Fluorescence Experiments Research Articles

Topologically Driven Pore/Surface Engineering in a Recyclable Microporous Metal-Organic Vessel Decorated with Hydrogen-Bond Acceptors for Solvent-Free Heterogeneous Catalysis.

The use of metal-organic frameworks (MOFs) comprising custom-designed linkers/ligands as efficient and recyclable heterogeneous catalysts is on the rise. However, the topologically driven bifunctional porous MOFs for showcasing a synergistic effect of two distinct activation pathways of substrates (e.g., involving hydrogen bonding and a Lewis acid) in multicomponent organic transformations are very challenging. In particular, the novelty of such studies lies in the proper pore and/or surface engineering in MOFs for bringing the substrates in close proximity to understand the mechanistic aspects at the molecular level. This work represents the topological design, solid-state structural characterization, and catalytic behavior of an oxadiazole tetracarboxylate-based microporous three-dimensional (3D) metal-organic framework (MOF), {[Zn2(oxdia)(4,4'-bpy)2]·8.5H2O}n (1), where the tetrapodal (4-connected) 5,5'-(1,3,4-oxadiazole-2,5-diyl)diisophthalate (oxdia4-), the tetrahedral metal vertex (Zn(II)), and a 2-connected pillar linker 4,4'-bipyridine (4,4'-bpy) are unique in their roles for the formation, stability, and function. As a proof of concept, the efficient utilization of both the oxadiazole moiety with an ability to provide H-bond acceptors and the coordinatively unsaturated Zn(II) centers in 1 is demonstrated for the catalytic process of the one-pot multicomponent Biginelli reaction under mild conditions and without a solvent. The key steps of substrate binding with the oxadiazole moiety are ascertained by a fluorescence experiment, demonstrating a decrease or increase in the emission intensity upon interaction with the substrates. Furthermore, the inherent polarizability of the oxadiazole moiety is exploited for CO2 capture and its size-selective chemical fixation to cyclic carbonates at room temperature and under solvent-free conditions. For both catalytic processes, the chemical stability, structural integrity, heterogeneity, versatility in terms of substrate scope, and mechanistic insights are discussed. Interestingly, the first catalytic process occurs on the surface, while the second reaction occurs inside the pore. This study opens new ways to catalyze different organic transformation reactions by utilizing this docking strategy to bring the multiple components close together by a microporous MOF.

Dicyanomethylene-4H-pyran based Schiff base for turn on NIR fluorescence sensing of Fe3+, Al3+ and Cr3+ and its application in molecular logic gate

A novel Schiff base derivative HMA containing dicyanomethylene-4H-pyran unit was designed and synthesized. It can be used to detect Fe3+, Al3+, Cr3+ accompanied with near-infrared (NIR) fluorescence enhancement. Fluorescence experiments show that it has a large Stokes shift (λex = 480 nm, λem = 670 nm), low detection limit and large binding constant. The interaction between HMA and Fe3+, Al3+, Cr3+ was investigated by NMR, FT-IR, as well as density functional theoretical calculations. It shows that the emission of HMA experiences an excited state intramolecular proton transfer (ESIPT) process, while the addition of Fe3+, Al3+ and Cr3+causes their coordination, increases the rigidity of HMA and enhances the fluorescence. Additionally, the reversible experiment was carried out by adding F− ion. For Al3+ and Cr3+, the corresponding solutions can return to their original state but not for Fe3+. On the basis of the reversible experiment, a molecular logic gate was designed with Al3+/Cr3+ and F− as input and the fluorescence signal at 670 nm of HMA as output.

