Symbiodinium communities in corals are inherited versus environmentally acquired is fundamental to understanding coral resilience and to predicting coral responses to stressors like warming oceans that disrupt this critical endosymbiosis. We examined the fidelity with which Symbiodinium communities in the brooding coral Seriatopora hystrix are vertically transmitted and the extent to which communities are genetically regulated, by genotyping the symbiont communities within 60 larvae and their parents  using high-throughput sequencing of the ITS2 locus. Unexpectedly, Symbiodinium communities associated with brooded larvae were distinct from those within parent colonies, including the presence of types not detected in adults. Bayesian heritability (h2) analysis revealed that 33% of variability in larval Symbiodinium communities was genetically controlled. Results highlight flexibility in the establishment of larval symbiont communities and demonstrate that symbiont transmission is not exclusively vertical in brooding corals. Instead, we show that Symbiodinium transmission in S. hystrix involves a mixed-mode strategy, similar to many terrestrial invertebrate symbioses. Also, variation in the abundances of common Symbiodinium types among adult corals suggests that microhabitat differences influence the structure of in hospite Symbiodinium communities. Partial genetic regulation coupled with flexibility in the environmentally acquired component of Symbiodinium communities implies that corals with vertical transmission, like S. hystrix, may be more resilient to environmental change than previously thought.Determining the extent to which  Symbiodinium has allowed corals to thrive in oligotrophic tropical seas through the utilization of symbiont photosynthates. Similar nutritional facilitation has been described for sap-sucking insects that rely on microbial partners to supplement their diets  or passed maternally into eggs or larvae , with the latter thought to be the most prevalent mode of transmission in brooding scleractinian corals  or associate with multiple partners, and superinfections of multiple Symbiodinium types and subtypes of varying abundances are common among species  is the parameter typically used to describe the degree to which variability in a trait is explained by genetic factors. Assuming that the Symbiodinium community associated with a coral can be represented as a complex trait, then an h2 value of 1 would imply that variability of the community is mostly due to host genetics. Conversely, an h2 value estimated at 0 would imply no genetic basis for variability in the community, thus the community would not be under selection and could not evolve  of this trait, we quantified the in hospite Symbiodinium communities of individual planula larvae and their parents across a spectrum of relatedness using high-throughput sequencing. Relatedness was based on a population genetic parentage analysis that assigned the likely paternal identity of each larva. In light of results on heritability and fidelity of symbiont transfer, we discuss the potential of larvae from brooding corals like S. hystrix to acclimate to novel environments.To examine Seriatopora hystrix broods sexually produced larvae following internal fertilization of eggs by sperm from surrounding colonies   were sequenced with the primers ITS2alg-F and ITS2alg-R  were analyzed using the USEARCH and UPARSE pipelines  and multiplicative (log2 fold) terms between or among treatments ) that incorporates OTU richness, evenness, and sequence similarity , adult colonies were divided into five size classes based on their mean diameter , 8 – <14 cm (n = 19), 14 – <20 cm (n = 13), 20 – <26 cm (n = 11), and 26–32 cm (n = 1). Differential abundance testing of Symbiodinium OTUs among size classes was performed as for larval communities.To determine if Symbiodinium OTUs associated with the 45 adult colonies of S. hystrix (ShA) that were genotyped across the 16 m × 40 m sampling area. Gradient-boosted models and linear models were run in the package “gbm”  . On average, adults contained 29.9 ± 0.6 (SE) OTUs and larvae had 22 ± 0.4 OTUs  Fig. . OverallTUs Fig. . Of the TUs Fig. . Unique ity Fig. . Of the ity Fig. . AlthougSymbiodinium, 8 of putative Symbiodinium taxonomy, Fig. Symbiodinium communities were characterized by up to 4–5.7 (2–2.5 log2 fold) times more D-types , and A-types (A1_OTU10 and A3_OTU8) compared to larvae  significantly higher abundances in adults  and the remaining C1 types had between 0.02–0.13 (−5.4 to −2.9 log2 fold)  although 95% probability regions overlapped  larvae was 0.43 ± 0.21 SD, with a posterior mode of 0.33  0.1–0.8; Fig. Leinster and Cobbold estimates of ies Fig. . The posp > 0.05; Fig. Symbiodinium C120/C120a were greatest in colonies that were closest to the lower left of the sampling area : p = 0.019), consistent with a gradual increase in distance down the reef slope, this pattern was not consistent along the reef slope (GBM: p = 0.00841). The abundance of D1 was significantly higher in the top-right and lower left side of the sampling area than in other aspects . D1a was least abundant in the top left and inner portion of the sampling area . Finally, although the variance normalized abundances of all three OTUs were significantly positively correlated overall , extremely low abundances of C120/C120a at x-coordinates >15 contrast markedly with high abundances of the two D-types in the same region   differ from those associated with their parents, providing experimental evidence that at least a portion of the Symbiodinium community is horizontally transmitted in a brooding coral. These results further demonstrate that coral-Symbiodinium symbioses align with well-characterized models of invertebrate symbioses. Results from previous studies suggesting exclusively vertical transmission in brooding corals may have been due to the lower detection capability of earlier methods and will no doubt change with the continued application of NGS technology to more brooding coral species. Overall, Symbiodinium communities were found to be moderately heritable, with only 33% of variability in larval symbiont communities under genetic regulation. Model selection also showed that, for larvae released from the same colony, sharing the same maternal environment did not significantly explain variability in Symbiodinium communities found among larvae from the same brood. This result, combined with the moderate heritability estimate, indicates that similarities in Symbiodinium communities among larvae of the same maternal brood were due to gene(s) inherited by these larvae.The availability of a full larval pedigree for Symbiodinium communities in S. hystrix (ShA). Much greater heritability of the Symbiodinium community was expected in this vertically transmitting coral, and in comparison with what is known of other important reproductive and fitness traits. For example, fertilization success, larval heat tolerance, protein content, settlement success, settlement substrate preferences, and juvenile growth and survivorship are all heritable traits  has some capacity to respond to changing environmental conditions. Thus, intervention efforts to facilitate such phenotypic change may be possible  could also be candidates for assisted Symbiodinium uptake.Heritability estimates reveal important information about the evolvability of a trait, such as the capacity of brooding corals to vary their symbiont communities in response to changing environmental conditions. If levels of heritability and genetic variance are low, then responses to natural or artificial selection (evolvability) would be limited  have a mixed-mode transmission strategy, in which dominant symbionts are transmitted vertically but additional background strains are acquired from environmental sources. Although adult diversity may have been under-sampled by only sequencing one branch of each parental colony, unique larval OTUs were not detected in any of the 45 adult colonies that were genotyped. Environmental uptake of novel Symbiodinium by larvae of this species is further supported by the appreciable amount of variation in the composition of larval Symbiodinium communities that was not under genetic control, according to our heritability model. These results validate the hypothesis of potential mixed-mode transmission initially raised by Byler et al.  contradicts previous assumptions that maternally transmitted symbiont communities are transferred to offspring with high fidelity in corals  larvae mirrors increased diversity of Symbiodinium communities detected in eggs of Montipora capitata and M. digitata compared to adults , vertical transmission of Symbiodinium that are locally adapted to the parental environment is likely to provide benefits for a species that is able to self-fertilize (Sherman S. hystrix (ShA), combined with a functional, established symbiosis upon release, may facilitate both local and long-distance dispersal . Theory suggests that competition among symbionts may preclude transmission of an exact replica of the parental symbiont community because conditions promoting growth for some symbionts may differ between life stages  larvae may provide benefits similar to those observed in insect symbioses, for example to provide larvae with the flexibility to host optimal symbiont types for the changing conditions through ontogeny  immune-related genes are being transmitted from parents to offspring and whether novel symbionts are a function of an under-developed immune response.Conversely, the greater variation and diversity found in larval compared to adult Symbiodinium communities in larvae versus adults found here further indicate that the re-shaping of the Symbiodinium community through ontogeny is an important developmental process in corals. Ontogenetic variability in microbial communities (both Symbiodinium and bacteria) is common in both vertically and horizontally transmitting cnidarian species   than in broadcast-spawning corals  are only partially genetically regulated by their host and that larvae retain the flexibility to associate with novel symbionts across generations. Our results reveal a mixed-mode transmission strategy for establishing Symbiodinium communities in larvae of a brooding coral, based on demonstrations that a subset of novel and unique Symbiodinium types are found in brooded larvae but not in adults . Importantly, this information aligns symbiosis transmission ecology in corals with well-known terrestrial invertebrate symbioses that typically exhibit mixed-mode transmission strategies. Advances in the understanding of heritable genetic mechanisms quantified here provide important insights into how Symbiodinium communities may be targeted for intervention strategies to increase reef resilience.On the basis of novel heritability and paternity analyses, we show that DNA sequences: All sequencing data will be made available through NCBI SRA SRP082130.Supporting Information


