bims-enlima 2024-12-22 papers

bims-enlima

Biomed News

on Engineered living materials

Issue of 2024–12–22
24 papers selected by
Rahul Kumar, Tallinna Tehnikaülikool

Turing patterns with cellular computers.
3D extrusion bioprinting of microbial inks for biomedical applications.
Evaluating the Antibacterial Potential of Distinct Size Populations of Stabilized Zinc Nanoparticles.
Hyperballistic intercellular signaling through growth assisted positive feedback.
Extracellular Peptide-Ligand Dimerization Actuator Receptor Design for Reversible and Spatially Dosed 3D Cell-Material Communication.
Natural nanofibers embedded in the seed mucilage envelope: composite hydrogels with specific adhesive and frictional properties.
Materials Nanoarchitectonics for Advanced Devices.
Polyacrylamide Hydrogel Containing Starch and Sugarcane Bagasse Ash: Synthesis, Characterisation, and Application in Cement Pastes and Mortars.
Utilizing AI algorithms to model and optimize the composite of nanocellulose and hydrogels via a new technique.
Cutting-edge developments in plastic biodegradation and upcycling via engineering approaches.
Self-healing, 3D printed bioinks from self-assembled peptide and alginate hybrid hydrogels.
Mechanochemical topological defects in an active nematic.
3D printing of bacterial poly(3-hydroxybutyrate-co-3-hydroxyvalerate)/poly(lactide-co-glycolide) blend loaded with β-tricalcium phosphate for the development of scaffolds to support human mesenchymal stromal cell proliferation.
Quantifying Biomass and Visualizing Cell Coverage on Fibrous Scaffolds for Cultivated Meat Production.
Photocrosslinkable Biomaterials for 3D Bioprinting: Mechanisms, Recent Advances, and Future Prospects.
Polymer Sorbent Design for the Direct Air Capture of CO2.
Protocol for the synthesis and activation of hydrogels with photocaged oligonucleotides.
Efficient Enzymatic Glycan Engineering of Extracellular Vesicles Using Nanomaterial-Interfaced Microfluidics.
Development and application of hydrogels in pathogenic bacteria detection in foods.
Counterfactual rewards promote collective transport using individually controlled swarm microrobots.
Influence of Internal Architecture and Ink Formulation on the Thermal Behavior of 3D-Printed Cementitious Materials.
Bioprinting extracellular vesicles as a "cell-free" regenerative medicine approach.
Spatially defined microenvironment for engineering organoids.
Harnessing genetically engineered cell membrane-derived vesicles as biotherapeutics.

Cell Syst. 2024 Dec 18. pii: S2405-4712(24)00351-X. [Epub ahead of print]15(12): 1105-1106

Turing patterns with cellular computers.

Lewis Grozinger, Ángel Goñi-Moreno.

Turing patterns are a key theoretical foundation for understanding organ development and organization. While they have been found to occur in natural systems, implementing new biological systems that form Turing patterns has remained challenging. To address this, Tica et al.1 used synthetic genetic networks to engineer living cellular computers that successfully generate Turing patterns within growing bacterial populations.

DOI: https://doi.org/10.1016/j.cels.2024.11.015
Adv Drug Deliv Rev. 2024 Dec 17. pii: S0169-409X(24)00327-2. [Epub ahead of print] 115505

3D extrusion bioprinting of microbial inks for biomedical applications.

Nicolas Burns, Arjun Rajesh, Avinash Manjula-Basavanna, Anna Duraj-Thatte.

  In recent years, the field of 3D bioprinting has witnessed the intriguing development of a new type of bioink known as microbial inks. Bioinks, typically associated with mammalian cells, have been reimagined to involve microbes, enabling many new applications beyond tissue engineering and regenerative medicine. This review presents the latest advancements in microbial inks, including their definition, types, composition, salient characteristics, and biomedical applications. Herein, microbes are genetically engineered to produce 1) extrudable bioink and 2) life-like functionalities such as self-regeneration, self-healing, self-regulation, biosynthesis, biosensing, biosignaling, biosequestration, etc. We also discuss some of the promising applications of 3D extrusion printed microbial inks, such as 1) drugs and probiotics delivery, 2) metabolite production, 3) tissue engineering, 4) bioremediation, 5) biosensors and bioelectronics, 6) biominerals and biocomposites, and 7) infectious disease modeling. Finally, we describe some of the current challenges of microbial inks that needs to be addressed in the coming years, to make a greater impact in health science and technology and many other fields.

Keywords:  3D Printing; Additive Manufacturing; Bioinks; Bioprinting; Engineered Living Materials; Extrusion; Microbial Inks

DOI:  https://doi.org/10.1016/j.addr.2024.115505
ACS Appl Mater Interfaces. 2024 Dec 16.

Evaluating the Antibacterial Potential of Distinct Size Populations of Stabilized Zinc Nanoparticles.

Dinny Stevens, Amanda K Charlton-Sevcik, W Evan Braswell, Christie M Sayes.

