Clinical outcomes of guided tissue regeneration with carbonate apatite granules and poly(lactic acid/caprolactone) membrane for the treatment of intrabony defects and mandibular Class II furcation involvements: A 12-month prospective pilot clinical study.

Introduction: For deep intrabony defects or Class II furcation involvements (FI), periodontal tissue regenerative therapy combined with bone graft materials and a barrier membrane is recommended. The objective of this study was to assess the safety and efficacy of using carbonate apatite (CO3Ap) granules and absorbable poly(lactic acid/caprolactone) (PLCL) membranes for periodontal regeneration in the treatment of intrabony defects and mandibular Class II FI. Methods: This prospective pilot clinical study, conducted at a single center with a single-arm design, aimed to assess the safety and efficacy of CO3Ap and PLCL membranes in patients with periodontitis. A total of 9 patients with 10 teeth, including seven deep intrabony defects and three Class II FI, were treated with CO3Ap granules and PLCL membranes. Clinical parameters such as probing pocket depth (PPD), clinical attachment level (CAL), bleeding on probing (BOP), tooth mobility (Mo), Plaque Index (PI), and Gingival Index (GI) were assessed at baseline, 6 and 12 months post-surgery. Radiographic analysis was performed using dental X-rays and cone beam computed tomography (CBCT) images taken at baseline, 6, and 12 months post-surgery. Results: Postoperative healing was uneventful in most of the cases. In some cases, membrane exposures were observed. However, there were no signs of inflammation, such as abnormal bleeding, pain, swelling, or pus. These exposures eventually healed well in the end. The mean reductions in PPD at 6 and 12 months were 4.5 ± 1.6 mm and 4.9 ± 1.4 mm, respectively, while the mean gains in CAL were 4.4 ± 1.7 mm at 6 months and 4.6 ± 1.2 mm at 12 months. Radiographic analysis showed improvements in linear bone height within intrabony defects and in the vertical subclassification of FI in Class II FI. Conclusions: Despite the limitations of this study, periodontal regenerative therapy using CO3Ap granules and a PLCL membrane demonstrated promising clinical safety and efficacy for treating intrabony defects and mandibular Class II furcation involvement.

Understanding Excipient-Induced Crystallization of Spray-Dried Amorphous Solid Dispersion.

This study investigates the compatibility of excipients with the model system SDI-X and their role in the induced crystallization of the amorphous compound-X in tablet formulations. We aimed to establish a straightforward and practical screening approach for evaluating excipient-induced crystallization of SDI in tablet matrices. Three methodologies-binary powder mixture, binary compact, and bilayer tablets-were employed to qualitatively and quantitatively evaluate the recrystallization of SDI-X with various excipients under accelerated storage conditions. The results demonstrated that binary compacts, providing direct physical contact between SDI-X and excipients, are superior in reflecting realistic drug-excipient contact within pharmaceutical tablets, enabling a more accurate assessment of excipient-induced crystallization for SDI-X. In contrast, the broadly used conventional binary blends can significantly underestimate this risk due to insufficient proximity. In addition, the bilayer tablets further confirmed that crystallization initiates at the contact surface between SDI-X and the excipients. The study highlighted that not only hygroscopicity but also the type of excipient and its physical contact with SDI-X significantly influence the recrystallization extent and rate of SDI-X. Interestingly, less hygroscopic diluents such as mannitol and lactose induced much higher levels of crystallization of SDIs, contrary to expectations based on moisture content alone. This suggests that the excipient type and contact surface are more critical in inducing recrystallization than just the level of moisture. The findings emphasize the need for careful excipient selection, study design, and sample preparation to enable appropriate assessments of SDI-excipient compatibility.

Development and characterization of bilayer chitosan/alginate cling film reinforced with essential oil based nanocomposite for red meat preservation.