Altered expression of many genes and proteins is essential for cancer development and progression. Recently, the affected expression of metadherin (MTDH), also known as AEG-1 (Astrocyte Elevated Gene 1) and Lyric, has been implicated in various aspects of cancer progression and metastasis. Elevated expression of MTDH/AEG-1 has been reported in many cancers including breast, prostate, liver, and esophageal cancers, whereas its expression is low or absent in non-malignant tissues. These expression studies suggest that MTDH may represent a potential tumor associated antigen. MTDH also regulates multiple signaling pathways including PI3K/Akt, NF-κB, Wnt/β-catenin, and MAPK which cooperate to promote the tumorigenic and metastatic potential of transformed cells. Several microRNA have also been found to be associated with the increased MTDH expression in different cancers. Increased MTDH levels were linked to the tumor chemoresistance making it an attractive novel therapeutic target. In this review, we summarize data on MTDH function in various cancers. Chen etin situ (DCIS) of the breast  leads to increased MTDH and acts as a prognostic marker for breast cancer outcome  play important, if subtle, roles in the regulation of gene expression and translation in cellular differentiation and proliferation. Unsurprisingly, aberrant expression of oncogenic miRs (onco-miRs) and tumor suppressor miRs is implicated in cancer pathogenesis , 50. MTDmiR-21 . In contt cancer   tumor suppressor via NF-κB mediated pathways and contributes to the HER2-targeting therapy resistance . Thus, M+ mediated immune response inhibited cancer cell proliferation and lung metastases in a mouse breast cancer xenograft model (Immunotherapy targeting the tumor antigens has been an area of intense research. Expression of MTDH is increased in many different cancer types. Importantly, auto-antibodies against the MTDH protein have been detected in cancer patients confirming its immunogenicity and supports MDTH as a potential immunotherapy target . We receft model . Given tMTDH is pro-oncogenic factor playing multifaceted and diverse roles in cancer progression. Its association and central role in regulating signaling pathways such a MAPK, wnt/β-catenin, PI3K/AkT, NF-κβ pathways in various cancers shows that it plays a vital role in metastasis. MTDH contribution to chemo and radiotherapy resistance provides a new direction for the development of anticancer therapeutics. Multiple mechanisms converge to promote expression of MTDH in cancers. Further studies are therefore warranted to determine whether the elevated MTDH expression has prognostic value for development of the malignancy. Given its increased expression in many cancer types it would be important to determine whether MTDH represents a feasible target for cancer therapy, including immune-therapy. For these reasons there is now an urgent need to determine the clinical and therapeutic significance of increased MTDH expression in cancer.GD originated the idea for writing the review and wrote about MTDH association in different cancers. NS and MG wrote the following sections: micro RNAs regulation of MTDH expression and MTDH target for cancer therapy. ER, JL-R, and NM were involved in writing the MTDH regulation and molecular functions and preparing the original figures. RM and SK contributed to the clinical significance of MTDH expression in cancer. AR managed the different collaborations during the writing of the review and contributed to editing the manuscript. MB was overall responsible for coordinating and managing multisite collaboration, and writing the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


This AIE process was further configured into an immunoassay for analyzing drug residues in foodstuffs. In this approach, glucose oxidase (GOx) was used as an enzyme label for the immunoassay and triggered GOx/glucose-mediated H2O2 generation, which caused oxidation of TPE-HPro and a “turn-on” fluorescence response at 540 nm. To quantitatively analyze the drug residues in foodstuffs, we used amantadine (AMD) as an assay model. By combining the AIE-active “turn-on” fluorescent signal generation mechanism with conventional ELISAs, quantifying AMD concentrations in chicken muscle samples was realized with an IC50 (50% inhibitory concentration) value of 0.38 ng/mL in buffer and a limited detection of 0.06 μg/kg in chicken samples. Overall, the conceptual integration of AIE with ELISA represents a potent and sensitive strategy that broadens the applicability of the AIE-based fluorometric assays.A new fluorescent “turn-on” probe-based immunosensor for detecting drug residues in foodstuffs was established by combining the mechanism of aggregation-induced emission (AIE) and an indirect competitive enzyme-linked immunosorbent assay (ELISA). In this study, a luminogen, with negligible fluorescence emission (TPE-HPro), aggregated in the presence of H The enzyme-linked immunosorbent assay (ELISA) is an extensively used immunoassay to detect the concentration of protein biomarkers and small molecules for clinical diagnosis, environmental monitoring, and food analysis . AMD was purchased from Tokyo Chemical Industry Co., Ltd . Bovine serum albumin (BSA) was purchased from Amresco Inc. . Glucose oxidase-labeled goat anti-mouse IgG (gtAm-GOx) was purchased from Abcam , and horseradish peroxidase-labeled goat anti-mouse IgG (gtAm-HRP) was purchased from Jackson ImmunoResearch . Phosphate-buffered saline (PBS) and Tris-HCl were purchased from Solaribo . TMB substrate was purchased from Beyotime Biotechnology . TPE-HPro was produced in our laboratory as previously described  were mixed with 42 μL Tris-HCl buffer . Then the mixtures were added into the TPE-HPro probe solutions (400 μM in acetonitrile) at a 9:1 volume ratio. The solutions were kept to react at 37°C for 30 min. The fluorescence spectra of the solutions were recorded using a microplate reader.The cleavage of the phenyl boronic ester occurs only in high pH environment. In our previous work, we evaluated and optimized the pH effect on the AIE process  was mixed with the gtAm-GOx solutions at a 3:1 volume ratio and incubated at 37°C for 30 min. Subsequently, 120 μL of these solutions were mixed with 42 μL Tris-HCl buffer . Finally, 135 μL of the mixture was incubated with 15 μL TPE-HPro probe solutions (400 μM in acetonitrile). Fluorescence intensity was measured after incubation at 37°C for 30 min.For AIE-based gtAm-GOx sensing, gtAm-GOx was serially diluted with deionized water 0–14 μg/mL. Then, glucose . Samples were analyzed by the AIE-based and conventional methods after the following pretreatment: 1.00 g of the chicken muscle sample was exactly weighed and spiked with AMD at 0.5, 1.0, and 2.0 μg/kg. Then, 5 mL 1% acetic acid in acetonitrile was added and well homogenized for 2 min. Subsequently, the homogenate was centrifuged for 5 min at 4,000 g. Finally, 3 mL of the supernatant was evaporated under nitrogen to dryness at 40°C, and the residue was dissolved in PBS and analyzed using an indirect competitive ELISA  and incubated it for 30 min at 37°C. Finally, 42 μL Tris-HCl buffer  was added and the mixtures were added into the TPE-HPro probe solutions (400 μM in acetonitrile) at a 9:1 volume ratio. FL intensity was recorded by the plate reader after incubation at 37°C for 30 min. The calibration curve was analyzed by a four-parameter logistic equation using OriginPro 9.1 .For the AIE-based fluorometric immunoassays, 100 μL AMD-OVA conjugates were added to a 96-well plate and incubated at 4°C overnight. The plate was then blocked with 150 μL 1% BSA in PBS at 37°C for 1 h. The plates were washed three times with washing buffer (PBS containing 0.025% Tween-20) then 50 μL mAb 3F2 was added with 50 μL AMD of varying concentration or samples for competitive reactions. After incubation at 37°C for 30 min, the plate was washed four times and 100 μL gtAm-GOx in PBS (containing 1% BSA and 0.01% Tween-20) was added and incubated for 30 min at 37°C. The plate was washed three times with washing buffer and three times with deionized water. Then, we added 120 μL glucose  and a cis-trans isomerization process of C = N  can even be sensed with the fluorescence spectrophotometer. The sensitivity suggested that H2O2-triggered “turn-on” fluorescence could be further implemented to design a highly sensitive AIE-active “turn-on” sensor.As a proof of principle, we first demonstrated that Hntration , the calresponse  inset. M2O2-generating enzyme that can modulate the AIE process for fluorescent measurements. To achieve this goal, we employed commercially available gtAm-GOx to effectively generate H2O2, as in our previous studies  H2O2 was efficiently formed by the GOx-catalyzed oxidation reaction; (II) the AIEgen could be hydrolyzed by H2O2 even at 5 μM to produce strongly emissive AIE aggregates. Therefore, we expect to achieve highly sensitive detection by combining the AIE-active “turn-on” fluorescent sensor with conventional immunoassay technology.When developing an enzyme-mediated immunoassay, the main challenge is finding a H process . The FL 2O2 formation and subsequent of AIE induction and fluorescent “turn-on” signal generation. Compared with traditional ELISAs, the only difference was that glucose rather than TMB was introduced into the signal generation step. Therefore, the procedures of our newly established AIE-based fluorescence “turn-on” immunoassay were nearly the same, and thus were fully compatible and comparable with current ELISA platforms. Instead of the absorbance values of a traditional ELISA, the fluorescence “turn-on” from the AIE-based sensor further amplified the signal output and enhanced the detection sensitivity.We further employed this AIE-active “turn-on” fluorescent sensor for highly sensitive indirect competitive ELISAs . AMD-OVA2O2 was generated. Thus, no noticeable H2O2-induced “turn-on” fluorescence was obtained in the presence of high concentration of AMD. With decreasing AMD concentration of, fluorescence intensity gradually increased (blk)/(FL0-FLblk)%] was used to quantitatively evaluate AIE process level .To evaluate the sensitivity of our AIE-based indirect competitive ELISA, various concentrations of AMD were tested (0.06–13.5 ng/mL). Because of the competitive nature of this immunoassay platform, an increased AMD concentration led to intense competition, such that less primary and GOx-labeled secondary antibodies were bound on the ELISA plate; and subsequently less Hncreased . Relativ50 value of a conventional TMB/HRP-based ELISA was 0.93 ng/mL, while our newly established AIE-based fluorescence “turn-on” immunoassay provided a higher sensitivity; the IC50 value was 0.38 ng/mL. This suggests approximately 2.5-fold improvement in sensitivity. The limit of detection  was determined to be 0.06 ng/mL. This improved sensitivity relied on the following two principles: first, H2O2 was efficiently generated by the enzyme-catalyzed oxidation reaction; and second, the production of strong emissive AIE aggregates could be triggered by low concentrations of H2O2. These unique features make this “turn-on” fluorescent immunosensor an attractive immunoassay platform for highly sensitive detection.We next compared our newly designed AIE-based ELISA with a conventional ELISA . These r2O2 was generated by GOx, which decreased the AIE-based fluorescence “turn-on.” Because of the complexity of chicken muscle samples, a pretreatment step was required. The color intensity change obtained for negative and positive chicken samples could be visually distinguished using a hand-held UV lamp (Finally, having successfully shown that the assay could quantify AMD, the “turn-on” fluorescent immunosensor was further evaluated using real food samples. AMD-contaminated chicken samples were used as a model. Chicken muscle samples were spiked with AMD at concentrations of 0.5, 1.0, and 2.0 μg/kg. In the indirect competitive immunoassay, spiked AMD in chicken samples competed with coating antigen for binding of the primary antibody. Thus, as the level of competition increased, less H UV lamp . To quan UV lamp . Therefo2O2-stimulated TPE-HPro aggregation. This cascade reaction makes use of the catalytic ability of the enzyme and the AIE characteristics, which lead to the signal amplification of the immunoassay. The AIE-based fluorogenic ELISA has much higher sensitivity due to the efficient generation of H2O2, which leads to a high fluorescence emission at low concentrations of H2O2. Compared with conventional HRP/TMB-based ELISAs, this newly designed immunoassay offers a 2.5-fold enhancement in sensitivity. Furthermore, this novel sensing strategy can be directly adapted to the current indirect competitive conventional ELISA format using an H2O2-triggered AIE fluorescence “turn-on” as the signal generation mechanism. This represents an alternative to the existing commercial immunoassay. More importantly, this approach can be applied to quantifying AMD in real food samples. Therefore, the developed AIE-based fluorescence “turn-on” ELISA has excellent potential for use in the testing of other drug residues in food samples.In conclusion, we developed an AIE-based fluorescence “turn-on” immunoassay by combining a GOx-triggered AIE process with an indirect competitive ELISA for the highly sensitive detection of drug residues in foodstuffs. In this approach, the fluorescence “turn-on” is triggered by the GOx-catalyzed oxidation of glucose and HWY planned and supervised the experiments, processed the raw data, and wrote the manuscript. YL synthesized TPE-HPro. BX performed the experiments. MM, CC, CL, and XY discussed the results and processed the data. ZW, KW, BT, and JS revised and approved the manuscript. All authors reviewed the manuscript and approved it for submission.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


