Crosslinking ionic oligomers as conformable precursors to calcium carbonate.

Inorganic materials have essential roles in society, including in building construction, optical devices, mechanical engineering and as biomaterials1-4. However, the manufacture of inorganic materials is limited by classical crystallization5, which often produces powders rather than monoliths with continuous structures. Several precursors that enable non-classical crystallization-such as pre-nucleation clusters6-8, dense liquid droplets9,10, polymer-induced liquid precursor phases11-13 and nanoparticles14-have been proposed to improve the construction of inorganic materials, but the large-scale application of these precursors in monolith preparations is limited by availability and by practical considerations. Inspired by the processability of polymeric materials that can be manufactured by crosslinking monomers or oligomers15, here we demonstrate the construction of continuously structured inorganic materials by crosslinking ionic oligomers. Using calcium carbonate as a model, we obtain a large quantity of its oligomers (CaCO3)n with controllable molecular weights, in which triethylamine acts as a capping agent to stabilize the oligomers. The removal of triethylamine initiates crosslinking of the (CaCO3)n oligomers, and thus the rapid construction of pure monolithic calcium carbonate and even single crystals with a continuous internal structure. The fluid-like behaviour of the oligomer precursor enables it to be readily processed or moulded into shapes, even for materials with structural complexity and variable morphologies. The material construction strategy that we introduce here arises from a fusion of classic inorganic and polymer chemistry, and uses the same cross-linking process for the manufacture the materials.

Biomimetic Mineral Synthesis by Nanopatterned Supramolecular-Block Copolymer Templates.

Supramolecular structures of matrix proteins in mineralizing tissues are known to direct the crystallization of inorganic materials. Here we demonstrate how such structures can be synthetically directed into predetermined patterns for which functionality is maintained. The study employs block copolymer lamellar patterns with alternating hydrophilic and hydrophobic regions to direct the assembly of amelogenin-derived peptide nanoribbons that template calcium phosphate nucleation by creating a low-energy interface. Results show that the patterned nanoribbons retain their ß-sheet structure and function and direct the formation of filamentous and plate-shaped calcium phosphate with high fidelity, where the phase, amorphous or crystalline, depends on the choice of mineral precursor and the fidelity depends on peptide sequence. The common ability of supramolecular systems to assemble on surfaces with appropriate chemistry combined with the tendency of many templates to mineralize multiple inorganic materials implies this approach defines a general platform for bottom-up-patterning of hybrid organic-inorganic materials.

Adaptive Laboratory Evolution of Halomonas bluephagenesis Enhances Acetate Tolerance and Utilization to Produce Poly(3-hydroxybutyrate).

Acetate is a promising economical and sustainable carbon source for bioproduction, but it is also a known cell-growth inhibitor. In this study, adaptive laboratory evolution (ALE) with acetate as selective pressure was applied to Halomonas bluephagenesis TD1.0, a fast-growing and contamination-resistant halophilic bacterium that naturally accumulates poly(3-hydroxybutyrate) (PHB). After 71 transfers, the evolved strain, B71, was isolated, which not only showed better fitness (in terms of tolerance and utilization rate) to high concentrations of acetate but also produced a higher PHB titer compared with the parental strain TD1.0. Subsequently, overexpression of acetyl-CoA synthetase (ACS) in B71 resulted in a further increase in acetate utilization but a decrease in PHB production. Through whole-genome resequencing, it was speculated that genetic mutations (single-nucleotide variation (SNV) in phaB, mdh, and the upstream of OmpA, and insertion of TolA) in B71 might contribute to its improved acetate adaptability and PHB production. Finally, in a 5 L bioreactor with intermittent feeding of acetic acid, B71 was able to produce 49.79 g/L PHB and 70.01 g/L dry cell mass, which were 147.2% and 82.32% higher than those of TD1.0, respectively. These results highlight that ALE provides a reliable method to harness H. bluephagenesis to metabolize acetate for the production of PHB or other high-value chemicals more efficiently.

Environmental microplastics: Classification, sources, fates, and effects on plants.