With a goal to finding suitable alternatives to plastic packaging in the food industry, we developed a multifunctional bio-based active packaging film to enhance the shelf life of red meat. A chitosan/alginate (Chi + Alg) bilayer film was developed through layer-by-layer (LBL) assembly and an active material i.e. lemongrass nanoemulsion with silver nanoparticles-based nanocomposite (NC1) was loaded into the alginate layer to improve the quality of the bio-based film (Chi + Alg + NC1). The Chi + Alg + NC1 film was characterized in terms of its microstructure, mechanical strength, thermal stability, and antimicrobial activity. Scanning electron microscopy (SEM) revealed a film (22.5 ± 1.44 µm thickness) with a smooth and even surface and a cross-sectional structure. The incorporation of NC1 improves the quality of the film by enhancing its mechanical strength and thermal stability. FT-IR spectra showed the successful interaction between chitosan and alginate in the LBL assembly and the incorporation of NC1 in the alginate layer. The red meat preservation test demonstrated that the shelf life improved when the meat was covered with the fabricated bio-based film. The color of the meat was retained for up to 7 days compared to that of the control (Chi alone and Chi + Alg). Additionally, a reduction in the microbial count in the Chi + Alg + NC1 film was observed, corroborating the shelf-life improvement. In addition to its inherent antimicrobial properties, NC1 induced hydrophilic properties to the film, which further aids in its antimicrobial activity against E. coli. These findings suggest that Chi + Alg + NC1 film could be a potent alternative to plastic packaging and can be used as a cling film to prolong the shelf life of red meat.

Engineering Phosphatidylserine Containing Asymmetric Giant Unilamellar Vesicles.

The plasma membrane lipid distribution is asymmetric, with several anionic lipid species located in its inner leaflet. Among these, phosphatidylserine (PS) plays a crucial role in various important physiological functions. Over the last decade several methods have been developed that allow for the fabrication of large or giant unilamellar vesicles (GUVs) with an asymmetric lipid composition. Investigating the physicochemical properties of PS in such asymmetric lipid bilayers and studying its interactions with proteins necessitates the reliable fabrication of asymmetric GUVs (aGUVs) with a high degree of asymmetry that exhibit PS in the outer leaflet so that the interaction with peptides and proteins can be studied. Despite progress, achieving aGUVs with well-defined PS asymmetry remains challenging. Recently, a Ca2+-initiated hemifusion method has been introduced, utilizing the fusion of symmetric GUVs (sGUVs) with a supported lipid bilayer (SLB) for the fabrication of aGUVs. We extend this approach to create aGUVs with PS in the outer bilayer leaflet. Comparing the degree of asymmetry between aGUVs obtained via Ca2+ or Mg2+ initiated hemifusion of a phosphatidylcholine (PC) sGUVwith a PC/PS-supported lipid bilayer, we observe for both bivalent cations a significant number of aGUVs with near-complete asymmetry. The degree of asymmetry distribution is narrower for physiological salt conditions than at lower ionic strengths. While Ca2+ clusters PS in the SLB, macroscopic domain formation is absent in the presence of Mg2+. However, the clustering of PS upon the addition of Ca2+ is apparently too slow to have a negative effect on the quality of the obtained aGUVs. We introduce a data filtering method to select aGUVs that are best suited for further investigation.

HIV-1 Gag Polyprotein Affinity to the Lipid Membrane Is Independent of Its Surface Charge.

The binding of the HIV-1 Gag polyprotein to the plasma membrane is a critical step in viral replication. The association with membranes depends on the lipid composition, but its mechanisms remain unclear. Here, we report the binding of non-myristoylated Gag to lipid membranes of different lipid compositions to dissect the influence of each component. We tested the contribution of phosphatidylserine, PI(4,5)P2, and cholesterol to membrane charge density and Gag affinity to membranes. Taking into account the influence of the membrane surface potential, we quantitatively characterized the adsorption of the protein onto model lipid membranes. The obtained Gag binding constants appeared to be the same regardless of the membrane charge. Furthermore, Gag adsorbed on uncharged membranes, suggesting a contribution of hydrophobic forces to the protein-lipid interaction. Charge-charge interactions resulted in an increase in protein concentration near the membrane surface. Lipid-specific interactions were observed in the presence of cholesterol, resulting in a two-fold increase in binding constants. The combination of cholesterol with PI(4,5)P2 showed cooperative effects on protein adsorption. Thus, we suggest that the affinity of Gag to lipid membranes results from a combination of electrostatic attraction to acidic lipids, providing different protein concentrations near the membrane surface, and specific hydrophobic interactions.

Lipid Selectivity of Membrane Action of the Fragments of Fusion Peptides of Marburg and Ebola Viruses.