The paraffin-enabled transfer process could open realms for the development of high-performance ubiquitous electronics based on large-area two-dimensional materials.The performance and reliability of large-area graphene grown by chemical vapor deposition are often limited by the presence of wrinkles and the transfer-process-induced polymer residue. Here, we report a transfer approach using paraffin as a support layer, whose thermal properties, low chemical reactivity and non-covalent affinity to graphene enable transfer of wrinkle-reduced and clean large-area graphene. The paraffin-transferred graphene has smooth morphology and high electrical reliability with uniform sheet resistance with ~1% deviation over a centimeter-scale area. Electronic devices fabricated on such smooth graphene exhibit electrical performance approaching that of intrinsic graphene with small Dirac points and high carrier mobility (hole mobility = 14,215 cm The transfer process of as-grown graphene limits its electrical performance and reliability. Here, the authors develop a transfer approach using paraffin as a support layer and obtain wrinkle-reduced and clean large-area graphene retaining high mobility. Although mobility values as high as 200,000 cm2 V−1 s−1 have been reported for freely suspended graphene exfoliated from bulk crystal at 5 K2, the mobility values reported for large-area graphene is several orders of magnitude lower, regardless of the growth method or substrate employed7. This significant decrease in carrier mobility in large-area graphene can be attributed to four factors: (1) polycrystalline nature of graphene, (2) effect of surrounding medium, (3) contamination from transfer support layer, and (4) wrinkles present in graphene. Any of these factors can reduce the carrier mean free path in graphene, which could act as a source of extrinsic scattering, thus limiting the carrier mobility8. To address the first factor, there are considerable advances in the synthesis of wafer-scale single-crystal graphene domains10, with the latest achievement of 1.5-inch-large graphene monolayer synthesized on Cu–Ni alloys11. As for the second factor, the effects of the surrounding medium on graphene can be minimized by fully encapsulating the graphene layer within insulating substrates through van der Waals assembly approach12. For instance, a mobility value of 110,000 cm2 V−1 s−1 at 1.6 K was reported for an as-synthesized chemical vapor deposition (CVD) graphene flake sandwiched between hexagonal boron nitride (h-BN) flakes13. Furthermore, to fabricate graphene-based functional electronics, graphene must be transferred from the growth substrate to a destination substrate. Since graphene is only one atom thick, a support layer is required during the transfer process to prevent cracks from appearing and propagating in the graphene film. To date, after one decade of research efforts, the most widely used support layer for graphene transfer is still polymethylmethacrylate (PMMA). However, the PMMA-assisted transfer process engenders two major issues that severely degrade the carrier mobility in graphene: polymer contamination and graphene wrinkling (third and fourth factor mentioned above). To overcome these two issues, support layers made of other polymers and organic molecules have been attempted21, but none of these solutions fully solved both issues.Charge carriers in graphene exhibit ultrahigh mobility owing to their zero-mass nature in the linear E−k dispersion at low energy, where the conduction and valence bands meet at the Dirac points22. Here, we propose to use paraffin as a transfer support layer based on two rationales: (1) paraffin is an alkane with a simple unreactive chemical structure and (2) it has a high thermal expansion coefficient. Wrinkles form in graphene either natively during the growth process23  or during the graphene transfer process, where water (including vapor) can be trapped between the graphene and the destination substrate24. To address the graphene wrinkling issue, we scoop up the paraffin-supported graphene layer with a destination substrate from deionized water at a higher temperature of 40 °C, rather than the usual room temperature. At 40 °C, the paraffin support layer undergoes thermal expansion due to the small amount of heat from the water25. This expansion can stretch the graphene film underneath and thus effectively reduces wrinkles in graphene.We have shown earlier that paraffin can be used as a flexible substrate to transfer graphene onto itOur paraffin-enabled transfer technique can concurrently addresses both support layer contamination and wrinkling in graphene, enabling transfers of large-area graphene with homogeneous and enhanced electrical properties. Compared to PMMA, paraffin is found to leave much less residue on graphene, as it does not contain carbonyl (C=O) functional groups that react with electrophiles or nucleophiles. Moreover, paraffin radicals are not expected to form covalent bonds with graphene. Our density functional theory (DFT) calculations confirm that paraffin has lower adsorption energy with graphene compared to PMMA, which supports the effective curtailing of contamination observed in our paraffin-transferred graphene. Through atomic force microscopy (AFM) and Raman studies, we confirm that paraffin-assisted transfers substantially reduce contamination and wrinkling in graphene compared to PMMA, and moreover, the paraffin-transferred graphene has near-intrinsic levels of doping and strain. Consequently, field-effect transistors fabricated on paraffin-transferred graphene exhibit near-zero Dirac voltage and the electron mobility that is ~fourfold higher than that of our PMMA-transferred graphene. Our paraffin-enabled transfer technique presented in this work offers a solution for the development of high-performance large-area graphene-based electronics by minimizing the charge carrier scattering centers in graphene.26. As can be seen in Fig. f and Li denote the final and initial lengths of a material; α is the linear thermal expansion coefficient of a material 25; and Tf and Ti are the final and initial temperatures of a material, respectively. Thermal expansion of paraffin translated to tensile strain on the graphene film underneath, which stretched and diminished wrinkles in the graphene film . Subsequently, resistivity and Hall measurements were made on each paraffin-transferred graphene film in the presence of 2800 Gauss magnetic field to extract its sheet resistance, carrier concentration, and Hall mobility values. During the Hall measurements, the magnetic field was applied by attaching a piece of magnet underneath the sample with desired polarity. For comparison, PMMA-transferred graphene film on Si/SiO2 substrates were also fabricated with the same dimensions (see Methods for details) and measured. To avoid run-to-run variations, the graphene used in this study was synthesized on the same Cu foil in the same CVD growth cycle and transferred on SiO2/Si substrates that were diced from the same SiO2/Si wafer.We observed homogeneous and improved electrical properties in the polycrystalline CVD graphene prepared with our paraffin-enabled transfer technique. To determine the electrical properties, CVD graphene was cut into 15 × 15 mm2 V−1 s−1, which was 2.6 times higher than the PMMA-transferred graphene (1698 ± 377 cm2 V−1 s−1). Furthermore, the carrier concentration of paraffin-transferred graphene ranged from 0.9 to 2.6 × 1012 cm−2, which was lower than that of PMMA (7.2 to 12 × 1012 cm−2). The lower carrier concentration indicated that the paraffin did not dope graphene as heavily as the PMMA. We note that no annealing was performed on any transferred graphene in this study. For the same group of graphene samples, sheet resistance was also extracted using the van der Pauw method and plotted in Fig. 2, with 1 mm step size in both the x- and y-directions. The spatial sheet resistance maps of both PMMA- and paraffin-transferred graphene are plotted in Fig. 14.In Fig. 2 substrates and tested ; Lch and Wch the channel length and width; IDS the drain-to-source current; VDS the drain voltage; and VG the back-gate voltage. For the typical transistor fabricated on paraffin-transferred graphene in Fig. 2 V−1 s−1, while for the PMMA-transferred graphene, the extracted hole and electron mobilities were much lower, at 3,719 and 1,653 cm2 V−1 s−1, respectively. Transferring graphene with paraffin greatly enhanced the effective hole and electron mobilities by 3.8 and 4.5 times, respectively, due to reduced charge carrier scattering centers in the paraffin-transferred graphene. Figure 28.To examine the electrical performance of devices fabricated with paraffin-transferred graphene, more than 100 graphene back-gated field-effect transistors (FETs) were fabricated on Si/SiOted Fig. . SimilarVDirac) of the device fabricated on paraffin-transferred graphene was much smaller and closer to zero compared to that of PMMA. The VDirac of a graphene device is the specific back-gate voltage where the device operates with minimum conductivity, corresponding to the Dirac point or charge neutrality point where the graphene film’s conduction and valence bands meet, and intrinsic graphene exhibits minimum conductivity when gate voltage equals to zero. Hence, VDirac is a good indicator showing how intrinsic a graphene film is. Figure VDirac distribution of 100 graphene FETs fabricated with both PMMA- and paraffin-enabled transfer process. The VDirac of PMMA-transferred graphene had a spread from 24 to 56 V with a mean value of 38.7 V, while that of paraffin-transferred graphene had a much narrower distribution between 4 and 12 V with a mean value of 7.4 V, which is five times smaller than the PMMA-transferred graphene. The lower mean value for VDirac signified that the paraffin-transferred graphene exhibited electrical properties that were closer to intrinsic graphene compared to that of PMMA, although the paraffin-transferred graphene was also contaminated by some PMMA residues during the device fabrication process. The improved process of transferring graphene might play a vital role here in determining the electrical performance of large-area graphene devices, even though the device fabrication process was not optimized to prevent contamination by PMMA residues.Remarkably, the back-gate measurements also indicated that paraffin did not induce significant doping effects on graphene, consistent with the Hall measurements in Fig. 2 substrates. As can be seen, our paraffin-transferred graphene sample has no C=O and O=C–OH peaks, and the C–OH peak intensity is relatively weak, indicating lower levels of polymer contamination. Figure 2 substrate using PMMA and paraffin support layers, respectively. We note that all graphene samples were not annealed after the transfer process. Compared to PMMA-transferred graphene, paraffin-transferred graphene had a much cleaner surface  are not observable on the graphene surface. For the removal of graphene wrinkles, we inferred that the paraffin support layer could effectively smoothen these wrinkles through thermal expansion and releasing the compressive strain generated in graphene during the cooling step in the CVD growth process. To ascertain this inference, we collected 3600 Raman spectra from each type of the transferred graphene and analyzed the strain-doping relationship. Supplementary Fig. 2 substrate. The characteristic D band peak, typical in the presence of defects, was absent in paraffin-transferred graphene, indicating the transferred graphene was of high quality. In Fig. 2D) for the PMMA-transferred graphene. The black circle represents the G and 2D peak positions of an intrinsic graphene, where graphene has neither doping nor strain , we envision the integration of paraffin support layer in industrial scale production of wrinkle-free, pristine two-dimensional materials in near future.2 substrate) was then used to scoop the paraffin-supported graphene sample from the 40 °C deionized water from only one end. After