Microplastic (MP) pollution has become a global concern due to the generation of extensive plastic waste and products (370 million metric tons in 2020) that are difficult to biodegrade. Therefore, MPs have attracted a great deal of research attention, and many new findings regarding MPs (over 9000 papers published in the last 3 years) have been reported. MPs generally exert adverse effects on plants. As MPs accumulate in agricultural ecosystems, many studies have sought to understand the sources and fates of MPs and their effects on various plants. However, there have been few reviews of the properties of MPs, their effects on plants, and their interactions with other factors (e.g., drought, heat, ultraviolet light, plant hormones, heavy metals, and other pollutants) remain poorly understood. In this review, we performed scientometrics analyses of research papers (January 1, 2019, to September 30, 2022) in this field. We focused on the recent progress in the classification of MPs and their sources, circulation, and deposition in agricultural ecosystems. We review MP uptake and transport in plants, as well as factors (size, type, and environmental factors) that affect MP uptake, the positive and negative effects of MPs on plants, and the mechanisms of MP impacts on plants. We discuss current issues and future perspectives concerning research into plant interactions with MPs, along with some promising methods to manage the MP issue.

Controlling Mineralization with Protein-Functionalized Peptoid Nanotubes.

Sequence-defined foldamers that self-assemble into well-defined architectures are promising scaffolds to template inorganic mineralization. However, it has been challenging to achieve robust control of nucleation and growth without sequence redesign or extensive experimentation. Here, peptoid nanotubes functionalized with a panel of solid-binding proteins are used to mineralize homogeneously distributed and monodisperse anatase nanocrystals from the water-soluble TiBALDH precursor. Crystallite size is systematically tuned between 1.4 and 4.4 nm by changing protein coverage and the identity and valency of the genetically engineered solid-binding segments. The approach is extended to the synthesis of gold nanoparticles and, using a protein encoding both material-binding specificities, to the fabrication of titania/gold nanocomposites capable of photocatalysis under visible-light illumination. Beyond uncovering critical roles for hierarchical organization and denticity on solid-binding protein mineralization outcomes, the strategy described herein should prove valuable for the fabrication of hierarchical hybrid materials incorporating a broad range of inorganic components.

Electrochemical and microbiological response of exoelectrogenic biofilm to polyethylene microplastics in water.

Exoelectrogenic biofilm and the associated microbial electrochemical processes have recently been intensively studied for water treatment, but their response to and interaction with polyethylene (PE) microplastics which are widespread in various aquatic environments has never been reported. Here, we investigated how and to what extent PE microplastics would affect the electrochemistry and microbiology of exoelectrogenic biofilm in both microbial fuel cells (MFCs) and microbial electrolysis cells (MECs). When the PE microplastics concentration was increased from 0 to 75 mg/L in the MECs, an apparent decline in the maximum current density (from 1.99 to 0.74 A/m2) and abundance of electroactive bacteria (EAB) in the exoelectrogenic biofilm was noticed. While in the MFCs, the current output was not significantly influenced and the abundance of EAB lightly increased at 25 mg/L microplastics. In addition, PE microplastics restrained the viability of the exoelectrogenic biofilms in both systems, leading to a higher system electrode resistance. Moreover, the microbial community richness and the microplastics-related operational taxonomic units decreased with PE microplastics. Furthermore, the electron transfer-related genes (e.g., pilA and mtrC) and cytochrome c concentration decreased after adding microplastics. This study provides the first glimpse into the influence of PE microplastics on the exoelectrogenic biofilm with the potential mechanisms revealed at the gene level, laying a methodological foundation for the future development of efficient water treatment technologies.

Insights into the impact of polyethylene microplastics on methane recovery from wastewater via bioelectrochemical anaerobic digestion.