The life cycle of Ebola and Marburg viruses includes a step of the virion envelope fusion with the cell membrane. Here, we analyzed whether the fusion of liposome membranes under the action of fragments of fusion peptides of Ebola and Marburg viruses depends on the composition of lipid vesicles. A fluorescence assay and electron microscopy were used to quantify the fusogenic activity of the virus fusion peptides and to identify the lipid determinants affecting membrane merging. Differential scanning calorimetry of lipid phase transitions revealed alterations in the physical properties of the lipid matrix produced by virus fusion peptides. Additionally, we found that plant polyphenols, quercetin, and myricetin inhibited vesicle fusion induced by the Marburg virus fusion peptide.

Membrane Activity of Melittin and Magainin-I at Low Peptide-to-Lipid Ratio: Different Types of Pores and Translocation Mechanisms.

Antimicrobial peptides (AMPs) are believed to be a prominent alternative to the common antibiotics. However, despite decades of research, there are still no good clinical examples of peptide-based antimicrobial drugs for system application. The main reasons are loss of activity in the human body, cytotoxicity, and low selectivity. To overcome these challenges, a well-established structure-function relationship for AMPs is critical. In the present study, we focused on the well-known examples of melittin and magainin to investigate in detail the initial stages of AMP interaction with lipid membranes at low peptide-to-lipid ratio. By combining the patch-clamp technique with the bioelectrochemical method of intramembrane field compensation, we showed that these peptides interact with the membrane in different ways: melittin inserts deeper into the lipid bilayer than magainin. This difference led to diversity in pore formation. While magainin, after a threshold concentration, formed the well-known toroidal pores, allowing the translocation of the peptide through the membrane, melittin probably induced predominantly pure lipidic pores with a very low rate of peptide translocation. Thus, our results shed light on the early stages of peptide-membrane interactions and suggest new insights into the structure-function relationship of AMPs based on the depth of their membrane insertion.

De Novo Missense Variations of ATP8B2 Impair Its Phosphatidylcholine Flippase Activity.

P4-ATPases comprise a family of lipid flippases that translocate lipids from the exoplasmic (or luminal) to the cytoplasmic leaflet of biological membranes. Of the 14 known human P4-ATPases, ATP8B2 is a phosphatidylcholine flippase at the plasma membrane, but its physiological function is not well understood. Although ATP8B2 could interact with both CDC50A and CDC50B, it required only the CDC50A interaction for its exit from the endoplasmic reticulum and subsequent transport to the plasma membrane. Three de novo monoallelic missense variations of ATP8B2 were found in patients with intellectual disability. None of these variations affected the interaction of ATP8B2 with CDC50A or its localization to the plasma membrane. However, variations of either of two amino acid residues, which are conserved in all P4-ATPases, significantly reduced the phosphatidylcholine flippase activity of ATP8B2. Furthermore, mutations in the corresponding residues of ATP8B1 and ATP11C were found to decrease their flippase activities toward phosphatidylcholine and phosphatidylserine, respectively. These results indicate that the conserved amino acid residues are crucial for the enzymatic activities of the P4-ATPases.

Ultrasteep Slope Cryogenic FETs Based on Bilayer Graphene.

Cryogenic field-effect transistors (FETs) offer great potential for applications, the most notable example being classical control electronics for quantum information processors. For the latter, on-chip FETs with low power consumption are crucial. This requires operating voltages in the millivolt range, which are only achievable in devices with ultrasteep subthreshold slopes. However, in conventional cryogenic metal-oxide-semiconductor (MOS)FETs based on bulk material, the experimentally achieved inverse subthreshold slopes saturate around a few mV/dec due to disorder and charged defects at the MOS interface. FETs based on two-dimensional materials offer a promising alternative. Here, we show that FETs based on Bernal stacked bilayer graphene encapsulated in hexagonal boron nitride and graphite gates exhibit inverse subthreshold slopes of down to 250 µV/dec at 0.1 K, approaching the Boltzmann limit. This result indicates an effective suppression of band tailing in van der Waals heterostructures without bulk interfaces, leading to superior device performance at cryogenic temperature.

FCS videos: Fluorescence correlation spectroscopy in space and time.

