3D-bioprinted anisotropic bicellular living hydrogels boost osteochondral regeneration via reconstruction of cartilage-bone interface.

Reconstruction of osteochondral (OC) defects represents an immense challenge due to the need for synchronous regeneration of special stratified tissues. The revolutionary innovation of bioprinting provides a robust method for precise fabrication of tissue-engineered OCs with hierarchical structure; however, their spatial living cues for simultaneous fulfilment of osteogenesis and chondrogenesis to reconstruct the cartilage-bone interface of OC are underappreciated. Here, inspired by natural OC bilayer features, anisotropic bicellular living hydrogels (ABLHs) simultaneously embedding articular cartilage progenitor cells (ACPCs) and bone mesenchymal stem cells (BMSCs) in stratified layers were precisely fabricated via two-channel extrusion bioprinting. The optimum formulation of the 7% GelMA/3% AlgMA hydrogel bioink was demonstrated, with excellent printability at room temperature and maintained high cell viability. Moreover, the chondrogenic ability of ACPCs and the osteogenic ability of BMSCs were demonstrated in vitro, confirming the inherent differential spatial regulation of ABLHs. In addition, ABLHs exhibited satisfactory synchronous regeneration of cartilage and subchondral bone in vivo. Compared with homogeneous hydrogels, the neo-cartilage and neo-bone in ABLHs were augmented by 23.5% and 20.8%, respectively, and more important, a more harmonious cartilage-bone interface was achieved by ABLHs due to their well-tuned cartilage-bone-vessel crosstalk. We anticipate that such a strategy of tissue-mimetic ABLH by means of bioprinting is capable of spatiotemporal cell-driven regeneration, offering insights into the fabrication of anisotropic living materials for the reconstruction of complex organ defects.

3D-bioprinted BMSC-laden biomimetic multiphasic scaffolds for efficient repair of osteochondral defects in an osteoarthritic rat model.

Osteochondral defect repair in osteoarthritis (OA) remains an unsolved clinical problem due to the lack of enough seed cells in the defect and chronic inflammation in the joint. To address this clinical need, we designed a bone marrow-derived mesenchymal stem cell (BMSC)-laden 3D-bioprinted multilayer scaffold with methacrylated hyaluronic acid (MeHA)/polycaprolactone incorporating kartogenin and ß-TCP for osteochondral defect repair within each region. BMSC-laden MeHA was designed to actively introduce BMSCs in situ, and diclofenac sodium (DC)-incorporated matrix metalloproteinase-sensitive peptide-modified MeHA was induced on the BMSC-laden scaffold as an anti-inflammatory strategy. BMSCs in the scaffolds survived, proliferated, and produced large amounts of cartilage-specific extracellular matrix in vitro. The effect of BMSC-laden scaffolds on osteochondral defect repair was investigated in an animal model of medial meniscectomy-induced OA. BMSC-laden scaffolds facilitated chondrogenesis by promoting collagen II and suppressed interleukin 1ß in osteochondral defects of the femoral trochlea. Congruently, BMSC-laden scaffolds significantly improved joint function of the injured leg with respect to the ground support force, paw grip force, and walk gait parameters. Therefore, this research demonstrates the potential of 3D-bioprinted BMSC-laden scaffolds to simultaneously inhibit joint inflammation and promote cartilage defect repair in OA joints.

STAT1-Induced Upregulation lncRNA LINC00958 Accelerates the Epithelial Ovarian Cancer Tumorigenesis by Regulating Wnt/ß-Catenin Signaling.