Bioelectrochemical anaerobic digestion (BEAD) is a promising next-generation technology for simultaneous wastewater treatment and bioenergy recovery. While knowledge on the inhibitory effect of emerging pollutants, such as microplastics, on the conventional wastewater anaerobic digestion processes is increasing, the impact of microplastics on the BEAD process remains unknown. This study shows that methane production decreased by 30.71% when adding 10 mg/L polyethylene microplastics (PE-MP) to the BEAD systems. The morphology of anaerobic granular sludge, which was the biocatalysts in the BEAD, changed with microbes shedding and granule crack when PE-MP existed. Additionally, the presence of PE-MP shifted the microbial communities, leading to a lower diversity but higher richness and tight clustering. Moreover, fewer fermentative bacteria, acetogens, and hydrogenotrophic methanogens (BEAD enhanced) grew under PE-MP stress, suggesting that PE-MP had an inhibitory effect on the methanogenic pathways. Furthermore, the abundance of genes relevant to extracellular electron transfer (omcB and mtrC) and methanogens (hupL and mcrA) decreased. The electron transfer efficiency reduced with extracellular cytochrome c down and a lower electron transfer system activity. Finally, phylogenetic investigation of communities by reconstruction of unobserved states analysis predicted the decrease of key methanogenic enzymes, including EC 1.1.1.1 (Alcohol dehydrogenase), EC 1.2.99.5 (Formylmethanofuran dehydrogenase), and EC 2.8.4.1 (Coenzyme-B sulfoethylthiotransferase). Altogether, these results provide insight into the inhibition mechanism of microplastics in wastewater methane recovery and further optimisation of the BEAD process.

Indoor microplastics and bacteria in the atmospheric fallout in urban homes.

Humans may be exposed to microplastics (MPs) through food, drink, and air. Although several studies have examined indoor environmental MPs, none have yet compared atmospheric MP and bacterial deposition characteristics among rooms in homes. We investigated indoor airborne MPs and bacteria in five room types (bedroom, dining room, living room, bathroom, and study) based on the duration of usage of each room. We identified synthetic polymers (23,889 MP particles of 21 types) and bacterial communities (383 genera belong to 24 phyla) collected through atmospheric deposition in various rooms of 20 homes. The abundance and composition of MPs are related to the duration of usage, human activities, goods, cleanliness, and the composition of occupants (family members) in households. In addition, the homes of elderly families (age 68-81 years) showed higher bacterial concentrations than those of young families (age 28-35 years), indicating that age markedly affects the structure of household microbiota. Furthermore, a significant correlation between MP concentration and bacterial community structure was observed. The abundances of polyamide (PA), polyurethane (PU), and polyethylene (PE) showed positive correlations with the relative abundances of major bacterial phyla. Taken together, our results suggest that various rooms in the home exhibit distinct MP abundances and bacterial structures that may be affected by age, cleanliness, and human activities.

(S)-1-(5-(4-Methylpiperazin-1-yl)-2,4-dinitrophenyl)pyrrolidine-2-carboxylic acid as a derivatization reagent for ultrasensitive detection of amine enantiomers by HPLC-MS/MS and its application to the chiral metabolite analysis of (R)-1-aminoindan in saliva.

(S)-1-(5-(4-Methylpiperazin-1-yl)-2,4-dinitrophenyl)pyrrolidine-2-carboxylic acid (Pro-PPZ) was employed as a chiral derivatization reagent (CDR) for the efficient enantioseparation and ultrasensitive mass spectrometric detection of chiral amines. Pro-PPZ was prepared from the one-step reaction of 1-(5-fluoro-2,4-dinitrophenyl)-4-methylpiperazine (PPZ) and l-proline. Two amines and two amino acid methyl esters were selected as model chiral amines, which were easily labeled with Pro-PPZ under mild reaction conditions (35 °C for 10 min) generating Pro-PPZ-amine derivatives. The resulting diastereomers were completely separated by reversed-phase liquid chromatography (RP-LC) using an ODS column (Rs = 3.4-17.0 for amines). Ultrasensitive detection limits on femtomolar level were obtained for the tested amines using multiple reaction monitoring (MRM) chromatograms at a single monitoring ion, m/z 289 (0.1-5.0 fmol for amines). The practical metabolite analysis of (R)-1-aminoindan (R-AI) in saliva samples was performed by LC-MS/MS using the Pro-PPZ derivatization method. The method was validated in terms of precision, accuracy, and linearity. Using this method, R-AI concentrations in saliva were determined after a single oral administration of the drug rasagiline to healthy male and female subjects, but no (S)-1-aminoindan (S-AI) was detected, which suggesting that R-AI was not converted into S-enantiomer in the metabolic process. R-AI concentrations in four healthy volunteers ranged from 32.85 nM to 49.45 nM, with an average value of 43.76 nM. To date, there is no LC-MS (or MS/MS) method reported for the enantioselective determination of R-AI in human saliva samples.