Effect of silica nanoparticles on cell membrane fluidity: The role of temperature and membrane composition

The increasing production and application of silica nanoparticles (SiO2 NPs) raise public concern regarding their environmental and health risks. The fluidity of the cell membrane is essential for supporting membrane proteins and regulating membrane transport. Changes in membrane fluidity inevitably influence the physiological activities of cells and even cause biological effects. In this study, the effect of SiO2 NPs on membrane fluidity was studied at 25 °C and 37 °C, and the role of membrane components in SiO2 NP-membrane interactions was investigated using giant plasma membrane vesicles (GPMVs) isolated from RBL-2H3 cells. SiO2 NPs cause a more serious membrane fluidity decrease at 37 °C than at 25 °C, which is revealed by the shift of Laurdan fluorescence emission and further quantified via forster resonance energy transfer (FRET) experiments. In addition, after the removal of 75 % cholesterol from the membrane, SiO2 NPs caused a greater extent of membrane gelation. These results indicate that SiO2 NPs prefer to interact with membranes that are more dynamic and less densely packed. Moreover, fluorescent experiments confirmed that the existence of phosphatidyl ethanolamine (PE) and phosphoinositide (PI) can mitigate NP-induced membrane gelation. Molecular dynamics simulation further demonstrated that SiO2 NPs form hydrogen bonds with the terminal of PE or PI but with the -PO4−- group in the middle of phosphatidylcholine (PC). The bonding that occurs in the terminal gives less restriction of phospholipid movement and a weaker effect on membrane fluidity. This research provides both evidence and mechanisms of SiO2 NP-induced membrane fluidity changes, which are helpful for understanding cell membrane damage and the biological effects of NPs.

A new zinc-organic framework with 1D channel for constructing a ratiometric Al3+-selective sensor and four inputs INHIBIT logic gate

To develop Al3+ fluorescent sensor is significant because the abnormal levels of Al3+ in environment may pose great threat to human body. Herein, a novel metal–organic framework {Zn(Dpada)(Imdba)·H2O}n (Dpada = 3, 6-di(1H-imidazol-1-yl) pyridazine and Imdba = 2, 2′-iminodibenzoic acid), named Zn-MOF, has been architected with one-dimensional channel under hydrothermal conditions. Zn-MOF exhibits good thermal and solvent stability and can also keep structural integrity over the pH range of 5.0 – 9.0. Fluorescent experiments show that Zn-MOF has high selectivity and sensitivity towards Al3+ via ratiometric fluorescence signal changes (F470 nm/F390 nm) and the detection limit is evaluated to be 0.69 μM. In addition, Zn-MOF performs good recyclability in sensing of Al3+ with at least 5 cycles. Besides, an INHIBIT logic gate has been constructed with chemical ions (Al3+, Cr3+, Fe3+ and Hg2+) as input signals and emission ratio (F470 nm/F390 nm) as output signal. Significantly, Zn-MOF can be applied to tracing Al3+ using real water samples, presenting great potential in water quality monitoring application.

Microscope alignment using real-time Imaging FCS

Modern electron-multiplying charge-coupled device (EMCCD) and scientific complementary metal-oxide semiconductor (sCMOS) cameras read out fluorescence data with single-molecule sensitivity at thousands of frames per second. Exploiting these capabilities in full requires data evaluation in real time. The direct camera-read-out tool presented here allows access to the data while the camera is recording. This provides simplified and accurate alignment procedures for total internal reflection fluorescence microscopy (TIRFM) and single-plane illumination microscopy (SPIM), and simplifies and accelerates fluorescence experiments. The tool handles a range of widely used EMCCD and sCMOS cameras and uses imaging fluorescence correlation spectroscopy for its evaluation. It is easily extendable to other camera models and other techniques and is a base for automated TIRFM and SPIM data acquisition.

Preparation and Application of Novel Mesoporous Silica Microsphere-Based Thin Layer Chromatography Plate

With excellent mechanical properties and high specific surface area, mesoporous silica microspheres are widely applied in various fields. However, mesoporous silica has been relatively studied as a thin-layer chromatographic (TCL) material for preliminary separation and detection. In this study, we obtained mesoporous silica microspheres with different particle sizes and apertures by a simple synthesis method, and prepare mesoporous silica microspheres-based TCL. Excellent test results have been exhibited, the 4.5 μm mesoporous silica microspheres-based TCL exhibits better separation results in dye and fluorescent polymer separation experiments, thus expanding the application of mesoporous silica microspheres.