BACKGROUND: Growing studies have demonstrated that long noncoding RNAs (lncRNAs) play important roles in tumor progression. In this study, we aimed to explore the potential roles of lncRNA LINC00958 (LINC00958) and its biological functions in epithelial ovarian cancer (EOC). METHODS: The expression of LINC00958 in 11 cases of EOC and adjacent nontumor specimens and five cell lines was detected by qRT-PCR. CCK-8, colony formation, and flow cytometry assays were conducted to study the cell viabilities of EOC cells. Wound scratch and transwell analyses were carried out for the examination of cell invasion and migration of EOC cells. The targeting associations between LINC00958 and STAT1 were demonstrated by ChIP analyses combined with luciferase reporter assays. The related proteins of Wnt/ß-catenin signaling were determined using RT-PCR. RESULTS: Higher levels of LINC00958 were observed in EOC tissues and cell lines. Our data also revealed that high LINC00958 expression was partly induced by STAT1. Functionally, knockdown of LINC00958 suppressed the proliferation, migration, and invasion of EOC cells. Mechanistic investigation showed that the inhibitory effect of LINC00958 knockdown on EOC cells was mediated by the Wnt/ß-catenin signaling. CONCLUSION: Our findings suggested that STAT1-induced overexpression of LINC00958 promoted EOC progression by modulating Wnt/ß-catenin signaling.

Synthesis and aqueous anion recognition of an imidazolium-based nonacationic cup.

An imidazolium-based nonacationic cup (1·9X; X = PF6- or Cl-) was synthesized via step-by-step SN2 reactions without using any template. The water-soluble 1·9Cl- as a molecular container can encapsulate anionic nucleoside triphosphate and dinucleotide molecules (e.g., ATP and NADH) inside its cavity through hydrogen bonds and electrostatic interactions in aqueous solution.

Characterization of Volatile Compounds of Rosa roxburghii Tratt by Gas Chromatography-Olfactometry, Quantitative Measurements, Odor Activity Value, and Aroma Intensity.

Rosa roxburghii tratt (RRT), widely distributed in the southwest of China, is favored by consumers for its good taste and healthy functions. In this study, thirty-seven compounds of Rosa roxburghii Tratt (RRT) were identified and quantified by gas chromatography-olfactometry (G-O) and gas chromatography-mass spectrometry (GC-MS) analysis. Furthermore, ethyl 2-methylpropanoate, ethyl butanoate, ethyl 2-methylbutyrate, and ethyl hexanoate were present with much higher odor activity values (OAVs) than other compounds. The key notes were confirmed by omission tests. Possible interaction among key notes was investigated through odor intensity determination and sensory analysis. It showed fruity and woody notes had synergistic effects. Full factorial design was used to evaluate the notes contribution to the whole odor. One important finding is the major effect of order interactions, fruity note (X1) and woody note (X4) especially, emphasizing the existence of complex interactions occurring between odor notes. The interaction X1X4 was further investigated. The woody note has a positive effect when the fruity note is also in the mixture but tends to show a negative effect otherwise.

Hierarchical Two-Level Supramolecular Chirality of an Achiral Anthracene-Based Tetracationic Nanotube in Water.

Herein, we report an achiral anthracene-based tetracationic nanotube (1⋅4Cl- ) that shows two levels of supramolecular chirality: namely, conformationally adaptive host-guest complexation with nucleoside triphosphates (e.g. ATP, GTP, CTP, and UTP) and twisted packing of the chiral host-guest complexes in water. Interestingly, achiral 1⋅4Cl- exhibits chiral recognition for ATP/GTP and CTP/UTP through structural transformation of its intramolecular M- and P-twisted conformation as the first level of supramolecular chirality, which leads to adaptive chirality with opposite CD responses. Furthermore, the formation of chiral M-1⋅4Cl- ⊃ATP can promote an intermolecular P-twisted dimeric packing of anthracene rings as the second level of supramolecular chirality to achieve assembled chirality with strong circularly polarized luminescence arising from the excimer ((+)-CPL, glum ≈10-2 ) in water.

In Vivo bone tissue induction by freeze-dried collagen-nanohydroxyapatite matrix loaded with BMP2/NS1 mRNAs lipopolyplexes.