Cell Catalysis of Citrate to Itaconate by Engineered Halomonas bluephagenesis.

Itaconic acid (IA), an important five-carbon unsaturated dicarboxylic acid, is one of the top 12 renewable chemicals with an urgent need to reduce industrial production costs. Halomonas bluephagenesis, which possesses the potential for cost-effective bioproduction of chemicals and organic acids due to its ability to grow under open nonsterile conditions and high tolerance to organic acid salts, was genetically engineered and used to produce IA from citrate by a cell catalytic strategy. Here, two essential genes (cis-aconitate decarboxylase encoding gene cadA and aconitase (ACN) encoding gene acn) were introduced into H. bluephagenesis to construct an IA biosynthesis pathway. Further engineering modifications including coexpression of molecular chaperones GroESL, increasing the copy number of the gene encoding rate-limiting enzyme ACN, and weakening the competing pathway were implemented. Under the optimized condition for the cell catalytic system, the engineered strain TAZI-08 produced 451.45 mM (58.73 g/L) IA from 500 mM citrate, with 93.24% conversion in 36 h and a productivity of 1.63 g/(L h). An intermittent feeding strategy further increased the IA titer to 488.86 mM (63.60 g/L). The IA titer and citrate conversion in H. bluephagenesis are the highest among heterologous hosts reported so far, demonstrating that this strain is a suitable chassis for hyperproduction of IA.

Effect of aspartic acid on the crystallization kinetics of ACP and dentin remineralization.

Type I collagen and non-collagen proteins are the main organic components of dentin. This study aimed to investigate the biomimetic remineralization of demineralized dentin by aspartic acid (Asp), which is abundant in non-collagenous proteins (NCPs). Asp was added to a mineralizing solution containing polyacrylic acid (PAA) to explore the mechanism of Asp regulating the pure amorphous calcium phosphate (ACP) phase transition process. The remineralization process and superstructure of the remineralized layer of demineralized dentin were evaluated and analyzed by transmission electron microscope (TEM) and scanning electron microscope (SEM), and the biological stability of the remineralized layer was investigated by collagenase degradation experiment. It demonstrated that Asp promoted the crystallization kinetics of PAA-stabilized amorphous calcium phosphate to hydroxyapatite (HAP), and shortened the remineralization time of demineralized dentin from 7 days to 2 days. The newly formed remineralized dentin had similar morphology and biological stability to the natural dentin layer. The presence of a large number of Asp residues in NCPs promoted the phase transformation of ACP, and further revealed the mechanism of action of NCPs in dentin biomineralization. This experiment also showed that Asp promoted the biomimetic remineralization of dentin; the morphology and hierarchical structure of remineralized layer was similar to that of natural teeth, and had good biological properties.

Experimental Study on the Bone Morphogenetic Protein 1-Modified Bone Marrow Mesenchymal Stem Cell Sheets to Promote Mandibular Distraction Osteogenesis.

OBJECTIVE: The present study aims to increase the concentration of genetically modified bone marrow mesenchymal stem cells (BMSCs) in the distraction osteogenesis (DO) interstitial space and induce the conversion of BMSCs to osteoblasts to improve the osteogenic efficiency in DO and shorten the treatment period. METHODS: Bone morphogenetic protein 1 (BMP-1) and green fluorescent protein (GFP) gene-modified cell sheets of BMSCs were constructed by tissue engineering. Thirty-six New Zealand white rabbits were randomly divided into three groups: group A (the blank control group), group B (the GFP group) with the injection of GFP gene-modified BMSC sheets into the DO gap, and group C (the BMP-1 group) with the injection of BMP-1 gene-modified BMSC sheets into the DO gap. Rabbits in all three groups were distracted for 5 days at a distraction rate of 2.0 mm/d, once/day. After distraction, the above-mentioned cell sheet suspension was injected into the distraction gap to observe osteogenesis, which was observed by gross specimen observation, micro-computed tomography (Micro-CT) scanning, and histomorphology. RESULTS: The gross specimen observation showed that all animals had smooth and continuous bone cortex in the distraction region with relatively high hardness. The osteogenesis quality or hardness was ranked from the highest to the lowest, as Group C > Group B > Group A. Micro-CT and histomorphological observation revealed that group C had better maturation and bone volume of the new bone in the DO region at weeks 3 and 6 than groups B and A. CONCLUSION: BMP-1 gene-modified BMSC sheets could effectively promote the formation of new bone during rapid DO in the mandible, compensating for the poor osteogenesis caused by rapid distraction and providing a new approach to shorten the DO treatment period in clinical practice.