  Engineered nanoparticles are precisely synthesized to exploit unique properties conferred by their small size and high surface area for environmental, biomedical, and agricultural applications. While these physical properties dictate functionality, they can also have various intended and unintended implications for biological systems. Both the particle size and shape influence cellular uptake. Because of zinc's antibacterial properties and role as a plant micronutrient, polyvinylpyrrolidone stabilized zinc nanoparticles (ZnNP) were selected for this study. Four synthesis methods were tested to produce distinct size populations of polymer-coated ZnNP, and all utilized water as the solvent to promote sustainable, green chemistry. The antibacterial activity of ZnNP was assessed in two agriculturally relevant bacteria strains: Escherichia coli and Bacillus cereus. To further examine the effects of ZnNP on bacterial cells, reactive oxygen species (ROS) generation was measured via hydrogen peroxide (H2O2) production. The bacteria's incubation temperature was also altered to assess bacterial growth and susceptibility after exposure to ZnNP. The ZnNP from the smaller size population inhibited the most growth across bacterial strains, assays, and incubation temperatures. Increased antibacterial effects and ROS production were observed after incubation at a higher temperature. These results indicate that the deliberately designed nanoparticles are potentially valuable in microbial control and offer promising solutions for the future of healthy agricultural systems.

Keywords:  B. cereus; E. coli; bacterial growth inhibition; incubation temperature; reactive oxygen species

DOI:  https://doi.org/10.1021/acsami.4c15245
bioRxiv. 2024 Dec 05. pii: 2024.12.04.626899. [Epub ahead of print]

Hyperballistic intercellular signaling through growth assisted positive feedback.

Meidi Wang, Louis González, Soutick Saha, Krešimir Josić, Andrew Mugler, Matthew R Bennett.

Intercellular signaling in bacteria is often mediated by small molecules secreted by cells. These small molecules disperse via diffusion which limits the speed and spatial extent of information transfer in spatially extended systems. Theory shows that a secondary signal and feedback circuits can speed up the flow of information and allow it to travel further. Here, we construct and test several synthetic circuits in Escherichia coli to determine to what extent a secondary signal and feedback can improve signal propagation in bacterial systems. We find that positive feedback-regulated secondary signals propagate further and faster than diffusion-limited signals. Additionally, the speed at which the signal propagates can accelerate in time, provided the density of the cells within the system increases. These findings provide the foundation for creating fast, long-range signal propagation circuits in spatially extended bacterial systems.

DOI: https://doi.org/10.1101/2024.12.04.626899
ACS Synth Biol. 2024 Dec 20.

Extracellular Peptide-Ligand Dimerization Actuator Receptor Design for Reversible and Spatially Dosed 3D Cell-Material Communication.

Matthias Recktenwald, Ritankar Bhattacharya, Mohammed Mehdi Benmassaoud, James MacAulay, Varun M Chauhan, Leah Davis, Evan Hutt, Peter A Galie, Mary M Staehle, Nichole M Daringer, Robert J Pantazes, Sebastián L Vega.

  Transmembrane receptors that endow mammalian cells with the ability to sense and respond to biomaterial-bound ligands will prove instrumental in bridging the fields of synthetic biology and biomaterials. Materials formed with thiol-norbornene chemistry are amenable to thiol-peptide patterning, and this study reports the rational design of synthetic receptors that reversibly activate cellular responses based on peptide-ligand recognition. This transmembrane receptor platform, termed Extracellular Peptide-ligand Dimerization Actuator (EPDA), consists of stimulatory or inhibitory receptor pairs that come together upon extracellular peptide dimer binding with corresponding monobody receptors. Intracellularly, Stimulatory EPDAs phosphorylate a substrate that merges two protein halves, whereas Inhibitory EPDAs revert split proteins back to their unmerged, inactive state via substrate dephosphorylation. To identify ligand-receptor pairs, over 2000 candidate monobodies were built in silico using PETEI, a novel computational algorithm we developed. The top 30 monobodies based on predicted peptide binding affinity were tested experimentally, and monobodies that induced the highest change in protein merging (green fluorescent protein, GFP) were incorporated in the final EPDA receptor design. In soluble form, stimulatory peptides induce intracellular GFP merging in a time- and concentration-dependent manner, and varying levels of green fluorescence were observed based on stimulatory and inhibitory peptide-ligand dosing. EPDA-programmed cells encapsulated in thiol-norbornene hydrogels patterned with stimulatory and inhibitory domains exhibited 3D activation or deactivation based on their location within peptide-patterned hydrogels. EPDA receptors can recognize a myriad of peptide-ligands bound to 3D materials, can reversibly induce cellular responses beyond fluorescence, and are widely applicable in biological research and regenerative medicine.

Keywords:  biomaterials; computational protein design; peptides; synthetic biology; transmembrane receptors

DOI:  https://doi.org/10.1021/acssynbio.4c00482
Beilstein J Nanotechnol. 2024 ;15 1603-1618

Natural nanofibers embedded in the seed mucilage envelope: composite hydrogels with specific adhesive and frictional properties.

Agnieszka Kreitschitz, Stanislav N Gorb.

