Nanocomposite hyaluronic acid adhesive hydrogel with controllable drug release for bone regeneration.

Hyaluronic acid (HA) hydrogels have arisen as candidate materials to simulate the extracellular matrix and restore the functions of both cartilage and hard bones. However, integration of bone tissue adhesion and long-term osteogenic properties in one hydrogel is often ignored. Herein, a strategy to construct nanocomposite hydrogel with host tissue adhesive properties, enhanced mechanical strength, improved stability and osteogenic effects was developed. Simvastatin (SIM) was firstly incorporated into zeolitic imidazolate framework-8 (ZIF-8) and surface decoration with hydroxyapatite was realized to obtain SIM loaded and hydroxyapatite modified ZIF-8 particles (SP). As the inorganic strengthening component, SP could further cross-link the mixture of dopamine-hyaluronic acid (dHA) and tannic (TA) via coordination interaction to fabricate the hybrid adhesive hydrogel (dHA/TA/SP). Sufficient phenolic groups endowed dHA/TA/SP with excellent tissue adhesion and antibacterial properties, while incorporation of SP significantly improved the mechanical strength and stability of hydrogel. Further, due to the multiple protective effects of ZIF-8 and hydrogel, SIM was sustainably released from dHA/TA/SP. Together with the active Zn2+ and Ca2+, the expressions of ALP, OCN and RUNX2 were upregulated, and the mineralization was also promoted. With significant osteogenic effect in vitro and in vivo, this nanocomposite adhesive hydrogel holds great potential for bone defects repair.

[Effects of Different Proportions of Ammonium Sulfate Replacing Urea on Soil Nutrients and Rhizosphere Microbial Communities].

In order to investigate the effects of ammonium sulfate, an industrial by-product, on soil nutrients and microbial community when applied in different proportions instead of using urea as nitrogen fertilizer, a pot corn experiment was conducted. A completely randomized block experimental design was used, with a total of five treatments:CK (no fertilization), U10S0 (100 % urea), U8S2 (80 % urea + 20 % ammonium sulfate), U6S4(60 % urea + 40 % ammonium sulfate), and U0S10 (100 % ammonium sulfate). The basic physical and chemical properties of soil and the dry weight of maize plants were determined by conventional methods, and microbial sequencing was performed using the Illumina NovaSeq platform. The experiment results showed that:① In each growth stage of maize, the pH of soil treated with fertilization (7.85-8.15) was decreased compared with that of CK (8.1-8.21), and the pH showed a decreasing trend with the increase in ammonium sulfate content. ② The soil available nitrogen content increased gradually with the increase in the ammonium sulfate ratio at each growth stage of maize. Compared with that in the CK and U10S0 treatments, the ratio in the U0S10 treatment increased 30.56 % to 63.68 % and 13.22 % to 38.43 %, respectively. The variation trend of organic carbon content was opposite to that of available nitrogen (U8S2 > U6S4 > U0S10), and the addition of ammonium sulfate was still higher than that of U10S0 at other growth stages except for the seedling stage. ③ The protease activity of all fertilization treatments was higher than that of the control, and the protease activity was gradually enhanced with the continuous growth of corn and the increase in the ammonium sulfate ratio. The protease activity of the U0S10 treatment was higher than that of the U10S0 treatment at each growth stage of corn, which increased by 10.54 %-100 %. Soil sucrase activity ranged from 0.04 to 0.24 mg·(g·24 h)-1, and those in the U0S10 treatments were significantly higher than those in the U10S0 and CK treatments at all growth stages, increasing by 20.32 % to 99.16 % and 24.31 % to 79.33 %, respectively. ④ The species abundance of bacteria and fungi in maize rhizosphere under all fertilization treatments were lower than those under the CK treatment, followed by those under the U10S0 treatment. The species diversity trend of the bacterial community in the three treatments with ammonium sulfate replacing urea were U8S2 > U0S10 > U6S4, and that of fungi were U6S4 > U8S2 > U0S10. ⑤ The maize dry weight of the U10S0 treatment and U0S10 treatment was the highest, which was 39.47 % and 36.16 % higher than that of the CK treatment, respectively, but the difference was not significant. The Pearson model showed that the species abundance and diversity of soil rhizosphere fungi and bacteria were affected by relevant environmental variables, among which pH value and soil available nitrogen content were the most important factors affecting microbial diversity. In conclusion, when corn planting in calcareous brown soil, replacing urea with a certain proportion of ammonium sulfate can improve soil nutrients more than urea alone, which affects the growth and rhizosphere microbial community of corn to a certain extent and has a greater yield.