Polydopamine Promotes Dentin Remineralization via Interfacial Control.

Biomineralization has intrigued researchers for decades. Although mineralization of type I collagen has been universally investigated, this process remains a great challenge due to the lack of mechanistic understanding of the roles of biomolecules. In our study, dentine was successfully repaired using the biomolecule polydopamine (PDA), and the remineralized dentine exhibited mechanical properties comparable to those of natural dentine. Detailed analyses of the collagen mineralization process facilitated by PDA showed that PDA can promote intrafibrillar mineralization with a decreased heterogeneous nucleation barrier for hydroxyapatite (HAP) by reducing the interfacial energy between collagen fibrils and amorphous calcium phosphate (ACP), resulting in the conversion of an increasing amount of nanoprecursors into collagen fibrils. The present work highlights the importance of interfacial control in dentine remineralization and provides profound insight into the regulatory effect of biomolecules in collagen mineralization as well as the clinical application of dentine restoration.

Repair of tooth enamel by a biomimetic mineralization frontier ensuring epitaxial growth.

The regeneration of tooth enamel, the hardest biological tissue, remains a considerable challenge because its complicated and well-aligned apatite structure has not been duplicated artificially. We herein reveal that a rationally designed material composed of calcium phosphate ion clusters can be used to produce a precursor layer to induce the epitaxial crystal growth of enamel apatite, which mimics the biomineralization crystalline-amorphous frontier of hard tissue development in nature. After repair, the damaged enamel can be recovered completely because its hierarchical structure and mechanical properties are identical to those of natural enamel. The suggested phase transformation-based epitaxial growth follows a promising strategy for enamel regeneration and, more generally, for biomimetic reproduction of materials with complicated structure.

Citrate Improves Collagen Mineralization via Interface Wetting: A Physicochemical Understanding of Biomineralization Control.

Biological hard tissues such as bones always contain extremely high levels of citrate, which is believed to play an important role in bone formation as well as in osteoporosis treatments. However, its mechanism on biomineralization is not elucidated. Here, it is found that the adsorbed citrate molecules on collagen fibrils can significantly reduce the interfacial energy between the biological matrix and the amorphous calcium phosphate precursor to enhance their wetting effect at the early biomineralization stage, sequentially facilitating the intrafibrillar formation of hydroxyapatite to produce an inorganic-organic composite. It is demonstrated experimentally that only collagen fibrils containing ≈8.2 wt% of bound citrate (close to the level in biological bone) can reach the full mineralization as those in natural bones. The effect of citrate on the promotion of the collagen mineralization degree is also confirmed by in vitro dentin repair. This finding demonstrates the importance of interfacial controls in biomineralization and more generally, provides a physicochemical view about the regulation effect of small biomolecules on the biomineralization front.

[Development of artificial bone made from nacre and polylactic acid and property assessment].

OBJECTIVE: To explore a new bone substitute for bone grafting and evaluate its properties. METHODS: Pressurized molding of nacre, polylactic acid and NaCl that were proportionally mixed was performed to obtain the bone grafting material, and its properties as porosity, pore size and mechanical strength were compared with those of the material prepared by solvent casting. RESULTS: Compared with that of the material made by solvent casting, the mechanical strength of the new artificial bone material was 10-fold stronger with an average porosity rate of 47.65%, pore size of 218.83 micrometer compressive strength of 16.10 MPa and flexural strength of 60.18 MPa. CONCLUSION: The new technique adopted in this study can effectively enhance the biomechanics of the artificial bone material, which can be suitable for repairing bone defect.

[In vitro biocompatibility and degradation of nacre/polylactic acid composite artificial bone].