  The increasing interests in natural, biodegradable, non-toxic materials that can find application in diverse industry branches, for example, food, pharmacy, medicine, or materials engineering, has steered the attention of many scientists to plants, which are a known source of natural hydrogels. Natural hydrogels share some features with synthetic hydrogels, but are more easy to obtain and recycle. One of the main sources of such hydrogels are mucilaginous seeds and fruits, which produce after hydration a gel-like, transparent capsule, the so-called mucilage envelope. Mucilage serves several important biological functions, such as supporting seed germination, protecting seeds against pathogens and predators, and allowing the seed to attach to diverse surfaces (e.g., soil or animals). The attachment properties of mucilage are thus responsible for seed dispersal. Mucilage represents a hydrophilic, three-dimensional network of polysaccharides (cellulose, pectins, and hemicelluloses) and is able to absorb large amounts of water. Depending on the water content, mucilage can behave as an efficient lubricant or as strong glue. The current work attempts to summarise the achievements in the research on the mucilage envelope, primarily in the context of its structure and physical properties, as well as biological functions associated with these properties.

Keywords:  adhesion; cellulose; friction; hydrogel; mucilage envelope; seeds

DOI:  https://doi.org/10.3762/bjnano.15.126
Materials (Basel). 2024 Dec 03. pii: 5918. [Epub ahead of print]17(23):

Materials Nanoarchitectonics for Advanced Devices.

Katsuhiko Ariga.

  Advances in nanotechnology have made it possible to observe and evaluate structures down to the atomic and molecular level. The next step in the development of functional materials is to apply the knowledge of nanotechnology to materials sciences. This is the role of nanoarchitectonics, which is a concept of post-nanotechnology. Nanoarchitectonics is defined as a methodology to create functional materials using nanounits such as atoms, molecules, and nanomaterials as building blocks. Nanoarchitectonics is very general and is not limited to materials or applications, and thus nanoarchitecture is applied in many fields. In particular, in the evolution from nanotechnology to nanoarchitecture, it is useful to consider the contribution of nanoarchitecture in device applications. There may be a solution to the widely recognized problem of integrating top-down and bottom-up approaches in the design of functional systems. With this in mind, this review discusses examples of nanoarchitectonics in developments of advanced devices. Some recent examples are introduced through broadly dividing them into organic molecular nanoarchitectonics and inorganic materials nanoarchitectonics. Examples of organic molecular nanoarchitecture include a variety of control structural elements, such as π-conjugated structures, chemical structures of complex ligands, steric hindrance effects, molecular stacking, isomerization and color changes due to external stimuli, selective control of redox reactions, and doping control of organic semiconductors by electron transfer reactions. Supramolecular chemical processes such as association and intercalation of organic molecules are also important in controlling device properties. The nanoarchitectonics of inorganic materials often allows for control of size, dimension, and shape, and their associated physical properties can also be controlled. In addition, there are specific groups of materials that are suitable for practical use, such as nanoparticles and graphene. Therefore, nanoarchitecture of inorganic materials also has a more practical aspect. Based on these aspects, this review finally considers the future of materials nanoarchitectonics for further advanced devices.

Keywords:  advanced device; doping control of organic semiconductor; inorganic materials nanoarchitectonics; nanoarchitectonics; organic molecular nanoarchitectonics; structural control

DOI:  https://doi.org/10.3390/ma17235918
Materials (Basel). 2024 Dec 01. pii: 5889. [Epub ahead of print]17(23):

Polyacrylamide Hydrogel Containing Starch and Sugarcane Bagasse Ash: Synthesis, Characterisation, and Application in Cement Pastes and Mortars.

Ana Elizabete Nunes Pereira, Edson Araujo de Almeida, Fábio Rodrigo Kruger, Edson Cavalcanti da Silva-Filho, Edvani Curti Muniz.

  Internal curing is a process based on the addition of materials that function as water reservoirs in cementitious media. Superabsorbent hydrogels are an alternative that can be used as an internal curing agent, as they have the ability to absorb and release water in a controlled manner. In the present work, superabsorbent hydrogels based on crosslinked polyacrylamide in the presence of starch and sugarcane bagasse ash (SCBA) were developed and applied to mortars as an internal curing agent. The synthesized hydrogels were evaluated by SEM, FTIR, and swelling analysis. Cement pastes and mortars were produced using different amounts of hydrogel (0.03%, 0.06%, and 0.1% by weight). An analysis of the cement pastes and mortars revealed that hydrogel contributes to hydration, thus improving the quality of the product. Furthermore, the addition of 0.03% hydrogel by weight increased the mechanical resistance of the mortars in up to 26.8% at 28 days of curing as compared with reference (without hydrogel). To the best of our knowledge, this is the first study to use a hydrogel based on polyacrylamide crosslinked with starch and SCBA as a curing agent for mortars and cement pastes. This approach is environmentally friendly, because it uses a natural product (starch) and a byproduct from the sugarcane industry (SCBA).

Keywords:  hydrogel; hydrolysis; mechanical resistance; mortar; starch; sugarcane bagasse

DOI:  https://doi.org/10.3390/ma17235889
Int J Biol Macromol. 2024 Dec 17. pii: S0141-8130(24)09714-9. [Epub ahead of print] 138903

Utilizing AI algorithms to model and optimize the composite of nanocellulose and hydrogels via a new technique.

Baohua Shen, Bibo Qian, Ni Tu.