that, the sample was baked at 40 °C in an oven for more than 24 h to minimize the residual water. In the event of incomplete removal of residual water, minor cracks randomly generated in the graphene  was contacted with the PMMA-supported graphene film and the sample was initially dried with nitrogen gun blowing, followed by an oven baking at 80 °C for at least 8 h. Subsequently, the sample was soaked in acetone at room temperature for 6 h to remove the PMMA support layer. The sample was then rinsed with IPA followed by nitrogen blow dry. We note that all process parameters chosen for the PMMA-supported graphene transfer in this work were adopted from earlier reports26. A separate experiment was conducted to further verify the advantageous of paraffin support layers over PMMA  950 A5 (Microchem Inc.) was spun at 2500 rpm for 1 min on the graphene synthesized above Cu foil. The PMMA-coated graphene was then baked in an oven at 80 °C for 1 h. Next, the sample was floated on top of Cu etchant  for 30 min to remove the growth substrate. The PMMA-supported graphene film was then rinsed with deionized water multiple times. Subsequently, the destination substrate (i.e. Si/SiO2 using either paraffin or PMMA support layers. The sample was then spin-coated with a 300 nm thick layer of polymethylmethacrylate (PMMA) 950 A5 (Microchem Inc.) and baked at 180 °C on a hotplate for 180 s. The source/drain contacts on graphene were delineated using electron beam lithography (EBL) and metallized with 20/30 nm of Ni/Au, followed by a 12-hour lift-off process in acetone at room temperature. Subsequently, the graphene film was delineated into a 5 µm wide ribbon using EBL followed by oxygen plasma etching. Finally, the sample was soaked in acetone for more than 12 h at room temperature to remove the PMMA electron beam resist. The dimensions of all graphene devices were kept constant in this work. The graphene channel and contact widths are 5 µm, while the channel length ranges from 1 to 11 µm. Ni was chosen as electrode as it is one of the metals that has been shown to provide reliable metal contacts to graphene31. No annealing is performed on any graphene devices prior to electrical measurements.Graphene synthesized on Cu foil was first transferred on an oxidized degenerately p-doped silicon substrate with 285 nm thick SiO−1, respectively. The laser beam size was ~500 nm and the laser power on the sample was adjusted to around 0.75 mW to avoid laser-induced heating. Atomic force microscopy (AFM) topography image was obtained using Veeco Digital Instrument Nanoscope III in tapping mode.All Raman spectra and maps were taken using a WITec Alpha300R system. The laser excitation wavelength and grating density were 532 nm laser line and 600 mm−1 from 0 to 100 °C in a nitrogen gas atmosphere.The measurement was run at a constant heating rate of 10 °C min32. The B3LYP functional was employed together with the 6-31 G* basis set to compute the optimized geometries, energies, and orbitals. All calculations were performed in implicit solvent using the conductor-like polarizable continuum (CPCM) model and the basis set superposition error (BSSE) was corrected with the geometrical counterpoise (gCP) method.DFT calculations were performed using the ORCA computational packageThe dual descriptor of the Fukui function, a local conceptual DFT reactivity descriptor, was determined to spatially locate where eventual chemical reactions would take place. Originally introduced by Parr and Yang, the Fukui function can be interpreted in two ways: the sensitivity of the chemical potential of a system to an external perturbation at a particular point, or the change of the electron density at each point around the molecule when the total number of electrons is altered. For our purposes, the dual descriptor of the Fukui function detailed the different locations where each molecule would undergo nucleophilic or electrophilic attacks upon chemical reaction. The Fukui function was determined from electron density differences between the neutral and the positively- and negatively-charged PMMA or paraffin molecule, according to the following expression:The chemical hardness, a global conceptual DFT descriptor, characterizes the polarizability of molecules as a measure of the resistance of a chemical species to changes in its electronic configuration. It can be interpreted as the global tendency of a molecule to give or accept electrons. Molecules with low polarizability are considered to be “hard” while molecules with high polarizability are conversely “soft”. The chemical hardness is formally calculated from the ionization energy and the electron affinity. Based on Koopman’s theorem, the first ionization energy can be approximated to the energy of the HOMO, while the electron affinity can be approximated to the energy of the LUMO. Thus, the chemical hardness was approximated from the energy “gap” between the HOMO and LUMO of the molecule,Supplementary InformationPeer Review File


We use large area chemical vapor deposited (CVD) graphene and MoS2, enabling large scale realization of these devices. These devices can emulate the most vital properties of a neuron, including the all or nothing spiking, the threshold driven spiking of the action potential, the post-firing refractory period of a neuron and strength modulated frequency response. These results show that the developed artificial neuron can play a crucial role in neuromorphic computing.With the ever-increasing demand for low power electronics, neuromorphic computing has garnered huge interest in recent times. Implementing neuromorphic computing in hardware will be a severe boost for applications involving complex processes such as image processing and pattern recognition. Artificial neurons form a critical part in neuromorphic circuits, and have been realized with complex complementary metal–oxide–semiconductor (CMOS) circuitry in the past. Recently, metal-insulator-transition materials have been used to realize artificial neurons. Although memristors have been implemented to realize synaptic behavior, not much work has been reported regarding the neuronal response achieved with these devices. In this work, we use the volatile threshold switching behavior of a vertical-MoS SNNs, the third generation of neural network models, have been found to be more hardware friendly and energy efficient; thus making them more biologically realistic compared to other neural networks2. The key components of an SNN are artificial neurons and synapses where the synapses connect the pre-neurons and the post-neurons. SNN uses discrete rather than continuous spikes along with the incorporation of the concept of time. It involves efficient transfer of information based on precise timing of a sequence of spikes. Important synaptic behaviors such as synaptic plasticity, long term potentiation and depression (LTP and LTD), and Spike Time dependent Plasticity (STDP) have been studied using emerging devices such as memristors for information processing, pattern recognition and learning methods of SNN5. But, neuronal response has mostly been realized using complex CMOS circuitry, which requires several transistors to mimic a single neuron. These circuits involve a large number of active components, increasing the power dissipation and making high density integration difficult7. To overcome these issues, various emerging devices have been used to emulate a biological neuron behavior. Insulator to metal transition (IMT) in materials such as vanadium dioxide (VO2)9 and niobium dioxide (NbO2)10, chalcogenide-based phase change materials11 and magnetoelectric switching of ferromagnets such as bismuth ferrite (BiFeO3)1, have been considered to develop various types of artificial neurons. VO2 has a very low critical temperature of ~340 K above which its switching behavior disappears, severely limiting the operational range of devices and making them unsuitable for on-chip integration12. In case of phase change materials-based neurons, the degree to which the neuronal characteristics can be tuned with nominal circuitry is limited by the fact that switching properties arise from the physics of crystallization11. For the same reason, the switching speed in phase change neuron is slow. Threshold switching in memristors can be a natural choice for realizing artificial neurons. However, the exploitation of threshold switching in memristors to realize artificial neurons is relatively unexplored. Recently, a Ag/SiO2/Au threshold switching memristor (TSM) was used to demonstrate an integrate-and-fire neuron13. The observation of volatile threshold switching in MoS2 stimulates the possibility of employing this phenomenon for the realization of artificial neurons14. However, the switching in a lateral MoS2 film mediated by the grain boundaries occludes scaling of lateral device dimensions. Hence, realizing such characteristics using a vertical structure can play a critical role in improving scaling opportunities.Understanding the complex and overwhelming functioning of the human brain has always fascinated scientists and researchers for biological fields and beyond. The way the human brain can process huge amounts of data and handle pattern recognition with relative ease is immensely compelling and its complexity is continuously evolving. The building blocks in the brain consist of interconnected neurons and synapses, which cooperate efficiently to process incoming signals and decide on outgoing actions. Inspired by the operation of the human brain, various neuromorphic devices, circuits, and systems have been developed that can work analogous to the brain. Owing to the low power dissipation and characteristics that are similar to biological neurons, spiking neural networks (SNN) have garnered huge interests for mimicking and closely resembling the neuro-biological system2 (v-MoS2) layers. A novel device structure was fabricated by growing v-MoS2 on a CVD-graphene monolayer as the bottom electrode and with Ni as the top contact to v-MoS2. The key characteristics of a biological neuron including an all or nothing spiking, a threshold driven spiking, a post firing refractory period and an input strength modulated frequency response were observed. The developed v-MoS2/graphene TSM artificial neuron can operate over a wide range of temperature and can potentially be scaled down to nanometer scale cross bar structures. This platform provides an attractive prospect for neuromorphic circuits, which can be seamlessly integrated with existing fabrication technologies enabling large scale realization of such devices.In this paper, we have constructed an integrate-and-fire artificial neuron by exploiting the threshold switching behavior in CVD-grown vertical MoS+, K+, Cl− and Ca2+ in and out of the neuron cell. The potential difference between the interior and the exterior of the neuron is referred to as the membrane potential1. As the membrane potential (VM) builds up, the neuron produces an output spike (Iout) when the potential reaches a particular threshold voltage (Vth)16. This phenomenon of neuron spiking is shown in Fig. 17. After the refractory period, the neuron starts integrating the input again and prepares for the following spike.The schematic of a biological neuron connected to multiple synapses is shown in Fig. 18. An IF artificial neuron fires as a function of the electrical potential developed across it, thus mimicking the crucial behavior of a biological neuron. Here, this behavior has been realized with a v-MoS2/graphene TSM as shown in the conceptual scheme in Fig. 2/graphene neuron integrates the input signals received with the help of a capacitor. The capacitor integrates the charge and as soon as the voltage across the capacitor increases beyond the threshold value of the TSM, the neuron fires and an output spike is produced.One of the most commonly used neuron models in computational neuroscience is the integrate-and-fire (IF) model2/graphene device is shown in Fig. 2 substrate19, followed by patterned growth of v-MoS2 on graphene. Nickel contacts were deposited on the graphene and on the v-MoS2. The optical image of the device is shown in Fig. 2/graphene Threshold Switching Memristors (TSMs) is shown in Fig. 2 film over graphene in the device. Two bands corresponding