Bionic double-crosslinked hydrogel of poly (γ-glutamic acid)/poly (N-(2-hydroxyethyl) acrylamide) with ultrafast gelling process and ultrahigh burst pressure for emergency rescue.

Injectable adhesive hydrogels combining rapid gelling with robust adhesion to wet tissues are highly required for fast hemostasis in surgical and major trauma scenarios. Inspired by the cross-linking mechanism of mussel adhesion proteins, we developed a bionic double-crosslinked (BDC) hydrogel of poly (γ-glutamic acid) (PGA)/poly (N-(2-hydroxyethyl) acrylamide) (PHEA) fabricated through a combination of photo-initiated radical polymerization and hydrogen bonding cross-linking. The BDC hydrogel exhibited an ultrafast gelling process within 1 s. Its maximum adhesion strength to wet porcine skin reached 254.5 kPa (9 times higher than that of cyanoacrylate (CA) glue) and could withstand an ultrahigh burst pressure of 626.4 mmHg (24 times higher than that of CA glue). Notably, the BDC hydrogel could stop bleeding within 10 s from a rat liver incision 10 mm long and 5 mm deep. The wound treated with the BDC hydrogel healed faster than the control groups, underlining the potential for emergency rescue and wound care scenarios.

Trichoderma-secreted anthranilic acid promotes lateral root development via auxin signaling and RBOHF-induced endodermal cell wall remodeling.

Trichoderma spp. have evolved the capacity to communicate with plants by producing various secondary metabolites (SMs). Nonhormonal SMs play important roles in plant root development, while specific SMs from rhizosphere microbes and their underlying mechanisms to control plant root branching are still largely unknown. In this study, a compound, anthranilic acid (2-AA), is identified from T. guizhouense NJAU4742 to promote lateral root development. Further studies demonstrate that 2-AA positively regulates auxin signaling and transport in the canonical auxin pathway. 2-AA also partly rescues the lateral root numbers of CASP1pro:shy2-2, which regulates endodermal cell wall remodeling via an RBOHF-induced reactive oxygen species burst. In addition, our work reports another role for microbial 2-AA in the regulation of lateral root development, which is different from its better-known role in plant indole-3-acetic acid biosynthesis. In summary, this study identifies 2-AA from T. guizhouense NJAU4742, which plays versatile roles in regulating plant root development.

Degradable Nanohydroxyapatite-Reinforced Superglue for Rapid Bone Fixation and Promoted Osteogenesis.

Bone glue with robust adhesion is crucial for treating complicated bone fractures, but it remains a formidable challenge to develop a "true" bone glue with high adhesion strength, degradability, bioactivity, and satisfactory operation time in clinical scenarios. Herein, inspired by the hydroxyapatite and collagen matrix composition of natural bone, we constructed a nanohydroxyapatite (nHAP) reinforced osteogenic backbone-degradable superglue (O-BDSG) by in situ radical ring-opening polymerization. nHAP significantly enhances adhesive cohesion by synergistically acting as noncovalent connectors between polymer chains and increasing the molecular weight of the polymer matrix. Moreover, nHAP endows the glue with bioactivity to promote osteogenesis. The as-prepared glue presented a 9.79 MPa flexural adhesion strength for bone, 4.7 times that without nHAP, and significantly surpassed commercial cyanoacrylate (0.64 MPa). O-BDSG exhibited degradability with 51% mass loss after 6 months of implantation. In vivo critical defect and tibia fracture models demonstrated the promoted osteogenesis of the O-BDSG, with a regenerated bone volume of 75% and mechanical function restoration to 94% of the native tibia after 8 weeks. The glue can be flexibly adapted to clinical scenarios with a curing time window of about 3 min. This work shows promising prospects for clinical application in orthopedic surgery and may inspire the design and development of bone adhesives.

Effect of application of short-form video health education on the health knowledge and satisfaction with nursing care of patients with lower extremity fractures.

