Liquid-embedded (bio)printing of alginate-free, standalone, ultrafine, and ultrathin-walled cannular structures.

While there has been considerable success in the three-dimensional bioprinting of relatively large standalone filamentous tissues, the fabrication of solid fibers with ultrafine diameters or those cannular featuring ultrathin walls remains a particular challenge. Here, an enabling strategy for (bio)printing of solid and hollow fibers whose size ranges could be facilely adjusted across a broad spectrum, is reported, using an aqueous two-phase embedded (bio)printing approach combined with specially designed cross-linking and extrusion methods. The generation of standalone, alginate-free aqueous architectures using this aqueous two-phase strategy allowed freeform patterning of aqueous bioinks, such as those composed of gelatin methacryloyl, within the immiscible aqueous support bath of poly(ethylene oxide). Our (bio)printing strategy revealed the fabrication of standalone solid or cannular structures with diameters as small as approximately 3 or 40 µm, respectively, and wall thicknesses of hollow conduits down to as thin as <5 µm. With cellular functions also demonstrated, we anticipate the methodology to serve as a platform that may satisfy the needs for the different types of potential biomedical and other applications in the future, especially those pertaining to cannular tissues of ultrasmall diameters and ultrathin walls used toward regenerative medicine and tissue model engineering.

Biofabrication of Living Actuators.

The impact of tissue engineering has extended beyond a traditional focus in medicine to the rapidly growing realm of biohybrid robotics. Leveraging living actuators as functional components in machines has been a central focus of this field, generating a range of compelling demonstrations of robots capable of muscle-powered swimming, walking, pumping, gripping, and even computation. In this review, we highlight key advances in fabricating tissue-scale cardiac and skeletal muscle actuators for a range of functional applications. We discuss areas for future growth including scalable manufacturing, integrated feedback control, and predictive modeling and also propose methods for ensuring inclusive and bioethics-focused pedagogy in this emerging discipline. We hope this review motivates the next generation of biomedical engineers to advance rational design and practical use of living machines for applications ranging from telesurgery to manufacturing to on- and off-world exploration.

Microengineering 3D Collagen Matrices with Tumor-Mimetic Gradients in Fiber Alignment.

Collagen fibers in the 3D tumor microenvironment (TME) exhibit complex alignment landscapes that are critical in directing cell migration through a process called contact guidance. Previous in vitro work studying this phenomenon has focused on quantifying cell responses in uniformly aligned environments. However, the TME also features short-range gradients in fiber alignment that result from cell-induced traction forces. Although the influence of graded biophysical taxis cues is well established, cell responses to physiological alignment gradients remain largely unexplored. In this work, fiber alignment gradients in biopsy samples are characterized and recreated using a new microfluidic biofabrication technique to achieve tunable sub-millimeter to millimeter scale gradients. This study represents the first successful engineering of continuous alignment gradients in soft, natural biomaterials. Migration experiments on graded alignment show that HUVECs exhibit increased directionality, persistence, and speed compared to uniform and unaligned fiber architectures. Similarly, patterned MDA-MB-231 aggregates exhibit biased migration toward increasing fiber alignment, suggesting a role for alignment gradients as a taxis cue. This user-friendly approach, requiring no specialized equipment, is anticipated to offer new insights into the biophysical cues that cells interpret as they traverse the extracellular matrix, with broad applicability in healthy and diseased tissue environments.

[n]Cycloparaphenylenes as Compatible Fluorophores for Melt Electrowriting.