to the in-plane E12g mode and the out-of-plane A1g mode were observed at 383 cm−1 and 411 cm−1 respectively. The presence of these two distinct high intensity peaks and the difference of about 28 cm−1 between their two positions signifies the presence of good quality multilayer MoS220. On pristine graphene, as shown in the inset of Fig. viz. the G band at 1590 cm−1, and the 2D band at 2690 cm−1. The peak intensity ratio I2D/IG is about ~2, indicating that the monolayer graphene used for our devices is of good quality21. After sulfurization, the spectrum, presented in Fig. −1, the G band at 1590 cm−1 and the 2D band at 2690 cm−1. A significant decrease in intensity of the 2D band with respect to the G band is revealed. This reduced value of the peak intensity ratio of the G and 2D peak and the large D band indicate the introduction of defects in the graphene layer during sulfurization22. The detailed analysis of the v-MoS2/graphene 2D-2D van der Waal’s heterojunction was carried out by High Resolution Transmission Electron Microscopy (HRTEM). Figure 2 on graphene/Si/SiO2. It can be clearly observed that MoS2 grows vertically on the graphene surface with high density of exposed edge planes. The vertical orientation of 2D MoS2 atomic layers was obtained by the sulfurization of thick Mo films, which essentially minimizes the strain energy arising as a result of volume expansion during Mo to MoS2 conversion23. This eventually leads to the formation of polycrystalline MoS2 structure with vertically orientated grains perpendicular to the substrate. The thickness of the CVD grown v-MoS2, as determined by using Atomic force microscopy (AFM)  until a voltage higher than the threshold voltage is applied. At the onset of the threshold voltage (V1), the device undergoes an abrupt transition from the HRS to a low resistance state (LRS). During the reverse sweep, the device reverts to HRS as the voltage reduces below a particular value (V2). It is interesting to note that the device retains the volatile threshold switching behavior for all the applied current compliances.To assess the switching characteristics with respect to the current compliance, the v-MoS2/graphene device, we now concentrate on the demonstration of an artificial neuron exploiting the volatile switching. The schematic of the circuit used for realizing the artificial neuron with the v-MoS2/graphene TSM is shown in Fig. p = 1 MΩ and a capacitor Co = 100 nF, which are connected in series to a resistor Ro = 10 kΩ. A load resistor RL = 1 MΩ is connected in series with the TSM to measure the output voltage. The resistors and the capacitor are connected to the device externally through a breadboard. The parallel resistor ‘Rp’ is connected to drain excessive current away from the device, thereby protecting the device. Its value is chosen to be 1 MΩ which is less than the RLRS = ~3 MΩ and greater than Ro such that Co does not discharge through Rp. The input source is a train of constant amplitude voltage pulses applied to the left node of the circuit with frequency 5 kHz and TON = 100 µs. The output voltage is measured across the RL. In Fig. ON = 100 µs are applied to the circuit, the capacitor starts charging and the voltage starts increasing at node A (black line) till the capacitor gets completely charged. Initially, the TSM is in its HRS and the device resistance ‘RHRS’ is high (~10 MΩ) compared to Ro. The charging time constant (RoC) is ~100 times lower than that for the discharging time constant [(RP||RHRS|| RL) Co], hence the leakage through the device is almost negligible. Therefore, on application of the input pulses to the circuit, the capacitor (Co) gets charged. As the charge across the capacitor increases and the voltage at node A reaches the threshold voltage of the TSM, the device switches from the HRS to the LRS. Since the TSM is now in the LRS state, the capacitor starts discharging through the device and an abrupt increase in the current appears at the rightmost node (output). This corresponds to the increase in voltage at t ≈ 300 ms, observed in Fig. After confirming the volatile threshold switching behavior of v-MoS13. Once the refractory period is over, the device reverts from the LRS state to its HRS and the capacitor ‘Co’ starts integrating again. The refractory period from the v-MoS2/graphene TSM is shown in Fig. During the timeframe when the neuron is firing, the net charge integration across the capacitor is essentially zero. This is because any input voltage pulse applied is being directly drained by the TSM device (it being turned on). This period emulates the post firing refractory period of a biological neuron13. To emulate this behavior using the developed artificial neurons, a train of pulses with the same frequency 5 kHz (Ton = 100 µs) but two different amplitudes are applied to the v-MoS2/graphene neurons. On application of input pulses of 7.5 V to the circuit  and from the LRS to the HRS (V2) are plotted over the 40 cycles shown in Fig. 1 and V2 is maintained over the cycling period. Also, it is observed that V1 and V2 are probabilistic in nature and follow a stochastic distribution. Figure 2/graphene TSM, which, in turn, produces the neuron spiking probability. Additionally, the stochastic switching behavior of the v-MoS2/graphene TSM originating from device level stochasticity makes these probabilistic spiking neurons viable for applications in the field of hardware security, such as Random Number Generators (RNGs)25.We studied the DC characteristics of the TSM for over 40 cycles to shed light on the nature of the switching voltages. It is observed that the device retained the abrupt transition from the HRS to the LRS over this period as shown in Fig. 2 can be attributed to the grain boundaries (GBs)14. The switching behavior of our v-MoS2/graphene devices can be attributed to the multiple grain boundaries in the polycrystalline films of v-MoS2. To further comprehend the switching mechanism in the v-MoS2/graphene devices, the DC measurements were carried out in vacuum. It is interesting to note that in the absence of oxygen, the fabricated devices do not show the volatile switching behavior  demanding high input voltages for the circuit operation, the v-MoS2 stack can be engineered in the future by modifying its thickness and growth conditions to allow larger currents to flow through the circuit. This will consequently reduce the voltage requirements of the circuit as well.In summary, we have demonstrated a novel 2D material-based TSM comprising v-MoS2 (300 nm SiO2) substrate. The graphene was then patterned by photolithography and etched by oxygen plasma. The sample was again patterned and 10 nm thick Mo films were deposited on the etched graphene using an electron beam (e-beam) evaporation system . The Mo films were subsequently sulfurized to MoS2 in a low-pressure CVD furnace. 50 nm Ni contacts on both graphene and MoS2 were patterned, e-beam evaporated and lifted off.First, commercially purchased CVD-grown monolayer graphene was wet transferred over a Si/SiO2 substrate were loaded in the center of a quartz tube CVD furnace with a ceramic boat having sulfur powder at the upstream of Mo sample. The furnace was pumped down to a base pressure of ≤1 mTorr and then purged with argon (Ar) gas. The furnace was heated up to a temperature of ∼650−700 °C with a constant flow of 100 SCCM (standard cubic centimeters per minute) Ar gas at a pressure of ∼100 mTorr. After a 30-min reaction time, the furnace was naturally cooled down and the substrate was taken out of the furnace. The change of color of the substrate from blue to dark green, indicates the sulfurization of Mo on the graphene.The patterned Mo film on graphene/Si/SiOThe Raman spectra were obtained using a confocal Raman system (WITEC Alpha 300RA) in ambient conditions with a 532 nm laser source for excitation. The laser was focused with a Zeiss 50× objective lens  with power ~5.7 mW and integration time 5 s. Each raman spectra were taken over 10 scans. The AFM topography was acquired on an Anasys NanoIR2 system in tapping mode using Anasys tapping mode AFM probes (Model No. PR-EX-T125-10). The TEM imaging was carried out using JEOL ARM-200F equipped with an aberration corrector which provides spatial resolution down to ~1 Å at an operating voltage of 200 kV.The electrical measurements were carried at room temperature in Micromanipulator 6200 probe station using a Keysight B1500A semiconductor device analyzer along with WGFMUs for pulse I-V measurements. The vacuum measurements were carried out in a Janis cryo-probe station. A Tektronix DPO 2024B oscilloscope was used to measure the node voltage.Supplementary Information


Graphene and graphene oxide (GO) structures and their reduced forms, e.g., GO paper and partially or fully reduced three-dimensional (3D) aerogels, are at the forefront of materials design for extensive biomedical applications that allow for the proliferation and differentiation/maturation of cells, drug delivery, and anticancer therapies. Various viability tests that have been conducted in vitro on human cells and in vivo on mice reveal very promising results, which make graphene-based materials suitable for real-life applications. In this review, we will give an overview of the latest studies that utilize graphene-based structures and their composites in biological applications and show how the biomimetic behavior of these materials can be a step forward in bridging the gap between nature and synthetically designed graphene-based nanomaterials. Carbon materials form one of the most abundant groups of materials, and are used in applications that range from electronics to biotechnology ,2. Not o3 bonds return back to a sp2 configuration [Graphene oxide (GO) is a chemically modified graphene that is typically prepared by oxidation and exfoliation of graphite-bearing oxygen functional groups, such as carboxyl (-OOH), hydroxyl (-OH), or epoxy (-O), on their basal planes and edges  with theguration . By chooguration . Robust 2·g−1) [2·v−1·s−1) [−1·K−1) [Graphene (G) exhibits unique thermal, electrical, and mechanical properties arising from its strictly two-dimensional 2D) structure, and offers immense potential for technical applications. Graphene has a large theoretical specific surface area 2630 m2·g−1) , high inv−1·s−1) , a high v−1·s−1) , high th[−1·K−1) , high op[−1·K−1) , and goo[−1·K−1) . The var[−1·K−1) , tissue [−1·K−1) ,20,21, dD structu[−1·K−1) ,23, bios[−1·K−1) ,25, gene30 m2·g−1[−1·K−1) ,28,29,30[−1·K−1) ,32,33.3 [3 are 7.5 × lighter than air [Aerogels are porous solids systems (solid foams) with a predominance of open pores in which the dispersed phase is a gas . Aerogel3 , which a3 . Graphenthan air , possessthan air , a lowerthan air  or nuclethan air . The porthan air . Typicalthan air  to tissuthan air .With regard to the extent of the large possibilities of graphene-based structures, we endeavor to give a brief overview of their use in drug delivery for anticancer therapies, in tissue engineering, and as imaging agents. We start by briefly exploring the toxicity of these materials.The growing biomedical field of applications of graphene-based materials raises questions about their short- and long-term (cyto)toxicity ,45. It i4 cells) did not weaken the cell activity significantly, as shown by an MTT assay [3 plasma [It is worth mentioning that cytotoxicity was observed to be cell-dependent, e.g., a loss in viability was observed for the human lung adenocarcinoma cell line A549) and an increase in viability for the human colon cancer (Caco2) and monkey kidney (Vero) cell lines ,50. The 9 and an TT assay ,56. InstTT assay . Moreove3 plasma . The gro3 plasma . Interes3 plasma .The cytotoxicity aspects of graphene-based materials should not be considered alone. Another important aspect to consider is the degradability of the formed structures, which will also intrinsically depend on the type and size of the used raw material. This issue is still being actively pursued in the field, for example by Mukherjee et al. , who shoTo ultimately reach final applications, graphene-based materials need to be properly assessed in vivo. One of the most basic in vivo studies shows that GO administered at a dose of 0.25 mg via tail-vein injection does not affect the lifespan of mice, while