MG53 preserves mitochondrial integrity of cardiomyocytes during ischemia reperfusion-induced oxidative stress

Ischemic injury to the heart induces mitochondrial dysfunction due to increasing oxidative stress. MG53, also known as TRIM72, is highly expressed in striated muscle, is secreted as a myokine after exercise, and is essential for repairing damaged plasma membrane of many tissues by interacting with the membrane lipid phosphatidylserine (PS). We hypothesized MG53 could preserve mitochondrial integrity after an ischemic event by binding to the mitochondrial-specific lipid, cardiolipin (CL), for mitochondria protection to prevent mitophagy. Fluorescent imaging and Western blotting experiments showed recombinant human MG53 (rhMG53) translocated to the mitochondria after ischemic injury in vivo and in vitro. Fluorescent imaging indicated rhMG53 treatment reduced superoxide generation in ex vivo and in vitro models. Lipid-binding assay indicated MG53 binds to CL. Transfecting cardiomyocytes with the mitochondria-targeted mt-mKeima showed inhibition of mitophagy after MG53 treatment. Overall, we show that rhMG53 treatment may preserve cardiac function by preserving mitochondria in cardiomyocytes. These findings suggest MG53's interactions with mitochondria could be an attractive avenue for developing MG53 as a targeted protein therapy for cardioprotection.

Effect of Fatty Acids and Uremic Toxins on the Binding of Nateglinide, an Insulin Secretagogue, to Site II on Human Serum Albumin.

Nateglinide (NAT) is used to treat diabetes, stimulating pancreatic islet β-cells with residual insulin secretory capacity to increase insulin secretion. NAT has been reported to bind to human serum albumin (HSA), but the detail is still unclear. In the current study, we investigated the location and the affinity for the binding of NAT to HSA. Quantitative analysis data from the ultrafiltration experiment indicated that NAT binds strongly to a primary site on HSA with a high affinity. The presence of diazepam (DZP) or ibuprofen (IB), the specific site II ligands of HSA, decreased the binding constants of NAT respectively, without the significant changes in the number of binding sites. Whereas warfarin (WF), a site I specific ligand, did not affect the binding of NAT. Fluorescent replacement experiment showed that NAT replaced dansylsarcosine (DNSS), a site II probe of HSA, but not WF. An increasing level of myristate and uremic toxins, indoxyl sulphate (IS), indoxyl acetate (IA) and p-cresyl sulphate (PCS), during renal disease significantly increased the concentration of unbound NAT. These findings suggest that NAT specifically binds to site II of HSA and the binding capacity and pharmacokinetics of NAT change in renal diseases.

Synthesis and Ultrafast Broadband Optical Limiting Properties of a Two-Branched Twistacene

A novel two-branched twistacene (PyDN) has been designed and synthesized for application on ultrafast optical limiting. This twistacene exhibits excellent two photon absorption and two photon absorption-induced excited singlet state absorption, which was systematically investigated with a femtosecond Z-scan experiment, transient absorption spectrum, and two-photon excited fluorescence experiments. The admirable two photon absorption is attributed to the high degree of π electron delocalization in twistacene which is caused by introduction of two strong donors. The excited singlet state absorption cooperates with two-photon absorption to provide an excellent ultrafast optical limiting behavior with high linear transmittance, where the thresholds are 2.3–5.3 mJ/cm2 in the spectral region of 532–800 nm of femtosecond laser and 133 mJ/cm2 for picosecond pulse at 532 nm. These thresholds are lower than that of most of the optical limiters reported previously, which indicates PyDN is a promising candidate for ultrafast optical limiting.