Fluorescent probes are an indispensable tool in the realm of bioimaging technologies, providing valuable insights into the assessment of biomaterial integrity and structural properties. However, incorporating fluorophores into scaffolds made from melt electrowriting (MEW) poses a challenge due to the sustained, elevated temperatures that this processing technique requires. In this context, [n]cycloparaphenylenes ([n]CPPs) serve as excellent fluorophores for MEW processing with the additional benefit of customizable emissions profiles with the same excitation wavelength. Three fluorescent blends are used with distinct [n]CPPs with emission wavelengths of either 466, 494, or 533 nm, identifying 0.01 wt% as the preferred concentration. It is discovered that [n]CPPs disperse well within poly(ε-caprolactone) (PCL) and maintain their fluorescence even after a week of continuous heating at 80 °C. The [n]CPP-PCL blends show no cytotoxicity and support counterstaining with commonly used DAPI (Ex/Em: 359 nm/457 nm), rhodamine- (Ex/Em: 542/565 nm), and fluorescein-tagged (Ex/Em: 490/515 nm) phalloidin stains. Using different color [n]CPP-PCL blends, different MEW fibers are sequentially deposited into a semi-woven scaffold and onto a solution electrospun membrane composed of [8]CPP-PCL as a contrasting substrate for the [10]CPP-PCL MEW fibers. In general, [n]CPPs are potent fluorophores for MEW, providing new imaging options for this technology.

Biofabrication of engineered blood vessels for biomedical applications.

To successfully engineer large-sized tissues, establishing vascular structures is essential for providing oxygen, nutrients, growth factors and cells to prevent necrosis at the core of the tissue. The diameter scale of the biofabricated vasculatures should range from 100 to 1,000 µm to support the mm-size tissue while being controllably aligned and spaced within the diffusion limit of oxygen. In this review, insights regarding biofabrication considerations and techniques for engineered blood vessels will be presented. Initially, polymers of natural and synthetic origins can be selected, modified, and combined with each other to support maturation of vascular tissue while also being biocompatible. After they are shaped into scaffold structures by different fabrication techniques, surface properties such as physical topography, stiffness, and surface chemistry play a major role in the endothelialization process after transplantation. Furthermore, biological cues such as growth factors (GFs) and endothelial cells (ECs) can be incorporated into the fabricated structures. As variously reported, fabrication techniques, especially 3D printing by extrusion and 3D printing by photopolymerization, allow the construction of vessels at a high resolution with diameters in the desired range. Strategies to fabricate of stable tubular structures with defined channels will also be discussed. This paper provides an overview of the many advances in blood vessel engineering and combinations of different fabrication techniques up to the present time.

This review covers several aspects and advancements of engineered blood vessel biofabrication, which are essential for establishment of large-sized tissues in different areas of biomedical applications.

Microbes-mediated synthesis strategies of metal nanoparticles and their potential role in cancer therapeutics.

Past few years have seen a paradigm shift towards ecofriendly, green and biological fabrication of metal nanoparticles (MNPs) for diverse nanomedicinal applications especially in cancer nanotheranostics. Besides, the well-known green synthesis methods of plant materials, the potential of the microbial world (bacteria, fungi, alga, etc.) in biofabrication is equally realized. Biomolecules and enzymes in the microbial cells are capable of catalyzing the biosynthesis process. These microbial derived inorganic nanoparticles have been frequently evaluated as potential agents in cancer therapies revealing exciting results. Through, cellular and molecular pathways, these microbial derived nanoparticles are capable of killing the cancer cells. Considering the recent developments in the anticancer applications of microbial derived inorganic MNPs, a dire need was felt to bring the available information to a single document. This manuscript reviews not only the mechanistic aspects of the microbial derived MNPs but also include the diverse mechanisms that governs their anticancer potential. Besides, an updated literature review is presented that includes studies of 2019-onwards.

Bio-fabricated bismuth-based materials for removal of emerging environmental contaminants from wastewater.

Although rapid industrialization has made life easier for humans, several associated issues are emerging and harming the environment. Wastewater is regarded as one of the key problems of the 21st century due to its massive production every year and requires immediate attention from all stakeholders to protect the environment. Since the introduction of nanotechnology, bismuth-based nanomaterials have been used in variety of applications. Various techniques, such as hydrothermal, solvo-thermal and biosynthesis, have been reported for synthesizing these materials, etc. Among these, biosynthesis is eco-friendly, cost-effective, and less toxic than conventional chemical methods. The prime focuses of this review are to elaborate biosynthesis of bismuth-based nanomaterials via bio-synthetic agents such as plant, bacteria and fungi and their application in wastewater treatment as anti-pathogen/photocatalyst for pollutant degradation. Besides this, future perspectives have been presented for the upcoming research in this field, along with concluding remarks.