Messenger RNA (mRNA) activated matrices (RAMs) are interesting to orchestrate tissue and organ regeneration due to the in-situ and sustained production of functional proteins. However, the immunogenicity of in vitro transcribed mRNA and the paucity of proper in vivo mRNA delivery vector need to be overcome to exert the therapeutic potential of RAM. We developed a dual mRNAs system for in vitro osteogenesis by co-delivering NS1 mRNA with BMP2 mRNA to inhibit RNA sensors and enhance BMP-2 expression. Next, we evaluated a lipopolyplex (LPR) formulation platform for in vivo mRNA delivery and adapted the LPRs for RAM preparation. The LPR formulated BMP2/NS1 mRNAs were incorporated into an optimized collagen-nanohydroxyapatite scaffold and freeze-dried to prepare ready-to-use RAMs. The loaded BMP2/NS1 mRNAs lipopolyplexes maintained their spherical morphology in the RAM, thanks to the core-shell structure of LPR. The mRNAs release from RAMs lasted for 16 days resulting in an enhanced prolonged transgene expression period compared to direct cell transfection. Once subcutaneously implanted in mice, the BMP2/NS1 mRNAs LPRs containing RAMs (RAM-BMP2/NS1) induced significant new bone tissue than those without NS1 mRNA, eight weeks post implantation. Overall, our results demonstrate that the BMP2/NS1 dual mRNAs system is suitable for osteogenic engagement, and the freeze-dried RAM-BMP2/NS1 could be promising off-the-shelf products for clinical orthopedic practice.

A tetraphenylethene-based Pd2L4 metallacage with aggregation-induced emission and stimuli-responsive behavior.

A tetraphenylethene-based Pd2L4 metallacage was self-assembled from four TPE-pyridine ligands with two Pd2+ ions. This metallacage with D4 symmetry exhibited a classical aggregation-induced emission property in different solvents and reversible stimuli-responsive behaviour with chloride ions and silver ions, successively.

Tetraphenylethene-Based Supramolecular Coordination Frameworks with Aggregation-Induced Emission for an Artificial Light-Harvesting System.

Supramolecular coordination is an efficient strategy to construct supramolecular coordination frameworks with predesigned structures, assembled shapes, and specific function. In this work, we report the synthesis, structural characterization, and photophysical property of two tetraphenylethene-based supramolecular coordination frameworks 1a and 1b formed from 1,1,2,2-tetrakis(4-(pyridin-4-yl)phenyl)ethene (2a) or 1,1,2,2-tetrakis(4-((E)-2-(pyridin-4-yl)vinyl)phenyl)ethene (2b) and a linear difunctional platinum(II) ligand (3a) via coordination-driven self-assembly. Controlled by the specific angularity and geometry of tetraphenylethene (with 60° and 120°) and difunctional Pt(II) linker (with 180°), these supramolecular coordination frameworks possess a well-defined and two-dimensional (2D) rhombic network-type topology with good periodicity and porosity. Given the aggregation-induced emission (AIE) property of tetraphenylethene units and the porosity of frameworks, 1a and 1b have been successfully used as fluorescent platforms and energy donors to fabricate efficient artificial light-harvesting materials with two fluorescent acceptors (Nile Red and Sulforhodamine 101) via noncovalent interactions in aqueous solution. Furthermore, these light-harvesting materials have been applied for promoting cancer cell imaging with a full shift of imaging channels from blue/green channels to the red channel. Thus, this study provides an effective approach to fabricate functional frameworks as fluorescent platforms for developing more fluorescent materials.

Co-delivery of NS1 and BMP2 mRNAs to murine pluripotent stem cells leads to enhanced BMP-2 expression and osteogenic differentiation.

