Bioengineering approaches for the endometrial research and application.

The endometrium undergoes a series of precise monthly changes under the regulation of dynamic levels of ovarian hormones that are characterized by repeated shedding and subsequent regeneration without scarring. This provides the potential for wound healing during endometrial injuries. Bioengineering materials highlight the faithful replication of constitutive cells and the extracellular matrix that simulates the physical and biomechanical properties of the endometrium to a larger extent. Significant progress has been made in this field, and functional endometrial tissue bioengineering allows an in-depth investigation of regulatory factors for endometrial and myometrial defects in vitro and provides highly therapeutic methods to alleviate obstetric and gynecological complications. However, much remains to be learned about the latest progress in the application of bioengineering technologies to the human endometrium. Here, we summarize the existing developments in biomaterials and bioengineering models for endometrial regeneration and improving the female reproductive potential.

Expression of filaments of the Geobacter extracellular cytochrome OmcS in Shewanella oneidensis.

The physiological role of Geobacter sulfurreducens extracellular cytochrome filaments is a matter of debate and the development of proposed electronic device applications of cytochrome filaments awaits methods for large-scale cytochrome nanowire production. Functional studies in G. sulfurreducens are stymied by the broad diversity of redox-active proteins on the outer cell surface and the redundancy and plasticity of extracellular electron transport routes. G. sulfurreducens is a poor chassis for producing cytochrome nanowires for electronics because of its slow, low-yield, anaerobic growth. Here we report that filaments of the G. sulfurreducens cytochrome OmcS can be heterologously expressed in Shewanella oneidensis. Multiple lines of evidence demonstrated that a strain of S. oneidensis, expressing the G. sulfurreducens OmcS gene on a plasmid, localized OmcS on the outer cell surface. Atomic force microscopy revealed filaments with the unique morphology of OmcS filaments emanating from cells. Electron transfer to OmcS appeared to require a functional outer-membrane porin-cytochrome conduit. The results suggest that S. oneidensis, which grows rapidly to high culture densities under aerobic conditions, may be suitable for the development of a chassis for producing cytochrome nanowires for electronics applications and may also be a good model microbe for elucidating cytochrome filament function in anaerobic extracellular electron transfer.

Enhanced corrosion of 2205 duplex stainless steel by Acetobacter aceti through synergistic electron transfer and organic acids acceleration.

Acetobacter aceti is a microbe that produces corrosive organic acids, causing severe corrosion of industrial equipment. Previous studies have focused on the organic acid corrosion of A. aceti, but neglected the possibility that it has electron transfer corrosion. This study found that electron transfer and organic acids can synergistically promote the corrosion of 2205 duplex stainless steel (DSS). Electrochemical measurement results showed that corrosion of 2205 DSS was more severe in the presence of A. aceti. Surface analysis indicated a thick biofilm formed on the steel surface, with low pH and dissolved oxygen concentrations under the biofilm. Corrosion intensified when A. aceti lacked a carbon source, suggesting that A. aceti can corrode metals by using metallic substrates as electron donors, in addition to its acidic by-products.

Accelerated corrosion of 316L stainless steel in a simulated oral environment via extracellular electron transfer and acid metabolites of subgingival microbiota.

316L stainless steel (SS) is widely applied as microimplant anchorage (MIA) due to its excellent mechanical properties. However, the risk that the oral microorganisms can corrode 316L SS is fully neglected. Microbiologically influenced corrosion (MIC) of 316L SS is essential to the health and safety of all patients because the accelerated corrosion caused by the oral microbiota can trigger the release of Cr and Ni ions. This study investigated the corrosion behavior and mechanism of subgingival microbiota on 316L SS by 16S rRNA and metagenome sequencing, electrochemical measurements, and surface characterization techniques. Multispecies biofilms were formed by the oral subgingival microbiota in the simulated oral anaerobic environment on 316L SS surfaces, significantly accelerating the corrosion in the form of pitting. The microbiota samples collected from the subjects differed in biofilm compositions, corrosion behaviors, and mechanisms. The oral subgingival microbiota contributed to the accelerated corrosion of 316L SS via acidic metabolites and extracellular electron transfer. Our findings provide a new insight into the underlying mechanisms of oral microbial corrosion and guide the design of oral microbial corrosion-resistant materials.

Harnessing active biofilm for microbial corrosion protection of carbon steel against Geobacter sulfurreducens.

