This review provides an overview of recent advancements in applying graphene-based materials as sorbents for liquid chromatography (LC) analysis. Graphene-based materials are promising for analytical chemistry, including applications as sorbents in liquid chromatography. These sorbents can be functionalized to produce unique extraction or stationary phases. Additionally, graphene-based sorbents can be supported in various materials and have consequently been applied to produce various devices for sample preparation. Graphene-based sorbents are employed in diverse applications, including food and environmental LC analysis. This review summarizes the application of graphene-based materials in food and environmental water analysis in the last five years (2019 to 2023). Offline and online sample preparation methods, such as dispersive solid phase microextraction, stir bar sorptive extraction, pipette tip solid phase extraction, in-tube solid-phase microextraction, and others, are reviewed. The review also summarizes the application of the columns produced with graphene-based materials in separating food and water components and contaminants. Graphene-based materials have been reported as stationary phases for LC columns. Graphene-based stationary phases have been reported in packed, monolithic, and open tubular columns and have been used in LC and capillary electrochromatography modes. Graphene and graphene derivates are emerging materials in research and development , includiDespite the potential for graphene and its derivates as a highly suitable material, the manufacturing obstacle hinders its adoption on an industrial scale. The mechanical exfoliation of graphite through adhesive tape is a practical way to produce graphene-based materials ,10. ThisThe diversity of potential functionalizations of graphene-based materials allows for a wide range of applications in separation science. These applications include the use of techniques such as stir bar sorptive extraction (SBSE) ,26, dispThe present review investigates the applications of graphene-based materials in food and water analysis, including their utilization as sorbents for sample preparation and analytical columns for LC, in a time frame from 2019 until 2023. The escalating necessity for enhanced analytical processes in food and water analysis has stimulated interest in using graphene-based materials as efficient and trustworthy solutions. Therefore, this review article aims to illuminate the current state and potential prospects for graphene-based sorbents in food and water analysis.Offline sample preparation techniques are the most common strategy in the analytical chemistry field. They consist of preparing the sample for analysis while disconnected from the instrument or application. Some advantages of this strategy are the simple procedure, which is usually not dependent on complex instrumentation set-up, and the possibility of analyzing sample fractions prepared with different analytical techniques. This section summarizes offline extraction techniques in which graphene-based sorbents have been employed recently.Dispersive solid phase extraction (d-SPE) and microextraction (d-SPME) are examples of sample preparation techniques that involve graphene-based materials as dispersive sorbents . This sa3O4/TBA) nanocomposite was successfully applied in the determination of herbicides in vegetables and water samples [3O4/TBA sorbent was prepared by co-precipitating GO and Fe3O4 particles. After the obtention of the GO/Fe3O4 particles, they were functionalized with TBA using ethylene glycol to form covalent bonds. A d-SPME method using the GO/Fe3O4/TBA to quantify 2,4-dichlorophenoxyacetic acid presented limit of detection (LOD) and limit of quantitation (LOQ) values lower than 0.02 μg·mL−1, and the recovery range was between 88.0 and 94.0%. Another graphene-based magnetic phase that has been successfully applied to the analysis of contaminants in vegetables is the GO-Fe3O4 nanoparticles functionalized with maltodextrin (MD) and β-cyclodextrin (β-CD), which was used for the investigation of triazole and triazine pesticides in corn, tomato, and potato [3O4 magnetic particles produced this magnetic extraction phase. After the obtention of the GO-Fe3O4, this material was functionalized with MD and β-CD using epichlorohydrin as a linker. The LODs obtained for determining the pesticides in the vegetables ranged from 0.01 to 0.08 μg·L−1 and recovery was between 88.4 and 112.0%. Metal-organic frameworks (MOF) have also been used to functionalize graphene-based sorbents. MOFs are porous crystalline materials formed by assembling metal ions and organic ligands with large surface areas, high porosity, and controllable pore structures. As a recent example, in d-SPME, Fe3O4 particles functionalized with GO sheets and UiO-66 (a Zr-based MOF) have been used as a sorbent for the determination of food dyes in soft drinks, candies, and pastilles [3O4, followed by the addition of ZrOCl2·8H2O and terephthalic acid into a GO-Fe3O4 suspension in order to produce the magnetic GO/UIO-66 sorbent. The s-SPME method employing the GO/UIO-66 sorbent presented LODs less than 0.218 ng·mL−1, LOQs lower than 9.368 ng·mL−1, and recovery ranging from 95.01 to 106.33%. The interaction mechanism between the MOF-based sorbent and the food dye analytes was attributed to multiple mechanisms, including n–π/π–π interactions, Yoshida/dipole–dipole hydrogen bonds, and electrostatic interactions. Though magnetic particles are the primary support for sample preparation in d-SPME and d-SPE in food and water analysis, no magnetic materials have been used. For example, GO-coating polystyrene (PS) microspheres were applied to extract bisphenol endocrine disruptors in environmental water samples [−1 and LOQs of 0.07–0.37 μg·L−1 and recovering in a range from 71.1% to 104.8%. The retention mechanism of this GO-PS extraction phase was attributed to the π–π interactions and hydrogen interactions between the sorbent and the analytes. Another interesting variation of the d-SPME using graphene-based sorbents applied in food analysis is the “rotating flat surface solid phase microextraction (RFS-SPME)” method. This strategy consists of suspending and stirring foams of the stationary phase, instead of powder material, in the matrix, and after the end of the stirring the foam can be removed with forceps [−1, LOQs lower than 0.475 µg·L−1, and recoveries over 90.0% The evaluated 3D GO/La Ni foams’ extraction phase presented an extraction mechanism sorbent–sorbate combining π–π interaction, hydrogen bonding, and electrostatic interaction.Graphene-based materials have been employed in d-SPE and d-SPME, and mainly supported in magnetic nanoparticles . Magneti samples . The GO/d potato . Co-precastilles . This in samples . To prod forceps . The RFS forceps . To prodCoating techniques involve sample preparation strategies where the sorbent is coated over, typically, a non-sorptive surface. For example, stir bar sorptive extraction (SBSE) and fiber solid phase microextraction (fiber-SPME) are techniques in which the sorbent is fixed over an inert surface. Usually, the sample preparation process using these techniques involves (i) immersing the device coated with sorbent in a matrix solution, (ii) stirring the solution for a specific time, (iii) removing and washing the device to remove the matrix, and finally (iv) desorbing the analyst in an appropriated solvent before analysis.−1 and recovery in a range of 83% to 112%. The rGO-Ni SBSE bar presented an attributed extraction mechanism based on π–π interaction and hydrophobic interactions [−1, LOQs of 0.5 to 1.0 ng·mL−1, and a recovery range of 89.38–108.54%. To produce this device, it was first necessary to activate the SS wire surface to allow the chemical binding of the GO. To do that, the SS wire was submitted to a sequence of reactions until the polydopamine-SS wire was formed. The GO was suspended in a solution and then reacted with the polydopamine-SS wire forming the GOF-SS wire SBSE bar [3O4 particles’ sorbent (G-Fe3O4) in the determination of seven pesticides in water [3O4 particles were produced by the co-precipitation method, similar to that previously described in the d-SPME section. This SBSDμE method presented a recovery range between 20% and 75%, with a LOQ between 5 ng·mL−1 and 9.5 ng·mL−1. Another interesting sample preparation technique based on the sorbent phase coating a surface was fiber solid phase microextraction (fiber-SPME). Fiber-SPME consists of the sorbent coating a capillary’s inner or outer surface, forming the extraction fiber. Commonly, fiber-SPME devices are filled with monolithic extraction phases, including the ones modified with graphene-based materials. Graphene-based materials as fiber-SPME sorbents have also been explored for food and water LC analysis in the last five years. For example, a composite prepared with GO, MOF zeolitic imidazolate framework 8 (ZIF-8), and molecularly imprinted polymers (MIP) deposited in fiber was used to evaluate sterol and steroid hormones in white meat, egg yolks, and vegetables [−1 and recovery ranging from 95.0% to 101.0%. This sorbent presented a mixed interaction mechanism, primarily attributed to the MIP efficient imprinting effect and the differences in size, functional group type, and position of the analytes and polar interactions, hydrogen bonds, and electrostatic interactions [−1 and LOQs lower than 2.27 μg·L−1 were obtained, and a recovery of range between 70.0 and 117% was reported using this sample preparation method.SBSE and related techniques are simple and effective ways to extract analytes from the matrix. SBSE devices are usually produced by chemically or physically adhering the sorbent to an inert material . Graphenractions . Moreoveractions , presentSBSE bar . AnotherSBSE bar . This baSBSE bar . A recenin water . The maggetables . The fibractions . Anotherractions . Using mractions . LODs lo−1 and LOQs lower than 0.91 μg·L−1. This extraction phase presented an interesting proposed interaction mechanism in which hydrogen bonding participates. However, the adsorption process also influences the interaction of π-electrons of the analyte with the Cr and back donation from the sorbent material into the molecular anti-bonding π∗ orbital [Hollow fiber solid phase microextraction (HF-SPME) is a variation of the fiber-SPME in which the sorbent coats the inner wall of a capillary, forming an open channel in the device’s core . Unfortu orbital .Packed sorbent techniques are sample preparation techniques that use a solid adsorbent material packed in a cartridge, a column, or a pipette. The sample preparation procedure of these techniques comprises the following steps: (i) equilibrate the sorbent by flushing a suitable solvent through the cartridge, (ii) load the sorbent with the sample by flushing it through the cartridge (for SPE), (iii) wash the sorbent to remove the matrix, and (iv) elute and collect the analytes using a proper solvent.