  Plants, various biological organisms, and certain marine organisms typically provide biopolymers, like cellulose. Some things that make them unique are that they are non-toxic, biodegradable, have high specific strength and specific modulus, are easy to change the surface of, are highly hydrophilic, and are biocompatible. Significantly, nanocellulose has emerged as a prominent development in the 21st century. The objective of this work was to create a model that can accurately predict and optimize the viscosity, storage modulus (G'), and loss modulus (G″) of sulfate nanocellulose (S-NC) hydrogen materials. These properties were analyzed in different experimental settings. To do this, the researchers used the RSM and multi-layer perceptron (MLP)-ANN techniques to accurately represent and optimize the viscosity, G', and G″ properties. Ultimately, the researchers conducted RSM optimization to identify the optimal patterns of viscosity, G', and G″ characteristics for a new method of producing nanocellulose materials. The results showed that the ANN and RSM methods were very good at predicting how nanocellulose hydrogels would behave while nanocellulose products were being made. Moreover, the ANN technique exhibited superior accuracy in forecasting processes' G' and G' behavior compared to the RSM method. Ultimately, the ideal viscosity state was attained by using a shear rate value of 95 S-1 and including 1.5 wt% of S-NC. The optimal mode for G' and G″ was achieved at a frequency of 14.532 Hz and an S-NC concentration of 1.468 wt%.

Keywords:  Biopolymers; Hydrogels; Modeling; Nano-cellulose; Optimization

DOI:  https://doi.org/10.1016/j.ijbiomac.2024.138903
Metab Eng Commun. 2024 Dec;19 e00256

Cutting-edge developments in plastic biodegradation and upcycling via engineering approaches.

Zeinab Rezaei, Amir Soleimani Dinani, Hamid Moghimi.

  The increasing use of plastics has resulted in the production of high quantities of plastic waste that pose a serious risk to the environment. The upcycling of plastics into value-added products offers a potential solution for resolving the plastics environmental crisis. Recently, various microorganisms and their enzymes have been identified for their ability to degrade plastics effectively. Furthermore, many investigations have revealed the application of plastic monomers as carbon sources for bio-upcycling to generate valuable materials such as biosurfactants, bioplastics, and biochemicals. With the advancement in the fields of synthetic biology and metabolic engineering, the construction of high-performance microbes and enzymes for plastic removal and bio-upcycling can be achieved. Plastic valorization can be optimized by improving uptake and conversion efficiency, engineering transporters and enzymes, metabolic pathway reconstruction, and also using a chemo-biological hybrid approach. This review focuses on engineering approaches for enhancing plastic removal and the methods of depolymerization and upcycling processes of various microplastics. Additionally, the major challenges and future perspectives for facilitating the development of a sustainable circular plastic economy are highlighted.

Keywords:  Biodegradation; Chemo-biological upcycling; Metabolic engineering; Plastic upcycling; Plastic wastes

DOI:  https://doi.org/10.1016/j.mec.2024.e00256
Biomater Adv. 2024 Dec 08. pii: S2772-9508(24)00388-1. [Epub ahead of print]169 214145

Self-healing, 3D printed bioinks from self-assembled peptide and alginate hybrid hydrogels.

Emily H Field, Julian Ratcliffe, Chad J Johnson, Katrina J Binger, Nicholas P Reynolds.

  There is a pressing need for new cell-laden, printable, biomaterials that are rigid and highly biocompatible. These materials can mimic stiffer tissues such as cartilage, fibrotic tissue and cancer microenvironments, and thus have exciting applications in regenerative medicine, wound healing and cancer research. Self-assembled peptides (SAPs) functionalised with aromatic groups such as Fluorenyl-9-methoxycarbonyl (Fmoc) show promise as components of these biomaterials. However, the harsh basic conditions often used to solubilise SAPs leads to issues with toxicity and reproducibility. Here, we have designed a hybrid material comprised of self-assembled Fmoc-diphenylalanine (Fmoc-FF) assemblies dispersed throughout a sodium alginate matrix and investigated the influence of different organic solvents as peptide solubilising agents. Bioinks fabricated from peptides dissolved in 1,1,1,3,3,3-Hexafluoro-2-propanol (HFIP) showed improved biocompatibility compared to those made from Dimethyl Sulfoxide (DMSO) peptide stocks, due to the increased volatility and reduced surface tension of HFIP, allowing for more efficient expulsion from the system. Through optimisation of assembly and solvent conditions we can generate hybrid bioinks with stiffnesses up to 8 times greater than sodium alginate alone that remain highly printable, even when laden with high concentrations of cells. In addition, the shear-thinning nature of the self-assembled peptide assemblies gave the hybrid bioinks highly desirable self-healing capabilities. Our developed hybrid materials allow the bioprinting of materials previously considered too stiff to extrude without causing shear induced cytotoxicity with applications in tissue engineering and biosensing.

Keywords:  3D bioprinting; 3D cell culture; Bioengineering; Biomaterials; Fmoc-FF; Hydrogel

DOI:  https://doi.org/10.1016/j.bioadv.2024.214145
Phys Rev E. 2024 Nov;110(5-1): 054605

Mechanochemical topological defects in an active nematic.

Michael M Norton, Piyush Grover.