a dose of 0.4 mg is too much and four out of nine mice died within 7 days post-injection. The autopsy showed GO granuloma localized in the lung, liver, spleen, and kidney, but no GO was detected in the brain . FurtherThe biocompatibility of pure-graphene-based structures can be further increased by G/GO surface functionalization or coatings. One of the most frequently used polymers is poly(ethylene glycol) (PEG), which has been shown to decrease cytotoxicity both in vitro  and in iThe aforementioned results emphasize the important role of the surface physicochemical characteristics of graphene and graphene-based materials in their interactions with biomolecules and cells . In partGraphene (G) is highly hydrophobic, whereas GO is decorated by oxygen-containing hydrophilic groups . This unAn ageing population, smoking, stress, and a lack of physical activity are major contributors to the rapid increase in the cancer mortality rates of many types of tumors . It remaN-isopropylacrylamide) (PNIPAM) was used as a polymer matrix for GO nanosheets, which resulted in a hybrid, self-healable, supramolecular hydrogel [+CD133+ cells, which are characterized by high tumorigenicity [Bxl-2 [Another strategy that is utilized in cancer treatment is the formation of injectable hydrogels with GO-based nanofillers that can bind and subsequently release typical hydrophobic anticancer drugs . One exahydrogel . Subsequhydrogel  preparedgenicity . These cy [Bxl-2 .2) functionalized GO nanosheets on a patterned gold surface. Interestingly, the authors observed that the same chip without the GO did not yield the same positive results; however, no discussion about the importance of GO was given. Nevertheless, such studies demonstrate a significant accomplishment towards the development of graphene-based diagnostic chips that can be used as sensors for targeting biomolecules from cancer patients’ samples, eventually leading to cheap cancer sensors.Functionalized GO chips for the highly sensitive capture of circulating tumor cells (CTCs) in the blood of cancer patients were also presented in another study . CTCs arGraphene quantum dots (GQDs) are another form of G used as carriers for cancer drug delivery applications. In one of the studies, GQDs were functionalized by a commonly used arginylglycylaspartic acid (RGD) peptide sequence, which binds to the “αvβ3” integrin site in a cancer cell. When Dox drugs were loaded onto the GQDs-RGD, their enhanced uptake by the PC-3 and DU-145 human prostate cell lines was observed in comparison to pure Dox . Ko et aOverall, many multimodal approaches that combine G/GO structures have been proposed for anticancer therapies. However, we still lack the full in vivo characteristics of the proposed G/GO nanomaterials, and it remains a challenge to assess their biodistribution, biodegradability, and the targeted effects in many types of cancers. Future work in the field will need to focus not only on multimodal approaches but mainly on the applications and feasible sites. In particular, all proposed structures will need to explicitly prove their safety in extensive trials prior to their proper use. Taking into consideration the fact that these materials could one day save someone’s life, their purity, firstly, will need to be of the highest level. To do so, the characterization techniques will need to be properly standardized and regulations will need to ensure that there are proper definitions for the materials and their properties.Engler et al. revolutionized the tissue engineering field in 2006 by showing explicitly the effect of substrate stiffness on the differentiation of mesenchymal stem cells (MSCs) into different cell lineages . They acOne of the interesting approaches for forming composite GO structures is their covalent crosslinking with biopolymers. One such approach was the formation of GO–chitosan hydrogel scaffolds that were prepared by covalent linkage of chitosan amino groups with carboxylate groups of GO . These G2 substrates [Currently, tissue damage remains one of the most crucial aspects that contributes to human death. In this regard, a number of studies have been performed that explore the use of graphene for stem cell engineering and musculoskeletal tissue engineering ,99,105. bstrates . One of bstrates .Human-adipose-derived stem cells (hASCs) have also been extensively studied in tissue engineering due to their being easy to obtain from patients. Chung et al. have investigated the effects of GO-coated substrates on the fate of hASCs . As othe2), poly-m-aminobenzene sulfonic acid- (–NH2/–SO3H), and methoxyl- (–OCH3)) on neurite outgrowth and branching [2 groups) was found to be more beneficial for neurite outgrowth and branching.Graphene-based materials were also explored in musculoskeletal tissue engineering using mouse myoblast C2C12 cell lines . The oxyranching . SurprisOverall, the abovementioned studies highlight the importance of understanding the physical interactions in biomaterials and substrates for biological purposes and emphasize the possibilities of modulating biological responses for tissue engineering by both chemical and physical approaches. Even though there exist a large number of studies on the effect of graphene-based materials, the exact elucidation mechanisms, due to the many combinations of protein–G/GO interactions or the endless chemical functionalization of G/GO, are yet to be revealed. Perhaps the growing power of artificial intelligence and multi-algorithm-based approaches will soon be able to screen the whole library of cell-influencing factors simultaneously to generate ideal tissue engineering platforms for regenerative medicine.Two-dimensional (2D) graphene-based material coatings on substrates and layered materials are insufficient for tissue engineering, and these need to be further developed in order to mimic a proper, functional 3D extracellular matrix (ECM) environment. Indeed, recent advances in this field include the exploitation of graphene foams (GFs) as three-dimensional (3D) scaffolds for a neural stem cell (NSC) culture  and humaAnother approach in tissue engineering is to use pure 3D G foams that are produced via metal-free methods . Pure G Another approach is to use hybrid 3D GFs with additional coatings. One such case was reported in which a commercially available hydrophobic pristine 3D GF was coated by a well-established biocompatible collagen that was cross-linked with genipin to form a hybrid 3D GF porous, hydrophilic, and conductive scaffold for the culture and differentiation of mouse mesenchymal stem cells . The obtObviously, 3D structures that mimic the tissue’s extracellular matrix are highly desirable in the tissue engineering field. Due to their intrinsic structural nature (very high aspect ratios), G and GO have so far mainly been used as coatings and substrates of flat surfaces and materials. However, there is a large trend to scale the results to the 3D world, either by simply incorporating the 2D flakes in a 3D biopolymer matrix or by forming hybrid foam-like structures that can better mimic the desired properties and characteristics of the chosen tissue. We envisage the use of such materials in real-world applications and wait for the future outlook, which will most likely consist of changes to the mass production of large quantities due to the increasing use of additive manufacturing techniques. In particular, we see a large rise in the 3D printing of graphene-based inks that, in the near future, may replace the complex 3D structure of tissues with enhanced properties ,116,117.Bio-imaging, which focuses on imaging selected biological entities, remains a crucial aspect of regenerative medicine . ImagingThe most often used imaging technique involves fluorescence. This has become the gold standard technique that is used in biology and regenerative medicine. In particular, Forster Resonance Energy Transfer (FRET) imaging  is most 2 [Some techniques, including computer tomography (CT), PET and MRI, and ultrasound imaging are already well-established and can be used in human studies. The EPRI technique is a relatively new technique under strong development. By using this method, one is able to detect and quantify multiple biological parameters in a tumor microenvironment in vivo in small animal models, including: pO2 ,135,136,2 ,137, and2 ,138. Usu2  or other2  are used2 . Raman s2 , ex vivo2 .Structures such as G, GO, and the related G-based composites that are used in bio-imaging are usually treated as platforms for the modification of existing contrast agents ,145. HowThe next and simultaneously higher level of bio-imaging development is the fabrication of multimodal materials (nanoparticles) that can simultaneously be used in more than one way, e.g., in photothermal and photodynamic therapy and tracking (imaging) ,156,157.Graphene-based materials in the form of flakes, layers, foams, nanofillers, nanodots, and other structured materials have had a profound impact on the regenerative medicine and biomedical fields. The cytotoxicity of these materials is strongly dependent on the intrinsic flake sizes that are used in the final material, the structure and shape of the material, its surface chemistry, and the type of cultured cells. The fundamental interactions between the surface and surface edges of materials with the biomolecules, drugs, and cells are driving the biological pathways, in many cases in still-unknown ways. The toxicity aspects of the structure of graphene/graphene oxide are to this day debatable and remain of high interest to the scientific community. Likewise, the structure of graphene/graphene oxide-based materials influences their possible application fields, which were briefly reviewed here and include anticancer drug delivery therapies, tissue engineering, and multimodal bio-imaging.Almost all of the drug delivery anticancer approaches rely on the successful adsorption of the drug on the surface of the graphene/graphene oxide structure. Many efforts have been focused on the use of a well-established anticancer drug, such as doxorubicin (Dox), and the use of versatile graphene-based and graphene-based hybrid vehicles for its delivery or co-delivery and imaging. However, to this day, the in vivo work has shown virtually no improvement in using such vehicles in comparison with pure drugs, or there have been no attempts to perform in vivo evaluations. This shows a challenging problem with anticancer therapeutics where the drug/material’s biodistribution, biodegradability, and the targeted effects are still unknown. Moreover, little has been done to derive new drugs using graphene-based materials as platforms. We see particular promise in the graphene quantum dots that have been used in multimodal therapies, which may, simultaneously, have selective sensitivity, be effective contrast agents for fluorescence imaging with a high quantum yield, and be used for photothermal and photodynamic therapy. Indeed, small doses of materials with proper characteristics were shown to be acceptable for cell culture growth and bio-imaging in terms of the cytotoxic effects.Interestingly, graphene-based materials remain highly successful in the tissue engineering field for the regeneration of a variety of tissues. However, most of the successful work has been performed on 2D substrate systems, which could prove that G/GO-based flake coatings are useful for various biomedical devices. On the other hand, G/GO have been used as nanofillers in a variety of hydrogels or soft-matter materials that add a desired functionality to the native materials, such as the possibility of adsorption for certain proteins or an electrically active percolation network and/or surface that triggers cell proliferation, adhesion, or differentiation. There is a lot of work that focuses these days on using G/GO in 3D systems, either as a pure network or as a hybrid with other materials, which holds great promise for the use of these materials in real-world biomedical applications. Based on the literature survey presented in this review, we believe that there remain many crucial aspects to consider before this field can progress further, namely: nanomaterial purity and characterization and proper definitions to ensure data reproducibility and appropriate data correlation.