−1 and a recovery range from about 55 to 90%. This stationary phase presents a complex interaction mechanism between the sorbent and the analytes due to the participation of π–π stacking, hydrophobic interaction, and size complementarity interactions. Another interesting graphene-based material packed in an SPE sample preparation cartridge is a zwitterionic sorbent modified with graphene (G) and fixed over Si particles [−1 and a recoveries range from 48 to 85%. Though graphene-based materials are usually fixed over support, SPE cartridges can be packed with graphene sheets that are not fixed in support. As mentioned above, this kind of packing presents limitations, such as blocking the frits caused by the accumulation of the graphene sheets, but it is still possible. For example, it was reported that rGO sheets packed on SPE cartridges were used to extract aflatoxins from food samples [−1, LOQs lower than 2.83 ng·g−1, and relative recoveries ranging from 70 to 113% using this cartridge. Another interesting example of an SPE sorbent not supported in particles is 3D reduced graphene oxide (3D-rGO), a self-supported material [Sorbents packed on cartridges are a usual approach for producing SPE devices. Silica particles are frequently used as a support for graphene-based sorbents due to their capacity to provide a stable structure and prevent blockages in cartridges . Additioarticles . This soarticles , present samples . It was material . In thismaterial . A sorbe−1 for juice and 5.0–25.0 ng·g−1 for candy and syrup. Additionally, recoveries ranging from 48 to 69%, 34 to 89%, and 28 to 76% were obtained for juice, candy, and syrup, respectively. MOF materials modified with graphene-base materials have also been reported as sorbents for PT-SPME. Si microspheres covalently bonded to fluorinated graphene (FG) and functionalized with ZIF-8 nanocrystals were packed in a PT-SPME device and applied to determine chlorophenols in tap water, honey, and black tea [−1 and LOQs of 15.9 µg·kg−1. This extraction phase presented multi-interaction mechanisms, including π-stacking and hydrophobic and hydrogen-bonding interactions between the adsorbent and the compounds. Though Si particles are standard, other types of particles can be used as supports for graphene-based materials in PT-SPME and have been applied in LC analyses of foods. For example, the nanocomposite GO-starch-polyacrylamide was packed in PT-SPME devices and used to determine antibiotic residues in cow’s milk [3 to form the porous material. The PT-SPME method presented LODs of 2.7–5.0 μg·kg−1 and a recovery range of 88–102%. This extraction phase presented an adsorption mechanism mainly based on hydrogen bond interactions between the analytes and the sorbent [Pipet tip SPME (PT-SPME) can be considered a variation of the SPE because the sorbent packed on a pipet tip forms a device similar to a cartridge. Si particles have also been used as a sorbent support for PT-SPME sample preparation. As a recent example, GO sheets fixed on Si particles have reportedly been packed in PT-SPME devices and applied to determine herbicides in sugarcane-derived foods . The SiGlack tea . About 8w’s milk . The sor sorbent .In the last five years, graphene-based sorbents have successfully been employed in diverse sample preparation techniques for food and water analysis. Water, milk, vegetables, cereals, and meat are examples of the diversity of samples that graphene-based sorbents could prepare recently . The divThe online sample preparation strategy presents a significant advantage over offline techniques; the automation process minimizes the chance of human error, one of the most common error sources in analytical methods ,62. Thou−1 and recovery between 70.4 and 91.6%. In-tube SPME also utilized SiGO particles’ functionalization with other bindings. For example, SiGO particles functionalized with C18 and end-capped were used for the online sample preparation of coffee samples, targeting the determination of xanthines [−1 and recoveries between 73 and 109% were obtained. Both works above employed a similar route to prepare the SiGO particle or its functionalized derivate with C18 and end-capping. Briefly, GO sheets, produced by Hummer’s method, were covalently bonded to the Si particle’s surface. For the functionalization with C18 and end-capping, the SiGO particles formed were reacted with chlorodimethyl-n-octadecylsilane and trimethylchlorosilane in separate steps [−1 for milk and 0.6 μg·kg−1 in chicken muscle and recoveries of about 70.3 to 98.5% and 79.0 to 108.0% for each matrix, respectively, were obtained [−1.Graphene-based materials fixed over silica particles are an interesting sorbent for this application. For example, GO supported into the Si particles’ (SiGO) sorbent was packed inside a fused silica capillary to produce a miniaturized extraction column employed in the extraction of β-lactam antibiotics from environmental water samples . This meanthines . LOQs ofte steps . Monolitte steps . After pte steps . The in obtained . Coated obtained . An inteobtained . This ITobtained  This IT-−1, a LOQ of 0.3 ng·mL−1, and a recovery range between 90.0 and 106.5%.Headspace in-tube solid phase microextraction (HS-SPME) using a graphene-based sorbents is another sample preparation strategy that has been applied recently, hyphenated to LC. This strategy consists of introducing the sample in a vail and inserting the HS-SPME device into the space above the liquid surface; after the extraction procedure, the HS-SPME system is connected online to LC equipment. A recently reported application of graphene-based sorbents in the HS-SPME technique for food analysis was performed, utilizing a stainless steel tubing coated with a composite formed by GO and ionic liquid (IL) for the analyses of the headspace of naphthalene, a volatile PAH, in honey samples . The GO-In short, a trend exists to employ GO as the graphene-based sorbent in online sample preparation . The fixGraphene-based materials have also been used as stationary phases for liquid chromatography columns. These sorbents have been employed in packed, monolithic, and open tubular (OT) columns, the three most common types of LC columns . These sPacked columns are the standard types of columns in LC. They comprise particles of the stationary phase packed inside a tubing (or capillary tubing). This type of column is easy to produce given its production procedures demand only a pump that allows the packing of the stationary phase in the column hardware . The simGraphene-based packed LC columns are still underexplored for food and water analysis. However, some applications reported in the last five years are described in the literature. In seed samples, a capillary column packed with a SiGO-C18ec stationary phase, produced by the functionalization of SiGO particles with C18 followed by end-caping (as previously mentioned), was employed to separate lecithin, a common phospholipid component in foods. This study evaluated the viability of applying an online liquid extraction (OLE) system coupled with an LC-MS to evaluate the hyphenated extraction and separation of lecithin from the pounder of seeds . The SiGMonolithic columns are another popular type of LC column composed of a monolith of the stationary phase, usually inside a capillary tubing. These columns are, conventionally, produced by the in situ polymerization of polymer-based  and siliSchisandra, a purple-red berry typically used in traditional Chinese medicine [Graphene-based monolithic columns have been used in LC and CEC modes, but recent food and water analysis applications present few reported examples. In LC, a monolithic column prepared by the polymerization of 3-chloro-2-hydroxypropylmethacrylate (HPMA-Cl) and ethylene dimethacrylate (EDMA), followed by the fixation of GO, was used recently for the determination of chloramphenicol (CAP), a veterinarian antibiotic, and chloramphenicol glucuronide (CAPG), a metabolite from CAP, in honey and milk samples . Anothermedicine . For thimedicine . The CECmedicine . These cmedicine .Open tubular columns have gained popularity in liquid chromatography due to their intrinsic advantages over packed columns, such as low pressures, and their theoretically higher performance ,93,94. TNevertheless, the separation of small molecules on OT columns has also been described ,104,105.2) as stationary phases for determining sulfonamides in environmental water [2 dispersion into a fused-silica capillary and leaving it overnight for the GO-MoS2 to bond in the fused-silica capillary wall [Graphene-based OT-LC columns have not received much attention for food and environmental water analysis. Very few works explore graphene-based OT columns in food and water analysis. An example reported in the last five years is the separation of casein protein variants from milk samples using an OT column produced with Poly-L-Lysine (PLL) grafted on a methacryloyl graphene oxide nanoparticles (MGONPs) stationary phase . In CEC,al water . The colary wall . The separy wall . For exaary wall . The enaary wall .Graphene sorbents have been employed in diverse analytical column configurations in the last five years. Packed, monolithic, and open tubular are the three most common column types, and all present examples are of graphene-based materials as stationary phases . In foodGraphene-based sorbents are becoming more popular year after year in diverse applications. In food and environmental water LC analysis, graphene-based materials have been underexplored in the last five years. However, a trend in the diversification of the application of those materials is also being observed. In offline sample preparation, dispersive solid phase extraction receives the most attention for the application of graphene-based sorbents in food and environmental water LC analysis, but other techniques, such as SBSE, Fiber-SPME, SPE, HF-SPME, and others, could employ graphene-based sorbents with success. Hence, graphene-based sorbents are in an early stages of development, and the number and variety of sample preparation techniques using these materials is tending to increase. In online sample preparation methods, IT-SPME is the most common application of graphene sorbents in food and environmental water analysis. This kind of online sample preparation allows an easy connection to LC analysis, which has been a great advantage of these techniques.Additionally, the extraction column might be packed with a wide variety of graphene-based sorbents and can deal with a high diversity of samples. Graphene-based sorbents have also been used for analytical LC columns. These sorbents have been utilized in packed, monolithic, and open tubular columns. Graphene oxide bonded to Si particles is the most common graphene-based sorbent used as stationary phases. Graphene-based sorbents have been used as stationary phases in the analysis of food components such as caseins, lecithin, and schizandrols. Graphene-based sorbents have also been employed in separating water contaminants such as alkylbenzenes and antibiotics. The future of graphene-based stationary phases in food and water analyses is promising, with packed columns becoming more frequent for LC and OT columns for CEC analysis. GO should remain the most preferred graphene-based material due to its molecular characteristics, allowing it to be functionalized. Functionalizing these materials with different bindings should be encouraged to find solutions to specific problems.Graphene-based sorbents present advantages and disadvantages in analyzing food and water samples through liquid chromatography. One significant advantage is the versatility of these sorbents in extracting various analytes while being compatible with different sample preparation techniques. Furthermore, functionalizing GO, and other graphene-based materials, offers a range of possibilities for online approaches. In contrast, as its main disadvantage, these phases are relatively new and may present a lower scope of applications than conventional commercially available extraction phases. In other words, graphene-based materials are not multipurpose, which means that for some applications it is crucial to select an adequate material to extract specific analytes. Another advantage of graphene-based materials is that they can be used as stationary phases in packed, monolithic, and open tubular chromatographic columns. Conversely, detachment followed by the accumulation of graphene sheets in the column outlet can again hinder effectiveness and limit the column’s lifetime. Nonetheless, the functionalization of graphene-based materials with different bindings for specific applications presents intriguing opportunities for improving the diversity of applications for food and water analysis.