We propose a reaction-diffusion system that converts topological information of an active nematic into chemical signals. We show that a curvature-activated reaction dipole is sufficient for creating a system that dynamically senses topology by producing a concentration field possessing local extrema coinciding with ±1/2 defects. The enabling term is analogous to polarization charge density seen in dielectric materials. We demonstrate the ability of this system to identify defects in both passive and active nematics. Our results illustrate that a relatively simple feedback scheme, expressed as a system of partial differential equations, is capable of producing chemical signals in response to inherently nonlocal structures in anisotropic media. We posit that such coarse-grained systems can help generate testable hypotheses for regulated processes in biological systems, such as morphogenesis, and motivate the creation of bio-inspired materials that utilize dynamic coupling between nematic structure and biochemistry.

DOI: https://doi.org/10.1103/PhysRevE.110.054605
Int J Biol Macromol. 2024 Dec 12. pii: S0141-8130(24)09555-2. [Epub ahead of print]288 138744

3D printing of bacterial poly(3-hydroxybutyrate-co-3-hydroxyvalerate)/poly(lactide-co-glycolide) blend loaded with β-tricalcium phosphate for the development of scaffolds to support human mesenchymal stromal cell proliferation.

Gianni Pecorini, Marco A N Domingos, Stephen M Richardson, Leonardo Carmassi, Diego Li Vecchi, Gianluca Parrini, Dario Puppi.

  Polyhydroxyalkanoates (PHAs) are microbially produced aliphatic polyesters investigated for tissue engineering thanks to their biocompatibility, processability, and suitable mechanical properties. Taking advantage of these properties, the present study investigates the development by 3D printing of bacterial poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) scaffolds loaded with β-tricalcium phosphate (β-TCP) for bone tissue regeneration. PHBV blending with poly(lactide-co-glycolide) (PLGA) (30 wt%) was exploited to enhance material processability via an optimized computer-aided wet-spinning approach. In particular, PHBV/PLGA blends were loaded with different β-TCP percentages (up to 15 wt%) by suspending the ceramic particles into the polymeric solution. The composite materials were successfully fabricated into 3D scaffolds with a fully interconnected porous architecture, as demonstrated by scanning electron microscopy. The ceramic phase had a significant effect on PHBV crystallization as shown by differential scanning calorimetry, as well as on scaffold mechanical properties with a significant increase of compressive modulus in the case of 5 wt% β-TCP loading. In addition, in vitro cell culture experiments demonstrated that β-TCP loading led to a significant increase of the viability of human mesenchymal stem/stromal cells grown on the scaffolds. Taken together, our data suggest that microbial PHBV processability, biomechanical performance, and bioactivity can be improved through combined PLGA blending and β-TCP loading.

Keywords:  3D printing; Bioactive ceramic; Polyhydroxyalkanoate

DOI:  https://doi.org/10.1016/j.ijbiomac.2024.138744
Curr Protoc. 2024 Dec;4(12): e70076

Quantifying Biomass and Visualizing Cell Coverage on Fibrous Scaffolds for Cultivated Meat Production.

Xinxin Li, Khin Thu Lwin, Hirunika U Kumarasinghe, Lilianne Iglesias-Ledon, Eesha Bethi, Yushu Wang, Colin Fennelly, Ryan Sylvia, Sonja Hatz, Timothy Olsen, Thomas Herget, Ying Chen, David Kaplan.

  Cultivated meat represents a transformative solution to environmental and ethical concerns of traditional meat industries, replicating livestock meat's texture and sensory attributes in vitro with a focus on cost, safety, and nutritional quality. Central to this process are biomaterial scaffolds that support tissue development from isolated animal cells grown in or on these matrices. Understanding scaffold interactions with cells, including scaffold degradation and biomass production, is crucial for process design and for scaling-up goals. In this article, we outline comprehensive methods to quantify scaffold-cell interactions for such scenarios, focusing on biomaterial scaffold degradation and changes in cell biomass [measured by cell weight, extracellular matrix (ECM) deposition, and cell coverage] during cell culture. We introduce two methodologies for assessing cell coverage: fixation and staining for detailed imaging analysis, and non-invasive, real-time evaluation across scaffolds. Here we focus on fiber-based scaffolds, while the assessments can be extrapolated to 2-dimensional (2D; films), and in part to 3-dimensional (3D; sponge) systems. Utilizing the C2C12 mouse myoblast cell line as a gold standard, the protocols deliver precise, step-by-step instructions for preparing fiber scaffolds (using silk proteins here), seeding cells, and monitoring key parameters for cultivated meat production, providing a framework for advancing cellular agriculture techniques. © 2024 Wiley Periodicals LLC. Basic Protocol 1: Fabrication and preparation of silk fiber scaffolds for cell seeding Support Protocol 1: Cultivation of C2C12 cells and seeding onto fibrous scaffolds Basic Protocol 2: Quantification of decellularized yarn scaffold degradation during cell culture Basic Protocol 3: Quantification of biomass variation and ECM deposition on yarn scaffolds during C2C12 cell culture Basic Protocol 4: Visualization of cell-laden yarn scaffolds and determination of cell coverage ratio using confocal microscopy Support Protocol 2: Real-time imaging of cell-laden yarn scaffolds using Celigo system Support Protocol 3: Applying green CellTracker fluorescent probes to C2C12 cells seeded on yarn scaffolds.

Keywords:  biomass quantification; cellular agriculture; cultivated meat; fibers; scaffolds

DOI:  https://doi.org/10.1002/cpz1.70076
Int J Mol Sci. 2024 Nov 22. pii: 12567. [Epub ahead of print]25(23):

Photocrosslinkable Biomaterials for 3D Bioprinting: Mechanisms, Recent Advances, and Future Prospects.