Fluorescence Correlation Spectroscopy (FCS), invented more than 50 years ago is a widely used tool providing information on molecular processes in a variety of samples from materials to life sciences. In the last two decades FCS was multiplexed and ultimately made into an imaging technique that provided maps of molecular parameters over whole sample cross-section. However, it was still limited by a measurement time on the order of minutes. With the improvement of FCS time resolution to seconds using deep learning, we extend here FCS to so-called FCS videos that can provide information how the molecular parameters determined by Imaging FCS change in space and time. This opens up new possibilities for the investigation of molecular processes. Here, we demonstrate the feasibility of the approach and show FCS video applications to lipid bilayers and cell membranes.

A molecular mechanism for how pressure induces interdigitation of phospholipid bilayer membranes.

The phase transition from the ripple gel phase to the interdigitated gel phase of bilayers of phosphatidylcholines (PCs) with two saturated long-chain fatty acids under high pressure was investigated by pressure-scanning microscopy, fluorometry, and dynamic light scattering (DLS) measurements. Microscopic observation for giant vesicles (GVs) of distearoyl-PC (DSPC) under high pressure showed that spherical GVs transforms significantly into warped and distorted spherical ones instantaneously at the pressure-induced interdigitation. The fluorescence intensities of amphiphilic probe Prodan and hydrophobic probe Laurdan in the dipalmitoy-PC (DPPC) bilayer steeply decreased and increased, respectively, at the interdigitation, suggesting that the conformational change of the polar head group of DPPC molecule in the bilayer transiently occurred at the interdigitation. Further, it was found from the high-pressure DLS measurements that the size of the vesicle particles of the DPPC and DSPC transiently increases near the interdigitation pressure, whereas the chemically induced interdigitation by adding ethanol to the DSPC bilayer membrane under atmospheric pressure produce no such change in the particle size. Taking account of the critical packing parameter of the PC molecule, the above experimental results would lead us to the conclusion that the pressure-induced interdigitation is attributable to the increase in repulsive interaction between the polar head groups of the PC molecules resulting from the orientational change of the head group from a parallel alignment to a perpendicular one with respect to the bilayer surface by applying pressure, namely the transient state: it occurs when the repulsive interaction exceeds a threshold value for the balance between the repulsive interaction and the attractive interaction among the hydrophobic acyl chains.

Development of cost-effective cellulose-based bilayer hybrid composite membranes for CO2 separation in biogas purification.

CO2 is the main pollutant in biogas, reducing its calorific value. Among various technological methods to eliminate carbon dioxide from biogas, membrane separation technology stands out for large-scale industrial biogas purification due to its advantages. The selection of membrane material and preparation process are key factors in membrane separation technology. In this study, a premixing process was initially used to blend different masses of packed molecular sieves TS-1 (or ZSM-5) and cellulose derivatives (ethyl cellulose, cellulose acetate) separately. These mixtures were then coated onto porous PVDF substrates using a coating process to create various bilayer hybrid composite membranes. Among these, PVDF/EC-ZSM-5 (containing 15 % ZSM-5) bilayer hybrid composite membrane is the most fitting. For CO2/CH4 gas mixtures, the gas selectivity of this membrane surpassed Robeson's 1991 standard line (the CO2 permeability was 597.48 Barrer, and the CO2/CH4 selectivity was 9.14). Overall, this composite membrane, made for the first time from PVDF, ZSM-5, and EC, is expected to be a promising CO2 selective separation membrane material for biogas purification in large-scale industrial processes due to its simple production process.

Characterization and biocompatibility of a bilayer PEEK-based scaffold for guiding bone regeneration.

BACKGROUND: Polyetheretherketone (PEEK) is well known for its excellent physical-chemical properties and biosafety. The study aimed to open up a new method for clinical application of PEEK to reconstruct large-scale bone defects. METHODS: A bilayer scaffold for bone regeneration was prepared by combining a sulfonated PEEK barrier framework (SPEEK) with a hydrogel layer loaded with aspirin (ASA) and nano-hydroxyapatite (nHAP) by the wet-bonding of Polydopamine (PDA). RESULTS: The hydrogel was successfully adhered to the surface of SPEEK, resulting in significant changes including the introduction of bioactive groups, improved hydrophilicity, and altered surface morphology. Subsequent tests confirmed that the bilayer scaffold exhibited enhanced compression resistance and mechanical compatibility with bone compared to a single hydrogel scaffold. Additionally, the bilayer scaffold showed stable and reliable bonding properties, as well as excellent biosafety verified by cell proliferation and viability experiments using mouse embryo osteoblast precursor (MC3T3-E1) cells. CONCLUSION: The bilayer bone regeneration scaffold prepared in this study showed promising potential in clinical application for bone regeneration.