BACKGROUND: Video health education has been increasingly adopted in the clinic to expand patient guidance and improve perioperative outcomes. To date, however, it is unclear whether the use of short-form videos and web-based clinician-created forums actually improve the perioperative experience of patients. OBJECTIVE: To evaluate the effect of application of short-form video health education on the health knowledge and satisfaction with nursing care of patients with lower extremity fractures. METHODS: This study is a quasi experimental study, using convenient sampling method and grouping according to historical control., one hundred and twenty-two patients admitted with lower limb fractures to the orthopedic ward of a tertiary first-class general hospital in Fujian, China were enrolled in this study. Based on their admission order, patients admitted from September 2021 to November 2021 were enrolled into the control group (n = 61) and patients admitted from December 2021 to March 2022 were enrolled in the intervention group (n = 61). Both groups received identical admission-based patient education, perioperative care, and discharge rehabilitation instructions. The control group received the traditional education method while the intervention group received a short-form educational video in addition to the traditional education method. Patient perioperative health knowledge and patient satisfaction with nursing care during treatment of lower limb fractures were compared across the two groups. RESULTS: Preoperative health knowledge in the intervention group was 95.1%, compared to 82.0% in the control group (χ2 = 3.954, P<0.05). The Newcastle satisfaction with nursing scales score in the intervention group was (80.902 ± 7.016) points, compared to (78.131 ± 8.213) points in the control group. These group differences were statistically significant (P < 0.05). CONCLUSION: The application of a short-form educational video during the perioperative care of patients with lower limb fractures may improve patients' understanding of perioperative health and increase satisfaction with nursing care.

Polydopamine-curcumin coating of titanium for remarkable antibacterial activity via synergistic photodynamic and photothermal properties.

Combined photothermal therapy (PTT) and photodynamic therapy (PDT) has emerged as a novel and effective antibacterial strategy. In order to endow titanium (Ti) with antibacterial properties, the Ti-PDA-Cur composite was prepared using the excellent adhesion properties of polydopamine (PDA) to load curcumin (Cur) on the surface of Ti. The Ti-PDA-Cur coating can produce singlet oxygen (1 O2 ) and heat under 405 + 808 nm light irradiation, which can effectively kill Staphylococcus aureus and Escherichia coli. Moreover, the cytotoxicity and hemolysis rate of Ti-PDA-Cur were low, indicating its good biocompatibility. Therefore, this study provided a new strategy for the development of new Ti implants.

Biomimetic, self-coacervating adhesive with tough underwater adhesion for ultrafast hemostasis and infected wound healing.

Massive bleeding and wound infection due to severe traumas pose a huge threat to the life and health of sufferers; therefore, it is of clinical importance to fabricate adhesives with rapid hemostatic and superior antibacterial capabilities. However, the weak wet adhesion and insufficient function of existing bioadhesives limits their practical application. In this study, a sandcastle worm protein inspired polyelectrolyte self-coacervate adhesive of poly-γ-glutamic acid (PGA) and lysozyme (LZM) was developed. The adhesive exhibited strong underwater adhesion to various surfaces (>250 kPa for solid plates and >50 kPa for soft tissues) and maintained a 80 kPa even when soaked in water for 7 days. Rat liver and tail defect bleeding models revealed that the hemostatic efficiency was superior to that of commercial samples. The in vitro antimicrobial tests showed that the bacterial inhibition to Staphylococcus aureus and Escherichia coli reached almost 100%. Additionally, the infected wound regeneration model demonstrated that the healing rate of the adhesive group was about 100% within 15 days, which was greater than that of the control group. In vitro and in vivo experiments proved that this facilely prepared adhesive will be a promising material to fulfil the integration functions for rapid wound closure and facilitating wound healing.

Tunable backbone-degradable robust tissue adhesives via in situ radical ring-opening polymerization.

Adhesives with both robust adhesion and tunable degradability are clinically and ecologically vital, but their fabrication remains a formidable challenge. Here we propose an in situ radical ring-opening polymerization (rROP) strategy to design a backbone-degradable robust adhesive (BDRA) in physiological environment. The hydrophobic cyclic ketene acetal and hydrophilic acrylate monomer mixture of the BDRA precursor allows it to effectively wet and penetrate substrates, subsequently forming a deep covalently interpenetrating network with a degradable backbone via redox-initiated in situ rROP. The resulting BDRAs show good adhesion strength on diverse materials and tissues (e.g., wet bone >16 MPa, and porcine skin >150 kPa), higher than that of commercial cyanoacrylate superglue (~4 MPa and 56 kPa). Moreover, the BDRAs have enhanced tunable degradability, mechanical modulus (100 kPa-10 GPa) and setting time (seconds-hours), and have good biocompatibility in vitro and in vivo. This family of BDRAs expands the scope of medical adhesive applications and offers an easy and environmentally friendly approach for engineering.

Aquatic Diatoms-Inspired Universal Adhesive Coacervates Triggered by Water.