Yushang Lai, Xiong Xiao, Ziwei Huang, Hongying Duan, Liping Yang, Yuchu Yang, Chenxi Li, Li Feng.

  Constructing scaffolds with the desired structures and functions is one of the main goals of tissue engineering. Three-dimensional (3D) bioprinting is a promising technology that enables the personalized fabrication of devices with regulated biological and mechanical characteristics similar to natural tissues/organs. To date, 3D bioprinting has been widely explored for biomedical applications like tissue engineering, drug delivery, drug screening, and in vitro disease model construction. Among different bioinks, photocrosslinkable bioinks have emerged as a powerful choice for the advanced fabrication of 3D devices, with fast crosslinking speed, high resolution, and great print fidelity. The photocrosslinkable biomaterials used for light-based 3D printing play a pivotal role in the fabrication of functional constructs. Herein, this review outlines the general 3D bioprinting approaches related to photocrosslinkable biomaterials, including extrusion-based printing, inkjet printing, stereolithography printing, and laser-assisted printing. Further, the mechanisms, advantages, and limitations of photopolymerization and photoinitiators are discussed. Next, recent advances in natural and synthetic photocrosslinkable biomaterials used for 3D bioprinting are highlighted. Finally, the challenges and future perspectives of photocrosslinkable bioinks and bioprinting approaches are envisaged.

Keywords:  3D bioprinting; hydrogel; photocrosslinkable biomaterials

DOI:  https://doi.org/10.3390/ijms252312567
ACS Appl Polym Mater. 2024 Dec 13. 6(23): 14169-14189

Polymer Sorbent Design for the Direct Air Capture of CO2.

Mark Robertson, Jin Qian, Zhe Qiang.

Anthropogenic activities have resulted in enormous increases in atmospheric CO2 concentrations particularly since the onset of the Industrial Revolution, which have potential links with increased global temperatures, rising sea levels, increased prevalence, and severity of natural disasters, among other consequences. To enable a carbon-neutral and sustainable society, various technologies have been developed for CO2 capture from industrial process streams as well as directly from air. Here, direct air capture (DAC) represents an essential need for reducing CO2 concentration in the atmosphere to mitigate the negative consequences of greenhouse effects, involving systems that can reversibly adsorb and release CO2, in which polymers have played an integral role. This work provides insights into the development of polymer sorbents for DAC of CO2, specifically from the perspective of material design principles. We discuss how physical properties and chemical identities of amine-containing polymers can impact their ability to uptake CO2, as well as be efficiently regenerated. Additionally, polymers which use ionic interactions to react with CO2 molecules, such as poly(ionic liquids), are also common DAC sorbent materials. Finally, a perspective is provided on the future research and technology opportunities of developing polymer-derived sorbents for DAC.

DOI: https://doi.org/10.1021/acsapm.3c03199
STAR Protoc. 2024 Dec 17. pii: S2666-1667(24)00667-1. [Epub ahead of print]6(1): 103502

Protocol for the synthesis and activation of hydrogels with photocaged oligonucleotides.

Danielle Herubin, Amanda Yang, Saanvi Gaddam, Katelyn Mathis, Brian Meckes.

  Hydrogels with spatial-temporal control over chemical and physical properties allow for the creation of cellular niches with controllable properties that better mimic tissue environments. Here, we present a protocol for synthesizing hydrogels incorporating photocaged oligonucleotides that can be activated with non-ultraviolet (UV) wavelengths. We detail the synthesis of bulk hydrogels and spatially defined hydrogels with different chemical functionalities that all share common photocaged DNA. Furthermore, we describe conditions for activating photocaged DNA to capture complementary oligonucleotides. For complete details on the use and execution of this protocol, please refer to Mathis et al.1.

Keywords:  Biotechnology and bioengineering; Chemistry; Material sciences; Tissue Engineering

DOI:  https://doi.org/10.1016/j.xpro.2024.103502
ACS Appl Mater Interfaces. 2024 Dec 19.

Efficient Enzymatic Glycan Engineering of Extracellular Vesicles Using Nanomaterial-Interfaced Microfluidics.

Xin Zhou, Mohit Jaiswal, Jingzhu Shi, Jiatong Guo, Sayan Kundu, Zhongwu Guo, Yong Zeng.

  Extracellular vesicles (EVs) present a promising modality for numerous biological and medical applications, including therapeutics. Developing facile methods to engineer EVs is essential to meeting the rapidly expanding demand for various functionalized EVs in these applications. Herein, we developed a technology that integrates enzymatic glycoengineering and microfluidics for effective EV functionalization. This method builds on a 3D nanostructured microfluidic device to streamline a multiple-step EV engineering process, which involves a step of enzymatic reaction to install azido-sialic acid residues to glycans on EVs using a sialyltransferase and an azide-tagged sialyl donor followed by the attachment of various functionalities, such as biotin and fluorescent labels, to the resulting azido-glycans on EVs through a biocompatible click reaction. Compared to traditional EV engineering methods, we show that our technology improves the efficiency of EV glycoengineering while simplifying and expediting the workflow. Furthermore, we demonstrated the applicability of this technology to EVs derived from the cell lines of different cancer types, including A549, PC3, and COLO-1 cells. Overall, this EV engineering technology could provide a potentially useful tool for broad applications.