Legal restrictions on alcohol availability have been used to address violence and injury in the world’s remote Indigenous communities. In Australia, alcohol management plans (AMPs) were implemented by the Queensland Government in 2002. This study reports changes in indicators of alcohol-related violence and injury in selected communities.Design and setting: A longitudinal observational study was conducted in four Aboriginal and Torres Strait Islander (Indigenous) communities in Cape York, far north Queensland. All communities are similarly-isolated from population centres where alcohol is available.Data: For 2000 to 2015 inclusive: 1019 Royal Flying Doctor Service aeromedical trauma retrievals; 5641 Queensland Police Service records of unique assault occurrences, including 2936 involving alcohol; and records for 2741 unique assault victims were examined.Data analysis:Rates (per 1000 population) of trauma retrievals, assault occurrences and assault victims (per 1000 population) were compared across three policy phases.Phase 1: 2000 to 2008. Initial restrictions on possession and consumption of alcohol in ‘restricted areas’ were implemented during 2002–2003.Phase 2: 2009 to 2012. All alcohol was prohibited in three study communities and its legal availability limited in the fourth from 2009.Phase 3: 2013 to 2015. Government reviews of AMP policies in light of legal challenges and community responses characterise this phase.Compared with Phase 1, in Phase 2 retrieval rates declined by − 29.4%, assault occurrences by − 34.1% with less than one-third involving alcohol, and assault victims by − 21.1%, reaching historically low levels in 2010–2012. These reductions did not continue consistently. Compared with Phase 1, in Phase 3 retrieval rates, assault occurrence rates and assault victim rates declined by somewhat lesser amounts, − 13.9%, − 15.0% and − 13.4%, respectively. In Phase 3, the proportion of assault occurrences involving alcohol in communities 2, 3 and 4 rose towards pre-2008 levels.Early successes of these controversial alcohol restrictions are jeopardised. Indicators of violence and injury appear to be rising once more in some AMP communities. Importantly, rates have not generally exceeded the highest levels seen in Phase 1. Fresh policy action is required with rigorous monitoring to prevent erosion of initial important successes. With 5.1% of the global burden of disease and injury attributable to alcohol . For theLiquor Act 1992 with controls on quantities and types of alcohol permitted in a restricted area designated under S173H [Liquor Regulation 2002 [In Australia, legal restrictions on alcohol’s physical availability in Indigenous communities became known as ‘Alcohol Management Plans’ (AMPs) towards the end of the twentieth century –14. AMPsion 2002 .all of the 19 AMP communities; bringing complete prohibition to seven of them [As already described in detail in a previous publication, these place-based controls were first introduced in 2002 . By 2006At the end of 2012, the Queensland Government implemented formal processes to review AMPs , 19. As Using data provided by Australia’s Royal Flying Doctor Service (RFDS), we have monitored rates of aeromedical retrievals for serious injury in four AMP communities over the 15 year period from 1995 to 2010 , 10. ThiCriminal Code 1995 with a local police presence in most remote communities, including the four in this study. This analytical approach ensures that the indicators examined reflect changing circumstances in these four communities, independent as far as practicable from external confounding factors, providing the opportunity to consider the local impacts of Queensland-wide policy initiatives and shifts.The communities, their populations and the reasons for choosing them as sentinel communities for monitoring have already been described in detail , 10. In This longitudinal observational study assesses the effects of a policy intervention which was developed and implemented with a changing focus over the past two decades. The very complex legislative and regulatory background for AMPs in all Queensland Indigenous communities has already been described and summarised . The fouSince the 1980s, each community’s elected Local Government Council (LGC) had been operating licensed premises, variously known as a ‘tavern’ or ‘canteen’ , from whFollowing Justice Fitzgerald’s 2000 inquiry , over a During 2007, legislation and regulation to control alcohol availability across the wider region saw ‘minimising harm’ conditions placed on the licences of liquor retailers in the ‘catchment’ areas of restricted area communities . RetaileLiquor Act 1992 was amended removing the option for LGCs to hold a liquor licence while also making it an offence to ‘attempt’ to bring prohibited quantities and types of liquor into a restricted area [In 2008, the ted area . For thrted area . The on-Significant legal challenges were mounted to the legitimacy of Queensland’s AMPs commencing during the previous Phase 2 , 35 and While this important period of review and legal challenge has unfolded, liquor restrictions in the four communities have not changed since 2008, with one exception. In mid-2014, in one community with prohibition, a venue formerly run by the LGC was permitted to operate once again, with limited trading hours and also with the on-premises purchase and consumption of liquor containing ≤4% alcohol by volume permitted under a ‘Restricted Liquor Permit’ .The causal mechanisms linking alcohol use and interpersonal violence are well known , 41, 42 In the general population, it is widely recognised that alcohol is involved in more than half of all occurrences of interpersonal violence with alcohol’s acute effects having the greatest impact on violent behaviour . For theIn addition, since QPS records also include information about individuals affected by violent occurrences, and since the victims of assault often require some kind of acute care medical intervention, to capture as wide a spectrum of relevant violent incidents as possible, we used the indicator of victims of ‘violence against the person’ recorded by QPS.A Government-funded clinic has provided local health care services in each study community throughout the observation period. The RFDS remains the principal provider of emergency medical transport to definitive care of all people presenting to the local clinic with serious injuries. There have been no substantive changes in RFDS aeromedical retrieval processes, logistics or management for these communities across the time frame of our previously published , 10 and As in these previous studies , 10, serThe QPS has a presence in each of the four communities in the form of a Division station. Two of the stations, i.e. those in the two largest communities, are staffed by the equivalent of eight permanent officers and, in the smaller communities, by the equivalent of two permanent officers. Some LGCs employ Community Police Officers to assist. In 2008, to support the ‘dry as possible’ phase, the Queensland Government committed funding for additional officers across all 19 AMP communities [Data were provided by QPS for all occurrences of person-to-person violent offences using Australian Standard Offence Classification Group Codes for ordering offences  includinFor harm observed by police in connection with an assault occurrence, or experienced or reported by victims, data for the same types of person-to-person violent offences  were useRates of RFDS trauma retrievals and QPS registered occurrences of assault and victims of assault (per 1000 population) were calculated for all years between 2000 and 2015 for all four communities , measures of statistical uncertainty  are unnecessary and would be factually misplaced.The Human Research Ethics Committees of James Cook University and the Cairns and Hinterland Health Services District provided relevant ethical approvals for the study to be conducted in these communities . The QPSA total of 1019 aeromedical trauma retrievals were extracted from the RFDS database and used in the analysis for the four communities for 2000 to 2015 inclusive. During the same period, data for 5641 unique assault occurrences were provided by QPS. In 2936 (52%) of these assault occurrences, alcohol was recorded as being involved. During the observation period 2741 individuals were victims of an assault, as recorded by QPS. The resulting rates for retrievals and assaults for the four communities as well as the average over all communities over the single years of the observation period are numerically detailed in Table Consistent with previous results , 10, TabFigure Table Table Communities 1 and 2 have not fared so well . This case required attention to international human rights conventions and the Australian Constitution [Liquor Act 1992 were invalid by reason of inconsistency with s 10 of Australia’s Racial Discrimination Act 1975 (Cth). Section 10 of this Commonwealth act provides that a State law cannot have the effect that persons of a particular race do not enjoy a right enjoyed by other races, or do not enjoy it to the same degree. In dismissing the appeal, the Court of Appeal held that s 10 did not apply, as it was a ‘special measure’ taken for the sole purpose of securing the adequate advancement of the Indigenous residents of Palm Island and to prevent human rights violations, such as violation of the right of vulnerable groups to live in a safe environment [In the background, during Phase 2, significant legal challenges to the legitimacy of AMPs were commenced , 36, onetitution , 38. An titution . The groThe issue of consultation for ‘special measures’ and the proportionality aspects of AMPs in Australia’s Northern Territory have recently raised concerns for the Australian Parliamentary Joint Committee on Human Rights . In its It is of great concern that rates of retrievals for serious injury, assault occurrences and victims show a generally increasing trend since 2012 in these four study communities. This is in the absence of any substantial policy change affecting alcohol’s legal availability. The underlying causes for these trends warrant urgent, refreshed policy action informed by rigorous objective monitoring to ensure the successes gained by AMPs in Queensland are not placed in further jeopardy.