Adhesives with strong underwater adhesion performance are urgently needed in diverse areas. However, designing adhesives with long-term stability to diverse materials underwater in a facile way is challenging. Here, inspired by aquatic diatoms, a series of novel biomimetic universal adhesives is reported that shows tunable performance with robust and long-lasting stable underwater adhesion to various substrates, including wet biological tissues. The versatile and robust wet-contact adhesives are pre-polymerized by N-[tris(hydroxymethyl)methyl]acrylamide, n-butyl acrylate, and methylacrylic acid in dimethyl sulfoxide and spontaneously coacervated in water triggered by solvent exchange. The synergistic interaction between hydrogen bonding and hydrophobic interaction allows the hydrogels with instant and strong adhesion to various substrate surfaces. The slowly formed covalent bonds enhance cohesion and adhesion strength in hours. The spatial and timescale-dependent adhesion mechanism endows the adhesives with strong and long-lasting stable underwater adhesion to be coupled with fault-tolerant convenient surgical operations.

Mussel-inspired polysaccharide-based sponges for hemostasis and bacteria infected wound healing.

Effective bleeding control and wound protecting from infection play critical roles in the tissue healing process. However, local hemostats are not involved in the whole healing processes to promote the final healing efficiency. Here, a multi-functional mussel-inspired polysaccharide-based sponge with hemostatic, antibacterial and adhesive properties was fabricated via cryopolymerization of oxidized dextran (OD), carboxymethyl chitosan (CC) and polydopamine nanoparticles (PDA-NPs), followed by lyophilization. Combining with the adsorbed thrombin, the sponges yielded a considerably lower amount of blood than the commercially available hemostatic dressings. Benefiting from the high photo-thermal transition efficiency of PDA-NPs, the sponges exhibited excellent antibacterial activity to both gram positive and negative bacteria. Owing to the rapid hemostatic activity and effective infection resistance, the sponges illustrated the significantly acceleratory wound healing efficiency compared with the control group. The thrombin-loaded OD/CC-PDA polysaccharide-based sponge has great potential for future clinical use as wound dressing.

[Isolation, identification, and degradation characteristics of three effective PAHs degradation strains]. / 3æ ªå¤šçŽ¯èŠ³çƒƒé«˜æ•ˆé™è§£èŒæ ªçš„åˆ†ç¦»é‰´å®šåŠé™è§£ç‰¹æ€§.

Isolating dominant strains for the degradation of polycyclic aromatic hydrocarbons (PAHs) is of great practical significance for the restoration of ecosystem polluted by PAHs. A total of 11 strains with capacity of degrading PAHs were obtained from soil polluted by PAHs around a coking plant, by enrichment culture, acclimation, and plate isolation. Three of them with effective PAH-degrading capability were identified and screened out by morphological observation, physiobiochemical characterization, and 16S rRNA gene sequencing, and respectively, named as DJ-3, DJ-8 and DJ-10. Based on the results of 16S rRNA gene sequencing, DJ-3, DJ-8, and DJ-10 were identified as Pseudomonas sp. Klebsiella sp., and Bacillus sp. The degradation rate of phenanthrene (200 mg·L-1), pyrene (200 mg·L-1), and naphthol (160 mg·L-1) by three strains (DJ-3, DJ-8 and DJ-10) after seven-day incubation were 48.9%-65.9%, 38.9%-43.1%, and 57.6%-64.9%, respectively. The degradation rates of mixed PAHs sample (1200 mg·L-1) by three strains were 49.1%, 44.5%, and 53.9%, which were significantly higher than other eight strains, indicating that they were highly effective in PAHs degradation. There was no antagonistic relationship among the three strains. This study would lay a foundation for building efficient PAHs degrading strains and improve the in situ bioremediation of PAHs contaminated soil.

Quantitatively mapping the interaction of HER2 and EGFR on cell membranes with peptide probes.

Human epidermal growth factor receptor-2 (HER2) is a member of the epidermal growth factor receptor (HER) family that is involved in various biological processes such as cell proliferation, survival, differentiation, migration and invasion. It generally functions in the form of homo-/hetero-dimers or oligomers with other HER family members. Although its essential roles in cellular activities have been widely recognized, questions concerning the spatial distribution of HER2 on the membranes and the interactions between it and other ErbB family members remain obscure. Here, we obtained a high-quality dSTORM image of HER2 nanoscale clusters recognized by peptide probes, and found that HER2 forms clusters containing different numbers of molecules on cell membranes. Moreover, we found that HER2 and EGFR formed hetero-oligomers on non-stimulated cell membranes, whereas EGF stimulation reduced the degree of heteromerization, suggesting that HER2 and EGFR hetero-oligomers may inhibit the activation of EGFR. Collectively, our work revealed the clustered distribution of HER2 and quantified the changes of the interaction between HER2 and EGFR in the resting and active states at the single molecular level, which promotes a deeper understanding of the protein-protein interaction on cell membranes.