Biofabrication of Mg-doped ZnO nanostructures for hemolysis and antibacterial properties.

The aim of this work was to synthesize 0.02 and 0.06 Mg-doped ZnO nanoparticles (NPs) using the aqueous extract of Plectranthus barbatus leaf. The structural integrity of the hexagonal phase was emphasized by X-ray diffraction analysis. The average crystallite size (D) of 0.02 and 0.06 Mg-doped ZnO NPs was found to be 23.83 and 26.95 nm, respectively. The scanning electron microscope images revealed a surface morphology of irregular nano-shapes of about 83 nm diameter with an elongated one-dimensional structure. The hemolysis activity demonstrated the safe nature of the synthesized materials at low doses. Antibacterial activity against S. aureus and E. coli, which assessed using the disc diffusion method, indicated that the prepared NPs could inhibit S. aureus but not E. coli. These findings suggest that the synthesized NPs could be explored for potential applications in biotechnology and medicine.

Emerging Strategies for the Biofabrication of Multilayer Composite Amniotic Membranes for Biomedical Applications.

The amniotic membrane (AM) is the innermost part of the fetal placenta, which surrounds and protects the fetus. Due to its structural components (stem cells, growth factors, and proteins), AMs display unique biological properties and are a widely available and cost-effective tissue. As a result, AMs have been used for a century as a natural biocompatible dressing for healing corneal and skin wounds. To further increase its properties and expand its applications, advanced hybrid materials based on AMs have recently been developed. One existing approach is to combine the AM with a secondary material to create composite membranes. This review highlights the increasing development of new multilayer composite-based AMs in recent years and focuses on the benefits of additive manufacturing technologies and electrospinning, the most commonly used strategy, in expanding their use for tissue engineering and clinical applications. The use of AMs and multilayer composite-based AMs in the context of nerve regeneration is particularly emphasized and other tissue engineering applications are also discussed. This review highlights that these electrospun multilayered composite membranes were mainly created using decellularized or de-epithelialized AMs, with both synthetic and natural polymers used as secondary materials. Finally, some suggestions are provided to further enhance the biological and mechanical properties of these composite membranes.

Bio-Fabrication of ZnONPs from Alkalescent Nucleoside Antibiotic to Control Rice Blast: Impact on Pathogen (Magnaporthe grisea) and Host (Rice).

In the traditional method of the bio-fabrication of zinc oxide nanoparticles (ZnONPs), bacterial strains face metal toxicity and antimicrobial action. In the current study, an alkalescent nucleoside antibiotic was mixed with zinc hexanitrate to fabricate the ZnONPs. An integrated approach of DIAION HP-20 macroporous resin and sephadex LH-20 column chromatography was adopted to separate and purify alkalescent nucleoside AN03 from Streptomyces koyanogensis. Alkalescent nucleoside was confirmed by the Doskochilova solvent system. The bio-fabricated ZnONPs were characterized by using Fourier transform infrared (FTIR), X-ray diffraction (XRD), and transmission electron microscopy (TEM) analyses. The XRD spectrum and the TEM images confirmed the crystallinity and the spherical shape of the ZnONPs with an average size of 22 nm. FTIR analysis showed the presence of functional groups, which confirmed the bio-fabrication of ZnONPs from alkalescent nucleoside ANO3. In-vitro studies showed that 75 µg/mL of ZnONPs had a strong inhibitory zone (28.39 mm) against the Magnaporthe grisea and significantly suppressed the spore germination. SEM and TEM observations respectively revealed that ZnONPs caused breakage in hyphae and could damage the cells of M. grisea. Greenhouse experiments revealed that the foliar spray of ZnONPs could control the rice blast disease by 98%. Results also revealed that ZnONPs had positive effects on the growth of the rice plant. The present study suggested that ZnONPs could be fabricated from microbe-derived nucleoside antibiotics without facing the problems of metal toxicity and antimicrobial action, thus overcoming the problem of pathogen resistance. This could be a potent biocontrol agent in rice blast disease management.