The ferroelectric domains are visualized through the imaging of the plasmonic response in a graphene monolayer adjacent to the moiré WSe2 bilayers. Specifically, we find that the ferroelectric polarization in moiré domains is imprinted on the plasmonic response of the graphene. Complementary nano-photocurrent measurements demonstrate that the optoelectronic properties of graphene are also modulated by the proximal ferroelectric domains. Our approach represents an alternative strategy for studying moiré ferroelectricity at native length scales and opens promising prospects for (opto)electronic devices.Ferroelectricity, a spontaneous and reversible electric polarization, is found in certain classes of van der Waals (vdW) materials. The discovery of ferroelectricity in twisted vdW layers provides new opportunities to engineer spatially dependent electric and optical properties associated with the configuration of moiré superlattice domains and the network of domain walls. Here, we employ near-field infrared nano-imaging and nano-photocurrent measurements to study ferroelectricity in minimally twisted WSe 2 by visualizing the plasmonic and photo-thermoelectric response of an adjacent graphene monolayer.Recent experiments have shown the formation of ferroelectric domains in twisted van der Waals bilayers. Here, the authors report near-field infrared nano-imaging and nano-photocurrent measurements to investigate ferroelectricity in minimally twisted WSe Moiré superlattices not only inherit characteristics of the constituent layers but also exhibit new emergent phenomena. Among these emergent effects, interfacial ferroelectricity was recently discovered in marginally twisted hexagonal boron nitride (h-BN)7 and transition metal dichalcogenides (TMDs)11. In minimally twisted bilayers, lattice reconstruction results in triangular domains with periodically alternating AB and BA stacking registries14. The charge transfer between the two layers creates moiré patterns with alternating out-of-plane polarization16. Moiré ferroelectricity in twisted TMDs and h-BN has been confirmed by local Kelvin probe force microscopy (KPFM) imaging17 and by area-averaged transport measurements8.Moiré superlattices of two-dimensional van der Waals materials have emerged as a capable platform to explore exotic electric and optical properties that can be controlled by selecting building blocks for the assembled atomic layers and manipulating the twist angles between them18 and tune photo-excited carrier dynamics19, among other effects. In transport experiments, modulation of the doping of graphene is detected and employed as a sensor for the ferroelectric polarizations20. However, in transport experiments, the ferroelectrically induced doping density was inferred from area-averaged analysis by assuming that the entire sample reveals a uniform polarization under an applied electric field in a gatable device8. In practice, such devices may possess domains with the opposite polarization even at high electric fields, as was recently revealed by backscattering electron imaging9 and KPFM5. This observation indicates that in stacked vdW devices, unlike conventional ferroelectric materials, aligning all the polarization in the same direction may require substantial energy for bending and eventually merging all the adjacent domain walls22. Therefore, in order to read out the doping from the ferroelectric potential, it is imperative to measure the carrier density in each moiré domain. Furthermore, novel optical and optoelectronic properties in devices integrated with ferroelectric have been investigated using far-field optical spectroscopy26. These measurements are diffraction-limited and thus cannot probe the rich optical or optoelectronic properties at the underlying moiré domain scale. Therefore, all existing results call for measurements capable of spatially resolving ferroelectric domains. Piezoresponse force microscopy (PFM) and KPFM can directly measure the domain structures of ferroelectric via electromechanical surface deformation and electrostatic force, respectively. Whereas the ferroelectric domains can be clearly visualized by PFM27, PFM results directly reflect the piezoelectric response, making it challenging to quantitatively evaluate ferroelectric properties. Usually, KPFM can provide a quantitative characterization of a ferroelectric by measuring the work function7. But KPFM has difficulty in quantitatively characterizing ferroelectric devices that include multiple materials because disentangling the electrostatic force from individual layers is a formidable task.Creating ferroelectricity by stacking vdW layers provides new opportunities to engineer materials and control their optoelectronic properties, as the ferroelectric polarization is expected to modulate the doping of an adjacent material2 (t-WSe2) revealing ferroelectricity. We first demonstrate that the plasmonic response of graphene is altered by the proximate twisted ferroelectric domains. Notably, by investigating the local plasmonic contrast in graphene residing underneath the ferroelectric domains, we obtain a reading of the local ferroelectric polarization. Moreover, we show that the proximity to the ferroelectric bilayer breaks the inversion symmetry and modulates the Seebeck coefficient of graphene, thereby allowing the generation of photocurrent. The nano-photocurrent mapping displays moiré patterns and further confirms the notion of the local modulation of the carrier density in graphene prompted by ferroelectric domains. These results uncover alternative approaches to controlling the optoelectronic response of graphene integrated with ferroelectric materials.Here, we utilize near-field infrared nano-imaging and nano-photocurrent to investigate the optical and optoelectronic properties of back-gated graphene encapsulated with a minimally twisted bilayer of WSe2 devices with the same general architecture. These devices are based on back-gated graphene structures, with graphene encapsulated by a minimally twisted bilayer of WSe2 on the top and h-BN at the bottom  the magnitude of the near-field scattering signal35 and (ii) the periodic oscillating patterns (fringes) arising from the propagating plasmon polaritons37. In principle, both observables are governed by the carrier density in graphene. In practice, for samples with moiré super-structures, all domain boundaries are potentially capable of launching and reflecting propagating plasmons38, leading to complex patterns that make it difficult to extract the local carrier density. Therefore, we will primarily focus on an analysis of the evolution of the near-field amplitude The experimental concept is outlined in Fig. er8 Fig. . The fermentclass2pt{minim−1 is shown in Fig. 39. The period of the moiré patterns is inhomogeneous as the twist angle between the WSe2 layers varies over the field of view due to strain and wrinkles. Indeed, across the wrinkles, the observed moiré periods prominently change. Intriguingly, the nearest-neighbor domains reveal clear contrast, while the next-nearest-neighbor domains show almost the same near-field intensity. We performed measurements for several devices, all of which consistently produced these moiré features  and graphene, a process governed by the Fermi energy43.  experiments45. Nano-photocurrent imaging has been extensively used to investigate domains and domain walls in twisted bilayer graphene46. The formation of photocurrent requires broken inversion symmetry, which can be accomplished by introducing p-n or n-n+ junctions in the graphene layer. In our devices, the staggered ferroelectric potential breaks the inversion symmetry, thereby allowing photocurrent formation.Now we use nano-photocurrent imaging to study how the moiré ferroelectricity of t-WSe2 displays moiré patterns marked by prominent sign flipping: red and blue colors denote positive and negative currents, respectively. In addition to the moiré regions, the photocurrent flips sign across wrinkles and terrace edges, as shown in Fig. 48.The nano-photocurrent mapping in Fig. 49, Now we elucidate how the PTE generates the photocurrent and interpret the above photocurrent features. In our experiment, when the tip is scanned over the devices, the tip-focused electric field increases the electron temperature of graphene. As a result, local thermoelectric current The first term on the right-hand side of Eq.  denotes The Seebeck coefficient of graphene depends on the Fermi energy Fig. . As the With the photocurrent mechanism in mind, we can gain a deeper understanding of the spatial features of the photocurrent. When graphene is locally heated, the heat spreads on a characteristic length referred to as the cooling length, which is hundreds of nanometers  and the use of a Bruker atomic force microscope (AFM). Monolayer WSe2 was first cut into two halves using a laser cauterization method and then underwent a dry stacking process in which an h-BN/graphene stack was used to pick up the two pieces of identical WSe2 without any intentional rotation. The final stack was flipped upside down to expose the WSe2/graphene upward using a flipped chip method55. Finally, we used low-temperature, high-vacuum annealing to remove the buried polypropylene carbonate (PPC). The final device geometry was defined by electron-beam lithography and reactive ion etching , followed by electron-beam evaporation (EBE) to deposit Cr/Au = 5/150 nm as the surface contacts. Piezoresponse force microscopy (PFM) imaging was achieved with the Bruker atomic force microscope.Monolayer WSeThe nano-infrared scattering experiments were performed using a home-built scattering-type scanning near-field optical microscope (s-SNOM) housed in an ultra-high vacuum chamber with a base pressure of ~The s-SNOM works by scattering tightly focused light from a sharp AFM tip. The spatial resolution of the s-SNOM is predominantly set by the tip radius of curvature The tapping amplitude is used as the z feedback. The tip excitation phase is controlled by a phase lock loop. The AFM also has the functions of a side-band Kelvin probe force microscope, and the feedback bias voltage is applied onto the AFM tip to compensate for the potential difference between the tip and the sample. This applied bias can ensure that the tip vibration is not influenced by the ferroelectric potential. Thus, the observed near-field contrast is the genuine near-field scattering signal, which is generated purely from the sample conductivity. During the near-field mapping, the z feedback, phase lock loop, and Kelvin probe force microscope are turned on simultaneously. We note that for a tip with a resonance frequency of 285 kHz, the large force constant of about 42 N/m makes it insensitive to the potential difference between the sample and the tip, so no discernible potential response is detected by the tip. Therefore, the KPFM function is not required when using tips with a stiff cantilever.-1 was used to avoid phonon resonances from substrate h-BN and SiO2. The laser beam was focused onto the metallized AFM tip using a parabolic mirror with a 12-mm focal length. The back-scattered light was registered by a mercury cadmium telluride (MCT) detector and demodulated following a pseudoheterodyne scheme. The signal was demodulated at the nth harmonic of the tapping frequency, yielding background-free images. To eliminate the far-field background, we chose n = 3 and 4 in this work.The laser source is a tunable quantum cascade laser (QCL) from Daylight Solutions. Photon energy of 860 ~ 920 cm7 and corresponding bandwidth >1 MHz. In order to isolate the photocurrent generated by the near fields underneath the tip, the photocurrent was sent to a lock-in amplifier and demodulated at the nth harmonic of the tip tapping frequency. In this work, n = 2 was used.The nano-photocurrent measurements were performed in a home-built s-SNOM housed in an ultra-high vacuum chamber with a base pressure of ~Supplementary InformationPeer Review File


Fixed dose combination (FDC) dolutegravir (DTG) plus rilpivirine (RPV) is an approved antiretroviral treatment regimen for people with HIV. The steady-state PK of FDC DTG+RPV in those requiring hemodialysis (HD) has not been previously studied.We performed a single-center, prospective evaluation of the steady-state PK of FDC DTG (50mg)+RPV(25mg) in HIV-negative adults either requiring HD  or with normal renal function, defined as CrCl ≥ 75mL/min . All participants received DTG+RPV daily for 10-14 days with food before undergoing an intensive 24-hour PK evaluation . Plasma drug and metabolite concentrations were measured using a validated LC/MS/MS assay method (QTRAP 6500+LC-MS/MS system) with turboelectrospray source operating in both positive (confirmation) and negative (quantification) modes. We did not evaluate dialysis extraction of DTG+RPV. Descriptive PK parameters were calculated.2). The Table shows the PK parameters assessed in the study population for circulating plasma DTG, DTG-glucuronide (DTG’s primary metabolite), and RPV.No participant experienced serious or grade 3-4 adverse events; there were no study discontinuations. The 4 HD and 2 normal renal function participants were of similar ages  and BMI  and RPV (12ng/mL). These data suggest no dosing modifications are needed for the FDC DTG+RPV regimen in HD.In this study, HD did not lead to clinically appreciable differential exposures to DTG and RPV; the markedly increased exposure to DTG-glucoranide (which is considered inert) in HD suggests increased UGT1A1 activation. All participants maintained exposures throughout the dosing interval greater than the reported ICSamir K. Gupta, MD, Gilead Sciences: Advisor/Consultant|ViiV Healthcare: Advisor/Consultant|ViiV Healthcare: Grant/Research Support Allon Friedman, MD, Eli Lilly: Advisor/Consultant|Eli Lilly: Stocks/Bonds|GI Dynamics: Advisor/Consultant|Gila Therapeutics: Advisor/Consultant