Microbiologically influenced corrosion (MIC) caused by corrosive microorganisms poses significant economic losses and safety hazards. Conventional corrosion prevention methods have limitations, so it is necessary to develop the eco-friendly and long-term effective strategies to mitigate MIC. This study investigated the inhibition of Vibrio sp. EF187016 biofilm on Geobacter sulfurreducens on carbon steel. Vibrio sp. EF187016 biofilm reduced the corrosion current density and impeded pitting corrosion. A thick and uniform Vibrio sp. EF187016 biofilm formed on the coupon surfaces, acting as a protective layer against corrosive ions and electron acquisition by G. sulfurreducens. The pre-grown mature Vibrio sp. EF187016 biofilms, provided enhanced protection against G. sulfurreducens corrosion. Additionally, the extracellular polymeric substances from Vibrio sp. EF187016 was confirmed to act as a green corrosion inhibitor to mitigate microbial corrosion. This study highlights the potential of active biofilms for eco-friendly corrosion protection, offering a novel perspective on material preservation against microbial corrosion.

Burning question: Are there sustainable strategies to prevent microbial metal corrosion?

The global economic burden of microbial corrosion of metals is enormous. Microbial corrosion of iron-containing metals is most extensive under anaerobic conditions. Microbes form biofilms on metal surfaces and can directly extract electrons derived from the oxidation of Fe0 to Fe2+ to support anaerobic respiration. H2 generated from abiotic Fe0 oxidation also serves as an electron donor for anaerobic respiratory microbes. Microbial metabolites accelerate this abiotic Fe0 oxidation. Traditional strategies for curbing microbial metal corrosion include cathodic protection, scrapping, a diversity of biocides, alloys that form protective layers or release toxic metal ions, and polymer coatings. However, these approaches are typically expensive and/or of limited applicability and not environmentally friendly. Biotechnology may provide more effective and sustainable solutions. Biocides produced with microbes can be less toxic to eukaryotes, expanding the environments for potential application. Microbially produced surfactants can diminish biofilm formation by corrosive microbes, as can quorum-sensing inhibitors. Amendments of phages or predatory bacteria have been successful in attacking corrosive microbes in laboratory studies. Poorly corrosive microbes can form biofilms and/or deposit extracellular polysaccharides and minerals that protect the metal surface from corrosive microbes and their metabolites. Nitrate amendments permit nitrate reducers to outcompete highly corrosive sulphate-reducing microbes, reducing corrosion. Investigation of all these more sustainable corrosion mitigation strategies is in its infancy. More study, especially under environmentally relevant conditions, including diverse microbial communities, is warranted.

Microbially Influenced Corrosion of Steel in Marine Environments: A Review from Mechanisms to Prevention.

Microbially influenced corrosion (MIC) is a formidable challenge in the marine industry, resulting from intricate interactions among various biochemical reactions and microbial species. Many preventions used to mitigate biocorrosion fail due to ignorance of the MIC mechanisms. This review provides a summary of the current research on microbial corrosion in marine environments, including corrosive microbes and biocorrosion mechanisms. We also summarized current strategies for inhibiting MIC and proposed future research directions for MIC mechanisms and prevention. This review aims to comprehensively understand marine microbial corrosion and contribute to novel strategy developments for biocorrosion control in marine environments.

The fate of arsenic during the crystallization process of FeIII oxyhydroxides: Effect of reaction media, pH value, and Fe/As molar ratio under relatively low arsenic loading.

Understanding the nature of arsenic (As) adsorbed on FeIII oxyhydroxides, and the subsequent behavior of As during the crystallization process, is critical to predicting its fate in a range of natural and engineered settings. In this work, As adsorbed on FeIII oxyhydroxides formed in the different reaction media at different pH values were characterized with X-ray diffraction (XRD), Raman spectra, transmission electron microscopy (TEM), and extended X-ray absorption fine structure spectroscopy (EXAFS) to determine how As is redistributed during the crystallization process. Results showed that at pH 12, a quarter of the added As was still left in the liquid phase with the formation of goethite and hematite as the major and minor product. The concentration of As was found to be the lowest at pH 4 which is independent of the reaction media, indicating the importance of pH value in the crystallization process of the As adsorbed FeIII oxyhydroxides. Under acidic conditions, sulfate and chloride media favored the formation of goethite and hematite, respectively. Arsenic can indeed be incorporated into the structure of the formed goethite at pH 4. The morphology of the formed products changed to rhombus-like particles if both goethite and hematite appeared as the later as the dominant product.

Accelerated Microbial Corrosion by Magnetite and Electrically Conductive Pili through Direct Fe0 -to-Microbe Electron Transfer.