From Static to Dynamic: Smart Materials Pioneering Additive Manufacturing in Regenerative Medicine.

The emerging field of regenerative medicine holds immense promise for addressing complex tissue and organ regeneration challenges. Central to its advancement is the evolution of additive manufacturing techniques, which have transcended static constructs to embrace dynamic, biomimetic solutions. This manuscript explores the pivotal role of smart materials in this transformative journey, where materials are endowed with dynamic responsiveness to biological cues and environmental changes. By delving into the innovative integration of smart materials, such as shape memory polymers and stimulus-responsive hydrogels, into additive manufacturing processes, this research illuminates the potential to engineer tissue constructs with unparalleled biomimicry. From dynamically adapting scaffolds that mimic the mechanical behavior of native tissues to drug delivery systems that respond to physiological cues, the convergence of smart materials and additive manufacturing heralds a new era in regenerative medicine. This manuscript presents an insightful overview of recent advancements, challenges, and future prospects, underscoring the pivotal role of smart materials as pioneers in shaping the dynamic landscape of regenerative medicine and heralding a future where tissue engineering is propelled beyond static constructs towards biomimetic, responsive, and regenerative solutions.

Microbased biorefinery for gold nanoparticle production: recent advancements, applications and future aspects.

Multifaceted utility of nanomaterials is indispensable to meet the environmental challenges across the globe. Nanomaterials substantially contribute in delineating the rapidly advancing field of nanotechnology. Recently, primary emphasis has been laid down on augmenting the biological methodologies for the synthesis of nanomaterials. In this aspect, green nanotechnology has revolutionized the entire process of nanosynthesis. Essentially biofabrication of nanoparticles have long-range applications, primarily in the field of medical applications such as drug delivery, cancer diagnostics and genetic engineering processes. Biocompatible and stable nanoparticles synthesized from biological source can be an effective approach against the chemically synthesized owing to their non-expensive and eco-friendly attributes. Biological systems including bacteria, yeasts, fungi and plants have already been exploited in the field of nanotechnology. Use of fungi seems to be a very effective and economical approach for the synthesis of gold nanoparticles. Gold nanoparticles possess anti-oxidation activity, are highly stable and biocompatible in nature. Fungi-mediated nanoparticle biosynthesis is more advantageous as compared to bacterial synthesis. Fungi secrete large amounts of enzymes, whereas the enzyme secretion of yeasts is weak. Here, we have reported the recent advancements and future implications in the field of gold nanoparticle production and applications.

Green Synthesis, Characterization and Bioactivity of Mangifera indica Seed-Wrapped Zinc Oxide Nanoparticles.

In the realm of nanoparticles, metal-based nanoparticles have traditionally been regarded as the pioneering category. Compared to other nanoparticles, zinc oxide nanoparticles have several advantages, including optical and biological properties, which provide them a significant competitive advantage in clinical and biological applications. In the current investigation, we used an aqueous Mangifera indica seed extract to synthesize nanoparticles of zinc oxide (ZnO NPs). UV-Vis spectroscopy, Fourier transform infrared spectroscopy analysis, atomic force spectroscopy, X-ray diffraction, scanning electron microscopy, and transmission electron microscopy were used to characterize the synthesized ZnO NPs. The nanoparticles were assessed for their potential to inhibit bacterial growth and protect cells from free radical damage. According to the current study's findings, zinc oxide nanoparticles that had been modified with the aid of mango seeds were very efficient in preventing the development of the tested bacteria and were also powerful antioxidants.

Biofabrication of Functional Pullulan by Aureobasidium pullulans under the Effect of Varying Mineral Salts and Sugar Stress Conditions.