Stimuli-responsive materials, also known as smart materials, can change their structure and, consequently, original behavior in response to external or internal stimuli. This is due to the change in the interactions between the various functional groups. Graphene, which is a single layer of carbon atoms with a hexagonal morphology and has excellent physiochemical properties with a high surface area, is frequently used in materials science for various applications. Numerous surface functionalizations are possible for the graphene structure with different functional groups, which can be used to alter the properties of native materials. Graphene-based hybrids exhibit significant improvements in their native properties. Since functionalized graphene contains several reactive groups, the behavior of such hybrid materials can be easily tuned by changing the external conditions, which is very useful in biomedical applications. Enhanced cell proliferation and differentiation of stem cells was reported on the surfaces of graphene-based hybrids with negligible cytotoxicity. In addition, pH or light-induced drug delivery with a controlled release rate was observed for such nanohybrids. Besides, notable improvements in antimicrobial activity were observed for nanohybrids, which demonstrated their potential for biomedical applications. This review describes the physiochemical properties of graphene and graphene-based hybrid materials for stimuli-responsive drug delivery, tissue engineering, and antimicrobial applications. N-vinylcaprolactam), poly(N-isopropylacrylamide), poly, and other copolymers are frequently used as a smart material for various biomedical applications [2-bonded carbon atoms with a honeycomb morphology. The higher surface area of graphene facilitates effective binding with several drugs through various interactions and is frequently used in targeted and controlled drug delivery applications [Nowadays, on-demand release of active materials in desired areas has drawn tremendous attention in the rapidly developing field of materials science. For this purpose, stimuli-responsive materials (SRMs), which are also known as smart materials, are frequently used. They can change their shapes or dimensions in the presence of external stimuli such as electric field , 2, magnications . For bioications . The proications –29. Accoications –32. Grapications , 34. Theications . The strications . Extraorications . Graphenications . Enhanceications . Moreoveications .  Figureications .This review describes the salient features of graphene and its biomedical applications such as stimuli-responsive drug delivery, tissue engineering, and antibacterial materials in the presence of different polymer matrices. Different techniques such as in situ polymerization, solution casting, and extrusion were used to fabricate graphene-based nanohybrids for desired applications.2-hybridized carbon atoms arranged in a honeycomb structure. It was initially described by Boehm et al. in 1986 followed by identification and isolation by Geim and Novoselov in 2004 [σ bonds with a delocalized π electron network. These delocalized π electrons provide a high electron density above and below the 2D planar structure of graphene. Because of the planar structure and delocalized π electrons, graphene undergoes various reactions such as cycloadditions, click reactions, and carbine insertions [Among various nanomaterials, graphene has a variety of advantages and gained tremendous attention from the scientific community. Graphene is a 2D single atomic layer of graphite with sp in 2004 , 43. In sertions . Pure grsertions . In addisertions . Some saSeveral techniques such as force displacement, force volume, nanoindentation atomic force microscopy (AFM) –48, and σ bonding and delocalized π electrons makes graphene a unique electrically and thermally conductive material with a low thermal expansion coefficient. The thermal conductivity of defect-free graphene is much higher than that of other carbon nanomaterials. Its thermal conductivity is approximately in the order of 4.5–5.5 × 103 W/mK, whereas it is approximately 2 × 103, 3 × 103, and 3.5 × 103 W/mK for graphene oxide, multiwalled carbon nanotubes, and single-walled carbon nanotubes, respectively [4 S/cm. The electrical mobility of GO is 10−1 S/cm [The presence of strong ectively , 57, 58.ectively –61. The 0−1 S/cm . With it0−1 S/cm , 64.In addition to exceptional mechanical strength and thermal and electrical properties, graphene has an excellent optical property. Single layer defect-free graphene has shown 97.7% incident light transmission over a wide range of wavelengths . This prπ-π stacking interactions between the drug and the polymer matrix [π-π stacking and hydrogen bonding interactions. A faster drug release from the nanohybrids was observed at pH 5.3 than at pH 7.4, which indicated its potential as a targeted and pH-mediated anticancer drug delivery vehicle [π-π stacking and hydrophobic interactions between the drug and the nanohybrids. However, drug release was observed only in an acidic medium but not in basic and neutral media, which are found in a tumor environment; this suggests the formation of a suitable site-specific drug carrier [N-isopropylacrylamide (pNIPAAm) polymer matrices. They noted that a faster drug (doxorubicin hydrochloride) release from the hydrogel occurred in the acidic medium than in the neutral and alkaline media due to the protonation of polar groups. Furthermore, the developed hydrogels exhibited temperature-mediated drug release, which was more controlled at higher temperatures due to dehydration of the hydrogel leading to a more compact structure that hinders the diffusion of the drug. On-demand or targeted drug release from biomedical devices has attracted great attention in the field of medical science. It has been noted that targeted drug release from carrier molecules exhibits high efficiency with a controllable release and minimum side effects. Several factors such as light, heat, pH of the medium, ultrasound waves, and electric or magnetic fields are responsible for the controlled release of drugs –72. For r matrix . In anot vehicle . Kurapat vehicle . Further carrier . Hydroge carrier . In anotγ (INF γ) biomolecules, a larger population of neuron cells occurred, while in the presence of a platelet-derived growth factor (PDGF), a larger population of oligodendrocytes occurred, suggesting its potential for controlling the NSC differentiation tendency for therapeutic applications [ε-caprolactone) (PCL) matrix to evaluate the cellular response toward stem cells. They observed that the composite with diamine-modified GO showed a higher proliferation and differentiation of human MSCs (hMSCs) followed by the GO composite. This was due to better interactions between the amine moiety and the cells [For tissue engineering applications, materials should be biocompatible, nontoxic, and biodegradable in nature. In addition, materials should not show any negative response in biological conditions –84. Tissications . In anotications . Chemicahe cells . Zhang ahe cells . Notablyhe cells . Furtherhe cells . Sayyar he cells . Hydroxyhe cells , 98. Thehe cells . A similhe cells . In anothe cells . Degradahe cells . In addiE. coli and S. aureus (~100% of E. coli and ~95.3% of S. aureus) by 10 μg/mL for 2.5 h. The higher antibacterial activity of GO-PEG-Ag composites was due to the damage of the bacterial structure and the production of reactive oxygen species, which led to cytoplasm leakage and decrease in metabolism [L-lysine) (PLL) composites through electrostatic interactions and covalent bonding between the graphene derivatives and PLL and evaluated their cytotoxicity and antibacterial behavior. They observed that the composites showed a strong antibacterial nature and biocompatibility toward human adipose-derived stem cells and non-small-cell lung carcinoma cells (A549), which indicated its dual functionality that can be used to inhibit bacterial growth as well as enhance human cell growth [E. coli (ATCC 25922) and Gram-positive S. aureus (ATCC 6538) by the plate count method and disk diffusion method. Significant antibacterial activity was observed for the nanocomposites, which suggested its potential use in biomedical applications [E. coli (~97.9% reduction) after 3 h of contact, indicating its multidynamics applications with strong antibacterial and separation performances. The antibacterial activity of the PDA-rGOC-modified membrane is shown in N-vinylcarbazole) (PVK) and graphene/GO. The PVK-GO-modified membrane exhibited a strong antibacterial activity toward B. subtilis and E. coli. This was due to the production of reactive oxygen species by the nanoparticles, which influenced the metabolic activity of the microorganisms [S. aureus [E. coli and S. aureus bacteria, respectively, were killed by the developed hybrids under irradiation of 660 nm light due to the formation of radical oxygen species; this indicated the strong photocatalytic activity of the hybrid toward microorganisms [S. aureus, E. coli, and C. albicans. A significant improvement in antimicrobial activity was observed for the GIO-based hydrogel film compared to chitosan-GO and chitosan-iron oxide films [2/Ag3PO4/graphene composite synthesized by an ion-exchange method and a hydrothermal approach [E. coli and S. aureus with negligible cytotoxicity was observed for silver-incorporated ZnO-chemically converted graphene nanocomposites synthesized by a low-temperature technique using zinc acetate dehydrate, silver nitrate, and GO [Nowadays, several antibiotics and antimicrobial agents have been developed for the treatment of various infectious diseases. However, lethal microorganisms remain a challenge for public health, causing several infectious diseases annually . Antibiotabolism . Some etl growth . In anotications . Microbiications . Severalications . Nanofilications –121. Zhurganisms . Liu et . aureus . It is w. aureus . A similrganisms . Konwar de films . Antibacapproach . Materiaapproach . Further, and GO .Nanomaterials with unique intrinsic physiochemical and biological properties, which are absent in their macro forms, have drawn significant attention in materials science for various applications. Several nanomaterials such as a graphene, carbon nanotubes, fullerenes, zeolite, and metals in different forms are frequently used to improve the native properties of materials for desired applications. Nowadays, graphene, an allotrope of carbon with excellent thermal, electrical, optical, mechanical, and biological properties and a higher surface area, is intensively used to enhance material properties. Moreover, properties of graphene can be tuned by surface functionalization with various groups. The higher surface area of graphene and the high charge density on the graphene surface facilitate the loading of several drug molecules, and they consequently act as a nanocarrier with tune rate in the biological medium. In addition, the excellent physical property of the graphene surface facilitates the proliferation and differentiation of cells. Moreover, its light-absorbing behavior plays an essential role in light-triggered drug delivery or cellular response. A significant improvement in antibacterial activity without cytotoxicity was observed for various graphene-based hybrids, suggesting its potential as a biomaterial for various applications. Hence, a discovery of wonder graphene nanomaterial has opened a new area of research to produce lightweight, high-performance hybrid materials for various biomedical applications.