Simulative Minimization of Mass Transfer Limitations Within Hydrogel-Based 3D-Printed Enzyme Carriers

Schmieg, Barbara; Nguyen, Mai Thanh; Franzreb, Matthias

doi:10.3389/fbioe.2020.00365

Cited by 17 publications

(17 citation statements)

References 23 publications

(29 reference statements)

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“…As already mentioned, there has been a need for developing 3D printed hydrogels with smaller and finer structures to reduce mass transfer limitations and improve the effectiveness factor of entrapped fast-acting enzymes [ 85 ]. Lahann and co-workers [ 80 ] developed a new 3D printing technique named “3D jet writing” to create micro-sized polymer hydrogel fibers oriented in a self-supporting 3D scaffold.…”

Section: Immobilization By Physical Entrapment During 3d Printingmentioning

confidence: 99%

“…The scope of this review focused only on studies that directly immobilized enzymes on or in the 3D printed objects and does not include many other innovations that use 3D printing to make customizable tools and devices, such as labware [ 88 ], mass spectrometry microcolumns [ 89 ], and biomedical devices [ 90 ], where enzymes are not directly included. The review emphasized the most recent techniques and chemistry for achieving successful enzyme immobilization by 3D printing, especially by gelation mechanisms that favor high retention of enzyme activity and longevity; however, the reader is referred elsewhere for more information on the basic physics of mass transfer limitations that may occur in these systems and how they are currently being studied through modeling [ 85 ].…”

Section: Limitation Of the Reviewmentioning

confidence: 99%

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Advances in 3D Gel Printing for Enzyme Immobilization

Shen

Zhang

Fang

et al. 2022

Gels

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Incorporating enzymes with three-dimensional (3D) printing is an exciting new field of convergence research that holds infinite potential for creating highly customizable components with diverse and efficient biocatalytic properties. Enzymes, nature’s nanoscale protein-based catalysts, perform crucial functions in biological systems and play increasingly important roles in modern chemical processing methods, cascade reactions, and sensor technologies. Immobilizing enzymes on solid carriers facilitates their recovery and reuse, improves stability and longevity, broadens applicability, and reduces overall processing and chemical conversion costs. Three-dimensional printing offers extraordinary flexibility for creating high-resolution complex structures that enable completely new reactor designs with versatile sub-micron functional features in macroscale objects. Immobilizing enzymes on or in 3D printed structures makes it possible to precisely control their spatial location for the optimal catalytic reaction. Combining the rapid advances in these two technologies is leading to completely new levels of control and precision in fabricating immobilized enzyme catalysts. The goal of this review is to promote further research by providing a critical discussion of 3D printed enzyme immobilization methods encompassing both post-printing immobilization and immobilization by physical entrapment during 3D printing. Especially, 3D printed gel matrix techniques offer mild single-step entrapment mechanisms that produce ideal environments for enzymes with high retention of catalytic function and unparalleled fabrication control. Examples from the literature, comparisons of the benefits and challenges of different combinations of the two technologies, novel approaches employed to enhance printed hydrogel physical properties, and an outlook on future directions are included to provide inspiration and insights for pursuing work in this promising field.

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Section: Immobilization By Physical Entrapment During 3d Printingmentioning

confidence: 99%

Section: Limitation Of the Reviewmentioning

confidence: 99%

Advances in 3D Gel Printing for Enzyme Immobilization

Shen

Zhang

Fang

et al. 2022

Gels

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“…For the application of the scaffolds, differing strand diameters have a huge impact on the scaffold geometry as well as the diffusive distances in the strands. [36,37] If uniform extrusion is achieved, the stackability of the ink is high (see Figure 3C and D). To ensure continuous material extrusion two strategies were used for GelMA.…”

Section: Materials Printing Behaviormentioning

confidence: 99%

Magnetic resonance imaging as a tool for quality control in extrusion‐based bioprinting

Schmieg

Gretzinger

Schuhmann

et al. 2022

Biotechnology Journal

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Bioprinting is gaining importance for the manufacturing of tailor‐made hydrogel scaffolds in tissue engineering, pharmaceutical research and cell therapy. However, structure fidelity and geometric deviations of printed objects heavily influence mass transport and process reproducibility. Fast, three‐dimensional and nondestructive quality control methods will be decisive for the approval in larger studies or industry. Magnetic resonance imaging (MRI) meets these requirements for characterizing heterogeneous soft materials with different properties. Complementary to the idea of decentralized 3D printing, magnetic resonance tomography is common in medicine, and image data processing tools can be transferred system‐independently. In this study, a MRI measurement and image analysis protocol was evaluated to jointly assess the reproducibility of three different hydrogels and a reference material. Critical parameters for object quality, namely porosity, hole areas and deviations along the height of the scaffolds are discussed. Geometric deviations could be correlated to specific process parameters, anomalies of the ink or changes of ambient conditions. This strategy allows the systematic investigation of complex 3D objects as well as an implementation as a process control tool. Combined with the monitoring of metadata this approach might pave the way for future industrial applications of 3D printing in the field of biopharmaceutics.

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“…One would expect that the restricted diffusion together with deactivation of a fraction of the enzymes due to the coupling chemistry would decrease the efficiency. [31,32] This counter intuitive result may be the result of the increased buffering effect of the CS polymer, maintaining better control of pH during the reaction as compared to the case of Urs in solution. In the absence of CS, the pH was observed to increase from 5.5 to 9 at the end of the reaction, while the presence of CS controlled the pH such that it increased from 5.5 to 6.5 (Figure S5a).…”

Section: Monitoring Enzyme Activitymentioning

confidence: 99%

Microfluidics Featuring Multilayered Hydrogel Assemblies Enable Real-Time NMR-Monitoring of Enzyme Cascade Reactions

Nordin¹,

Bordonali²,

Davoodi³

et al. 2020

Preprint

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Compartmentalized chemical reactions at the microscale are interesting from many perspectives including (multi)functional surfaces and biotechnology. Monitoring the molecular content as a measure of functional performance at these small scales is challenging. As a means to address this challenge, we leverage microtechnology and biocompatible materials to integrate a compact, reconfigurable reaction cell featuring electrochemical functionality with high-resolution nuclear magnetic resonance spectroscopy (NMR). We demonstrate the operation of this system by monitoring the activity of enzymes immobilized in chemically distinct layers within a multi-layered chitosan hydrogel assembly. As a benchmark, we observed the parallel activities of urease (Urs), catalase (Cat), and glucose oxidase (GOx) by recording NMR spectra to extract reagent and product concentrations in real-time. As a result, simultaneous monitoring of a cooperative enzymatic process (GOx + Cat) together with an independent process (Urs) is achieved. Using Michaelis-Menten progress curve analysis of the NMR data, kinetic data is extracted: in the case of GOx, the Michaelis constants (K<sub>M</sub>) are consistent with previous reports, while for Urs, deviations are observed, attributed to an inhibitory effect under our reaction conditions. The system therefore enables the construction of complex reaction cascades with spatial control, as would be interesting in, for example, metabolic engineering and multiplexed sensing applications.

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Simulative Minimization of Mass Transfer Limitations Within Hydrogel-Based 3D-Printed Enzyme Carriers

Cited by 17 publications

References 23 publications

Advances in 3D Gel Printing for Enzyme Immobilization

Advances in 3D Gel Printing for Enzyme Immobilization

Magnetic resonance imaging as a tool for quality control in extrusion‐based bioprinting

Microfluidics Featuring Multilayered Hydrogel Assemblies Enable Real-Time NMR-Monitoring of Enzyme Cascade Reactions

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