Keywords:  click chemistry; enzymatic glycan engineering; extracellular vesicle; fluorescent labeling; microfluidics

DOI:  https://doi.org/10.1021/acsami.4c20294
J Mater Chem B. 2024 Dec 18.

Development and application of hydrogels in pathogenic bacteria detection in foods.

Shuxiang Liu, Md Rashidur Rahman, Hejun Wu, Wen Qin, Yanying Wang, Gehong Su.

Hydrogels are 3D networks of water-swollen hydrophilic polymers. It possesses unique properties (e.g., carrying biorecognition elements and creating a micro-environment) that make it highly suitable for bacteria detection (e.g., expedited and effective bacteria detection) and mitigation of bacterial contamination in specific environments (e.g., food systems). This study first introduces the materials used to create hydrogels for bacteria detection and the mechanisms for detection. We also summarize different hydrogel-based detection methods that rely on external stimuli and biorecognition elements, such as enzymes, temperature, pH, antibodies, and oligonucleotides. Subsequently, a range of widely utilized bacterial detection technologies were discussed where recently hydrogels are being used. These modifications allow for precise, real-time diagnostics across varied food matrices, responding effectively to industry needs for sensitivity, scalability, and portability. After highlighting the utilization of hydrogels and their role in these detection techniques, we outline limitations and advancements in the methods for the detection of foodborne pathogenic bacteria, especially the potential application of hydrogels in the food industry.

DOI: https://doi.org/10.1039/d4tb01341g
Sci Robot. 2024 Dec 18. 9(97): eado5888

Counterfactual rewards promote collective transport using individually controlled swarm microrobots.

Veit-Lorenz Heuthe, Emanuele Panizon, Hongri Gu, Clemens Bechinger.

Swarm robots offer fascinating opportunities to perform complex tasks beyond the capabilities of individual machines. Just as a swarm of ants collectively moves large objects, similar functions can emerge within a group of robots through individual strategies based on local sensing. However, realizing collective functions with individually controlled microrobots is particularly challenging because of their micrometer size, large number of degrees of freedom, strong thermal noise relative to the propulsion speed, and complex physical coupling between neighboring microrobots. Here, we implemented multiagent reinforcement learning (MARL) to generate a control strategy for up to 200 microrobots whose motions are individually controlled by laser spots. During the learning process, we used so-called counterfactual rewards that automatically assign credit to the individual microrobots, which allows fast and unbiased training. With the help of this efficient reward scheme, swarm microrobots learn to collectively transport a large cargo object to an arbitrary position and orientation, similar to ant swarms. We show that this flexible and versatile swarm robotic system is robust to variations in group size, the presence of malfunctioning units, and environmental noise. In addition, we let the robot swarms manipulate multiple objects simultaneously in a demonstration experiment, highlighting the benefits of distributed control and independent microrobot motion. Control strategies such as ours can potentially enable complex and automated assembly of mobile micromachines, programmable drug delivery capsules, and other advanced lab-on-a-chip applications.

DOI: https://doi.org/10.1126/scirobotics.ado5888
Materials (Basel). 2024 Nov 23. pii: 5736. [Epub ahead of print]17(23):

Influence of Internal Architecture and Ink Formulation on the Thermal Behavior of 3D-Printed Cementitious Materials.

Michael Kosson, Lesa Brown, Garrett Thorne, Florence Sanchez.

  Cement-based 3D printing provides an opportunity to create cement-based elements with a hierarchy of structures and patterns that are not easily achievable using traditional casting techniques, thereby providing new possibilities for improving thermal control and energy storage in cement-based materials. In this study, the influence of internal architecture and ink formulation on the thermal behavior of 3D-printed cement composite beams was investigated using infrared thermal imaging and a conceptual one-dimensional heat transfer model based on cooling fins in convective media. Three-dimensional printed beams with rectilinear, three-dimensional honeycomb, and Archimedean chord infill patterns and cement ink formulations with and without 5% halloysite nanoclay were exposed to a heating source at one end. The thermal behavior of the beams was found to be predominantly influenced by their internal architecture rather than the cement ink formulation, with differences in void structures and heat transfer pathways among the different architectures resulting in a hierarchy of apparent thermal conductivity. The internal architecture resulted in a reduction in apparent thermal conductivity by up to 75%, while the incorporation of halloysite nanoclay in the cement ink led to a reduction of up to 14%. Among the tested internal architecture, the rectilinear architecture showed a 10-15% higher apparent thermal conductivity compared to the three-dimensional honeycomb architecture and a 35-40% higher apparent thermal conductivity than the Archimedean architecture. The research demonstrates a promising strategy for fabricating and evaluating cement-based materials with thermal management capabilities using 3D printing methods.

Keywords:  cement paste; extrusion 3D printing; halloysite nanoclay; internal architecture; thermal properties

DOI:  https://doi.org/10.3390/ma17235736
Extracell Vesicles Circ Nucl Acids. 2023 ;4(2): 218-239

Bioprinting extracellular vesicles as a "cell-free" regenerative medicine approach.