Previous research has indicated altered attentional processing in individuals with experiences of maltreatment or victimization in childhood and adolescence. The present study examined the impact of child and adolescent experiences of relational peer victimization on attentional processes in adulthood when confronted with emotional facial expressions.As part of an online study, a community sample of adults completed a facial dot-probe task. In the present task, pictures of facial expressions displaying four different emotions  were used.The results of the hierarchical regression analyses showed that retrospective reports of peer victimization made a significant contribution to the prediction of facilitated orienting processes for sad facial expressions. Experiences of emotional child maltreatment, on the other hand, made a significant contribution to the prediction of attentional biases for angry facial expressions.Our results emphasize the relevance of experiences of emotional and relational maltreatment in childhood and in adolescence for the processing of social stimuli in adulthood. The findings regarding emotional child maltreatment are more indicative of attentional biases in the context of threat detection, whereas the altered attentional processes in peer victimization are more indicative of mood-congruent biases. These altered processes may be active in social situations and may therefore influence future social situations, behavior, feelings, and thus mental health. There are a variety of studies demonstrating the negative impact of maltreatment experiences in a peer context in childhood and adolescence on psychosocial adjustment and particularly mental health  and Psychotherapeutische Ambulanz der Universität Bielefeld (PAdUB)] were recruited for participation. The flyer contained information about the aims and methods of the study as well as a notice about the anonymity of the participation. At the beginning of the experiment information on general sociodemographic variables such as age, gender, educational level, and family status were requested. Furthermore, the instrument assessed the presence of mental illnesses, the use of medication, and other physical and psychological health questions in addition to the questionnaires used in this study. The sociodemographic and psychopathological characteristics of the 90 participants who were included in the analyses can be found in Participants were recruited through the distribution of the participation link or QR code with access to the study Questionnaires were administered using the Qualtrics survey platform. For the experiment, the web version of the program Inquisit 6 (Millisecond software) was used. At the beginning of the study, participants were informed that participation was voluntary and that it was possible to quit the study at any time without penalty. They were also informed that participation would not be remunerated and that confidential information about mental health symptoms and stressful life experiences would be collected. Participation was only possible after participants had given their consent to participate by clicking on a box. After answering the questionnaires, participants were redirected to the Inquisit homepage, from where the Inquisit application could be downloaded. The experiment was designed in such a way that it could be carried out on both computers and mobile devices. The procedure was approved by the Ethics Committee of Bielefeld University.To assess general symptoms of psychopathology the Symptom Check List-27 . In addition, individual trials were removed in which the reaction time deviated more or less than 2 SD from the individual mean reaction time . For measuring the attentional bias scores for each trial type  the overall attentional bias score was calculated by subtracting the reaction times for congruent trials  from the reaction time for incongruent trials . Attention biases are reflected in shorter reaction times for the dot when attention was focused on this area and longer reaction times for the dot when attention was not focused there. Based on the calculation of the score, positive values for the attentional bias score indicated that the attention was on the emotional faces, whereas negative values indicated that the attention of the subjects was not on the emotional face, but the neutral face. To specify altered attentional processes more precisely with respect to the different types of attentional biases, the orientation score and the disengaging score were calculated in addition to the attentional bias score to capture processes of facilitated attention or difficulties of disengagement . Thus, with α = 0.05, power = 0.95, and the initially planned inclusion of seven predictors  the required sample size was N = 86.For sample size estimation a statistical power analysis was calculated for the multiple regression analyses using G Power 3.1  correction  and the STAI scores , only the SCL-27 was included in the regression analyses. In a second step, the sum scores of the CTQ subscales were included in the model . In a final step, the FBS sum score  was included as the last predictor in the model. These regression analyses were conducted separately for the individual bias indices and the respective emotion presented. Participants who scored on all three items of the minimization/denial subscale of the CTQ (n = 3) were excluded from the analyses . For all analyses, a significance level of t-tests of the absolute index scores for the presentation of one emotion each are shown in t-tests showed that the orienting scores for disgusted and happy faces differed significantly from zero and were negative, indicating attentional avoidance of happy and disgusted facial expressions.A detailed description of the attentional bias index scores for the different emotions, the mean values, standard deviations, and one-sample p's > 0.05). The analyses showed a positive correlation between the emotional abuse score and the disengaging score for sad-neutral trials as well as a positive correlation between the emotional neglect score and the disengaging score for sad-neutral trials.The bivariate Pearson correlation coefficients between maltreatment experiences and the different index scores for each trial type can be found in F = 2.34, adjusted R2 = 0.08, p = 0.039]. There was no significant relationship between the index scores and the level of peer victimization experiences for angry faces  = 0.91, adjusted R2 = –0.01, p = 0.494], the orienting score  and the disengaging score . In addition, there were no significant effects for trials in which happy facial expressions were presented for the attentional bias score, the orienting score, and the disengaging score  = 3.08, adjusted R2 = 0.12, p = 0.009]. The associations with the attentional bias score behaved in opposite ways. Higher scores on the emotional abuse subscale were associated with higher attentional bias scores, whereas higher scores on emotional neglect were associated with lower attentional bias scores on angry faces. For the orienting score, emotional abuse was a significant predictor, with an overall non-significant final model . Associations between emotional maltreatment experiences and the orienting score for happy faces could also be found in happy-neutral trials  = 1.86, adjusted R2 = 0.06, p = 0.098].Regarding the prediction of attentional biases in angry-neutral trials see , the regials see . This reGiven the ambiguous findings on attentional biases in the context of peer victimization, the present work provided new insights into the relationship between relational peer victimization and attentional biases beyond the influence of child maltreatment experiences. In this context, the present study was designed to provide differentiated accounts of attentional biases in the context of maltreatment and peer victimization experiences, thus extending previous research. Consistent with our hypothesis we found altered attentional processes in individuals reporting higher levels of victimization experiences in the present sample. However, this influence was found in sad faces and not, as previously hypothesized, in emotions that were expected to be relevant as threatening stimuli in the context of peer victimization. Furthermore, altered attention processes were found in individuals reporting experiences of emotional maltreatment when confronted with angry facial expressions.In the present study, the results indicated evidence for facilitated attention to sad facial expressions in individuals with higher levels of relational peer victimization experiences beyond the influence of experiences of child maltreatment. This effect was seen even when controlling for symptoms of psychopathology. These findings were consistent with the results of Günther et al. , who alst-tests suggest that participants generally showed significant avoidance of happy and disgusted faces. Therefore, the findings for the overall sample are not reflected in the analyses for peer victimization. The present results contradict the findings of Iffland et al.  had an influence on the results. In addition, the online study could not control the situational conditions under which the performance took place. However, by adjusting the experimental data, we were able to minimize the influence that the behavior would have had if the instructions had not been followed or if the subjects had been unfocused or distracted. Moreover, results were consistent with the findings of several studies that have used reaction times, and also neural measures (Günther et al., Limitations of the present study must be considered when interpreting the results. One-third of the participants stated that they were currently suffering from a mental disorder. For reasons of anonymity, no information could be collected on whether participants were patients of the outpatient clinics. It cannot be excluded that treatment or current medication influenced the attentional processes or reaction times. However, by including psychopathological symptom severity in the regression models, and by adjusting the reaction time data, we were able to reduce the potential influence on our results. In addition, the data did not allow us to assert causal relationships due to the cross-sectional design of our study. Longitudinal studies for analyzing the relationship to mental health should be addressed in the future. The retrospective assessment of experiences of child maltreatment and peer victimization as self-reports also limits the interpretability of the results as they may be affected by distortions (Baldwin et al., In line with previous results, our study showed that experiences of relational peer victimization and emotional child maltreatment in childhood and adolescence influence attentional processes in adulthood. Higher levels of peer victimization were associated with facilitated attention to sad facial expressions in our sample. The results are thus indicative of mood-congruent attentional biases in individuals who have experienced relational peer violence. In addition, altered attentional processes for angry faces were present in participants with higher levels of emotional child maltreatment experiences. Adverse childhood experiences, particularly experiences of emotional maltreatment and relational peer victimization, can thus be considered relevant to the development of cognitive schemata that continue to be activated in adulthood, and therefore can potentially influence new experiences, feelings, thoughts in social situations, and thus presumably mental health.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by Ethics Committee of Bielefeld University. Written informed consent was not provided because of the design as an online study. The participants gave their informed consent by clicking on a box.KB contributed to the conception and design of the work, the acquisition, analysis, interpretation of the data, drafted, revised, approved the manuscript, and ensures the accuracy and integrity of any part of the work. BI was the chief investigator for this study, contributed to the conception of the study, supervised data analyses, participated in the interpretation of the data, and critically revised the manuscript for important intellectual content. All authors contributed to the article and approved the submitted version.