Electrobiocorrosion, the process in which microbes extract electrons from metallic iron (Fe0 ) through direct Fe0 -microbe electrical connections, is thought to contribute to the costly corrosion of iron-containing metals that impacts many industries. However, electrobiocorrosion mechanisms are poorly understood. We report here that electrically conductive pili (e-pili) and the conductive mineral magnetite play an important role in the electron transfer between Fe0 and Geobacter sulfurreducens, the first microbe in which electrobiocorrosion has been rigorously documented. Genetic modification to express poorly conductive pili substantially diminished corrosive pitting and rates of Fe0 -to-microbe electron flux. Magnetite reduced resistance to electron transfer, increasing corrosion currents and intensifying pitting. Studies with mutants suggested that the magnetite promoted electron transfer in a manner similar to the outer-surface c-type cytochrome OmcS. These findings, and the fact that magnetite is a common product of iron corrosion, suggest a potential positive feedback loop of magnetite produced during corrosion further accelerating electrobiocorrosion. The interactions of e-pili, cytochromes, and magnetite demonstrate mechanistic complexities of electrobiocorrosion, but also provide insights into detecting and possibly mitigating this economically damaging process.

Microbially mediated metal corrosion.

A wide diversity of microorganisms, typically growing as biofilms, has been implicated in corrosion, a multi-trillion dollar a year problem. Aerobic microorganisms establish conditions that promote metal corrosion, but most corrosion has been attributed to anaerobes. Microbially produced organic acids, sulfide and extracellular hydrogenases can accelerate metallic iron (Fe0) oxidation coupled to hydrogen (H2) production, as can respiratory anaerobes consuming H2 as an electron donor. Some bacteria and archaea directly accept electrons from Fe0 to support anaerobic respiration, often with c-type cytochromes as the apparent outer-surface electrical contact with the metal. Functional genetic studies are beginning to define corrosion mechanisms more rigorously. Omics studies are revealing which microorganisms are associated with corrosion, but new strategies for recovering corrosive microorganisms in culture are required to evaluate corrosive capabilities and mechanisms. Interdisciplinary studies of the interactions among microorganisms and between microorganisms and metals in corrosive biofilms show promise for developing new technologies to detect and prevent corrosion. In this Review, we explore the role of microorganisms in metal corrosion and discuss potential ways to mitigate it.

Synergistic Antibacterial Activity of Benzalkonium Bromide and Cu-Bearing Duplex Stainless Steel against Pseudomonas aeruginosa.

The bactericide benzalkonium bromide is widely used to kill Pseudomonas aeruginosa, which causes microbiologically influenced corrosion (MIC). However, the extensive use of benzalkonium bromide will enhance bacterial drug resistance and cause environmental pollution. In this study, benzalkonium bromide combined with Cu-bearing 2205 duplex stainless steel (2205-Cu DSS) was used to kill Pseudomonas aeruginosa; the germicidal rate of the combination of benzalkonium bromide and 2205-Cu DSS was 24.2% higher than that of using benzalkonium bromide alone, after five days. The antibacterial efficacy was evaluated using an antibacterial test and biofilm observation. The results showed that, in the presence of P. aeruginosa, the combination of 23.44 ppm benzalkonium bromide and 2205-Cu DSS showed the best antibacterial efficacy.

3D printed Ti-5Cu alloy accelerates osteogenic differentiation of MC3T3-E1 cells by stimulating the M2 phenotype polarization of macrophages.

Ti-5Cu alloy has been proved to have excellent mechanical properties and cell compatibility and has certain antibacterial properties due to the addition of Cu. However, there are few studies on the effects of Ti-5Cu alloy on macrophage polarization and immune-related bone formation. In this study, we prepared Ti-5Cu alloy by three-dimensional printing technology and found that Ti-5Cu alloy presented a much smoother surface compared with Ti. In addition, the CCK-8 results indicated the Ti-5Cu alloy had no cytotoxicity to RAW264.7 cells by co-culture. The results of inductively coupled plasma mass spectrometry showed that the concentration of Cu2+ was 0.133 mg/L after 7 days of co-culture, and the CCK-8 results proved that Cu2+ had no cytotoxicity to RAW264.7 at this concentration. Then, we studied the effects of Ti-5Cu alloy on macrophage polarization; it was shown that the Ti-5Cu alloy is more prone to modulate the RAW264.7 polarization towards the M2 phenotype and the conditioned medium derived from Ti-5Cu alloy significantly promoted the proliferation and osteogenic differentiation of MC3T3-E1 cells. However, when the expression of Oncostatin M (OSM) gene of RAW264.7 was knocked down, the osteogenic differentiation of MC3T3-E1 cells was decreased. This suggests that the OSM secreted by RAW264.7 co-cultured with Ti-5Cu alloy could accelerate the osteogenic differentiation of MC3T3-E1 cells by acting on OSMR/gp130 receptors.