The miR-200 Family Targeting amh Affects the Gonadal Development of Japanese Flounder

Four members of the miR-200 family in Japanese flounder (Paralichthys olivaceus) have sex-biased expression patterns, but their target genes and how they work in the development of the gonads are rarely known. Anti-Müllerian hormone (AMH) can inhibit the development of Muller’s duct in female mammals and regulate the formation of gametes after sexual maturity. There is no Muller’s duct in teleosts, but the amh gene still exists. Knockout of amh results in sex reversal from male to female. Therefore, it is essential to explore the relationship between the miR-200 family and amh to clarify what role miR-200 plays in the development of the gonads. In Japanese flounder, the two binding sites for the miR-200 family in the 3′UTR of amh were found through bioinformatic prediction. Double luciferase and green fluorescent protein reporter experiments demonstrated amh to be directly targeted by miR-200a and miR-200b. Moreover, miR-200a and miR-200b reduced the expression of amh through site 1 rather than site 2. To explore the regulatory role of miR-200a in gonadal development, we further overexpressed miR-200a in the primary Sertoli cells of the testis. With the overexpression of miR-200a, the expression of amh decreased, while the expression of the other two male sex-related genes, dmrt1 (doublesex and mab-3 related transcription factor 1) and gsdf (diagonal soma driven factor), increased significantly. This result indicates that the miR-200 family regulates the gonadal differentiation and development by targeting amh in Japanese flounder.

Coumarin-based fluorescent probes toward viscosity in mitochondrion/lysosome

Viscosity is an important microenvironmental indicator that plays an important role in the process of information transmission in various regions. Herein, two coumarin-based viscosity-sensitive fluorescent probes (CHB, CHN) were synthesized and the photophysical properties of the two probes were studied. The fluorescence quantum yields of CHB and CHN in glycerol can be as high as 25.2% and 18.3% respectively. The two probes can linearly detect viscosity in the viscosity logarithm range of 0.83–2.07, which is not interfered with pH, metal ions, anions and biomolecules. Fluorescent confocal cell experiments show CHB and CHN have good targeting ability to mitochondrion, lysosome, Endoplasmic reticulum and Golgi apparatus, and can be used to detect viscosity in mitochondrion/lysosome.

LcCCL28-25, Derived from Piscine Chemokine, Exhibits Antimicrobial Activity against Gram-Negative and Gram-Positive Bacteria In Vitro and In Vivo.

ABSTRACTAntimicrobial peptides (AMPs) are currently recognized as potentially promising antibiotic substitutes. Fish are an important seawater/freshwater medicinal biological resource, and the antimicrobial peptides and proteins that are key components of their innate immune systems are potential candidates for the development of novel antibacterial agents. The rainbow trout Oncorhynchus mykiss chemokine CK11 (omCK11), classified in the C-C motif chemokine ligand 27/28 (CCL27/28) family, is the only CC-type chemokine reported to play a direct antibacterial role in the immune response; however, its antibacterial domain remains unknown. In this study, we analyzed the structure-activity relationship of omCK11 and identified the antibacterial C-terminal domain. Additionally, we performed structure-function analyses of CCL27/28 proteins from different, representative freshwater and seawater fishes, revealing their shared C-terminal antibacterial domains. Surprisingly, a synthesized cationic peptide (named lcCCL28-25), derived from the large yellow croaker Larimichthys crocea CCL28, exhibited broad-spectrum and the most acceptable bactericidal activity, as well as antibiofilm activity and negligible hemolytic and cytotoxic activity in vitro. Additionally, lcCCL28-25 conferred a protective effect in the thighs of neutropenic mice infected with Staphylococcus aureus. SYTOX green fluorescence and electron microscopy experiments revealed that lcCCL28-25 was capable of rapidly destroying the integrity and permeability of the bacterial cell membrane. Overall, this study aided in the advancement of antibacterial CC-type chemokine research and also suggested a new strategy for exploring novel AMPs. Additionally, the efficacy of lcCCL28-25 in in vivo antibacterial activity in a mammalian model revealed that this compound could be a promising agent for the development of peptide-based antibacterial therapeutics.IMPORTANCE The primary function of chemokines has been described as recruiting and activating leukocytes to participate in the immune response. Some chemokines are also broad-spectrum antibacterial proteins in mammals. The Oncorhynchus mykiss chemokine CK11 (omCK11) is the first reported and currently the only CC-type antibacterial chemokine. The present study identified the antibacterial domain of omCK11. Structure-function analysis of various fish CCL27/28 proteins identified a novel antibacterial peptide (lcCCL28-25) from Larimichthys crocea CCL28 that exhibited broad-spectrum and the most acceptable bactericidal activity in vitro, as well as a protective effect in a Staphylococcus aureus infection mouse model. The antibacterial mechanisms included membrane disruption and permeation. This study advanced the field of antibacterial chemokine research in fish and also suggested a new strategy for exploring novel AMPs. The novel peptide lcCCL28-25 may prove to be an effective antibacterial agent.