Kexin Jiao, Chun Liu, Saraswat Basu, Nimal Raveendran, Tamaki Nakano, Sašo Ivanovski, Pingping Han.

  Regenerative medicine involves the restoration of tissue or organ function via the regeneration of these structures. As promising regenerative medicine approaches, either extracellular vesicles (EVs) or bioprinting are emerging stars to regenerate various tissues and organs (i.e., bone and cardiac tissues). Emerging as highly attractive cell-free, off-the-shelf nanotherapeutic agents for tissue regeneration, EVs are bilayered lipid membrane particles that are secreted by all living cells and play a critical role as cell-to-cell communicators through an exchange of EV cargos of protein, genetic materials, and other biological components. 3D bioprinting, combining 3D printing and biology, is a state-of-the-art additive manufacturing technology that uses computer-aided processes to enable simultaneous patterning of 3D cells and tissue constructs in bioinks. Although developing an effective system for targeted EVs delivery remains challenging, 3D bioprinting may offer a promising means to improve EVs delivery efficiency with controlled loading and release. The potential application of 3D bioprinted EVs to regenerate tissues has attracted attention over the past few years. As such, it is timely to explore the potential and associated challenges of utilizing 3D bioprinted EVs as a novel "cell-free" alternative regenerative medicine approach. In this review, we describe the biogenesis and composition of EVs, and the challenge of isolating and characterizing small EVs - sEVs (< 200 nm). Common 3D bioprinting techniques are outlined and the issue of bioink printability is explored. After applying the following search strategy in PubMed: "bioprinted exosomes" or "3D bioprinted extracellular vesicles", eight studies utilizing bioprinted EVs were found that have been included in this scoping review. Current studies utilizing bioprinted sEVs for various in vitro and in vivo tissue regeneration applications, including angiogenesis, osteogenesis, immunomodulation, chondrogenesis and myogenesis, are discussed. Finally, we explore the current challenges and provide an outlook on possible refinements for bioprinted sEVs applications.

Keywords:  3D bioprinting; bioprinted sEVs; regenerative medicine; small extracellular vesicles

DOI:  https://doi.org/10.20517/evcna.2023.19
Biophys Rev (Melville). 2024 Dec;5(4): 041302

Spatially defined microenvironment for engineering organoids.

Yilan Zhang, Fukang Qi, Peng Chen, Bi-Feng Liu, Yiwei Li.

In the intricately defined spatial microenvironment, a single fertilized egg remarkably develops into a conserved and well-organized multicellular organism. This observation leads us to hypothesize that stem cells or other seed cell types have the potential to construct fully structured and functional tissues or organs, provided the spatial cues are appropriately configured. Current organoid technology, however, largely depends on spontaneous growth and self-organization, lacking systematic guided intervention. As a result, the structures replicated in vitro often emerge in a disordered and sparse manner during growth phases. Although existing organoids have made significant contributions in many aspects, such as advancing our understanding of development and pathogenesis, aiding personalized drug selection, as well as expediting drug development, their potential in creating large-scale implantable tissue or organ constructs, and constructing multicomponent microphysiological systems, together with functioning at metabolic levels remains underutilized. Recent discoveries have demonstrated that the spatial definition of growth factors not only induces directional growth and migration of organoids but also leads to the formation of assembloids with multiple regional identities. This opens new avenues for the innovative engineering of higher-order organoids. Concurrently, the spatial organization of other microenvironmental cues, such as physical stresses, mechanical loads, and material composition, has been minimally explored. This review delves into the burgeoning field of organoid engineering with a focus on potential spatial microenvironmental control. It offers insight into the molecular principles, expected outcomes, and potential applications, envisioning a future perspective in this domain.

DOI: https://doi.org/10.1063/5.0198848
Extracell Vesicles Circ Nucl Acids. 2024 ;5(1): 44-63

Harnessing genetically engineered cell membrane-derived vesicles as biotherapeutics.

Xiaohong Li, Yuting Wei, Zhirang Zhang, Xudong Zhang.

  Cell membrane-derived vesicles (CMVs) are particles generated from living cells, including extracellular vesicles (EVs) and artificial extracellular vesicles (aEVs) prepared from cell membranes. CMVs possess considerable potential in drug delivery, regenerative medicine, immunomodulation, disease diagnosis, etc. owing to their stable lipid bilayer structure, favorable biocompatibility, and low toxicity. Although the majority of CMVs inherit certain attributes from the original cells, it is still difficult to execute distinct therapeutic functions, such as organ targeting, signal regulation, and exogenous biotherapeutic supplementation. Hence, engineering CMVs by genetic engineering, chemical modification, and hybridization is a promising way to endow CMVs with specific functions and open up novel vistas for applications. In particular, there is a growing interest in genetically engineered CMVs harnessed to exhibit biotherapeutics. Herein, we outline the preparation strategies and their characteristics for purifying CMVs. Additionally, we review the advances of genetically engineered CMVs utilized to target organs, regulate signal transduction, and deliver biomacromolecules and chemical drugs. Furthermore, we also summarize the emerging therapeutic applications of genetically engineered CMVs in addressing tumors, diabetes, systemic lupus erythematosus, and cardiovascular diseases.

Keywords:  EVs; biomedicine; drug delivery; genetically engineering

DOI:  https://doi.org/10.20517/evcna.2023.58