values possible to be reached in the WAVE 25: the dimensional correlation defining the kLa coefficient, as well as the dimensionless correlation that defines Sherwood number and integrating the originally-defined Reynolds number for the liquid phase that is subjected to wave-induced mixing. The validity of results predicted by both correlations has been verified by acceptable level of the relative errors. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • Mass transfer in single-use WAVE™ 25 bioreactor was investigated in details. • Relevant vs. irrelevant operating parameters influencing on k L a were screened with DoE. • Operating parameters impacting mass transfer in WAVE™25 were identified. • New and original form of Re number for wave-induced mixing of liquids was defined. • Correlations to estimate the k L a coefficient values in the WAVE™ 25 were proposed. Abstract The knowledge of a volumetric liquid-side mass transfer coefficient ( k L a ) characterizing the oxygen transfer in bioreactor working at defined operating parameters, is a fundamental principle for establishing the aeration strategy for aerobic bioprocesses. The design of experiments (DoE) methodology has been applied for distinguishing relevant from irrelevant operating parameters, and for prediction of characteristics of oxygen mass transfer effects, in the whole range of values of operating parameters accessible in setup of ReadyToProcess WAVE™ 25 (WAVE 25; GE Healthcare) bioreactor equipped with 2 dm3 disposable culture bag. Due to DoE-aided analysis, rocking speed ( ω ), rocking angle ( α ) and volumetric flow of gas phase through the culture bag ( Q G ) have been indicated as the operating parameters robustly impacting on the value of the k L a coefficient. All relevant operational parameters, i.e. ω , α and Q G , exerted monotonically increasing influence on k L a. Otherwise, the influence of volume of liquid poured into culture bag ( V L ) and oxygen partial pressure in applied gas phase ( p 1 ) on k L a proved to be negligible. Two original correlations have been proposed to generalize the experimental results and to estimate the k L a coefficient values possible to be reached in the WAVE 25: the dimensional correlation defining the k L a coefficient, as well as the dimensionless correlation that defines Sherwood number and integrating the originally-defined Reynolds number for the liquid phase that is subjected to wave-induced mixing. The validity of results predicted by both correlations has been verified by acceptable level of the relative errors. [ABSTRACT FROM AUTHOR]