Sus_circPAPPA2 Regulates Fat Deposition in Castrated Pigs through the miR-2366/GK Pathway.

CircRNAs play an important role in fat deposition, and testosterone-deficient boars exhibit significantly increased fat deposition; however, the mechanism by which testosterone regulates fat deposition through circRNAs remains unclear. In this study, circRNA-seq of backfat and abdominal fat from castrated and intact full-sib Yorkshire pigs was performed. The GO and KEGG enrichment analyses revealed that the host genes of the dorsal DE circRNAs were mainly involved in fatty acid transport, while in abdominal tissues, these genes were mainly involved in adipogenesis and inflammation. The interaction among sus_circPAPPA2, ssc-miR-2366 and GK was verified by dual fluorescence experiments and in porcine preadipocytes. The overexpression of sus_circPAPPA2 significantly inhibited the differentiation of preadipocytes. The expression of sus_circPAPPA2 was increased after adding 100 nM of testosterone, and preadipocyte differentiation was significantly inhibited. Testosterone can affect preadipocyte differentiation by upregulating the expression of sus_circPAPPA2, sponging miR-2366 and regulating the expression of genes, such as GK. These results indicate that testosterone can regulate the expression of adipocyte differentiation- and lipid metabolism-related genes by regulating the expression of circRNA, and ceRNA networks are different in the testosterone regulation of adipose deposition in different parts. This study provides basic data enhancing the understanding of the interaction between the hormone environment and mir-2366/GK to regulate trait performance in pigs.

Synthesis, Structure, and Properties of Complexes Based on 2,4-Bis-(triazol-1-yl)benzoic Acid as a Ligand.

Using 2,4-bis-(triazol-1-yl)-benzoic acid as the main ligand and terephthalic acid (TPA) as the auxiliary ligand, combined with Cd(NO3)2·4H2O and Zn(NO3)2·6H2O, self-assembly under solvothermal conditions gave three novel complexes: [Cd0.5(L)(H2O)] (1), [Cd(L)(TPA)0.5(H2O)]·H2O (2), and [Zn(L)(TPA)0.5]·H2O (3) (TPA = terephthalic acid). The crystal structure test showed that complex 1 belongs to the triclinic crystal system and the P1̅ space group and complexes 2 and 3 belong to the monoclinic crystal system and the P21/c space group. Solid-state fluorescence experiments show that complexes 1, 2, and 3 all have excellent optical properties: among them, complexes 1 and 3 can selectively detect MnO4– with low detection limits (0.96 μM and 0.232 μM, respectively) and complex 2 can detect Cr2O72– [limit of detection (LOD) = 0.035 μM], and the most interesting thing is that all three complexes can be used as sensors for detecting Fe3+ (LOD = 0.76 μM, 0.657 μM, and 0.11 μM, respectively). In addition, the detection capabilities of these three complexes for different amino acids and antibiotics were also analyzed, and the results showed that all three complexes can effectively detect tetracycline hydrochloride through the quenching effect and 2 and 3 can selectively detect tryptophan via the fluorescence enhancement effect.