Synergistic antibacterial and wound healing effects of chitosan nanofibers with ZnO nanoparticles and dual antibiotics.

One concern that has been considered potentially fatal is bacterial infection. In addition to the development of biocompatible antibacterial dressings, the screening and combination of new antibiotics effective against antibiotic resistance are crucial. In this study, designing hemostasis electrospun composite nanofibers containing chitosan (CS), polyvinyl pyrrolidone (PVP) and Gelatin (G) as the major components of hydrogel and natural nanofibrillated sodium alginate (SA)/polyvinyl alcohol (PVA) and ZnO nanoparticles (ZnONPs) combination as the nanofiller ingredient, has been investigated which demonstrated significant potential for accelerating wound healing. The hydrogels were developed for the delivery of the amikacin and cefepime antibiotics, along with zinc oxide nanoparticles that were applied to an electrospun layer. Amikacin is a highly effective aminoglycoside antibiotic, particularly for hospital-acquired infections, but its use is limited due to its toxicity. By utilizing it in low concentrations in the form of nanofibers and combining it with cefepime, which exhibits synergistic effects, enhanced efficacy against bacterial pathogens is achieved while potentially minimizing cytotoxicity compared to individual antibiotics. This dressing demonstrated efficient drug release, flexibility, and good swelling properties, indicating its suitable mechanical properties for therapeutic applications. After applying the biocompatible hydrogel to wounds, a significant acceleration in wound closure was observed within 14 days compared to the control group. Furthermore, the notable antibiotic and anti-inflammatory properties underscore its effectiveness in wound healing, making it a promising candidate for medical applications.

Electron Collimation in Twisted Bilayer Graphene via Gate-Defined Moiré Barriers.

Electron collimation via a graphene p-n junction allows electrostatic control of ballistic electron trajectories akin to that of an optical circuit. Similar manipulation of novel correlated electronic phases in twisted-bilayer graphene (tBLG) can provide additional probes to the underlying physics and device components toward advanced quantum electronics. In this work, we demonstrate collimation of the electron flow via gate-defined moiré barriers in a tBLG device, utilizing the band-insulator gap of the moiré superlattice. A single junction can be tuned to host a chosen combination of conventional pseudo barrier and moiré tunnel barriers, from which we demonstrate improved collimation efficiency. By measuring transport through two consecutive moiré collimators separated by 1 µm, we demonstrate evidence of electron collimation in tBLG in the presence of realistic twist-angle inhomogeneity.

Fatigue strength of bilayer yttria-stabilized zirconia after low-temperature degradation.

This study examined the impact of interfacial interactions on bilayer yttria-stabilized zirconia (YSZ) used in dental restorations. In-house bilayer structures of 3YSZ and 5YSZ composition underwent hydrothermal degradation to compare the properties of control and low-temperature degradation (LTD) treated groups. Biaxial flexural strength via piston-on-three-balls, staircase fatigue strength over 106 cycles at 15 Hz, phase characterization and quantification through XRD and Rietveld refinement, and fractography were conducted. Weibull analysis was employed to determine the Weibull modulus and characteristic strength. Results demonstrated an enhancement in the mechanical performance of 3YSZ composition after LTD treatment, whereas the mechanical properties of 5YSZ remained largely unaffected post-degradation. Fractographic analysis revealed that failure originated at the surface tensile location across all specimen groups. These findings offer insights into the mechanical behavior of bilayer zirconia structures and reinforce the significance of hydrothermal treatment in enhancing their performance, particularly in the case of 3Y compositions.

Versatile effects of galectin-1 protein-containing lipid bilayer coating for cardiovascular applications.