Graphene, as a promising material, holds the potential to significantly enhance the field of dental practices. Incorporating graphene into dental materials imparts enhanced strength and durability, while graphene-based nanocomposites offer the prospect of innovative solutions such as antimicrobial dental implants or scaffolds. Ongoing research into graphene-based dental adhesives and composites also suggests their capacity to improve the quality and reliability of dental restorations. This narrative review aims to provide an up-to-date overview of the application of graphene derivatives in the dental domain, with a particular focus on their application in prosthodontics and periodontics. It is important to acknowledge that further research and development are imperative to fully explore the potential of graphene and ensure its safe use in dental practices. Dental diseases are a worldwide concern due to their high prevalence and incidence . MoreovePeriodontitis is an infectious-inflammatory disease that, as it progresses, leads to the destruction of the tissues that support the tooth on the dental arch, which may result in tooth loss. In addition, periodontitis has been associated with the negative evolution and aggravation of some systemic conditions and diseases, such as diabetes mellitus , cardiovEdentulism represents a complex, impairing condition with profound implications for performing functions such as mastication, swallowing, phonation, and aesthetics . EdentulThus, establishing effective dental treatment as well as identifying appropriate therapeutic methods is a priority. Dental technology constantly evolves, and new materials have been developed that respond favorably to mechanical stress and oral bacterial conditions. In this regard, nanomaterials have been emphasized as biomaterials with superior physical, chemical, and biological properties .Nanotechnology is a prerogative of the 21st century and impacts the medical field. Any material used for medical purposes, especially those that have long-term contact with the human body, must have specific biological and mechanical properties, such as not interfering with the host environment, not allowing the growth of bacteria on its surface, and not initiating a local inflammatory response.Graphene fulfills these characteristics, excelling in antibacterial properties and gaining ground in most medical fields, especially dentistry. The antimicrobial effect of graphene-based materials (GBMs) is one of their most exciting properties. This effect is attributed to their physical interaction mechanism with bacteria, which is likely to prevent the development of antimicrobial resistance (AMR) ,18. UnliOne question arises: What is graphene? Graphene is a carbon-based material, first theorized by PR Wallace in 1947 and obtained by Andre Geim and Konstantin Novoselov in 2004 , for whiChemical modifications have produced graphene derivatives, including graphene oxide (GO) and reduced graphene oxide (rGO). These derivatives are highly versatile and more applicable .2 weighs less than 1 mg), and ecological [Among the properties that make graphene so special are: stability at extremely small dimensions , high level of cohesion, hexagonal configuration, and being the most robust material known so far—stronger than steel (200 times), with the same density as diamond (four layers of graphene would support an elephant) , elasticological .Many applications of this material have been implemented in various fields, starting with electronics , energy production and storage , aviation (improving the structure of aircraft wings), telephony, and computers .Medicine is another field that promises to be transformed by graphene. Because this material is thin, flexible, and resistant to the saline solutions that are part of living tissues, graphene is ideal for designing bionic devices. Unlike metal devices, which last only a few years in the human body, graphene can be used for life. Moreover, because graphene is an electrical conductor, it could be used to transmit electrical impulses to neurons, allowing paralyzed people to regain control of their limbs following an accident that resulted in spinal cord damage . The exaThe main fields in which graphene and its derivatives are applied are presented in Among the medical fields that have explored the potential of graphene and its derivatives is dentistry. Ge et al. and Desante et al. emphasized the existing research void regarding the potential applications of graphene in dentistry, which contrasts with its comprehensive utilization in various other medical domains such as drug delivery, imaging agents, biosensors, and tissue engineering scaffolds ,36 Figu. HoweverResearchers at Princeton University have conducted a noteworthy study where graphene is incorporated onto the tooth surface as a tattoo . As grapOur literature research aimed to provide readers with a “broad view” of the potential and applications of graphene-based materials (GBMs) for the treatment of oral conditions, with a specific focus on periodontal disease and edentulism.A narrative review was conducted aiming to answer the question, “Can graphene and its derivatives contribute to the improvement of periodontal and prosthetic treatment”?An electronic search was conducted in the Web of Science digital database using MeSH terms and unspecific free-text words with Boolean operators. The final search terms were defined after iterative trial and refinement processes. The strategy employed was as follows: “graphene periodontal” OR “graphene dental prosthesis” OR “graphene dental material” OR “graphene dental implant” OR “graphene dental restoration” OR “graphene dental scaffold” OR “graphene dental PMMA”, within the topic field, without imposing any year restrictions. The most recent search was carried out in July 2023, yielding a total of 288 studies. Review articles, book chapters, editorial materials, and studies unrelated to the medical field were excluded, resulting in 168 studies published in English between 2013 and 2023 . Followi2/g) [Graphene oxide (GO) and reduced graphene oxide (rGO) are the leading derivatives of graphene, as shown in 2/g) , and its2/g) . The GO 2/g) . Moreove2/g) . GO also2/g) .Reduced graphene oxide (rGO) is obtained by reducing graphene oxide. This process is ecological, and the resulting product is biocompatible and dispersible . rGO canGraphene quantum dots (GQDs) are produced from graphene oxide. They have strong quantum confinement and photoluminescence properties. These properties make GQDs successful in cell imaging .Nanocomposites that use graphene as a base are created by attaching other active agents to either GO or rGO through covalent or non-covalent bonds . For thiGraphene oxide can bond with silver nanoparticles, forming a stable product that favors a more efficient dispersion, which leads to increased antibacterial properties . It has 3O4), or titanium oxide (TiO2) have also been used. The addition of ZnO also enhanced the antibacterial effect of GO based on the membrane stress caused by sharp edges on microbial cells [In addition to metal ions, oxides such as zinc oxide (ZnO), iron oxide  , polyvinThe antimicrobial mechanisms of graphene derivatives are based on both their chemical and mechanical effects . In termAnother antimicrobial mechanism is based on cellular uptake. Following contact between graphene sheets and bacterial cells, the latter are trapped, isolated from the environment, and without access to nutrients . This efChemically, GO can induce lipid peroxidation in bacteria . This phPorphyromonas gingivalis or Fusobacterium nucleatum, are more susceptible to GO than facultative anaerobes (such as Streptococcus mutans) [There is a widely held belief that graphene oxide (GO) possesses a greater capacity to generate reactive oxygen species (ROS) and oxidize reduced glutathione than reduced graphene oxide (rGO) . However mutans) .It is commonly understood that the effects in question are attributable to a combination of mechanical and chemical processes that occur gradually over time. Oxidative stress can facilitate the damage of bacterial membranes by the sharp edges of graphene derivatives through ATP degradation .Candida albicans, Lactobacillus acidophilus, S. mutans, and Aggregatibacter actinomycetemcomitans of rGO and silver (rGp-NS-Ag) composites with those of silver nanoparticles (AgNP) and rGO nanosheets alone. rGp-NS-Ag generated enhanced antimicrobial effects [P. gingivalis in the percentage of 95.45%, as well as a low rate of bacterial adhesion (4.55%), and showed impacts by data related to microstructures, quantity, cell membrane disruption, bacterial cell apoptosis, and bacterial gene expression [The antibacterial effect of graphene derivatives on periodontopathogenic bacteria was investigated. Peng et al. compared the antimicrobial activities against  effects . Treatmepression .S. mutans, F. nucleatum, and P. gingivalis, was assessed. The approach involved using varying concentrations of nanosheets  to determine their impact on bacterial growth. The results of this study revealed that at a concentration of 40 μg/mL, both P. gingivalis and F. nucleatum exhibited complete inhibition of growth in the presence of GO nanosheets. However, a reduction of S. mutans was observed at a concentration of 80 μg/mL [The antibacterial efficacy of GO nanosheets on three distinct bacterial strains, 80 μg/mL . The mecS. mutans, F. nucleatum, and P. gingivalis) [P. gingivalis at 96 h. Moreover, the authors concluded it was broadly effective against multiple pathogens rather than just one strain. It is plausible that the transfer of electrons from bacterial biofilms to the graphene-reinforced titanium element is the underlying mechanism responsible for its anti-bacterial effect. This transfer has been observed to disrupt the bacterial respiratory chain, ultimately reducing microbial viability [P. gingivalis and F. nucleatum included [Graphene-reinforced titanium (Ti-0.125G) was evaluated against the same pathogens  . The deviability . Qin et included .P. gingivalis, F. nucleatum, and C. albicans). When coated with graphene, the researchers observed that the Ti-6Al-4V alloy exhibited enhanced resistance against oral pathogens compared to the uncoated Ti-6Al-4V alloy. Moreover, the graphene-coated Ti-6Al-4V alloy generated a higher concentration of ROS in the pathogens tested than the uncoated Ti-6Al-4V alloy [Wang et al. investigated the antimicrobial properties of graphene-coated Ti-6Al-4V against oral pathogens . The antibacterial mechanism involved is based on the generation of ROS [Pourhajibagher et al. demonstrated that curcumin-coupled GQDs suppress biofilm formation capacity by 76% for periodontopathogenic bacteria n of ROS . Trusek n of ROS . The maiFavorable osseointegration for ceramic and titanium implants and their alloys is still a major challenge in implant-prosthetic