Application of messenger RNA (mRNA) for bone regeneration is a promising alternative to DNA, recombinant proteins and peptides. However, exogenous in vitro transcribed mRNA (IVT mRNA) triggers innate immune response resulting in mRNA degradation and translation inhibition. Inspired by the ability of viral immune evasion proteins to inhibit host cell responses against viral RNA, we applied non-structural protein-1 (NS1) from Influenza A virus (A/Texas/36/1991) as an IVT mRNA enhancer. We evidenced a dose-dependent blocking of RNA sensors by NS1 expression. The co-delivery of NS1 mRNA with mRNA of reporter genes significantly increased the translation efficiency. Interestingly, unlike the use of nucleosides modification, NS1-mediated mRNA translation enhancement does not dependent to cell type. Dual delivery of NS1 mRNA and BMP-2 mRNA to murine pluripotent stem cells (C3H10T1/2), promoted osteogenic differentiation evidenced by enhanced expression of osteoblastic markers (e.g. alkaline phosphatase, type I collagen, osteopontin, and osteocalcin), and extracellular mineralization. Overall, these results support the adjuvant potentiality of NS1 for mRNA-based regenerative therapies. STATEMENT OF SIGNIFICANCE: mRNA therapy has the potential to improve the efficiency of nucleic acid based regenerative medicine. Up to now, the incorporation of expensive modified nucleotides is a common way to avoid IVT mRNA-induced detrimental immunogenicity. We here introduce co-delivery of Influenza virus immune evasion protein-NS1 coding mRNA as a strategy to suppress RNA sensors for maximizing IVT mRNA expression. An increased osteogenic commitment of pluripotent stem cells was observed after BMP2 mRNA and NS1 mRNA delivery. This study revealed how applying non-modified mRNA with NS1 could be a promising alternative as a therapeutic in bone regeneration.

Characterization of Odor-Active Volatiles and Odor Contribution Based on Binary Interaction Effects in Mango and Vodka Cocktail.

Thirty-six volatile compounds, composed of 18 esters, 10 terpenes, and 8 others, were detected by headspace-solid phase microextraction (HS-SPME) equipped with gas chromatography-mass spectrometry (GC-MS) in mango and vodka cocktail. Moreover, these compounds were detected by olfactometry using aroma intensities. Comparing these compounds revealed that the aroma intensities (AIs) of limonene, 3-carene, myrcene, ß-caryophyllene, and citronellyl propanoate were higher than others (AIs ≥ 4). In this context, limonene was selected as the reference compound on the basis of the strongest component model. The aim of this study was to determine the perceptual interaction between limonene and 3-carene, myrcene, ß-caryophyllene, citronellyl propanoate, respectively, in a binary mixture. In addition, feller's addition model revealed that limonene presented an addition effect when combined with 3-carene, myrcene, ß-caryophyllene, and citronellyl propanoate. It could be stated that these compounds played an important role in the aroma of mango and vodka cocktail. The results demonstrated that molecular structure and the ratio between compounds affected the synergistic effect, and compounds with similar structure and aroma were more prone to undergo addition and synergy.

Host-Guest Recognition and Fluorescence of a Tetraphenylethene-Based Octacationic Cage.

We report the synthesis and characterization of a three-dimensional tetraphenylethene-based octacationic cage that shows host-guest recognition of polycyclic aromatic hydrocarbons (e.g. coronene) in organic media and water-soluble dyes (e.g. sulforhodamine 101) in aqueous media through CH⋅⋅⋅π, π-π, and/or electrostatic interactions. The cage⊃coronene exhibits a cuboid internal cavity with a size of approximately 17.2×11.0×6.96 Å3 and a "hamburger"-type host-guest complex, which is hierarchically stacked into 1D nanotubes and a 3D supramolecular framework. The free cage possesses a similar cavity in the crystalline state. Furthermore, a host-guest complex formed between the octacationic cage and sulforhodamine 101 had a higher absolute quantum yield (ΦF =28.5 %), larger excitation-emission gap (Δλex-em =211 nm), and longer emission lifetime (τ=7.0 ns) as compared to the guest (ΦF =10.5 %; Δλex-em =11 nm; τ=4.9 ns), and purer emission (ΔλFWHM =38 nm) as compared to the host (ΔλFWHM =111 nm).

RNA-based therapy for osteogenesis.