Modulating inflammatory cells in an implantation site leads to severe complications and still unsolved challenges for blood-contacting medical devices. Inspired by the role of galectin-1 (Gal-1) in selective functions on multiple cells and immunomodulatory processes, we prepared a biologically target-specific surface coated with the lipid bilayer containing Gal-1 (Gal-1-SLB) and investigate the proof of the biological effects. First, lipoamido-dPEG-acid was deposited on a gold-coated substrate to form a self-assembled monolayer and then conjugated dioleoylphosphatidylethanolamine (DOPE) onto that to produce a lower leaflet of the supported lipid bilayer (SLB) before fusing membrane-derived vesicles extracted from B16-F10 cells. The Gal-1-SLB showed the expected anti-fouling activity by revealing the resistance to protein adsorption and bacterial adhesion. In vitro studies showed that the Gal-1-SLB can promote endothelial function and inhibit smooth muscle cell proliferation. Moreover, Gal-1- SLB presents potential function for endothelial cell migration and angiogenic activities. In vitro macrophage culture studies showed that the Gal-1-SLB attenuated the LPS-induced inflammation and the production of macrophage-secreted inflammatory cytokines. Finally, the implanted Gal-1-SLB reduced the infiltration of immune cells at the tissue-implant interface and increased markers for M2 polarization and blood vessel formation in vivo. This straightforward surface coating with Gal-1 can be a useful strategy for modulating the vascular and immune cells around a blood-contacting device.

Temperature-Dependent Activation of Thermosensitive Transient Receptor Potential Channels.

Temperature detection is essential for the survival and perpetuation of any species. Thermoreceptors in the skin sense the body temperature and also the temperatures of the ambient air and the objects. In 1997, Dr. David Julius and his colleagues found that a receptor expressed in small-diameter primary sensory neurons was activated by capsaicin (the pungent chemical in hot pepper). This receptor was also activated by temperature above 42 °C. That was the first time that a thermal receptor in primary sensory neurons has been identified. This receptor is named transient receptor potential vanilloid 1 (TRPV1). Now, 11 thermosensitive TRP channels are known. In this chapter, we summarize the reports and analyze thermosensitive TRP channels in a variety of ways to clarify the activation mechanisms by which temperature changes are sensed.

Unclonable Encryption-Verification Strategy Based on Bilayer Shape Memory Photonic Crystals.

The ability to reversibly exhibit structural color patterns has positioned photonic crystals (PCs) at the forefront of anti-counterfeiting. However, the security offered by the mere reversible display is susceptible to illicit alteration and disclosure. Herein, inspired by the electronic message captcha, bilayer photonic crystal (BPC) systems with integrated decryption and verification modules, are realized by combining inverse opal (IO) and double inverse opal (DIO) with polyacrylate polymers. When the informationized BPC is immersed in ethanol or water, the DIO layer displayed encrypted information due to the solvent-induced ordered rearrangement of polystyrene (PS) microspheres. The verification step is established based on the different structural colors of the IO layer pattern, which result from the deformation or recovery of the macroporous skeleton induced by solvent evaporation. Moreover, through the evaporation-induced random self-assembly of PS@SiO2 and SiO2 microspheres, unclonable structurally colored identifying codes are created in the IO layer, ensuring the uniqueness upon the verification. The decrypted code in the DIO layer is valid only when the IO layer displays the pattern with the predetermined structural color; otherwise, it is a pseudo-code. This structural color-based "decryption-verification" approach offers innovative anti-counterfeiting applications in nanophotonics.

Lignin-induced rapid polymerization of asymmetrical adhesion Janus gel for strain sensor.

Functional hydrogel sensors have shown explosive growth in the health and medical fields. However, the uniform adhesion and the complicated polymerization process of hydrogels seriously hinder their further development. Herein, inspired by the layered structure of human skin, we prepare a Janus gel using in-situ polymerization. Based on the lignin-Fe3+ dual catalytic system, the rapid polymerization of the gel was achieved at room temperature. By tailoring the mass ratio of lignin and Fe3+ in the precursor, the adhesion of the upper and bottom layers can be easily adjusted. In addition, hydrophobic association is introduced into the upper layer to improve the gel's mechanical properties. The obtained asymmetric bilayer gel has a significant difference in adhesion (7 times), and exhibits excellent mechanical properties in the elongation at break (1437 %) and the breaking strength (463.2 kPa). Moreover, the bilayer gel also has good freezing and UV resistance. We use the bilayer gel as a wearable strain sensor, which shows a wide strain detection range of 0-800 % (maximum gauge factor = 5.3). The proposed simple strategy avoids UV irradiation and heating processes, which provides a new idea for the rapid polymerization of multifunctional Janus hydrogels with adjustable performances.