Pullulan is a linear exopolysaccharide, produced in the fermentation media of Aureobasidium pullulans, with a variety of applications in the food and pharmaceutical industries. Pullulan derivatives have growing potential for biomedical applications, but the high cost of pullulan biofabrication currently restricts its commercial use. Better control over pullulan yield, molecular weight and melanin production by altering fermentation conditions could improve the economics. In this study, the effects of sugar and mineral salt stresses on the pullulan production of A. pullulans ATCC 42023 were examined in batch processes. The chemical structure of the recovered pullulan was characterized by FTIR and NMR spectroscopy, and the molecular weight distribution was obtained via SEC. Pullulan yield and melanin production varied when the conditions were adjusted, and pullulans with different molar masses were obtained. Higher-yield pullulan production and a lower polydispersity index were observed when CuSO4 was added to the fermentation in comparison with the control and with the addition of sugars and other salts. Biofabrication of pullulan under stress conditions is a promising strategy to enhance biopolymer yield and to obtain pullulan with a targeted molecular weight.

Biofabricated yeast: super-soldier for detoxification of heavy metals.

The advances in nanotechnology have shown enormous impacts in environmental technology as a potent weapon for degradation of toxic organic pollutants and detoxification of heavy metals. It is either by in-situ or ex-situ adaptive strategies. Mycoremediation of environmental pollutants has been a success story of the past decade, by employing the wide arsenal of biological capabilities of fungi. Recently, the proficiency and uniqueness of yeast cell surface alterations have encouraged the generation of engineered yeast cells as dye degraders, heavy metal reduction and its recovery, and also as detoxifiers of various hazardous xenobiotic compounds. As a step forward, recent trends in research are towards developing biologically engineered living materials as potent, biocompatible and reusable hybrid nanomaterials. They include chitosan-yeast nanofibers, nanomats, nanopaper, biosilica hybrids, and TiO2-yeast nanocomposites. The nano-hybrid materials contribute significantly as supportive stabilizer, and entrappers, which enhances the biofabricated yeast cells' functionality. This field serves as an eco-friendly cutting-edge cocktail research area. In this review, we highlight recent research on biofabricated yeast cells and yeast-based biofabricated molecules, as potent heavy metals, toxic chemical detoxifiers, and their probable mechanistic properties with future application perspectives.

[Bioprinting technologies in ophthalmology]. / Tekhnologii biopechati v oftal'mologii.

Bioprinting allows additive fabrication of bioengineered constructs with defined two- or three-dimensional organization using live cells, biopolymers and other materials. This article reviews main bioprinting technologies and their capabilities in clinical and experimental ophthalmology. Analysis of literature sources helped reveal and describe the main types of bioprinting technologies: inkjet, laser-assisted, and extrusion. Extrusion bioprinting is the most widely used method, providing the ability to use various types of bioinks and a wide range of cell concentrations. The following materials can be used as the base for bioinks: alginate, collagen, gelatin, hyaluronic acid, chitosan, fibrin, as well as their different combinations. These materials can be modified for best bioprinting properties by adding various functional groups. The major directions of application of bioprinting technologies in ophthalmology are tissue engineering for regenerative medicine and fabrication of model systems for fundamental and preclinical studies. Experiments in creating a bioprinted cornea are being conducted in the field of regenerative medicine. Furthermore, there are studies on fabricating retinal tissue equivalents, although tissue engineering of this structure is a task of great complexity. Model systems, which can be fabricated by bioprinting, are represented by tissue equivalents of ocular structures and the appendages of the eye, as well as by microphysiological organ-on-a-chip systems. Another promising application of bioprinting is fabrication of biocompatible implantable electrode arrays for visual neuroprostheses.

Advanced biomaterials for periodontal tissue regeneration.

The periodontium is a suitable target for regenerative intervention, since it does not functionally restore itself after disease. Importantly, the limited regeneration capacity of the periodontium could be improved with the development of novel biomaterials and therapeutic strategies. Of note, the regenerative potential of the periodontium depends not only on its tissue-specific architecture and function, but also on its ability to reconstruct distinct tissues and tissue interfaces, suggesting that the advancement of tissue engineering approaches can ultimately offer new perspectives to promote the organized reconstruction of soft and hard periodontal tissues. Here, we discuss material-based, biologically active cues, and the application of innovative biofabrication technologies to regenerate the multiple tissues that comprise the periodontium.