therapy. To increase the success and survival rate of implants while reducing potential complications following their placement, treatment of bioinert implant surfaces with various materials has been attempted, aiming for enhanced osseointegration through antimicrobial activity and functionalization of the tissue-implant interface. This interface is the site of interaction with the surrounding tissues where all the osseointegration processes occur—inflammatory reactions, cell recruitment, adsorption of proteins, or biofilm formation .Therefore, graphene is utilized in implants due to its exceptional attributes, including high biocompatibility, physical interaction with biomolecules such as proteins, enzymes, or peptides , effectiSeveral techniques, such as chemical vapor deposition , plasma GO-coated titanium implants stimulated cell proliferation, increased alkaline phosphatase (ALP) activity levels and gene expression levels of osteogenesis-related markers, and promoted BSP, Runx2, and OCN protein expression . Moreove2 nanotubes resulted in significant improvements in various cellular functions, including proliferation, adhesion, migration, and the expression of genes related to adhesion. These enhancements are critical for achieving successful soft tissue sealing [The findings of Cao et al. indicate that the growth of human gingival fibroblasts on TiO sealing .In their research, Gao and his team employed a sandwich-structured dental implant coating featuring graphene oxide encapsulated within mineralized collagen. This innovative structure demonstrated antibacterial properties and improved adhesion, cytoskeleton organization, and proliferation of human gingival fibroblasts, ultimately leading to superior soft tissue sealing .Enrichment of implant surfaces with GO and bioactive proteins was also attempted. Bone morphogenetic proteins (BMPs) are a class of proteins that can remarkably trigger bone growth. Among them, BMP-2 stands out as a potent factor that facilitates the differentiation of stem cells into bone cells, thereby augmenting the integration of implants by encouraging bone regeneration in the region between the implant and the recipient site . The impA study conducted by Ren et al. investigated the impact of GO and rGO dexamethasone-loaded titanium foils, specifically DEX-GO-Ti and DEX-rGO-Ti, on the proliferation and osteodifferentiation of rat bone mesenchymal stem cells (rBMSCs). The study results showed that DEX-GO-Ti significantly enhanced cell proliferation. At the same time, rBMSCs cultivated on DEX-GO-Ti demonstrated elevated expression levels of calcium, proteins, and mRNA, which are closely associated with osteogenic differentiation .Titanium implant abutment surfaces treated with minocycline hydrochloride (MH)-loaded graphene oxide films were placed in a canine model of peri-implantitis  and comp2) are attractive due to their mechanical, physical, and high chemical and thermal stability, combined with the absence of corrosion or toxicity, leading to decreased peri-implant inflammatory reactions [If the literature abounds in research on improving the surface condition of titanium implants with graphene-based materials, there are fewer studies on the association of these materials with zirconia-based implants . Zirconieactions  and higheactions ,118,119 eactions ,128,129.Two groups of graphene-based 2D nanomaterial (GBN) fillers for ceramic composites can be distinguished depending on the number of graphene sheets. Graphene nanoplatelets (GNP) have more than ten layers and a thickness lower than 100 nm, while multi-layered graphene (MLG) has fewer than ten layers. The latter are divided into two classes: reduced graphene oxide (rGO) and few-layer graphene (FLG), with two to about five layers . GraphenAs for coating materials for zirconia surfaces, few attempts were made. Kou et al. produced functionalized multi-walled carbon nanotubes (fMWCNTs) that improved Saos-2 cell attachment by increasing the surface roughness of coated zirconia-based ceramic surfaces . Further2/GNs) composite with a homogeneous distribution of GNs in the ZrO2 matrix using the plasma spraying technique [Li et al. produced a zirconia/graphene nanosheets  sheets through electrostatic forces, resulting in the collection of zirconia ions on the surface of the GO sheets. The characterization techniques employed to analyze the structure of the obtained materials confirmed that the synthesis route assured the successful bonding of zirconia nanoparticles to graphene oxide sheets. It allowed the efficient production of uniform zirconia/graphene nanopowders, both economically and practically.A homogeneous precursor powder must be obtained to produce zirconia-based graphene-containing composites, which are then compacted and sintered. The processing route of graphene-based ceramic composites is of paramount importance for the mechanical properties of the final material . In 2020O powder . The proLorusso et al. conducted a study aimed at assessing the efficacy of biomaterials derived from polymethylmethacrylate (PMMA) in terms of bone integration, with a specific focus on PMMA infused with graphene (GD-PMMA) . In theiPEEK material has become increasingly prevalent in dentistry due to its diverse applications. These applications encompass oral implant treatments such as oral implants and implant abutments, restorative dentistry procedures such as crowns, fixed and removable dentures, posts and cores, and maxillofacial prosthetics, as well as other oral applications such as orthodontic wires and retainers and scaffolds for cartilage repair . It is fIn recent years, advancements have been made in using materials such as carbon nanotubes (CNTs) and graphene to reinforce PEEK composites. The treatment of the surface of the CF/PEEK composite with concentrated sulfuric acid leads to the creation of a three-dimensional porous network. Notably, an increased CF content results in larger pore sizes within this porous layer. Following treatment with a solution of graphene oxide (GO), the sulfonated material exhibits the development of filamentous GO folds on its surface. It has been observed that the contact angles of all samples increase post-sulfonation. However, the GO functional wrinkles cause a significant reduction in the contact angle of the material, leading to an increase in surface hydrophilicity .biomediGraphene oxide is also used to make barrier membranes and scaffolds for periodontal tissue regeneration. During bone regeneration, a barrier membrane is utilized to prevent the infiltration of epithelial cells into the neo-formation site. This promotes optimal healing and ensures the development of new bone tissue. Moreover, such a membrane must possess several properties, which include biocompatibility, site protection and maintenance, cell occlusion, tissue attachment, and clinical sensitivity . In the S. oralis, Veilonella parvula, F. nucleatum, and P. gingivalis, the GO-Ti membrane did not produce toxic or inflammatory effects. Moreover, the multiplication of human gingival fibroblasts and osteoblastic promotion were observed [A titanium membrane was coated with GO in a study conducted by Radunovic. Besides the antibacterial effect exerted on observed . De Marcobserved .Scaffolds for use in periodontal tissue regeneration that have been investigated were made of materials including collagen , poly signaling pathways, which results in significant stimulation of bone growth in laboratory conditions. Furthermore, the use of these scaffolds has demonstrated a noteworthy ability to facilitate bone regeneration in calvarial defects in live subjects .Scaffolds based on chitosan and GO exhibited high water retention capacity, porosity, and hydrophilic nature . HermeneThe association between graphene derivatives and natural polymers, primarily collagen, has also been investigated. Nishida et al. fabricated a GO-coated collagen sponge scaffold and implanted it into the post-extraction alveolus in canine subjects. This scaffold resulted in a fivefold stimulation of bone neoformation compared to the control group .A bilayer composite was synthesized by incorporating silk fibroin into graphene oxide and reduced graphene oxide. This composite has been observed to exhibit beneficial properties in promoting cell proliferation and inducing osteoblastic and cementoblastic cell differentiation in periodontal ligament stem cells . AdditioMulti-composite structures have been developed to enhance the mechanical and biological characteristics of scaffolds. Wang et al. synthesized a scaffold of GO and a collagen/nanohydroxyapatite nanocomposite. The high porosity of the obtained product exhibited improved hydrophilic and mechanical properties and outstanding proliferation potential .Scaffolds based on hydroxyapatite, chitosan, and graphene oxide promoted cell adhesion and proliferation and improved osteogenesis in vitro tests ,176. TheA research team led by Zhang has successfully developed a method that utilizes water-soluble graphene oxide-copper (GO-Cu) nanocomposites to coat porous calcium phosphate (CaP) scaffolds. This innovative coating has demonstrated the ability to significantly enhance the adhesion and osteogenic differentiation of rat bone marrow-derived stem cells (BMSCs). Moreover, these scaffolds stimulated angiogenesis and osteogenesis when transplanted into rats with calvarial defects . A GO scGraphene-based 3D scaffolds have also been developed and investigated for stem cell growth and differentiation. Crowder et al. employed three-dimensional graphene foams as culture substrates for human mesenchymal stem cells. The cells maintained their viability for one week and strongly expressed the osteogenic markers osteocalcin and osteopontin, indicating their spontaneous osteogenic differentiation without extrinsic growth factors . The stuThe great potential of graphene derivates has allowed their use in various fields of dentistry. For direct or indirect restorations, restorative dentistry employs composites, adhesives, and cement types with aesthetic properties and high hardness. However, they face limitations due to high polymerization shrinkage and poor antibacterial properties. Graphene nanoplates (GNPs) are utilized as nanofillers in porous and prone-to-dissolution materials such as resins, cement, and adhesives to effectively reinforce commonly used dental composites and exert an anticaries effect.Adding graphene nanosheets to two different powders of bioactive calcium silicate cement (Biodentine and Endocem Zr) improved bonding time and hardness. However, Endocem Zr experienced significant impairment in bonding properties when adding GNPs. This indicates that while GNPs enhance the physical-mechanical properties of materials, they may not be suitable for all materials in terms of bonding . IncorpoStaphylococcus aureus and S. mutans while exhibiting a favorable release rate of