Nucleic acid-based therapy has shown great promise in accelerating bone regeneration as well as other diseases. Nucleic acids used in gene therapy mainly are either plasmid DNA (pDNA) or RNAs. Although pDNA therapy has been extensively studied for decades with encouraging preclinical and clinical results, side effects, and low efficiency associated with nuclear trafficking are hard to bypass. Unlike pDNA, RNAs (mRNA, siRNA, miRNA) exert their function in the cytoplasm, thereby being more efficient in hard-to-transfect cells such as primary osteoblasts. RNA interference-based gene silencing represents a negative regulation which knockdown the expression of antagonists that impair osteogenesis process. In contrary, mRNA therapy for osteogenesis represents a positive regulation which delivers mRNA encoding growth factors to accelerate bone regeneration. This review presents a comprehensive summary of the mRNA and siRNA-based therapies and the targets for bone regeneration in case of bone defect and osteoporosis.

Characterization of ester odorants of apple juice by gas chromatography-olfactometry, quantitative measurements, odour threshold, aroma intensity and electronic nose.

In this study, twenty ester compounds of juiced apples from three different varieties were identified and quantified by gas chromatography-olfactometry (GC-O) and gas chromatography-mass spectrometry (GC-MS) analysis. Six esters were selected as the key odour-active compounds by omission Tests. Perceptual interaction among esters was investigated through odour thresholds and intensity of six esters and fifteen binary mixtures. Most of these mixtures followed an addition behavior. Ethyl butanoate and butyl acetate had a synergistic effect. Sensory analysis revealed that fruity, sour and green notes were enhanced significantly by adding the binary mixture of ethyl butanoate and butyl acetate (E&B). Electronic nose (e-nose) data showed that, compared with the individual ester, the response values of E&B were increased on four sensors. Moreover, with regard to partial least squares regression (PLSR), fruity, sour and green notes attribute covaried well with five sensors of e-nose. The present study would provide a theoretical basis for understanding the aroma characteristic of apple juice, hoping to be helpful for the improvement of apple juice aroma and flavour.

Diamondoid Frameworks via Supramolecular Coordination: Structural Characterization, Metallogel Formation, and Adsorption Study.

Supramolecular coordination has been developed as an efficient tool to construct a variety of discrete metallacycles and metallacages with well-defined shapes and sizes. However, its application in framework construction has been barely exploited. In this paper, we report the direct synthesis of two diamondoid frameworks from a simple tetrahedral precursor, tetra(4-(4-pyridinyl)phenyl)methane, and two linear difunctional platinum(II) ligands via one-step supramolecular coordination. Controlled by the specific angularity and geometry of the tetrahedral and linear subunits, these frameworks possess a well-defined diamondoid topology with highly regulated periodicity and three-dimensional porosity. Moreover, these rigid frameworks can be directly changed into a metallogel when prepared in DMSO at high concentrations. Interestingly, these diamondoid frameworks exhibit a cationic nature and stimuli-responsive behavior, which potentially endow them with the selective adsorption and controlled release for anionic dyes and drugs in aqueous environments. Thus, this study demonstrates that supramolecular coordination is a facile and efficient approach for the preparation of functional framework materials containing predesigned and well-defined supramolecular coordination assemblies as molecular skeletons.

Evaluation of the perceptual interaction among ester aroma compounds in cherry wines by GC-MS, GC-O, odor threshold and sensory analysis: An insight at the molecular level.

The ester aroma profiles of five Chinese cherry wines and the perceptual interaction among esters were investigated in this study. 24 esters were identified by Gas chromatography-olfactometry (GC-O) and Gas Chromatography-Mass Spectrometer (GC-MS). According to addition/omission analysis, seven volatile compounds among them were selected and studied using sensory profiling and multivariate statistic methods such as Principal Component Analysis (PCA). In sensory analysis, a significant reduction of olfactory threshold for total aromatic reconstitution was induced by the addition among each of them in Feller's additive model, which demonstrated their synergistic effects. The σ/τ plot showed that most of them were followed by a partial addition behavior. Furthermore, PCA indicated that the addition among each of them had a significant effect on fruity, floral, sweet and fermentation aroma intensity. Specifically, ethyl decanoate and methyl salicylate at sub-threshold concentrations were also likely to contribute to overall aroma. The results of perceptual interaction were mainly influenced by chemical structure and molecular polarity.