Ruscogenin Ameliorated Sjögren's Syndrome by Inhibiting NLRP3 Inflammasome Activation.

This article investigated the role and the specific mechanism of Ruscogenin in Sjögren's syndrome (SS). NOD/ShiLtJ mice were treated with Ruscogenin, and acinar cells isolated from submandibular glands were treated with TNF-α, Ruscogenin and transfected with NLRP3 overexpression plasmid. Salivary flow rate (SFR) was measured at weeks 11, 13, 15, 17, and 20. Histological analysis of the submandibular glands was conducted by hematoxylin-eosin staining assay. IL-6, IL-17, TNF-α, and IL-1ß mRNA expression was detected through qRT-PCR. AQP 5, AQP 4, P2X7R, NLRP3, caspase 1, IL-1ß, Bax, and Bcl-2 protein levels were tested by western blot. Cell apoptosis was assessed through acridine orange and propidium iodide (AO/PI) staining assay and flow cytometry assay. Ruscogenin ameliorated the SFR and submandibular gland inflammation of NOD/ShiLtJ mice. Ruscogenin promoted the preservation of acinar cells and suppressed inflammation-related factors (P2X7R, NLRP3, caspase 1, and IL-1ß) in submandibular gland tissues of NOD/ShiLtJ mice. Ruscogenin inhibited acinar cell apoptosis in NOD/ShiLtJ mice and reversed TNF-α-induced apoptosis and inflammation of acinar cells. NLRP3 overexpression reversed the repressive effect of Ruscogenin on TNF-α-induced inflammation and apoptosis of acinar cells. Ruscogenin ameliorated SS by inhibiting NLRP3 inflammasome activation.

Accelerated biocorrosion of stainless steel in marine water via extracellular electron transfer encoding gene phzH of Pseudomonas aeruginosa.

Microbiologically influenced corrosion (MIC) constantly occurs in water/wastewater systems, especially in marine water. MIC contributes to billions of dollars in damage to marine industry each year, yet the physiological mechanisms behind this process remain poorly understood. Pseudomonas aeruginosa is a representative marine electro-active bacterium, which has been confirmed to cause severe MIC on carbon steel through extracellular electron transfer (EET). However, little is known about how P. aeruginosa causes corrosion on stainless steel. In this study, the corrosivity of wild-type strain, phzH knockout, phzH complemented, and phzH overexpression P. aeruginosa mutants were evaluated to explore the underlying MIC mechanism. We found the accelerated MIC on 2205 duplex stainless steel (DSS) was due to the secretion of phenazine-1-carboxamide (PCN), which was regulated by the phzH gene. Surface analysis, Mott-Schottky test and H2O2 measurement results showed that PCN damaged the passive film by forming H2O2 to oxidize chromium oxide to soluble hexavalent chromium, leading to more severe pitting corrosion. The normalized corrosion rate per cell followed the same order as the general corrosion rate obtained under each experimental condition, eliminating the influence of the total amount of sessile cells on corrosion. These findings provide new insight and are meaningful for the investigation of MIC mechanisms on stainless steel. The understanding of MIC can improve the sustainability and resilience of infrastructure, leading to huge environmental and economic benefits.

Direct microbial electron uptake as a mechanism for stainless steel corrosion in aerobic environments.

Shewanella oneidensis MR-1 is an attractive model microbe for elucidating the biofilm-metal interactions that contribute to the billions of dollars in corrosion damage to industrial applications each year. Multiple mechanisms for S. oneidensis-enhanced corrosion have been proposed, but none of these mechanisms have previously been rigorously investigated with methods that rule out alternative routes for electron transfer. We found that S. oneidensis grown under aerobic conditions formed thick biofilms (∼50 µm) on stainless steel coupons, accelerating corrosion over sterile controls. H2 and flavins were ruled out as intermediary electron carriers because stainless steel did not reduce riboflavin and previous studies have demonstrated stainless does not generate H2. Strain ∆mtrCBA, in which the genes for the most abundant porin-cytochrome conduit in S. oneidensis were deleted, corroded stainless steel substantially less than wild-type in aerobic cultures. Wild-type biofilms readily reduced nitrate with stainless steel as the sole electron donor under anaerobic conditions, but strain ∆mtrCBA did not. These results demonstrate that S. oneidensis can directly consume electrons from iron-containing metals and illustrate how direct metal-to-microbe electron transfer can be an important route for corrosion, even in aerobic environments.