fluoride ions [A graphene variant, Fluorinated Graphene (FG), has been developed and incorporated into glass ionomer cement. In dentistry, FG has emerged as a more desirable option than conventional gray GNPs owing to its visually appealing bright white color, which renders it an excellent choice for aesthetic applications. FG has been used to modify poly(acrylic acid)-based glass ionomer types of cement (GICs), offering advantages in mechanical, tribological, and antibacterial properties. Therefore, the GIC/FG composites exhibit increased Vickers microhardness and compression, flexural strength, and a decreased friction coefficient. This widens the application of glass ionomer cement in restorative dentistry for various procedures, such as restoration of non-carious and carious lesions, class III and class V restorations, and crown cementation. Furthermore, these compounds demonstrate potent antibacterial activity against ide ions .The advantages of GBMs have led to their application in enhancing the properties of adhesive materials . Dental S. mutans cells while simultaneously preserving their bonding properties [GNPs are frequently utilized as fillers in polymer-based dental adhesives owing to their potent antimicrobial and antibiofilm characteristics. These nanocomposites, filled with GNPs, exhibit efficacy in suppressing the activity of operties . Therefooperties . Accordioperties .A research team led by Nizami has successfully developed a composite material known as nHAP/MWCNT-GO that exhibits exceptional properties as a shield for dentin. The composite comprises nanohydroxyapatite, multi-walled carbon nanotubes, and graphene oxide. This material forms a surface film that effectively resists acid and minimizes dentin erosion .S. mutans. Furthermore, the composite demonstrated low cytotoxicity, except at higher concentrations of approximately 0.1 w/v% [In addition, graphene oxide (GO) has been modified by incorporating different nanoparticles, such as calcium fluoride, silver, and tricalcium phosphate, to prevent dentin decalcification. GO combined with silver and silver-calcium fluoride exhibited inhibition of 0.1 w/v% .2 [2, improves surface roughness, and slightly increases the water contact angle [The application of silane primer significantly influences the bonding of zirconia, with the adhesive layer proving to have low mechanical properties . Incorpo2 . Adding ct angle .Dentistry currently utilizes various types of medical materials, each with its own advantages and disadvantages. Regarding prosthodontics, there is a great diversity of materials for indirect restorations, especially those used in CAD/CAM systems . Due to For the past 80 years, polymethylmethacrylate (PMMA) resin has remained a highly favored material within the realm of prosthetic dentistry. This is particularly true for the manufacturing of complete and removable partial dentures. One of the reasons for this popularity is the material’s ease of manufacture, affordable cost, pleasing aesthetic properties, and low modulus of elasticity. Moreover, PMMA resin also boasts easy repair capabilities, making it a versatile choice for dental professionals. Polymethyl methacrylate (PMMA)-based resins find extensive application in provisional restorations due to their low mechanical properties that prevent their use for permanent restorations, considerable polymerization shrinkage, and poor inhibition of biofilms that restrict their use in permanent restorations . EffortsAzevedo et al. have achieved a remarkable result in the realm of maxillary full arch rehabilitation by introducing graphene oxide into PMMA resin, resulting in complete restoration . Recent A study was conducted by Abad-Coronel et al. to compare the fracture resistance of various materials used in temporary restorations. The materials included PMMA, graphene-modified PMMA (GRA), acetal resin (AR), and polysulfone (PS). The purpose of the study was to evaluate the effectiveness of different materials for temporary restorations and their ability to resist fractures. The materials were employed in the fabrication of a three-unit fixed dental prosthesis (FDP) using a milling process facilitated by a CAD/CAM system. During compression testing, it was observed that PMMA displayed substantially lower values in contrast to other materials, whereas PS exhibited the highest values. Moreover, GRA and AR showed similar values, which were still greater than those of PMMA. These findings suggest that GRA, AR, and PS are feasible options for interim milled restorative materials and can potentially serve as substitutes for PMMA. This study highlights the comparative fracture resistance of various materials utilized in temporary restorations and emphasizes the potential benefits of GRA, AR, and PS in this domain .S. aureus, Escherichia coli, and S. mutans [A study was conducted by Bacali et al. that focused on the incorporation of graphene-silver nanoparticles (Gr-Ag) in PMMA. The study aimed to assess the material’s mechanical properties, hydrophilic abilities, and morphology . Accordi. mutans .Candida albicans after 28 days of culturing, indicating a positive impact on hydrophilicity. These results suggest that incorporating nGO into PMMA may have potential applications in improving antimicrobial and antiadhesive properties for medical devices and implants.The utilization of graphene oxide nanosheets (nGO) by Lee and colleagues has been found to effectively enhance the antimicrobial and adhesive properties of PMMA resin . The finAdding less than 1% graphene nanoparticles to the component of PMMA improves the physical characteristics and confers antimicrobial activity . The conNumerous studies have investigated the intrinsic antimicrobial properties of graphene . These pAccording to previous research, the incorporation of graphene within PMMA did not yield a noteworthy effect on the adhesion of microorganisms. This is attributed to the intricate mechanics of microbial adhesion, which are subject to the topographical features and roughness of the material surface and the very low concentration of graphene present in the material .C. albicans growth displayed a proportional relationship with the GNPs content. The alteration did not trigger a toxic reaction, as its biocompatibility remained within the prescribed range for biomedical devices [Research has shown that the incorporation of graphene nanofibers into PMMA resin can significantly improve its mechanical and antimicrobial properties, making it a promising material for CAD/CAM applications. While further research is necessary to fully assess its potential, the findings from previous studies are highly encouraging. The use of graphene nano-reinforced biopolymer in G-CAM discs, designed explicitly for permanent dental structures, offers various benefits. These discs are available in different chromatic crowns, providing a natural aesthetic appearance. Moreover, they address the mechanical, physicochemical, and biological shortcomings associated with other materials currently used in the dental industry. The biopolymer discs nanoreinforced with G-CAM graphene  offer numerous properties for dental structures and meet the requirements to be considered a good material for prosthetic works utilizing CAD/CAM technology . Aati et devices .Despite the increasing research on the antimicrobial properties of graphene-based materials (GBMs), there remain uncertainties regarding the mechanisms underlying their behavior and short-term and long-term effects. GBMs exhibit antibacterial activity independent of antimicrobial resistance (AMR) and do not appear to induce long-term secondary resistance. This unique characteristic makes GBMs suitable for various antimicrobial applications; however, their potential toxicity to the environment and human health needs to be thoroughly understood . ConsideAs a catalyst in oxidative environments, graphene can penetrate cell membranes through its jagged edges and disrupt normal cellular functions ,61,62,69Dziewięcka et al. investigated the cytotoxicity of various graphene oxide nanoparticles based on their structure. They systematically produced and analyzed the particles regarding cell viability, oxidative stress, apoptosis stages, and DNA damage. Biomarkers were correlated with various physicochemical parameters of graphene oxide . The autThe stomatognathic system is a highly intricate and sophisticated physiological mechanism that remains in a perpetual state of flux due to countless environmental, supra-, and sub-systemic factors. Its primary function is to maintain a delicate equilibrium between a host of risk and protective factors. Therefore, it is imperative to precisely identify and quantify any factor that disrupts this equilibrium to determine the most efficacious treatment approach. However, the oral cavity is replete with numerous variables that pose a significant challenge to accurate measurement and prediction .The efficacy of dental treatments is contingent upon the biological and biomechanical responses of restorations, which are influenced by a plethora of factors. Within the dental industry, there exists a persistent demand to extend the longevity of dental restorations, driven by patients, public practitioners, and administrative bodies in the public health sector. Despite significant advancements in biomaterials and restorative techniques, absolute therapeutic success cannot be guaranteed. Nevertheless, there are ongoing initiatives aimed at enhancing existing biomaterials and developing novel ones that surpass current standards .Contemporary dental materials have undergone extensive development through advanced research conducted in diverse fields such as materials science, chemistry, physics, and engineering. For example, dental composites, which are used for aesthetically pleasing fillings and restorations, were developed based on composite materials that were initially employed in the aerospace and automotive industries. Furthermore, titanium, which is widely acclaimed for its exceptional properties such as its resistance to corrosion and biocompatibility, has found extensive use in dentistry thanks to research conducted in the aerospace industry. Moreover, restorative materials used in prosthodontics and CAD/CAM technology are the outcome of research conducted in the fields of engineering and IT. Additionally, groundbreaking research in biomaterial science has led to the development of bioactive materials that can facilitate dentin remineralization. The integration of technologies and materials from other fields has enabled the field of dentistry to make significant progress and offer more efficient, durable, and aesthetically pleasing dental treatments.The remarkable antimicrobial, chemical, physical, and mechanical attributes of GBMs justify the large interest in their application in the biomedical and dental fields , an inteNumerous factors, namely