Biofabrication approaches and regulatory framework of metastatic tumor-on-a-chip models for precision oncology.

The complexity of the tumor microenvironment (TME) together with the development of the metastatic process are the main reasons for the failure of conventional anticancer treatment. In recent years, there is an increasing need to advance toward advanced in vitro models of cancer mimicking TME and simulating metastasis to understand the associated mechanisms that are still unknown, and to be able to develop personalized therapy. In this review, the commonly used alternatives and latest advances in biofabrication of tumor-on-chips, which allow the generation of the most sophisticated and optimized models for recapitulating the tumor process, are presented. In addition, the advances that have allowed these new models in the area of metastasis, cancer stem cells, and angiogenesis are summarized, as well as the recent integration of multiorgan-on-a-chip systems to recapitulate natural metastasis and pharmacological screening against it. We also analyze, for the first time in the literature, the normative and regulatory framework in which these models could potentially be found, as well as the requirements and processes that must be fulfilled to be commercially implemented as in vitro study model. Moreover, we are focused on the possible regulatory pathways for their clinical application in precision medicine and decision making through the generation of personalized models with patient samples. In conclusion, this review highlights the synergistic combination of three-dimensional bioprinting systems with the novel tumor/metastasis/multiorgan-on-a-chip systems to generate models for both basic research and clinical applications to have devices useful for personalized oncology.

Micropore-Forming Gelatin Methacryloyl (GelMA) Bioink Toolbox 2.0: Designable Tunability and Adaptability for 3D Bioprinting Applications.

It is well-known that tissue engineering scaffolds that feature highly interconnected and size-adjustable micropores are oftentimes desired to promote cellular viability, motility, and functions. Unfortunately, the ability of precise control over the microporous structures within bioinks in a cytocompatible manner for applications in 3D bioprinting is generally lacking, until a method of micropore-forming bioink based on gelatin methacryloyl (GelMA) was reported recently. This bioink took advantage of the unique aqueous two-phase emulsion (ATPE) system, where poly(ethylene oxide) (PEO) droplets are utilized as the porogen. Considering the limitations associated with this very initial demonstration, this article has furthered the understanding of the micropore-forming GelMA bioinks by conducting a systematic investigation into the additional GelMA types (porcine and fish, different methacryloyl-modification degrees) and porogen types (PEO, poly(vinyl alcohol), and dextran), as well as the effects of the porogen concentrations and molecular weights on the properties of the GelMA-based ATPE bioink system. This article exemplifies not only the significantly wider range of micropore sizes achievable and better emulsion stability, but also the improved suitability for both extrusion and digital light processing bioprinting with favorable cellular responses.

Lithiophilic Carbon Nanofiber/Graphene Nanosheet Composite Scaffold Prepared by a Scalable and Controllable Biofabrication Method for Ultrastable Dendrite-Free Lithium-Metal Anodes.

Li metal is regarded as a promising anode for high-energy-density Li batteries, while the limited cycle life and fast capacity decay caused by notorious Li dendrite growth seriously impedes its application. Herein, a robust and highly lithiophilic bacterial cellulose-derived carbon nanofiber@reduced graphene oxide nanosheet (BC-CNF@rGO) composite scaffold is fabricated as a host for dendrite-free Li metal anode through an in situ biofabrication method. The abundant lithiophilic functional groups, conductive 3D network, and excellent mechanical property can effectively regulate uniform Li nucleation and deposition, enable fast reaction kinetics, and alleviate volume change. As a result, the BC-CNF@rGO skeleton achieves exceptional Li plating/stripping performance with a high average Coulombic efficiency of 98.3% over 800 cycles, and a long cycle life span of 5000 h at 2 mA cm-2 @1 mAh cm-2 with a low overpotential of ≈15 mV for lithium plating. Furthermore, full cells coupling BC-CNF@rGO-Li anode with LiFePO4 cathode achieves an unprecedented cycling stability with a long cycle life of 3000 cycles at 1 C. This work sheds light on a promising material design and fabrication strategy for realizing high performance Li metal batteries.