Engineering microbial systems for the production and functionalization of biomaterials.

A new trend in biomaterials synthesis is harnessing the production of microorganisms, owing to the low cost and sustainability. Because microorganisms use DNA as a production code, it is possible for humans to reprogram these cells and thus build living factories for the production of biomaterials. Over the past decade, advances in genetic engineering have enabled the development of various intriguing biomaterials with useful properties, with commercially available biomaterials representing only a few of these. In this review, we discuss the common strategies for the production of bulk and commodity biogenic polymers, and highlight several notable approaches such as modular protein engineering and pathway optimization in achieving these goals. We finally investigate the available synthetic biology tools that allow engineering of living materials, and discuss how this emerging class of materials has expanded the application scope of biomaterials.

Polyethyleneimine Functionalized Mesoporous Magnetic Nanoparticles with Enhanced Antibacterial and Antibiofilm Activity in an Alternating Magnetic Field.

Despite a lot of research on the antibacterial effect of Fe3O4 nanoparticles, their interactions with biofilm matrix have not been well understood. The surface charge of nanoparticles mainly determines their ability to adhere on the biofilm. In this work, negatively charged Fe3O4 nanoparticles were synthesized via a trisodium citrate-assisted solvothermal method and then the surfaces were functionalized using polyethyleneimine (PEI) to obtain positively charged Fe3O4 nanoparticles. The antibacterial and antibiofilm activities of both negatively and positively charged Fe3O4 nanoparticles in an alternating magnetic field were then systematically investigated. The positively charged Fe3O4 nanoparticles showed a strong self-adsorbed attachment ability to the planktonic and sessile cells, resulting in a better antibacterial activity and enhanced biofilm eradication performance compared to the conventional Fe3O4 nanoparticles with negative charges. Fe3O4@PEI nanoparticles produced physical stress and thermal damage in response to an alternating magnetic field, inducing the accumulation of intracellular reactive oxygen species into live bacterial cells, bacterial membrane damage, and biofilm dispersion. Utilizing an alternating magnetic field along with positively charged nanoparticles leads to a synergistic antibacterial approach to improve the antibiofilm performance of magnetic nanoparticles.

Bacterial biofilms as platforms engineered for diverse applications.

Historically, biofilms have been perceived as problematic or detrimental. However, biofilms possess favorable traits such as self-regeneration, sustainability, scalability, and tunability, which make them candidates for diverse applications. Traditional applications of biofilms, such as environmental remediation, bioleaching, microbial fuel cells, and corrosion protection, are often built on the basis of wild-type or metabolically engineered strains. In this review, we further comment on the design strategies for multiple innovative applications of living functional biofilms. With the integration of signaling pathways, engineering of metabolic pathways and modification of extracellular polymeric substances, living functional biofilms have been constructed by researchers through various strategies. Functional biofilms for diverse applications, including catalysis, electric conduction, bioremediation, and medical therapy have been demonstrated in the literature. The mechanical properties of biofilms can be tuned through genetic editing, metal ion curing and synthetic gene circuits, etc. In addition, the improvement of 3D printing to use bioinks has also achieved significant progresses in fabricating living functional biofilms with specific structures. In the future, the combination of synthetic biology and techniques from other disciplines will lead to practical large-scale applications of biofilms.

Enhanced antibacterial behavior of a novel Cu-bearing high-entropy alloy.

Contact infection of bacteria and viruses has been a critical threat to human health. The worldwide outbreak of COVID-19 put forward urgent requirements for the research and development of the self-antibacterial materials, especially the antibacterial alloys. Based on the concept of high-entropy alloys, the present work designed and prepared a novel Co0.4FeCr0.9Cu0.3 antibacterial high-entropy alloy with superior antibacterial properties without intricate or rigorous annealing processes, which outperform the antibacterial stainless steels. The antibacterial tests presented a 99.97% antibacterial rate against Escherichia coli and a 99.96% antibacterial rate against Staphylococcus aureus after 24 h. In contrast, the classic antibacterial copper-bearing stainless steel only performed the 71.50% and 80.84% antibacterial rate, respectively. The results of the reactive oxygen species analysis indicated that the copper ion release and the immediate contact with copper-rich phase had a synergistic effect in enhancing antibacterial properties. Moreover, this alloy exhibited excellent corrosion resistance when compared with the classic antibacterial stainless steels, and the compression test indicated the yield strength of the alloy was 1015 MPa. These findings generate fresh insights into guiding the designs of structure-function-integrated antibacterial alloys.