concentration, size, shape, and surface chemistry, exert a significant influence on the properties of graphene derivatives. It is noteworthy that functionalized graphene oxide and reduced graphene oxide exhibit the most robust antibacterial properties among graphene derivatives . FurtherTitanium- and zirconia-based implants are widely regarded as the optimal choice for dental root replacement due to their exceptional biocompatibility, impressive corrosion resistance, and exceptional long-term performance. However, it is worth noting that titanium does have a relatively low shear strength , and zirIt has been found that graphene and its derivatives positively impact the biocompatibility of materials and promote cell adhesion to the substrate. In comparison to the Gelatin-alginate scaffold, the Gelatin-alginate/Graphene oxide scaffold has exhibited superior compressive strength, enhanced cell attachment and proliferation, amplified expression of osteoblast transcription factors, and elevated activity of alkaline phosphatase . The incResearch has shown that a scaffold made from a combination of 3D-printed poly(ε-caprolactone) (PCL) and graphene exhibits improved protein adsorption, cell attachment, and dispersion, as well as increased cell viability and connective and mineralized tissue formation in both laboratory and animal testing. Furthermore, the levels of proinflammatory molecules TNFα- and IL-1β were observed to decrease . In compAnother study discovered that the inclusion of GO within collagen 3D sponges resulted in a noticeable enhancement of osteoblastic differentiation in vitro and an accelerated rate of new bone formation in vivo . TherefoLiterature data have demonstrated that GO can have a low cytotoxic effect, resulting in reduced cell viability . It is nIn prosthodontics, the conventional technology used for manufacturing resin-based restorations negatively affects the materials’ sustainability and their physical, chemical, and mechanical properties. This is primarily due to factors such as resin shrinkage, residual monomer, and porosity. To address and mitigate these shortcomings, 3D technologies have been adopted in conjunction with the incorporation of graphene additives.Nevertheless, the utilization of graphene derivatives poses certain challenges, including toxicity and a paucity of comprehensive knowledge concerning their biological characteristics and pathways. These challenges demand further in-depth examination.Currently, there is a limited amount of research available on the long-term effects of graphene derivatives, which has made their potential toxicity a primary concern. This lack of data has had a significant impact on the widespread use of these biomaterials in clinical settings, and as such, it is crucial to prioritize human safety when considering the use of these materials. To achieve the best possible clinical outcomes, it is necessary to develop solutions to the potential challenges posed by graphene derivatives, including their toxicity and biodegradability. This can be accomplished by developing standardized parameters for their use, as the toxicity and biological features of these materials are inherently linked to their physicochemical properties. Additionally, it is important to note that the effects of graphene derivatives are highly dependent on the dosage and time of exposure, which can significantly impact the potential for detrimental impact on cells through ROS scavenging or oxidative stress.Our research employed a narrative review approach, a method that, besides its scientific value, is also acknowledged for its inherent limitations within the scientific community. The selection criteria for our study have been thoughtfully established by our team, drawing upon our collective experience. Nevertheless, we acknowledge that this method may be prone to subjectivity, and therefore, we have implemented measures to ensure that only the most reliable sources are utilized. Specifically, we have meticulously curated articles from the Web of Science database. By doing so, we have taken the necessary steps to guarantee the quality of the sources used in our study.A comprehensive understanding of the potential use of graphene derivatives within the fields of periodontology and dental prosthodontics was provided. Graphene derivatives have been found to exhibit antimicrobial properties through a combination of mechanical and chemical mechanisms, including cellular uptake, the generation of reactive oxygen species, and sharp-edge-mediated actions. Incorporating derivatives of graphene into coatings for titanium and zirconia-based implants, as well as scaffolds for tissue engineering, has shown promising results in terms of biocompatibility and the formation of new tissue. The physical-mechanical properties of direct and indirect restorations were notably enhanced through the incorporation of various graphene types and concentrations. It has been observed that GO may potentially have a cytotoxic effect that could lead to reduced cell viability. It is important to note, however, that the response of cells to GO is influenced by the specific type of graphene derivative and the concentration utilized in the material.While we cannot definitively confirm whether graphene is currently viable for successful periodontal and dental prosthetic treatments due to the limitations of the investigated materials, our research highlights potential areas for further exploration. Extensive and rigorous research endeavors are imperative in the domains of implantology and tissue engineering to determine the safety and durability of employing graphene derivatives as a viable course of treatment. We maintain the conviction that the unwavering dedication and persistent efforts of researchers hold the potential to pave the way for the development of reliable, efficacious, and tailored therapeutic interventions, ultimately leading to optimal therapeutic outcomes.


Aggregation of the RNA-binding protein, TDP-43, is the unifying hallmark of amyotrophic lateral sclerosis and frontotemporal dementia. TDP-43-related neurodegeneration involves multiple changes to normal physiological TDP-43, which undergoes nuclear depletion, cytoplasmic mislocalisation, post-translational modification, and aberrant liquid–liquid phase separation, preceding inclusion formation. Along with toxic cytoplasmic aggregation, concurrent depletion and dysfunction of normal nuclear TDP-43 in cells with TDP-43 pathology is likely a key potentiator of neurodegeneration, but is not well understood. To define processes driving TDP-43 dysfunction, we used CRISPR/Cas9-mediated fluorescent tagging to investigate how disease-associated stressors and pathological TDP-43 alter abundance, localisation, self-assembly, aggregation, solubility, and mobility dynamics of normal nuclear TDP-43 over time in live cells. Oxidative stress stimulated liquid–liquid phase separation of endogenous TDP-43 into droplet-like puncta, or spherical shell-like anisosomes. Further, nuclear RNA-binding-ablated or acetylation-mimicking TDP-43 readily sequestered and depleted free normal nuclear TDP-43 into dynamic anisosomes, in which recruited endogenous TDP-43 proteins remained soluble and highly mobile. Large, phosphorylated inclusions formed by nuclear or cytoplasmic aggregation-prone TDP-43 mutants also caused sequestration, but rendered endogenous TDP-43 immobile and insoluble, indicating pathological transition. These findings suggest that RNA-binding deficiency and post-translational modifications including acetylation exacerbate TDP-43 aggregation and dysfunction by driving sequestration, mislocalisation, and depletion of normal nuclear TDP-43 in neurodegenerative diseases.The online version contains supplementary material available at 10.1007/s00018-023-04739-2. Pathological aggregation of TAR DNA-binding protein 43 (TDP-43) in neurons is a key hallmark of almost all amyotrophic lateral sclerosis (ALS) and approximately half of frontotemporal dementia (FTD) cases , 2. TDP-Post-translational modifications (PTMs) are a major feature of TDP-43 pathology in ALS and FTD, and likely play diverse roles in TDP-43 dysfunction, aggregation propensity, and neurotoxicity. For example, phosphorylation is a widely recognised hallmark of pathology progression –20. LessRNA deficiency promotes TDP-43 instability , 27, andAmong additional disease-associated cellular stressors, oxidative stress is observed in brain and spinal cord of ALS and FTD patients , 44–47, TARDBP expression is mediated by TDP-43 binding its own mRNA , mouse anti-FLAG MAb , and applied overnight at room temperature. Secondary antibodies were similarly prepared, including goat anti-Rabbit H+L AlexaFluor647 , and applied for 1.5 h at room temperature. Cells were counterstained with DAPI  for 10 min at room temperature, mounted onto microscope slides with ProLong Gold Antifade Mountant , and allowed to cure overnight at room temperature before microscopy.Confocal images were captured using either the Zeiss Laser Scanning Microscope (LSM) 510 META or Zeiss LSM 710. Both are equipped with a 63 × oil objective , blue laser diode for DAPI (405 nm), argon laser for AlexaFluor488 (488 nm), 561 nm laser for AlexaFluor555, and HeNe 633 nm laser for AlexaFluor647 excitation. Images were acquired using Zen software and all acquisition parameters were applied consistently for all samples in an experimental set. Image processing and intensity profile analyses were completed with FIJI. Automated and unbiased single-cell segmentation analyses were conducted using CellProfiler version 4.2.1  using cuP values < 0.05 deemed statistically significant. Live-cell imaging of nuclear TDP-43-mCherry expression over time included n = 3 independent repeats, recording 12 ROIs at 20 × magnification for each experimental condition. FRAP experiments also included n = 3 independent repeats, with at least 5 cells per replicate for each of the distinct TDP-43 assemblies studied. All immunoblot experiments included n = 3 independent repeats. Individual data points for each independent replicate are shown overlaid with a line at the mean. Statistically significant differences between the means were determined using a two-tailed paired t-test (where indicated in figure legends) for comparisons between only two groups, or a one-way ANOVA with a Tukey’s post hoc test for multiple comparisons in experiments with several groups.Data were represented as mean ± standard error about the mean (SEM). GraphPad Prism 9 was used for statistical analysis and preparation of graphs, with Supplementary file1 (XLSX 503 KB)Supplementary file2 (XLSX 79 KB)Supplementary file3 (XLSX 250 KB)Supplementary file4 (XLSX 76734 KB)Supplementary file5 (MP4 16010 KB)Supplementary file6 (MP4 9739 KB)Supplementary file7 (MP4 23411 KB)Supplementary file8 (PDF 9613 KB)Below is the link to the electronic supplementary material.