Characterization of humic acids from original coal and its oxidization production.

Five coal samples obtained from Chinese coal-producing areas were oxidized by hydrogen peroxide (H2O2), and humic acids (HAs) were derived from original coal and its oxidizition samples. HAs were characterized by physical and chemical methods, between which was also comparison. Yield, ash, aromaticity, molecular weight and functional group of HAs showed variance between original coals. While, yield, molecular weight, and the quantity of oxygen-containing groups of HAs increased more from coals oxidized with H2O2. However, the increase of oxygen-containing functional groups depended on original coals. For Yimin lignite, the oxidation of H2O2 could obviously improve the carboxyl group content of HAs, thus promoting the adsorption of nitrogen. This study demonstrated that oxidation of coal by using H2O2 was one pretreatment way to obtain and modify HAs which could be used as prerequisite and functional material in agricultural field.

Organization of Protein Tyrosine Kinase-7 on Cell Membranes Characterized by Aptamer Probe-Based STORM Imaging.

Protein tyrosine kinase-7 (PTK7), as an important membrane receptor, regulates various cellular activities, including cell polarity, movement, migration, and invasion. Although lots of research studies focused on revealing its functions from the aspect of the expression of the gene and protein are present, the relationship between the spatial distribution at the single-molecule level and the function remains unclear. Through combining aptamer probe labeling and super-resolution imaging technology, after verifying the specificity and superiority of the aptamer probe, a more significant clustering distribution of PTK7 is found on the MCF10A cell basal membrane than on the apical membrane, which is thought to be related to their specific functions on different membranes. By exploring the relationship between the assembly of PTK7 and lipid rafts, actin cytoskeleton, and carbohydrate chains on the membrane, the unique distribution of PTK7 on disparate membranes is revealed to be probably because of the varied dominant position of these three factors. These findings present the detailed spatial information of PTK7 and the related potential organization mechanism on the cell membrane, which will facilitate a better understanding of the relationship between the molecular assembly and its function, as well as the overall structure of the cell membrane.

Self-Adaptive Antibacterial Coating for Universal Polymeric Substrates Based on a Micrometer-Scale Hierarchical Polymer Brush System.

Surface-tethered hierarchical polymer brushes find wide applications in the development of antibacterial surfaces due to the well-defined spatial distribution and the separate but complementary properties of different blocks. Existing methods to achieve such polymer brushes mainly focused on inorganic material substrates, precluding their practical applications on common medical devices. In this work, a hierarchical polymer brush system is proposed and facilely constructed on polymeric substrates via light living graft polymerization. The polymer brush system with micrometer-scale thickness exhibits a unique hierarchical architecture consisting of a poly(hydroxyethyl methacrylate) (PHEMA) outer layer and an anionic inner layer loading with cationic antimicrobial peptide (AMP) via electrostatic attraction. The surface of this system inhibits the initial adhesion of bacteria by the PHEMA hydration outer layer under neutral pH conditions; when bacteria adhere and proliferate on this surface, the bacterially induced acidification triggers the cleavage of labile amide bonds within the inner layer to expose the positively charged amines and vigorously release melittin (MLT), allowing the surface to timely kill the adhering bacteria. The hierarchical surface employs multiple antibacterial mechanisms to combat bacterial infection and shows high sensitiveness and responsiveness to pathogens. A new paradigm is supplied by this modular hierarchical polymer brushes system for the progress of intelligent surfaces on universal polymer substrates, showing great potential to a promising strategy for preventing infection related to medical devices.

Relationship between the Molecular Coil Dimension and the Energy Storage Modulus of Polymer Solution Configured with Oilfield-Produced Sewage.