Calculating off-axis efficiency of coaxial HPGe detectors by Monte Carlo simulation

In beam geometries where a directed γ-ray beam hits the surface of a coaxial high purity germanium detector (HPGe), the detector efficiency is sensitive to the position where γ-rays initially hit the detector surface because the structure of the detector is nonuniform. This may cause inaccuracy of the detector efficiency when measured using standard sources that are point-like sources emitting γ-rays isotropically. Obtaining a precise estimation of the full energy peak efficiency of the coaxial HPGe detector in the beam geometry for on-axis and off-axis measurements requires a Monte Carlo simulation. We performed Monte Carlo simulations that calculate the detector efficiency in the beam geometry. The effects of the off-axis distance and γ-ray beam size on the efficiency are quantitatively analyzed. We found that the intrinsic efficiency in the beam geometry is maximized when the beam hits the detector at specific off-axis distances. Our Monte Carlo calculations have been supported by nuclear resonance fluorescence experiments using laser Compton scattering γ-ray beams.

On the physical nature of the turbulent/turbulent interface

The existence of a turbulent/turbulent interface (TTI) has recently been verified in the far wake of a circular cylinder exposed to free-stream turbulence (Kankanwadi & Buxton, J. Fluid Mech., vol. 905, 2020, p. A35). This study aims to understand the physics within the TTI. The wake boundary, approximately 40 diameters downstream of a circular cylinder subjected to grid-generated turbulence, was investigated through simultaneous cinematographic, stereoscopic particle image velocimetry and planar laser induced fluorescence experiments. With no grid placed upstream of the cylinder, the behaviour of the resultant interface, our closest approximation to a turbulent/non-turbulent interface, exactly matched what is observed in existing literature. When background turbulence is present, viscous action is no longer the only method by which enstrophy is transported to the background fluid, unlike for turbulent/non-turbulent interfaces. The presence of rotational fluid on both sides of the TTI allows the vorticity stretching term of the enstrophy budget equation to be the dominant actor in this process. The role of viscosity within a TTI is greatly diminished as the vorticity stretching term takes over responsibilities for enstrophy production. The turbulent strain rate normal to the TTI was found to be enhanced in the interfacial region. Decomposing the vorticity stretching term into components aligned with the three principal strain-rate directions, it was found that the term most aligned with the interface-normal direction contributed to the largest share of enstrophy production. This indicates that better ‘organised’ vorticity on the wake side of the interface yields the enstrophy amplification leading to the previously discovered enstrophy jump across the TTI by Kankanwadi & Buxton (J. Fluid Mech., vol. 905, 2020, p. A35).

A novel amino functionalized three-dimensional fluorescent Zn-MOF: The synthesis, structure and applications in the fluorescent sensing of organic water pollutants

An amino functionalized three-dimensional (3D) Zn-MOF with molecular formula Zn(NH2bdc)(mbib) (complex 1) has been synthesized by using solvothermal method. The structure, purity, stability and morphology of complex 1 was fully characterized with single-crystal X-ray diffraction, PXRD, FT-IR, TGA, TEM, et al. The characterization results revealed that complex 1 crystallizes in the orthorhombic space group Pna21. In the structure of complex 1, each NH2bdc2− ligand connected to two Zn(II) to form a 1D zigzag chain which was further connected by the mbib ligands to give a fourfold interpenetration 3D framework. The single-crystal X-ray diffraction analysis indicated that the 3D framework of complex 1 was functionalized with free amino groups both on the surface and in the channels. Characteristic fluorescent emission centered at 430 nm which could be remarkably enhanced in water was observed, indicating complex 1 might be a promising candidate for the fluorescent sensing of water pollutants. The functionalized amino groups remarkably enhanced the selectivity and sensitivity of complex 1. Gradually fluorescence quenching of complex 1 was recorded in the fluorescent titration experiments with good quantitative relation after addition of acetone or picric acid in water samples. The experimental detection limits for acetone and picric acid were 0.03% (%V) and 340 nM, respectively. Plausible mechanisms for the fluorescent quenching sensing of acetone and picric acid were proposed.

