Diffusion Studies of Glucose and Sucrose in Chitosan Membranes and Beads for Enzymatic Production Processes

Hartig, Dave; Hacke, Sandra; Ott, Lisanne; Gabrielczyk, Jakub; Müller, Clarissa; Jördening, Hans‐Joachim; Scholl, Stephan

doi:10.1002/ceat.201800075

Cited by 3 publications

(4 citation statements)

References 29 publications

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“…Across 42 studies from prominent research groups in disc cell culture, alginate beads were the most popular 3D culture system (45.2%), followed by cylindrical hydrogel constructs (31.0%) and then pellet or microaggregate cultures (23.8%), Figure 1D . The effective diffusion coefficient of oxygen, 39 , 40 , 41 , 42 glucose, 43 , 44 , 45 , 46 and lactate, 46 , 47 , 48 , 49 through a number of different hydrogels were gathered to experimentally inform the in silico models, Figure 1E . Within the bead model alginate‐specific diffusion coefficients for oxygen (1.8 × 10 −9 m 2 /s), glucose (5.1 × 10 −10 m 2 /s), and lactate (4.67 × 10 −10 m 2 /s) were used.…”

Section: Methodsmentioning

confidence: 99%

Section: Three-dimensional Cell Culture Modelsmentioning

confidence: 99%

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Two‐ and three‐dimensional in vitro nucleus pulposus cultures: An in silico analysis of local nutrient microenvironments

McDonnell

Buckley

2022

JOR Spine

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Background It is well established that the unique biochemical microenvironment of the intervertebral disc plays a predominant role in cell viability and biosynthesis. However, unless the effect of microenvironmental conditions is primary to a study objective, in vitro culture parameters that are critical for reproducibility are both varied and not routinely reported. Aims This work aims to investigate the local microenvironments of commonly used culture configurations, highlighting physiological relevance, potential discrepancies, and elucidating possible heterogeneity across the research field. Materials and Methods This work uses nutrient‐transport in silico models to reflect on the effect of often underappreciated parameters, such as culture geometry and diffusional distance (vessel, media volume, construct size), seeding density, and external boundary conditions on the local microenvironment of two‐dimensional (2D) and three‐dimensional (3D) in vitro culture systems. Results We elucidate important discrepancies between the external boundary conditions such as the incubator level or media concentrations and the actual local cellular concentrations. Oxygen concentration and cell seeding density were found to be highly influential parameters and require utmost consideration when utilizing 3D culture systems. Discussion This work highlights that large variations in the local nutrient microenvironment can easily be established without consideration of several key parameters. Without careful deliberation of the microenvironment within each specific and unique system, there is the potential to confound in vitro results leading to heterogeneous results across the research field in terms of biosynthesis and matrix composition. Conclusion Overall, this calls for a greater appreciation of key parameters when designing in vitro experiments. Better harmony and standardization of physiologically relevant local microenvironments are needed to push toward reproducibility and successful translation of findings across the research field.

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Section: Methodsmentioning

confidence: 99%

Section: Three-dimensional Cell Culture Modelsmentioning

confidence: 99%

Two‐ and three‐dimensional in vitro nucleus pulposus cultures: An in silico analysis of local nutrient microenvironments

McDonnell

Buckley

2022

JOR Spine

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“…Glucose oxidase and lactate oxidase were used as their reaction with the respective analytes produce H 2 O 2 , which can easily be detected on Prussian bluecoated carbon electrodes. Chitosan membranes were selected as protection membranes and scaffolds for enzyme immobilization due to their wide-spread and established performance in such enzyme sensors [1,2,[31][32][33][34][35][36][37]. Here, these wellestablished concepts were established on the LIG electrodes.…”

Section: Amperometric Glucose and Lactate Biosensorsmentioning

confidence: 99%

Electrochemical multi-analyte point-of-care perspiration sensors using on-chip three-dimensional graphene electrodes

et al. 2020

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Multi-analyte sensing using exclusively laser-induced graphene (LIG)-based planar electrode systems was developed for sweat analysis. LIG provides 3D structures of graphene, can be manufactured easier than any other carbon electrode also on large scale, and in form of electrodes: hence, it is predestinated for affordable, wearable point-of-care sensors. Here, it is demonstrated that LIG facilitates all three electrochemical sensing strategies (voltammetry, potentiometry, impedance) in a multi-analyte system for sweat analysis. A potentiometric potassium-ion-selective electrode in combination with an electrodeposited Ag/AgCl reference electrode (RE) enabled the detection of potassium ions in the entire physiologically relevant range (1 to 500 mM) with a fast response time, unaffected by the presence of main interfering ions and sweat-collecting materials. A kidney-shaped interdigitated LIG electrode enabled the determination of the overall electrolyte concentration by electrochemical impedance spectroscopy at a fixed frequency. Enzyme-based strategies with amperometric detection share a common RE and were realized with Prussian blue as electron mediator and biocompatible chitosan for enzyme immobilization and protection of the electrode. Using glucose and lactate oxidases, lower limits of detection of 13.7 ± 0.5 μM for glucose and 28 ± 3 μM for lactate were obtained, respectively. The sensor showed a good performance at different pH, with sweat-collecting tissues, on a model skin system and furthermore in synthetic sweat as well as in artificial tear fluid. Response time for each analytical cycle totals 75 s, and hence allows a quasi-continuous and simultaneous monitoring of all analytes. This multi-analyte all-LIG system is therefore a practical, versatile, and most simple strategy for point-of-care applications and has the potential to outcompete standard screen-printed electrodes.

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“…More specifically, GI as a main enzyme in production of high fructose corn syrup (HFCS) is used at much higher temperatures. Although diffusion in chitosan is significantly slower than free diffusion [8], the impact on overall reaction rate is small [9] and thermal stability of chitosan-based hybrid immobilizates is increased by a factor of 10 compared to native enzymes [7]. But glucose is needed for start of the reaction and kinetics requirements and for that reason, glucose is added at the beginning of the reaction [5].…”

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confidence: 99%

Continuous enzymatic production and adsorption of laminaribiose in packed bed reactors

Abi

Hartig

Vorländer

et al. 2018

Engineering in Life Sciences

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Bienzymatic production of laminaribiose from sucrose and glucose was combined with adsorption on zeolite BEA to introduce a first capture and purification step. Downstream processing including washing and desorption steps was characterized and optimized on a milliliter scale in batch mode. Results were then transferred to a packed bed system for enzymatic production and adsorption where the influence of adsorbent particle diameter on purity and productivity was evaluated. Finally, a continuous enzymatic production of laminaribiose was conducted over 10 days. The subsequent downstream processing of the loaded zeolites led to purities of over 0.5 gLaminaribiose gsugar−1 in the desorbate with a total productivity of 5.6 mgLaminaribiose Lenzyme bed−1 h−1 without the use of recycles.

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Diffusion Studies of Glucose and Sucrose in Chitosan Membranes and Beads for Enzymatic Production Processes

Cited by 3 publications

References 29 publications

Two‐ and three‐dimensional in vitro nucleus pulposus cultures: An in silico analysis of local nutrient microenvironments

Two‐ and three‐dimensional in vitro nucleus pulposus cultures: An in silico analysis of local nutrient microenvironments

Electrochemical multi-analyte point-of-care perspiration sensors using on-chip three-dimensional graphene electrodes

Continuous enzymatic production and adsorption of laminaribiose in packed bed reactors

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