Olfactory impact of esters on rose essential oil floral alcohol aroma expression in model solution.

This study focused on the impact of esters on the perception of floral aroma in rose essential oil. Various aromatic reconstitutions were prepared, consisting of 10 alcohols and 9 esters, all the concentrations found in rose essential oil. Sensory analysis by the triangular tests revealed the interesting behavior of certain compounds among the 9 esters following their addition or omission. The results tend to highlight the important role of ethyl octanoate, ethyl tetradecanoate, citronellyl acetate, geranyl acetate, and 2-phenethyl acetate of esters in rose essential oil. The "olfactory threshold" (OT) of the 5 esters, the floral reconstitution and the mixtures of ester and floral reconstitution were evaluated in alkanes solution. Through the Feller's additive model analysis, it was found that the presence of ethyl octanoate, ethyl tetradecanoate, and citronellyl acetate led to a significant in decrease the OT of the mixtures, whereas geranyl acetate raised the OT. The floral reconstitution in alkanes solution was supplemented with the 5 esters at high, medium, and low concentration, then analyzed by quantitative descriptive analysis. It was revealed that ethyl octanoate, ethyl tetradecanoate, and citronellyl acetate adding overall aroma, and geranyl acetate masking the overall aroma perception in a model floral mixture. Sensory profiles highlighted changes in the perception of aroma nuances in the presence of the 5 esters, with specific perceptive interactions, and reported on the graph based on two parameters [σ = f(τ)]. This paper provided a reference for the flavourists.

Olfactory Impact of Terpene Alcohol on Terpenes Aroma Expression in Chrysanthemum Essential Oils.

The key point of our work was evaluating the impact of terpene alcohols on the aroma expression of terpenes recombination in Chrysanthemum essential oils. Using pure commercial products, various aromatic recombinations were prepared, consisting of terpenes recombination and six terpene alcohols, all the concentrations found in Chrysanthemum essential oils. There were five groups of terpene alcohols mixtures performed very interesting with the addition or omission tests. The "olfactory threshold" of the terpenes recombination had a notable decrease when adding isoborneol, d-Fenchyl alcohol respectively through the Feller's additive model analysis. Furthermore, the descriptive test indicated that the addition of terpene alcohols mixture had the different effect on fruity, floral, woody, green, and herbal aroma intensity. Specifically, when isoborneol was added to the terpenes recombination in squalane solution, it was revealed that isoborneol had a synergy impact on herbal and green notes of the terpenes recombination and masked the fruity note.

Diamondoid Supramolecular Coordination Frameworks from Discrete Adamantanoid Platinum(II) Cages.

Recently, porous framework materials with various network-type structures have been constructed via several different approaches, such as coordination interactions, reversible covalent bonds, and non-covalent interactions. Here, we have combined the concepts of supramolecular coordination complex (SCC) and metal-organic framework to offer a new strategy to construct a diamondoid supramolecular coordination framework (SCF) from an adamantanoid supramolecular coordination cage as the tetrahedral node and a difunctional Pt(II) ligand as the linear linker via stepwise orientation-induced supramolecular coordination. The adamantanoid supramolecular coordination cage has four uncoordinated pyridyl groups, which serve as the four vertexes of the tetrahedral geometry in the diamondoid framework. As a result, this diamondoid SCF exhibits an adamantanoid-to-adamantanoid substructure with two sets of pores, including the interior cavity of the adamantanoid cage and the extended adamantanoid space between the individual cages in the framework. In addition, the shape-controllable and highly ordered self-assembly of nanometer-sized diamondoid SCF is observed as micrometer-sized regular octahedrons by evaporation under heating in DMSO. This study demonstrates the potential application of supramolecular coordination complexes in the precise construction of highly regulated porous framework materials.