Cytotoxicity of Methacrylate Dental Resins to Human Gingival Fibroblasts.

This study aimed to assess the acute and delayed cytotoxicity of three, popular light-cured methacrylate-based restorative resins (MRs): Charisma (C), Estelite (E), and Filtek (F), to human gingival fibroblasts in culture. Cells were grown for up to 24 h with light-cured (or pre-cured) resins. We evaluated resin cytotoxicity, redox imbalance, necrosis/apoptosis, miR-9, and heat shock protein 70 (HSP70). The role of resin-induced oxidative stress (damage) in HSP70-response (repair) was assessed using binary fluorescence labeling. All MRs decreased viable cell numbers and cell proliferation and damaged cell membranes, and their 24 h-delayed toxicity was lower (C), higher (F), or similar (E) to that induced by freshly-cured resins. Cell membrane damage induced by C and E decreased with time, while F produced a linear increase. All resins generated intracellular oxidative stress with the predominant necrotic outcome, and produced heterogeneous responses in miR-9 and HSP70. The double fluorescence (damage/repair) experiments pointed to common features of E and F but not C. In the subset of cells, the binary response induced by E and F was different from C, similar to each other, and positively interrelated. Experimental data show that selective MR cytotoxicity should be taken into account when considering repetitive use or massive reconstruction.

Reducing Endogenous Labile Zn May Help to Reduce Smooth Muscle Cell Injury around Vascular Stents.

Vascular stent service involves complex service environments and performance requirements, among which the histocompatibility of the stent could seriously affect the therapeutic effect. In the pathology of vascular disease, the thin fiber cap is easily ruptured, exposing the necrotic core below, and triggering a series of dangerous biochemical reactions. In contrast, the thin neointima, considered an essential structure growing on the stent, may evolve into vulnerable plaque structures due to lesions induced by the stent. Therefore, the reduction of necrosis around the stent below the thin neointima is indispensable. In this work, different cell model experiments suggested that the content of endogenous labile Zn positively correlated with cell injury. Zinquin-Zn fluorescence experiments and zinc ion channels research suggested that the change in the content of endogenous labile Zn in smooth muscle cells is affected by different stent coatings. The content of endogenous labile Zn in cells negatively correlated with cell viability. Animal experiments indirectly verified the increase in endogenous labile Zn by detecting the expression of Zn regulatory protein (metallothionein) in the necrotic tissues. Reducing the content of endogenous labile Zn may favor a reduction in smooth muscle cell injury and necrosis. This biochemical mechanism is effective in improving the therapeutic effect of vascular stents.

Porous Polymers as Universal Reversal Agents for Heparin Anticoagulants through an Inclusion-Sequestration Mechanism.

Heparins are widely used anticoagulants for surgical procedures and extracorporeal therapies. However, all of them have bleeding risks. Protamine sulfate, the only clinically approved antidote for unfractionated heparin (UFH), has adverse effects. Moreover, protamine can only partially neutralize low-molecular-weight heparins (LMWHs) and is not effective for fondaparinux. Here, an inclusion-sequestration strategy for efficient neutralization of heparin anticoagulants by cationic porous supramolecular organic frameworks (SOFs) and porous organic polymers (POPs) is reported. Isothermal titration calorimetric and fluorescence experiments show strong binding affinities of these porous polymers toward heparins, whereas dynamic light scattering and zeta potential analysis confirm that the heparin sequences are adsorbed into the interior of the porous hosts. Activated partial thromboplastin time, anti-FXa, and thromboelastography assays indicate that their neutralization efficacies are higher than or as high as that of protamine for UFH and generally superior to protamine for LMWHs and fondaparinux, which is further confirmed by tail-transection model in mice and ex vivo aPTT or anti-FXa analysis in rats. Acute toxicity evaluations reveal that one of the SOFs displays outstanding biocompatibility. This work suggests that porous polymers can supply safe and rapid reversal of clinically used heparins, as protamine surrogates, providing an improved approach for their neutralization.