Polymer viscoelastic solution is the non-Newtonian fluid and widely used in oil production. In the process of seepage, the mechanism of the polymer solution with different molecular coil dimensions (Dh) flooding on remaining oil is unknown. By using the dynamic light scattering instrument, the molecular coil dimension of the polymer solution is tested. By using the HAAKE rheometer, the creep recovery test data of the polymer solution under the same creep time condition are obtained. The effects of polymer solutions with different Dh on residual oil are observed, by using the visible pore model. The results show that the higher the molecular weight (M w) of the polymer, the larger the size of the molecular coil dimension. The elasticity characteristics of the polymer solution are sensitive to the molecular coil dimension. As Dh of polymer molecules becomes larger, the contribution of the elastic portion to the viscosity of the polymer solution increases. The higher the M w of polymer is, the longer the molecular chain is and the size of Dh is larger. On the condition of the polymer solution with different M w with 2.5 g/L, when Dh is between 320.0 nm and 327.8 nm, the ratio of the elastic part in the apparent viscosity exceeds the proportion of the viscous part, and the polymer solution composition after these data can be used as a comparative study of elasticity for residual oil use. In the visible pore model, the pore-throat ratio is 3.5, the E R of water flooding is 54.26%, the E R of the polymer solution with Dh = 159.7 nm is 75.28%, and the increase of E R is 21.02% than that of water flooding. With the increase of Dh to 327.8 nm, the final E R of the experimental polymer solution is 97.82%, and the increase of E R of the polymer solution than that of water flooding is 43.56%. However, in the model with a pore-throat ratio of 7.0 and the same polymer solution with Dh = 327.8 nm, the increase of E R of the polymer solution is only 10.44% higher than that of water flooding. The effect of the polymer solution with the same Dh is deteriorated with the increase of the pore-throat ratio.

Correlative dual-color dSTORM/AFM reveals protein clusters at the cytoplasmic side of human bronchial epithelium membranes.

The organization of a cell membrane is vital for various functions, such as receptor signaling and membrane traffic. However, the understanding of membrane organization remains insufficient, especially the localizations of specific proteins in the cell membrane. Here, we used correlative super-resolution fluorescence/atomic force microscopy to correlate the distributions of specific proteins Na+/K+-ATPase (NKA, an integral membrane protein) and ankyrin G (AnkG, a scaffolding protein) with the topography of the cytoplasmic side of human bronchial epithelium membranes. Our data showed that NKA and AnkG proteins preferred to localize in the protein islands of membranes. Interestingly, we also found that functional domains composed of specific proteins with a few hundreds of nanometers were formed by assembling protein islands with a few tens of nanometers.

Developing substrate-based small molecule fluorescent probes for super-resolution fluorescent imaging of various membrane transporters.

Super-resolution imaging technology has been a powerful tool for revealing fine biological structures and functions. Its high-quality imaging always needs highly accurate labeling. Here, by exploiting the high specificity and affinity of natural substrates to transporters, we developed one set of substrate-based small molecule fluorescent probes for labeling membrane transporters. A glucose-based probe (Glu-probe) and tyrosine-based probe (Tyr-probe) were synthesized as two examples. Confocal imaging showed that the Glu-probe could label glucose transporters on live cells by being stuck into the binding site. Compared with antibody-probe labeling, the labeling advantages of the Glu-probe were revealed. High specificity of the Glu-probe or Tyr-probe was examined by a colocalization experiment and glucose replacement or amino acid (AA) blocking. The synthetic probes were also tested on imaging HeLa cells to confirm their wide labeling application. Additionally, we found that membrane transporters were mostly in the clustered state on cellular membranes, changing their assembly pattern to regulate the transport effectiveness. These results suggest that the substrate-based probes can serve as valuable tools for investigating the spatial information of membrane transporters.

Using an RNA aptamer probe for super-resolution imaging of native EGFR.

Aptamers, referred to as "chemical antibodies", are short single-stranded oligonucleotides that bind to targets with high affinity and specificity. Compared with antibodies, aptamers can be designed, developed and modified easily. Since their discovery, aptamers have been widely used in in vitro diagnostics and molecular imaging. However, they are relatively less studied and applied in advanced microscopy. Here we used an RNA aptamer in dSTORM imaging and obtained a high-quality image of EGFR nanoscale clusters on live cell membranes. The results showed that the cluster number and size with aptamer labeling were almost the same as those with labeling with the natural ligand EGF, but the morphology of the clusters was smaller and more regular than that with cetuximab labeling. Meanwhile, dual-color imaging demonstrated sufficient fluorophore labeling, highly specific recognition and greatly accurate clustering information provided by aptamers. Furthermore, the aptamer labeling method indicated that active EGFR formed larger clusters containing more molecules than resting EGFR, which was hidden under the antibody labeling. Our work suggested that aptamers can be used as versatile probes in super-resolution imaging with small steric hindrance, opening a new avenue for detailed and precise morphological analysis of membrane proteins.