OBJECTIVE: To investigate the in vitro biocompatibility and degradation of nacre/polylactic acid composite artificial bone (NPCB) grafts. METHODS: Human osteoblast cells were cultured with NPCB discs for observation of the morphological features and cell proliferation by optical microscope and scanning electron microscope. The mass loss of NPCB discs and pH variations of the saline in which the discs were immersed were examined every 2 weeks in a course of 16 weeks in vitro. RESULT: Compared with the negative control group, NPCB showed no visible cytotoxicity and facilitated the growth and proliferation of the osteoblast cells. As the immersion prolonged, gradual decrease in the mass of NPCB was observed accompanied by regular pH alteration of the saline. CONCLUSION: NPCB possesses good in vitro biocompatibility and may be gradually degraded.

[Diaphyseal defect repair with nacre/polylactic acid composite artificial bone in rabbits].

OBJECTIVE: To evaluate the biocompatibility, degradation and bone formation activity of a new bone substitute for bone grafting, nacre/polylactic acid composite artificial bone (NPCB). METHODS: Radial bone defects 1.5 cm in length were induced in 32 New Zealand rabbits and immediately filled with NPCB or nothing. The animals' local and whole body responses to the implants were observed after the operation, and the serum calcium levels were detected 1 week and 4 weeks postoperatively. Tissue response, new bone formation in the defects and degradation of the implants were evaluated by X-ray, and examination of the bone mineral content (BMC) in the defects and histomorphological analysis were performed after the rabbits were sacrificed. RESULTS: All the rabbits survived the operation and the incisions healed smoothly. No significant difference was noted in the serum calcium level between 1 day before operation, 1 week and 4 weeks postoperation. BMC in the defects of the rabbits with NPCB grafts increased more rapidly than that in the control group 6-12 weeks after the operation and began to decrease after 12 weeks. Gross, X-ray and histological observations revealed that NPCB possessed osteoconductive activity and new bone ingrowth in the implants was found during the whole experiment. At 16 weeks, the defects grafts were almost completely repaired with NPCB while the defects in the control group remained nonunion. Biodegradation of the implants was observed early at 6 weeks, with numerous macrophage and multinuclear giant cells containing phagocytosed NPCB particles around the implants at 12 weeks, but NPCB remnants were still visible at 16 weeks. CONCLUSION: As a bone substitute, NPCB possesses good biocompatibility, biodegradability and osteoconductive activity.

Comparison of expansive pedicle screw and polymethylmethacrylate-augmented pedicle screw in osteoporotic sheep lumbar vertebrae: biomechanical and interfacial evaluations.

BACKGROUND: It was reported that expansive pedicle screw (EPS) and polymethylmethacrylate-augmented pedicle screw (PMMA-PS) could be used to increase screw stability in osteoporosis. However, there are no studies comparing the two kinds of screws in vivo. Thus, we aimed to compare biomechanical and interfacial performances of EPS and PMMA-PS in osteoporotic sheep spine. METHODOLOGY/PRINCIPAL FINDINGS: After successful induction of osteoporotic sheep, lumbar vertebrae in each sheep were randomly divided into three groups. The conventional pedicle screw (CPS) was inserted directly into vertebrae in CPS group; PMMA was injected prior to insertion of CPS in PMMA-PS group; and the EPS was inserted in EPS group. Sheep were killed and biomechanical tests, micro-CT analysis and histological observation were performed at both 6 and 12 weeks post-operation. At 6-week and 12-week, screw stabilities in EPS and PMMA-PS groups were significantly higher than that in CPS group, but there were no significant differences between EPS and PMMA-PS groups at two study periods. The screw stability in EPS group at 12-week was significantly higher than that at 6-week. The bone trabeculae around the expanding anterior part of EPS were more and denser than that in CPS group at 6-week and 12-week. PMMA was found without any degradation and absorption forming non-biological "screw-PMMA-bone" interface in PMMA-PS group, however, more and more bone trabeculae surrounded anterior part of EPS improving local bone quality and formed biological "screw-bone" interface. CONCLUSIONS/SIGNIFICANCE: EPS can markedly enhance screw stability with a similar effect to the traditional method of screw augmentation with PMMA in initial surgery in osteoporosis. EPS can form better biological interface between screw and bone than PMMA-PS. In addition, EPS have no risk of thermal injury, leakage and compression caused by PMMA. We propose EPS has a great application potential in augmentation of screw stability in osteoporosis in clinic.